Determination of UPLC-ESI MS/MS- and GC-MS-based altitudinal variations in the bioactive potential of traditional medicinal plants

Determination of UPLC-ESI MS/MS- and GC-MS-based altitudinal variations in the bioactive potential of traditional medicinal plants | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Determination of UPLC-ESI MS/MS- and GC-MS-based altitudinal variations in the bioactive potential of traditional medicinal plants Garima Singh, Ajit Kumar Passari, N Senthil Kumar, Brijesh Kumar, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4195244/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 08 Aug, 2024 Read the published version in Applied Biochemistry and Biotechnology → Version 1 posted 5 You are reading this latest preprint version Abstract Traditional medicinal plants have attracted scientific interest due to their bioactive compounds, and the levels of their constituents vary with location and altitude. The present study was designed to evaluate the pharmacological potential of two traditional medicinal plants, Mikania micrantha and Ageratum huostonianum; these were collected from two sites, Murlen National Park (MNP) and Dampa Tiger Reserve (DTR), which are located at different altitudes. Both plant species are used by local traditional healers in Mizoram, Northeast India, for the treatment of various health problems. We hypothesized that altitudinal variation would affect these plants' chemical composition and bioactive potential. Plant extracts were evaluated for antioxidant, antimicrobial, and cytotoxic activities. The results show that the plants located at a higher altitude, i.e., MNP, showed higher TPC (615.7 ± 0.58 and 453.80 ± 0.95 µg gallic acid equivalents/mg of plant extract dry weight (µg GAE/mg) for M. micrantha and A. huostonium , respectively) and TFC (135.4 ± 0.46 and 120.66 ± 1.93 µg quercetin equivalents/mg of plant extract dry weight (µg GE/mg) for M. micrantha and A. huostonium , respectively). The extract of A. houstonianum (MNP) exhibited significantly greater antioxidant activity against ABTS radicals (IC 50 241.6 µg/mL) as compared to the extract of A. houstonianum (DTR) (IC 50 371.2 µg/mL). The composition of the bioactive compounds present in the plants was determined using UPLC-ESI MS/MS and GC/MS, which detected five and ten compounds in the A. houstonianum and M. micrantha extracts, respectively. Plant species collected from the Murlen National Park site had high bioactivity potential and contained several bioactive compounds. A distinct variation between the volatile and non-volatile compounds was revealed. The collective data in this study show the influence of altitude on the biological compound production of selected medicinal plants. The findings will be utilized in determining the plant material needed for the development of bioactive formulations. Ageratum huostonianum Mikania micrantha UPLC-ESI MS/MS GC-MS traditional medicinal plants altitude variation Figures Figure 1 Figure 2 Introduction Plant species produce a tremendous range of primary and secondary metabolites such as alkaloids, terpenes, and polyphenols. Primary metabolites are essential for growth, while secondary metabolites are non-essential and mainly involved in functional roles such as the defense and protection of plants against herbivores, pathogens, and biotic and abiotic stress [ 1 ]. Plants contain diverse chemical entities that may exhibit novel and beneficial pharmacological activity. Medicinal plants with bioactive compounds can be beneficial to human health. For instance, they can function as antibiotics, antioxidants, mycotoxins, insecticides, herbicides, etc. [ 2 ]. Medicinal plants have been exploited for their phytochemicals and used for treating various prevalent diseases and common ailments such as malaria, tuberculosis, diarrhea, and asthma [ 3 ]. Some researchers have previously suggested that environmental factors such as light intensity, photoperiod, rainfall, soil properties, and temperature can affect the biosynthesis of secondary metabolites in plants [ 4 – 6 ]. Other factors, including plant age, season, nutritional status, grazing, or microbial attack, may also impact the composition and quantity of phytocompounds found in plants. Elevated levels of UV-B radiation, which occur at high altitudes, have been reported to induce the biosynthesis of secondary metabolites (phytochemicals) to mitigate radiation damage in plants [ 7 , 8 ]. In 2015, Nascimento et al. reported that UV-B radiation stimulates the biosynthesis of secondary metabolites [ 9 ]. Phenolic compounds are generally known to enhance the bioactivity of plants. Thus, we hypothesized that variations in altitude could affect the chemical composition and bioactivity of extracts derived from plants. Using the HPLC and GC-MS methods, we characterized the variations in the chemical composition of Mikania micrantha and Ageratum houstonianum collected at different altitudes. We also determined the antioxidant and anticancer properties of the extracts from both plants. M. micrantha and A. huostonianum , which are members of the Asteraceae family, are considered to be invasive weeds. Both plant species have a long history of being widely used in traditional medicine in many regions, specifically by the Mizo tribe in Mizoram, Northeast India [ 10 ]. The antitumor activity of M. micrantha and its ability to inhibit the growth of cancer cell lines have previously been reported [ 11 , 12 ]. M. micrantha is also supposed to have antifungal, antibacterial [ 13 , 14 ], and anti-inflammatory properties [ 15 ]. A. houstonianum has been examined for antimicrobial, mosquitocidal, and other types of bioactivity [ 16 , 17 ]. In the present study, the variation in the pharmacological properties of both species was evaluated in plants collected from sites at two different altitudes. One site was the Dampa Tiger Reserve (DTR), which is a 500 km 2 protected area of forest located at 23°32'42"- 23°41’36” N and 92°13’12”- 92°27’27” E. The second site, Murlen National Park (MNP), is also a protected area of forest that has an area of approximately 100 km 2 and is located at 23 o 32’ -23 o 42’ N and 92 o 13’ -92 o 27’ E. The climate in Mizoram is moderate, and temperatures vary from 10 o C to 30 o C throughout the year. Material and Methods Plant material Samples of Mikania micrantha and Ageratum houstonianum were collected from two different locations in June 2016 from the Dampa Tiger Reserve and Murlen National Park in Mizoram, India. The authenticity of the plants was confirmed by Dr. Kalidas Upadhyay from the Department of Forestry in Mizoram University, and voucher specimens were deposited in the Mizoram University herbarium. Genomic DNA was isolated from both the plant samples using PureLink Genomic Plant DNA Purification Kit (Invitrogen-K183001) according to manufacturer’s protocol and amplified maturase K (matK) gene using universal primer (matK Forward 5'-CGATCTATTCATTCAATATTTC-3‘and matK Reverse 5'-TCTAGCACACGAAAGTCGAAGT-3') a per Cuenoud et al. (2002). PCR reaction conditions for matK gene was initial denaturation at 95°C for 5 min, followed by 40 cycles of denaturation at 95°C for 30 sec, annealing at 49°C for 35 sec and extension at 72°C for 1 min with a final extension step at 72°C for 8 min. The obtained PCR products were analyzed using 1.2% of agarose gel and visualized under Bio-Rad Gel Doc XR + system (Hercules, CA, USA). Finally, the PCR products was purified using the In-vitrogen Pure Link kit (Catalogue No: K310001), and were sequenced at DBT-State Biotech Hub, Department of Biotechnology, Mizoram University, Aizawl, India and sequence deposited in NCBI GenBank with the accession number of Ageratum houstonianum (MF770205) and Mikania micrantha (MF770206) respectively. Preparation of plant extracts Collected plant samples was washed and plant parts were shade dried at room temperature (30 0 C ± 2 0 C). The dried plant material was grinded to powder by using domestic mixture grinder. Around 100 g of dried powder was extracted thrice in 1000 ml of methanol for 48 h with occasional stirring. The extracts were concentrated using rotary evaporator (Buchi, India) at 40 0 C under reduced pressure and the obtained crude extract was stored at -20 0 C until further use [ 18 ]. Determination of total phenolics and flavonoids The level of total phenolics in the plant extracts was determined using Folin–Ciocalteu reagent, as previously described by Singh et al. [ 18 ]. An aliquot of 10µl of extract was mixed with 90 µl folin reagent (1:10 v/v in water) and 100 µl of 15% Na2CO3 to make the 200 µl volume in a 96 well microplate and recorded using a UV/VIS microplate spectrophotometer (MultiscanTM GO, Thermo Scientific, MA, USA). Gallic acid was used as a standard, and total phenolic content was expressed in gallic acid equivalents/mg of plant extract. Similarly, the level of total flavonoids was evaluated using a modified version of the aluminum colorimetric method, as previously described in [ 18 ]. 150 µl of methanol extract is mixed with 150 µl of 2% ethanolic AlCl3 and allowed to incubate in dark for 1 h and the absorbance was recorded at 420 nm. The level of total flavonoids in the samples of plant extracts was expressed in µg quercetin equivalents (QE)/mg of plant dry weight, quercetin was used as a standard. Determination of free radical scavenging potential Free radical scavenging capability of methanolic leaves extract of both plants were determined by DPPH assay as described by previous studies [ 19 , 20 ]. Briefly, 200 µl of freshly prepared DPPH methanolic solution was added along with different concentrations of plant extracts and incubated for 30 minutes to observe the absorbance at 517nm. Likewise, the level of free radical scavenging potential present in the methanolic plant extracts was determined using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical assays [ 18 ]. Each experiment was repeated three times. The IC 50 concentration of each sample of plant extract was calculated as the amount that could scavenge 50% of the DPPH and ABTS free radicals. Ascorbic acid was used as a positive control. Determination of antiproliferative The antiproliferative potential of the plant extracts was calculated using an MTT assay. To evaluate cell viability, four cell lines (HepG2: liver hepatocellular carcinoma cell line; MCF7: human breast cancer cell line; AGS: human gastric adenocarcinoma cell line; HeLa: human cervical cancer cell line) were procured from the National Centre for Cell Science (NCCS) in Pune, India. The cells were cultured in DMEM medium supplemented with 10% inactivated fetal bovine serum (FBS), 100 µg/mL penicillin, 100 µg/mL streptomycin, and 5µg/mL amphotericin B, in a humidified atmosphere of 5% CO 2 at 37◦C. MTT Assay The viability of the cell lines treated with different concentrations of the plant extracts was first evaluated using the MTT assay, as previously described in [ 18 ]. All four selected cell lines were trypsinized and seeded in 96-well microtiter plates (approx. 1 × 10 4 cells/well) and incubated for 24h at 37°C. The cells were then treated with different concentrations of the plant extracts (10, 25, 50, 75, 100, 125, 150, 175, 200, 225, or 250 µg/mL) and incubated for 72 h. An appropriate blank and control were used in each assay. The culture medium was removed by aspiration, and 20µL of MTT was added to each well and incubated for 4 h. DMSO was added to each well to dissolve the purple formazan of MTT. Absorbance values were recorded at 570nm, and cell viability was calculated based on a standard curve. All MTT assays were carried out in triplicate. HPLC determination of phenolic compounds Instrumentation HPLC/QTOF-MS conditions The instrumentation conditions were the same as those reported by Singh et al. (Singh et al., 2020), with all parameters set identically for the standards and samples. Chromatographic acquisitions Chromatographic separation was carried out using an Xbridge® C18 column (100 mm × 4.6 mm, 3.5µm) operated at 25°C. The mobile phase consisted of a 0.1% formic acid aqueous solution (A) and methanol (B); it had a flow rate of 0.5 mL/min with an isocratic elution of 90% (B) and a duration of 15–25 minutes. The sample injection volume was 1µL. The UV spectra were obtained by scanning the samples in the range 200–600 nm. Mass spectrometry Mass spectrometric analysis was performed on an Agilent 6520 QTOF mass spectrometer in positive ESI mode, as reported by Bajpai et al. [ 22 ]. Appropriate measurements were taken using the auto mass calibration method with an external mass calibration solution, as reported by Kumar et al. [ 23 ]. GC/MS analysis of volatile compounds Volatile organic compounds (VOCs) in the plant extracts were identified using GC/MS spectrometry [ 24 ]. A Clarus 680 GC utilizing a fused silica column and packed with Elite-5MS (5% biphenyl 95% dimethylpolysiloxane, 30 m × 0.25 mm ID × 250µm df) was employed, and the components were separated using helium as a carrier gas at a constant flow of 1 mL/min. The injector temperature was set at 260°C during the chromatographic run. A 1µL sample of the plant extract was injected into the instrument, and the oven temperature was adjusted as follows: 60°C for 2 min; followed by 300°C at the rate of 10°C min − 1 ; and 300°C, where the sample was held for 6 min. The mass detector conditions were as follows: transfer line temperature, 240°C; ion source temperature, 240°C; ionization mode electron impact, 70 eV; scan time, 0.2 sec; scan interval, 0.1 sec. The fragments ranged from 40 to 600 Da, and the spectra of the components were compared with a database containing the spectra of known components stored in the GC-MS NIST (2008) library. Statistical analysis All of the experiments were performed in triplicate, and the obtained results were statistically analyzed via one-way ANOVA using SPSS 20.0 software [ 25 ]. The presented data represent the mean ± standard deviation. P-values less than 0.05 were considered significant. Results Total phenolic content (TPC) The total phenolic content in the plant extracts was calculated based on the use of a standard curve (R 2 = 0.988). Both A. houstonianum and M. micrantha plant samples collected from the MNP location had higher total phenolic contents than the corresponding plant samples collected from the DTR location. The obtained data are summarized and presented in Table 1 . Table 1 Total phenolic content (TPC) and Total flavonoids content (TFC) of methanolic extract of selected plants collected from two locations and their EC 50 values using DPPH and ABTS assay (n = 3) Name of the plant (Location) TPC, GAE (µg/ml) TFC, QE (µg/ml) EC 50 (µg/mL) DPPH assay ABTS assay Ageratum huostonianum (MNP) 453.80 ± 0.95 120.66 ± 1.93 388.00 241.60 Ageratum huostonianum (DTR) 414.30 ± 0.60 104.86 ± 1.27 265.10 371.20 Mikania micrantha (MNP) 615.70 ± 0.58 135.46 ± 0.46 109.80 164.80 Mikania micrantha (DTR) 440.30 ± 0.29 123.33 ± 1.28 194.10 201.30 Values were taken as mean of three replicates (± SD). QE: Querctin; GAE: Gallic acid equivalent; MNP: Murlen national park; DTR: Dampa tiger reserve. Total flavonoids Samples of the plant extracts of A. houstonianum and M. micrantha collected from the MNP site exhibited a higher concentration of total flavonoids than those collected from the DTR site (Table 1 ). Extracts of M. Micrantha collected from the MNP site had a higher total flavonoid content (135.4 ± 0.46) than M. Micrantha extracts from the DTR site (123.33 ± 1.28). Similarly, extracts of A. houstonianum plants collected from the MNP site exhibited a higher amount of total flavonoids (120.66 ± 1.93) than what was present in A. houstonianum collected from the DTR site (104.86 ± 1.27) (Table 1 ). Determination of free radical scavenging potential The antioxidant potential of the plant extracts was determined by calculating the IC 50 , which represents the concentration of the plant extract needed to scavenge 50% of the free radicals present in the test solution. Extracts of A. houstonianum collected from the DTR site exhibited good antioxidant potential against DPPH radicals (IC 50 265.10µg/mL) as compared to extracts of A. houstonianum plants collected from the MNP site (IC 50 388.00 µg/mL). The extracts of A. houstonianum plants collected from the MNP site exhibited better scavenging of ABTS free radicals (IC 50 241.60 µg/mL) than the extracts of A. houstonianum collected from the DTR site (IC 50 371.20 µg/mL) (Table 1 ). Antiproliferative potential of plant extracts The antiproliferative of the plant extracts was evaluated with the use of four cell lines (HepG2: liver hepatocellular carcinoma cell line; MCF7: human breast cancer cell line; AGS: human gastric adenocarcinoma cell line; HeLa: human cervical cancer cell line) and using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium] assay. Extracts obtained from both A. houstonianum and M. Micrantha plants collected from the MNP site were found to be more cytotoxic to the HepG2, MCF7, and HeLa cell lines than the corresponding extracts of both plant species collected from the DTR site (Table 2 ). In contrast, extracts of the A. houstonianum and M. micrantha plants collected from the DTR site were found to be more cytotoxic to AGS cell lines than the corresponding extracts collected from the MNP site (Table 2 ). Table 2 Cytotoxicity screening of selected plants collected from two locations against cancer cell lines. Name of the plant (Location) Cell lines (IC 50 in µg/ml) HepG2 HeLa MCF-7 AGS A. huostonianum (MNP) 34.13 23.06 83.65 35.83 A. huostonianum (DTR) 54.99 46.11 111.40 31.57 M. micrantha (MNP) 22.98 124.40 162.60 144.30 M .micrantha (DTR) 110.60 211.90 487.70 79.97 HepG2: liver hepatocellular carcinoma cell line; MCF7: human breast cancer cell line; AGS: human gastric adenocarcinoma cell line; HeLa: human cervical cancer cell line; MNP: Murlen national park; DTR: Dampa tiger reserve. Phenolic compounds identified via HPLC/QTOF-MS The methanolic extracts of A. huostonianum (from MNP and DTR) and M. micrantha (from MNP and DTR) were analyzed with the use of mobile phase consisting of aqueous formic acid (0.1% formic acid) and methanol. Parameters such as column type, column temperature, mobile phase, elution conditions, flow rate, and MS conditions were optimized. The base peak chromatograms (BPCs) of the methanolic extract of A. huostonianum (from MNP and DTR) are shown in Fig. 1 , whereas the base peak chromatograms of the methanolic extract of M. micrantha (from MNP and DTR) are shown in Fig. 2 . The retention time (calculated as m/z [M + H] + ), molecular formula, error (Δppm), and distribution, along with a compound assignment, are presented in Tables 3 and 4 . All of the compounds were identified based on their exact mass, molecular formula calculation, and database search results. Twenty-four compounds were identified in the A. huostonianum (MNP) and A. huostonianum (DTR) plant extracts. The presence of catechin, chlorogenic acid, p-coumaric acid, vanillic acid, ferulic acid, and rutin was also confirmed through a comparison with purified standards. All of the compounds were identified in the extract derived from A. huostonianum (MNP), with the exception of catechin (1) , heliohoustine (2) , agecorynin D (6) , and precocene 2 (21) . In comparison, extracts of A. huostonianum (DTR) contained all of the compounds, except for vanillic acid (5) , cinnamic acid (7) , ferulic acid (9) , agehoustin D (10) , benzoic acid (12) , ononin (13) , and penduletin (16) . Table 3 Determination of bioactive compounds in methanolic extract of A. huostonianum in positive ionization mode by using HPLC-QTOF-MS/MS collected from two different locations Peak No. t R (min) M. formula Cal. mass [M + H] + Obs. Mass [M + H] + Error (Δppm) Identification Distribution A. huostonianum (MNP) A. huostonianum (DTR) 1 7.5 C 15 H 14 O 6 291.0863 291.0859 0.96 Catechin* - + 2 7.8 C 14 H 23 NO 4 270.17 270.1700 -0.19 Heliohoustine - + 3 8.0 C 16 H 18 O 9 355.1024 355.1026 -0.83 Chlorogenic acid* + + 4 8.4 C 9 H 8 O 3 165.0546 165.0546 0.01 p -Coumaric acid* + + 5 9.1 C 8 H 8 O 4 169.0495 169.0495 0.57 Vanillic acid* + - 6 9.4 C 19 H 18 O 9 391.1024 391.1019 1.02 Agecorynin D - + 7 9.9 C 9 H 8 O 2 149.0597 149.0587 1.03 Cinnamic acid + - 8 10.3 C 9 H 8 O 3 165.0546 165.0546 0.01 o -Coumaric acid + + 9 11.0 C 10 H 10 O 4 195.0652 195.0653 -0.64 Ferulic acid* + - 10 11.2 C 21 H 22 O 10 435.1286 435.1281 1.29 Agehoustin D + - 11 12.0 C 27 H 30 O 16 611.1607 611.1607 -0.01 Rutin* + + 12 12.4 C 7 H 6 O 2 123.0441 123.0442 -0.94 Benzoic acid + - 13 14.2 C 22 H 22 O 9 431.1337 431.1327 1.73 Ononin + - 14 14.4 C 20 H 20 O 8 389.1231 389.1229 0.60 Agehoustin G + + 15 15.2 C 21 H 22 O 9 419.1337 419.1340 -0.68 Agehoustin E + + 16 15.6 C 18 H 16 O 7 345.0969 345.0962 1.98 Penduletin + - 17 15.7 C 20 H 20 O 7 373.1282 373.1279 0.95 Sinensetin + + 18 15.9 C 21 H 22 O 8 403.1387 403.1387 0.02 Nobiletin + + 19 16.4 C 20 H 18 O 8 387.1074 387.1074 -1.29 Linderoflavone B + + 20 16.5 C 21 H 22 O 8 403.1387 403.1384 0.79 5,6,8,3',4',5'-Hexamethoxyflavone + + 21 16.8 C 13 H 16 O 3 221.1172 221.1172 0.02 Precocene 2 - + 22 17.1 C 22 H 24 O 9 433.1493 433.1494 -0.28 Agecorynin C + + 23 18.4 C 21 H 20 O 9 417.118 417.1180 -0.01 Eupalestin + + 24 24.4 C 17 H 26 O 4 295.1904 295.1904 -1.39 1-O-Acetylageratriol + + *Detected as compared with standard Table 4 Determination of bioactive compounds in methanolic extract of M. micrantha in negative ionization mode by using HPLC-QTOF-MS/MS collected from two different locations Peak No. t R (min) M. formula Cal. mass [M + H] + Obs. Mass [M + H] + Error (Δppm) Identification Distribution M. micrantha (MNP) M. micrantha (DTR) 1 7.5 C 15 H 14 O 6 291.0863 291.0859 0.96 Catechin* - + 2 8.0 C 16 H 18 O 9 355.1024 355.1026 -0.83 Chlorogenic acid* + + 3 9.3 C 9 H 10 O 5 199.0601 199.0597 1.03 Syringic acid* + + 4 9.7 C 15 H 16 O 6 293.1020 293.1021 -0.48 Dihydromikanolide - + 5 10.0 C 15 H 14 O 5 275.0914 275.0912 0.88 Mikacynancholide + + 6 10.6 C 17 H 18 O 7 335.1125 335.1126 -0.33 Scandenolide - + 7 12.0 C 27 H 30 O 16 611.1607 611.1607 -0.01 Rutin* + - 8 12.9 C 18 H 16 O 6 329.102 329.1039 -1.29 Salvigenin + - 9 13.8 C 18 H 16 O 10 S 425.0537 425.0536 0.16 Mikanin 3-sulfate + + 10 13.9 C 16 H 12 O 7 317.0656 317.0656 0.04 Nodifloretin + + 11 14.0 C 18 H1 6 O 7 345.0969 345.0967 0.56 Eupatorin + + 12 17.1 C 15 H 20 O 3 249.1485 249.1482 1.39 Quadrangolide + + 13 17.7 C 20 H 30 O 287.2369 287.2366 1.05 Mikanifuran - + 14 17.8 C 18 H 16 O 7 345.0969 345.0966 0.71 Mikanin + + 15 18.5 C 15 H 22 O 2 235.1693 235.1693 -0.15 5-[3-(3,3-Dimethyloxiranyl)-1-methylenepropyl]-2-methyl-2-cyclohexen-1-one + + 16 27.1 C 20 H 32 O 2 305.2475 305.2466 1.83 ent-8(17),12E,14-Labdatriene-3beta,19-diol + + *Detected as compared with standard Similarly, sixteen compounds were identified in the methanolic extracts of M. micrantha (MNP) and M. micrantha (DTR). All of the compounds were identified in the extracts of M. micrantha (MNP), except for catechin (1) , dihydromikanolide (4) , scandenolide (6) , and mikanifuran (13) . In comparison, all of the compounds were identified in the extracts of M. micrantha (DTR), except for rutin (7) and salvigenin (8) . Catechin (1) , chlorogenic acid (2) , syringic acid (3) , and rutin (7) were also identified through a cross-evaluation with the spectra of purified standards. Volatile compounds identified in the extracts via GC-MS The volatile compounds found in the methanol extracts of A. huostonianum (MNP and DTR) and M. micrantha (MNP and DTR) are shown in Tables S1-S4. A total of nine volatile compounds were tentatively detected in the A. huostonianum (MNP and DTR) plant extracts. Notably, seven volatile compounds—Benzenamine, 2,4,6-tribromo; methyl trans-9-(2-butylcyclopentyl)nonanoate; 7-hexadecenoic acid, methyl ester, (Z)-; 1-hexyl-2-nitrocyclohexane; 1,2,3,4-butanetetrol, 1,4-diacetate 2,3-dibenzoate, [S-(R*,R*)]; Cyclotrisiloxane, hexamethyl; and trimethyl[4-(1,1,3,3,-tetramethylbutyl) phenoxy]silane—were found in the A. houstonianum (MNP) plant extracts ( Table S1 ), while only two volatile compounds—2H-1-Benzopyran, 6,7-dimethoxy-2,2-dimethyl and 2,6-DI-T-butyl-4-methylphenol acetate—were found in the extracts of A. houstonianum ( DTR) ( Table S2 ). A total of seven volatile compounds were detected in the M. micrantha (MNP and DTR) plant extracts. Notably, six volatile compounds—Decanal; Cyclotrisiloxane hexamethyl; 2,4,6-Cycloheptatrien-1-one, 3,5-bis-trimethylsilyl; Trimethyl[4-(2-methyl-4-oxo-2-pentyl) phenoxy]silane; 1,2-BIS(Trimethylsilyl)benzene; and Trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy] silane—were present in the M. micrantha MNP plant extracts ( Table S3 ), while four compounds—11-Tridecen-1-ol; Cyclotrisiloxane, hexamethyl; Trimethyl[4-(1,1,3,3,-tetramethylbutyl) phenoxy]silane; and 1,2-BIS(trimethylsilyl) benzene—were identified in the M. micrantha DTR plant extracts ( Table S4 ). The GC-MS peaks were identified by comparing the measured mass spectra with the known mass spectra of the compounds in the National Institute of Standards and Technology (NIST, USA) database. Discussion In the present study, the bioactive potential of methanolic plant extracts from A. houstonianum and M. Micrantha was determined using samples that had been collected from two locations at different altitudes. The total phenolic content, total flavonoid content, antioxidant potential, and cytotoxicity of the A. houstonianum and M. Micrantha plant extracts were evaluated. The presence of phenolic compounds in the plant extracts was determined using HPLC/QTOF-MS, and the VOCs were determined through the GC-MS analysis of the collected plant extracts. The results of this study clearly show variations in the bioactivity potential and composition of the phytochemical constituents of the plant extracts collected from the Dampa Tiger Reserve (23°34′ N, 92°22′ E) and Murlen National Park (23°37′01″ N, 93°18′00″ E) in Mizoram, India. Phenolics comprise a large group of secondary metabolites that are reported to provide a protective effect against UV-B radiation. Spitaler et al. [ 8 ] reported that the biosynthesis of phenolic compounds is enhanced in plants growing at high altitudes. Jin et al. [ 26 , 27 ] reported that the levels of total phenolics and antioxidant activity in red wine significantly increase with the altitude at which the grape plants are grown. This may occur in response to the reactive oxygen species (ROS) that are induced by the elevated UV-B radiation at high altitudes. Similarly, Rieger et al. [ 28 ] reported greater levels of flavonoids in C. vulgaris and S. nigra with increasing altitude. In the present study, both species of medicinal plants growing in the MNP location, which had a higher elevation than the DTR site, exhibited a high level of both total phenolics and total flavonoids. Phenolic compounds are excellent reducing agents that effectively scavenge free radicals. The ortho-dihydroxylated structure of flavonoids traps ultraviolet radiation and scavenges free radicals [ 29 ]. This may be the reason why the plant extracts with high total phenolics and flavonoids exhibited good antioxidant activity. Copaja et al. [ 30 ] reported elevated phytochemical production in plants collected from sites with adverse environmental conditions. The primary reason behind the higher production of phytochemicals may be the extreme fluctuations in temperature and the lack of nutrients. Antioxidants are the primary source of protection that organisms use against free radical molecules that induce oxidative stress [ 31 ]. Free radicals are an essential aspect of cellular biochemistry [ 32 ]. High levels of free radicals, however, can oxidize and damage cellular lipids, proteins, and DNA and cause a metabolic imbalance that induces aging [ 33 ]. In comparison to humans, who have an antioxidant system to protect them, plants contain a wide array of phenolic compounds that possess antioxidant properties [ 34 ]. Interestingly, the number of antioxidants produced by plants can vary between low and high altitudes. In our study, the plant extracts derived from A. houstonianum (MNP) exhibited significantly greater antioxidant activity against ABTS (IC 50 241.6 µg/mL) radicals as compared to the extracts of A. houstonianum (DTR) (IC 50 371.2 µg/mL). A higher level of DPPH (IC 50 109.8 µg/mL) and ABTS (IC 50 194.1 µg/mL) radical scavenging activity was exhibited by the extracts of Mikania micrantha from the MNP site than those from the DTR site. We found that the methanolic extracts of the M. micrantha and A. houstonianum plants growing at the higher altitude exhibited a greater level of DPPH and ABTS radical scavenging activity than the extracts of the same plants collected from the site at the lower altitude (DTR). Overall, the extracts of the M. micrantha plants collected from the MNP site exhibited higher scavenging activity against DPPH and ABTS free radicals as compared to the extracts of the M. micrantha plants collected from the DTR site. Kishore et al. [ 35 ] reported that the maximum amount of DPPH and beta-carotene radical scavenging potential had been detected in methanolic extracts of tartar buckwheat seed samples obtained from plants growing at high altitudes. In our study, however, the extracts of the A. houstonianum (DTR) plants growing at the lower altitude had significantly greater antioxidant activity against DPPH (IC 50 265.1 µg/mL) radicals than the extracts of A. houstonianum (MNP) obtained from plants growing at the higher altitude (IC 50 388.0 µg/mL). In the present study, the antiproliferative of the plant extracts of A. houstonianum and M. micrantha was evaluated using an MTT assay. The MTT assay shows the number of viable cells based on the change of the yellow tetrazolium MTT into purple formazan that results from the enzymatic activity of a mitochondrial dehydrogenase enzyme, which is present only in viable cells. Thus, the amount of formazan is what shows the amount of metabolically active viable cells [ 36 ]. The methanolic extracts of A. houstonianum and M. micrantha obtained from the MNP site exhibited strong antiproliferative against the HepG2 (liver hepatocarcinoma cells), MCF7 (human breast cancer cell), and HeLa (human cervical cancer cell) cell lines as compared to extracts obtained from plants growing at the DTR location. In contrast, the methanolic extracts of the A. houstonianum and M. micrantha plants collected from the DTR location exhibited greater antiproliferative against AGS (human adenoma gastric cancer) cell lines than the plant extracts collected from the MNP site. Bhagat et al. [ 37 ] investigated the anticancer activity of different extracts of Anisochilus carnosus (L.f.) (Lamiaceae) leaves collected from a high altitude. They reported that petroleum ether and ethanolic extracts of A. carnosus had greater antiproliferative against the BT-549 cancer cell line than plant extracts collected at a lower altitude. In another study, ethyl acetate and butanol extracts of the rhizomes of Paris vietnamensis obtained from a high altitude (1579.6 min) in Sapa, Laocai province, Vietnam, exhibited antiproliferative against three cancer cell lines, including the SK-LU-1 cell line (human lung carcinoma), HeLa cell line (human cervical carcinoma), and MKN7 cell line (human gastric carcinoma), with IC50 values ranging from 1.07 to 4.37 µM [ 38 ]. The methanolic extracts of A. houstonianum and M. Micrantha growing at two different locations showed the effect of altitude on the composition of phenolic compounds present in the plants, which was based on their mass, molecular weight, and retention time, along with the m/z ratio. A total of 24 phenolic compounds were identified in A. huostonianum extracts using HPLC-QTOF-MS/MS. Among them, 20 compounds were detected in extracts obtained from plants collected from the MNP location, and 17 compounds were present in extracts obtained from plants growing at the DTR location. A total of 16 phenolic compounds were identified in M. micrantha plant extracts, which included 12 phenolic compounds in extracts obtained from plants at the MNP site and 2 phenolic compounds in the extracts of plants growing at the DTR site. Notably, three compounds (catechin, chlorogenic acid, and rutin) were identified in the extracts of both A. houstonianum and M. micrantha. The compounds identified in the A. huostonianum extracts were catechin ( 1 ) [ 39 ], heliohoustine ( 2 ) [ 40 ], chlorogenic acid ( 3 ) [ 41 ], p -coumaric acid ( 4 ) [ 39 ], vanillic acid ( 5 ) [ 41 ], agecorynin D ( 6 ) [ 42 ], cinnamic acid ( 7 ) [ 41 ], o -coumaric acid ( 8 ) [ 43 ], ferulic acid ( 9 ) [ 41 ], agehoustin D ( 10 ) [ 40 ], rutin ( 11 ) [ 44 ], benzoic acid ( 12 ) [ 39 ], ononin ( 13 ), agehoustin G ( 14 ), agehoustin E ( 15 ), penduletin ( 16 ), sinensetin ( 17 ) [ 43 ], nobiletin ( 18 ) [ 43 ], linderoflavone B ( 19 ) [ 43 ], 5,6,8,3',4',5'-hexamethoxyflavone ( 20 ) [ 43 ], precocene 2 (2 1 ) [ 45 ], agecorynin C ( 22 ) [ 40 ], eupalestin ( 23 ) [ 43 ], and 1-O-acetylageratriol ( 24 ). The thephenolic compounds identified in the M. micrantha extracts were catechin ( 1 ), chlorogenic acid ( 2 ) [ 46 ], syringic acid ( 3 ), dihydromikanolide ( 4 ) [ 47 ], mikacynancholide ( 5 ), scandenolide ( 6 ) [ 47 ], rutin ( 7 ) [ 48 ], salvigenin ( 8 ), mikanin 3-sulfate ( 9 ) [ 49 ], nodifloretin ( 10 ), eupatorin ( 11 ), quadrangolide ( 12 ), mikanifuran ( 13 ), mikanin ( 14 ) [ 50 ], 5-[3-(3,3-Dimethyloxiranyl)-1-methylenepropyl]-2-methyl-2-cyclohexen-1-one ( 15 ), and ent-8(17),12E,14-Labdatriene-3-beta,19-diol ( 16 ). Zhao and Hu [ 51 ] reported that catechin has both antioxidant and anticancer properties. The phenolic compound rutin has antihypertensive, antiviral, and antiplatelet properties that can increase free radical scavenging activity and overall antioxidant capacity [ 52 ]. Chlorogenic acid has a wide array of biological properties, including antibacterial, antioxidant, and anti-carcinogenic activity [ 53 , 54 ]. To the best of our knowledge, this is the first study to report the five compounds in A. huostonianum plant extracts, namely ononin ( 13 ), agehoustin G ( 14 ), agehoustin E ( 15 ), penduletin ( 16 ), and 1-O-acetylageratriol ( 24 ). Similarly, ten compounds in the plant extracts of Mikania micrantha are reported for the first time, namely catechin ( 1 ), syringic acid ( 3 ), mikacynancholide ( 5 ), salvigenin ( 8 ), nodifloretin ( 10 ), eupatorin ( 11 ), quadrangolide ( 12 ), mikanifuran ( 13 ), 5-[3-(3,3-Dimethyloxiranyl)-1-methylenepropyl]-2-methyl-2-cyclohexen-1-one ( 15 ), and ent-8(17),12E,14-Labdatriene-3-beta,19-diol ( 16 ). Kurade et al. [ 16 ] and Zeeshan et al. [ 55 ] previously identified the presence of volatile organic compounds (VOCs) in A. huostonianum , while Ishak et al. [ 56 ] and Perez-Amador et al. [ 15 ] identified the presence of volatile compounds in M. Micrantha extracts. In the present study, we identified volatile compounds in these two plant species growing at two different altitudes. A total of nine VOCs were found in the extracts of A. houstonianum plants growing at the MNP and DTR sites. Seven volatile compounds—Benzenamine, 2,4,6-tribromo [ 1 ]; methyl trans-9-(2-butylcyclopentyl)nonanoate [ 2 ]; 7-hexadecenoic acid, methyl ester, (Z)- [ 3 ]; 1-hexyl-2-nitrocyclohexane [ 4 ]; 1,2,3,4-butanetetrol, 1,4-diacetate 2,3-dibenzoate, [S-(R*,R*)] [ 5 ]; Cyclotrisiloxane, hexamethyl; and trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy]silane [ 7 ]—were found in the A. houstonianum (MNP) plant extracts, while only two VOCs—2H-1-Benzopyran, 6,7-dimethoxy-2,2-dimethyl and 2,6-DI-T-butyl-4-methylphenol acetate—were detected in the A. houstonianum (DTR) plant extracts. A total of seven VOCs were identified in M. micrantha (MNP) and M. micrantha (DTR) plant extracts. Six VOCs—Decanal [ 1 ]; Cyclotrisiloxane, hexamethyl [ 2 ]; 2,4,6-Cycloheptatrien-1-one, 3,5-bis-trimethylsilyl [ 3 ]; Trimethyl[4-(2-methyl-4-oxo-2-pentyl)phenoxy]silane [ 4 ]; 1,2-BIS(Trimethylsilyl)benzene [ 5 ]; and Trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy]silane [ 7 ]—were detected in the M. micrantha (MNP) plant extracts, while four VOCs—11-Tridecen-1-ol [ 1 ]; Cyclotrisiloxane, hexamethyl [ 2 ]; Trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy]silane [ 3 ]; and 1,2-BIS(trimethylsilyl)benzene [ 4 ]—were detected in the M. micrantha (DTR) plant extracts. The effect of altitude on the phytochemical production of plants grown at a higher elevation has not been well-documented, especially for plants growing in their natural habitats. Therefore, there is a great need to investigate the differences in phytochemical composition, especially phenolics, in plants growing at low and high altitudes [ 57 ]. Zidorn [ 58 ] reported that the differences in plant secondary metabolites due to altitude are primarily dependent on ecological factors. Sampio et al. [ 5 ] reported that the seasonal variation in the production of secondary metabolites in Tithonia diversifolia was primarily due to changes in rainfall and temperature. The types of compounds detected in the present study and their respective bioactivities varied in both plant species. The observed variation may be due to several environmental factors, including differences in geographical location and soil fertility. Temperature and other environmental activities such as relative humidity, wind speed, water availability, and elevation are among the most critical factors that affect plant health. Notably, plant location and altitude can alter many eco-physiological factors as well as plant response to those factors [ 59 ]. Conclusions The geographical distribution of plants is one of the most important factors that determine their degree of adaptation to environmental conditions. Their production of primary and secondary metabolites is a result of the interaction between their internal constitution and living environment. In our study, the extracts obtained from the A. huostonianum and M. micrantha plants growing at the MNP site (high elevation) exhibited a higher level of antioxidant activity and cytotoxicity against cancer cell lines relative to the extracts obtained from those growing at the DTR site (low elevation). The plants of both species obtained from the MNP site possessed a larger number of phenolic compounds and volatile organic compounds than the corresponding extracts obtained from the DTR site. The results of this study clearly indicate that plants growing at higher elevations have a greater ability to harness bioactive potential and synthesize more phytochemicals than plants growing at lower elevations. The present study also demonstrated the phytochemical potential of selected plants for the preparation of traditional formulations. Declarations Author Contributions: GS, BPS – Conceptualization & editing; GS – Investigation & Original writing; AKP, NSK, BK & CN – Methodology; GDAQ-Editing. Ethics Approval Not applicable. Consent to Participate Not applicable. Consent for Publication Authors have read and give the permission to publish the current version of the manuscript. Conflict of Interest The authors declare no competing interests. Funding None. Data Availability The data will be provided with reasonable request. Acknowledgement Authors acknowledged the support of the department of Zoology, Jai Narain Vyas University, Jodhpur for providing the essential facilities. References Waterman, P. G. (1992). Roles for secondary metabolites in plants. Ciba Foundation symposium , 171 , 255–275. https://doi.org/10.1002/9780470514344.ch15 . Vining, L. C. (1992). Secondary metabolism, inventive evolution and biochemical diversity - a review. Gene , 115 (1–2), 135–140. https://doi.org/10.1016/0378-1119(92)90551-Y . Verpoorte, R. (1998). Exploration of nature’s chemodiversity: The role of secondary metabolites as leads in drug development. Drug Discovery Today , 3 (5), 232–238. https://doi.org/10.1016/S1359-6446(97)01167-7 . Chapin, F. S., Bloom, A. J., Field, C. B., & Waring, R. H. (1987). Plant Responses to Multiple Environmental Factors. BioScience , 37 (1), 49–57. https://doi.org/10.2307/1310177 . Sampaio, B. L., Edrada-Ebel, R., & Da Costa, F. B. (2016). Effect of the environment on the secondary metabolic profile of Tithonia diversifolia: A model for environmental metabolomics of plants. Scientific Reports , 6 (July), 1–11. https://doi.org/10.1038/srep29265 . Dixon, R. A., & Dickinson, A. J. (2024). A century of studying plant secondary metabolism—From what? to where, how, and why? Plant Physiology , 00 , 1–19. https://doi.org/10.1093/plphys/kiad596 . Chaves, N., Escudero, J. C., & Gutierrez-Merino, C. (1997). Role of ecological variables in the seasonal variation of flavonoid content of Cistus ladanifer exudate. Journal of Chemical Ecology , 23 (3), 579–603. https://doi.org/10.1023/B:JOEC.0000006398.79306.09 . Spitaler, R., Schlorhaufer, P. D., Ellmerer, E. P., Merfort, I., Bortenschlager, S., Stuppner, H., & Zidorn, C. (2006). Altitudinal variation of secondary metabolite profiles in flowering heads of Arnica montana cv. ARBO Phytochemistry , 67 (4), 409–417. https://doi.org/10.1016/j.phytochem.2005.11.018 . Nascimento, L. B. D. S., Leal-Costa, M. V., Menezes, E. A., Lopes, V. R., Muzitano, M. F., Costa, S. S., & Tavares, E. S. (2015). Ultraviolet-B radiation effects on phenolic profile and flavonoid content of Kalanchoe pinnata. Journal of Photochemistry and Photobiology B: Biology , 148 (3), 73–81. https://doi.org/10.1016/j.jphotobiol.2015.03.011 . Sharma, H. K., Chhangte, L., & Dolui, A. K. (2001). Traditional medicinal plants in Mizoram, India. Fitoterapia , 72 (2), 146–161. https://doi.org/10.1016/S0367-326X(00)00278-1 . Dou, X., Zhang, Y., Sun, N., Wu, Y., & Li, L. (2014). The anti-tumor activity of Mikania micrantha aqueous extract in vitro and in vivo. Cytotechnology , 66 (1), 107–117. https://doi.org/10.1007/s10616-013-9543-9 . Debaprotim, D., Suvakanta, D., & Jashabir, C. (2014). Evaluation of Anticancer Activity of Mikania micrantha Kunth (Asteraceae) Against Ehrlich Ascites Carcinoma in Swiss Albino Mice. International Journal of Pharmaceutical Research & Allied Sciences , 3 (2), 9–18. Ghosh, A., Das, B. K., Roy, A., Mandal, B., & Chandra, G. (2008). Antibacterial activity of some medicinal plant extracts. Journal of Natural Medicines , 62 (2), 259–262. https://doi.org/10.1007/s11418-007-0216-x . Li, Y., Li, J., Li, Y., Wang, X., & Cao, A. (2013). Antimicrobial Constituents of the Leaves of Mikania micrantha H. B. K. Plos One , 8 (10 (e76725)), 1–10. https://doi.org/10.1371/journal.pone.0076725 . Pérez-Amador, M. C., Muñoz Ocotero, V., Balcazar, I., R., & García Jiménez, F. (2010). Phytochemical and pharmacological studies on Mikania micrantha H.B.K. (Asteraceae). Phyton , 79 , 77–80. Kurade, N. P., Jaitak, V., Kaul, V. K., & Sharma, O. P. (2010). Chemical composition and antibacterial activity of essential oils of Lantana camara, Ageratum houstonianum and Eupatorium adenophorum. Pharmaceutical Biology , 48 (5), 539–544. https://doi.org/10.3109/13880200903193336 . Kumar Passari, A., Vineeth Leo, V., Singh, G., Samanta, L., Ram, H., Siddaiah, N., & Pratap Singh, C., B (2020). In Vivo Studies of Inoculated Plants and in Vitro Studies Utilizing Methanolic Extracts of Endophytic Streptomyces sp. Strain DBT34 Obtained from Mirabilis Jalapa L. Exhibit ROS-Scavenging and Other Bioactive Properties. International Journal Of Molecular Sciences , 21 , 7364. https://doi.org/10.3390/ijms21197364 . Singh, G., Passsari, A. K., Singh, P., Leo, V. V., Subbarayan, S., Kumar, B., & Kumar, N. S. (2017). Pharmacological potential of Bidens pilosa L. and determination of bioactive compounds using UHPLC-QqQLIT-MS/MS and GC/MS. BMC Complementary and Alternative Medicine , 17 (1), 1–16. https://doi.org/10.1186/s12906-017-2000-0 . Akpoveso, O. P., Ubah, E. E., & Obasanmi, G. (2023). Antioxidant Phytochemicals as Potential Therapy for Diabetic Complications. Antioxidants , 12 , 1–30. Al-dalahmeh, Y., Al-bataineh, N., Al-balawi, S. S., Lahham, J. N., Al-momani, I. F., Al-sheraideh, M. S., & Al-qudah, M. A. (2022). LC-MS/MS Screening, Total Phenolic, Flavonoid and Antioxidant Contents of Crude Extracts from Three Asclepiadaceae Species Growing in Jordan. Molecules , 27 , 1–11. Singh, P., Bajpai, V., Gond, V. K., Kumar, A., & Tadigoppula, N. (2020). and K. B. Determination of Bioactive Compounds of Fenugreek (Trigonella foenum-graecum) Seeds Using LC-MS Techniques. In LLegume Genomics: Methods and Protocols. Methods in Molecular Biology. doi:10.1007/978-1 -0716-0235-5 (pp. 377–392). Retrieved from http://www.springer.com/series/7651. Bajpai, V., Singh, A., Arya, K. R., Srivastava, M., & Kumar, B. (2015). Rapid screening for the adulterants of Berberis aristata using direct analysis in real-time mass spectrometry and principal component analysis for discrimination. Food Additives and Contaminants - Part A Chemistry, Analysis, Control . Exposure and Risk Assessment , 32 (6), 799–807. https://doi.org/10.1080/19440049.2015.1022885 . Kumar, S., Singh, A., & Kumar, B. (2019). Screening of monoterpene indole alkaloids in six Rauwolfia species by ultra-high performance liquid chromatography orbitrap velos pro mass spectrometer. Separation Science Plus , 2 (8), 300–308. https://doi.org/10.1002/sscp.201900029 . Shelke, V., & Bhot, M. (2019). GC-MS Analysis of Bio-active Compounds in Ethanolic Extract of Leaf and Stem of Asclepias curassavica L. Int J Pharm Investigation , 9 (2), 67–70. https://doi.org/10.5330/ijpi.2019.2.13 . Assaad, H. I., Zhou, L., Carroll, R. J., & Wu, G. (2014). Rapid publication-ready MS-Word tables for one-way ANOVA. SpringerPlus , 3 , 1–8. https://doi.org/doi:10.1186/2193-1801-3-474 . Jin, X., Wu, X., & Liu, X. (2017). Phenolic Characteristics and Antioxidant Activity of Merlot and Cabernet Sauvignon Wines Increase with Vineyard Altitude in a High-altitude Region. South African Journal of Enology & Viticulture , 38 (2), 132–143. https://doi.org/10.21548/38-2-1068 . Hagaggi, N. S. A., Abdul-Raouf, U. M., & Radwan, T. A. A. (2024). Variation of antibacterial and antioxidant secondary metabolites and volatiles in leaf and callus extracts of Phulai (Acacia Modesta Wall). BMC Plant Biology , 24 (1), 1–12. https://doi.org/10.1186/s12870-024-04747-9 . Rieger, G., Müller, M., Guttenberger, H., & Bucar, F. (2008). Influence of altitudinal variation on the content of phenolic compounds in wild populations of Calluna vulgaris, Sambucus nigra, and Vaccinium myrtillus. Journal of Agricultural and Food Chemistry , 56 (19), 9080–9086. https://doi.org/10.1021/jf801104e . Liu, W., Yin, D., Li, N., Hou, X., Wang, D., Li, D., & Liu, J. (2016). Influence of environmental factors on the active substance production and antioxidant activity in Potentilla fruticosa L. and its quality assessment. Scientific Reports , 6 (July), 1–18. https://doi.org/10.1038/srep28591 . Copaja, S. V., Blackburn, C., & Carmona, R. (2003). Variation of saponin contents in Quillaja saponica molina. Wood Science and Technology , 37 (2), 103–108. https://doi.org/10.1007/s00226-002-0150-8 . Halliwell, B., & Gutteridge, J. M. C. (2015). Free Radicals in Biology and Medicine . Free Radicals in Biology and Medicine . https://doi.org/10.1093/acprof:oso/9780198717478.001.0001 . Singh, R., Sharad, S., & Kapur, S. (2004). Free radicals and oxidative stress in neurodegenerative diseases: relevance of dietary antioxidants. J Indian Acad Clin Med , 5 (3), 218–225. Valko, M., Leibfritz, D., Moncol, J., Cronin, M. T. D., Mazur, M., & Telser, J. (2007). Free radicals and antioxidants in normal physiological functions and human disease. International Journal of Biochemistry and Cell Biology , 39 (1), 44–84. https://doi.org/10.1016/j.biocel.2006.07.001 . Gaikwad, P., Barik, A., Priyadarsini, K. I., & Rao, B. S. M. (2010). Antioxidant activities of phenols in different solvents using DPPH assay. Research on Chemical Intermediates , 36 (9), 1065–1072. https://doi.org/10.1007/s11164-010-0220-8 . Kishore, G., Ranjan, S., Pandey, A., & Gupta, S. (2010). Influence of altitudinal variation on the antioxidant potential of tartar buckwheat of Western Himalaya. Food Science and Biotechnology , 19 (5), 1355–1363. https://doi.org/10.1007/s10068-010-0193-9 . van de Loosdrecht, A. A., Beelen, R. H. J., Ossenkoppele, G. J., Broekhoven, M. G., & Langenhuijsen, M. M. A. C. (1994). A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. Journal of Immunological Methods , 174 (1–2), 311–320. https://doi.org/10.1016/0022-1759(94)90034-5 . Bhagat, J., Lobo, R., Kumar, N., Mathew, J. E., lizabet, & Pai, A. (2014). Cytotoxic potential of Anisochilus carnosus (L.f.) wall and estimation of luteolin content by HPLC. BMC complementary and alternative medicine , 14 (October), 421. https://doi.org/10.1186/1472-6882-14-421 . Vu, T. T. T., Vu, L. T. K., Nguyen, Q. H., Pham, K., Van, Nguyen, D. T., Nguyen, L. T. N., & Chu, M. H. (2019). Cytotoxic effects of steroidal glycosides isolated from the Paris vietnamensis plant on cancer cell lines and against bacterial strains. Biotechnology and Biotechnological Equipment , 33 (1), 1516–1524. https://doi.org/10.1080/13102818.2019.1676168 . Xuan, T. D., Shinkichi, T., Hong, N. H., Khanh, T. D., & Min, C. I. (2004). Assessment of phytotoxic action of Ageratum conyzoides L. (billy goat weed) on weeds. Crop Protection , 23 (10), 915–922. https://doi.org/10.1016/j.cropro.2004.02.005 . Pharmacognosy Magazine , 10 (37), 18–26. https://doi.org/10.4103/0973-1296.126653. Okereke, S., Chukwudoruo, C., & Nwaokezie, C. (2017). Phytochemical screening using GC-FID and sub- chronic assessment of Hydroethanolic leaf extract of Ageratum conyzoides Linn on albino rats. Journal of Medicinal Plants Studies , 5 (2), 282–287. Quijano, L., Gómez-Garibay, F., Garcia-Perez, G., & Rios, T. (1992). Highly oxygenated flavonoids from Ageratum corymbosum. Phytochemistry , 31 (8), 2859–2862. https://doi.org/10.1016/0031-9422(92)83647-H . Faqueti, L. G., Sandjo, L. P., & Biavatti, M. W. (2017). Simultaneous identification and quantification of polymethoxyflavones, coumarin and phenolic acids in Ageratum conyzoides by UPLC-ESI-QToF-MS and UPLC-PDA. Journal of Pharmaceutical and Biomedical Analysis , 145 , 621–628. https://doi.org/10.1016/j.jpba.2017.07.034 . Akhtar, N., Ihsan-ul-Haq, & Mirza, B. (2018). Phytochemical analysis and comprehensive evaluation of antimicrobial and antioxidant properties of 61 medicinal plant species. Arabian Journal of Chemistry , 11 (8), 1223–1235. https://doi.org/10.1016/j.arabjc.2015.01.013 . Herbert, N. N., & David, S. (1992). Phytochemical Resources for medicine and agriculture . Wen, J., Kang, L., Liu, H., Xiao, Y., Zhang, X., & Chen, Y. (2012). A validated UV-HPLC method for determination of chlorogenic acid in Lepidogrammitis drymoglossoides (Baker) Ching, Polypodiaceae. Pharmacognosy Research , 4 (3), 148–153. https://doi.org/10.4103/0974-8490.99076 . PLoS Neglected Tropical Diseases , 11 (9), 1–16. https://doi.org/10.1371/journal.pntd.0005929. Li, A., Hou, X., & Wei, Y. (2018). Fast screening of flavonoids from switchgrass and: Mikania micrantha by liquid chromatography hybrid-ion trap time-of-flight mass spectrometry. Analytical Methods , 10 (1), 109–122. https://doi.org/10.1039/c7ay02103h . Rufatto, L. C., Gower, A., Schwambach, J., & Moura, S. (2012). Genus Mikania: Chemical composition and phytotherapeutical activity. Brazilian Journal of Pharmacognosy , 22 (6), 1384–1403. https://doi.org/10.1590/S0102-695X2012005000099 . Kiang, A. K., Sim, K. Y., & Yoong, S. W. (1968). Constituents of Mikania cordata (burm. f.) B. L. Robinson (compositae)-II. Phytochemistry , 7 (6), 1035–1037. https://doi.org/10.1016/S0031-9422(00)82193-7 . Zhao, B., & Hu, M. (2013). Gallic acid reduces cell viability, proliferation, invasion and angiogenesis in human cervical cancer cells. Oncology Letters , 6 (6), 1749–1755. https://doi.org/10.3892/ol.2013.1632 . Yang, J., Guo, J., & Yuan, J. (2008). In vitro antioxidant properties of rutin. LWT - Food Science and Technology , 41 (6), 1060–1066. https://doi.org/10.1016/j.lwt.2007.06.010 . Kasai, H., Fukada, S., Yamaizumi, Z., Sugie, S., & Mori, H. (2000). Action of chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesis model. Food and Chemical Toxicology , 38 (5), 467–471. https://doi.org/10.1016/S0278-6915(00)00014-4 . Dos Santos, M. D., Almeida, M. C., Lopes, N. P., & De Souza, G. E. P. (2006). Evaluation of the anti-inflammatory, analgesic and antipyretic activities of the natural polyphenol chlorogenic acid. Biological and Pharmaceutical Bulletin , 29 (11), 2236–2240. https://doi.org/10.1248/bpb.29.2236 . Zeeshan, M., Rizvi, S. M. D., Khan, M. S., & Kumar, A. (2012). Isolation, partial purification and evaluation of bioactive compounds from leaves of Ageratum houstonianum. EXCLI Journal , 11 , 78–88. https://doi.org/10.17877/DE290R-4911 . Ishak, A. H., Shafie, N. H., Esa, N. M., Bahari, H., & Ismail, A. (2018). From weed to medicinal plant: Antioxidant capacities and phytochemicals of various extracts of Mikania micrantha. International Journal of Agriculture and Biology , 20 (3), 561–568. https://doi.org/10.17957/IJAB/15.0522 . Ganzera, M., Guggenberger, M., Stuppner, H., & Zidorn, C. (2008). Altitudinal variation of secondary metabolite profiles in flowering heads of Matricaria chamomilla cv. BONA. Planta Medica , 74 (4), 453–457. https://doi.org/10.1055/s-2008-1034326 . Zidorn, C. (2010). Altitudinal variation of secondary metabolites in flowering heads of the Asteraceae: Trends and causes. Phytochemistry Reviews , 9 (2), 197–203. https://doi.org/10.1007/s11101-009-9143-7 . Wong, S. K., Lim, Y. Y., & Chan, E. W. C. (2009). Antioxidant properties of hibiscus: Species variation, altitudinal change, coastal influence and floral colour change. Journal of Tropical Forest Science , 21 (4), 307–315. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4195244","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":288229841,"identity":"6b298138-87b8-4d7b-bc6d-5dd5b3eb22ca","order_by":0,"name":"Garima Singh","email":"","orcid":"","institution":"Pachhunga University College: Mizoram University Pachhunga University College","correspondingAuthor":false,"prefix":"","firstName":"Garima","middleName":"","lastName":"Singh","suffix":""},{"id":288229842,"identity":"d25a255b-bc39-450d-b99c-75540cb6d03c","order_by":1,"name":"Ajit Kumar Passari","email":"","orcid":"","institution":"Universidad Autonoma de la Ciudad de Mexico: Universidad Autonoma de la Ciudad de Mexico","correspondingAuthor":false,"prefix":"","firstName":"Ajit","middleName":"Kumar","lastName":"Passari","suffix":""},{"id":288229843,"identity":"4c071d30-20e2-460a-ba65-6d16af3954e1","order_by":2,"name":"N Senthil Kumar","email":"","orcid":"","institution":"CSIR-CDRI: Central Drug Research Institute","correspondingAuthor":false,"prefix":"","firstName":"N","middleName":"Senthil","lastName":"Kumar","suffix":""},{"id":288229844,"identity":"23db2101-89fa-4ca2-8780-b11d3c07d7d4","order_by":3,"name":"Brijesh Kumar","email":"","orcid":"","institution":"CDRI: Central Drug Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Brijesh","middleName":"","lastName":"Kumar","suffix":""},{"id":288229845,"identity":"b0f5a10e-5abb-4d48-bb25-6624528f09de","order_by":4,"name":"S Chandra Nayak","email":"","orcid":"","institution":"University of Mysore Department of Biotechnology","correspondingAuthor":false,"prefix":"","firstName":"S","middleName":"Chandra","lastName":"Nayak","suffix":""},{"id":288229846,"identity":"95670dd8-4760-4731-adfc-b81817c311d8","order_by":5,"name":"Heera Ram","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAuklEQVRIiWNgGAWjYBACAzBZwQbh8YARUVrOkKyFsY0BroUwMJc+wPzh4zy+PIMbCYwP3rYxyJgT0mLZl8BgOHMbWzFQC7Ph3DYGHssGQg47w8CQzLuNLXHmjAQ2aV6gFoMDRGg5zDsHrIX9N7FaGJt5G9gS+yUS2JiJ1MLYzDjjGFALz8NmyTnnJIjRwnz4w4eaY4lt7MkHP7wps7EnqAUYKQ1A4hiMIUFQPQzUEK1yFIyCUTAKRiAAABB3OCAsGr/wAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-6743-1321","institution":"Jai Narain Vyas University Faculty of Science","correspondingAuthor":true,"prefix":"","firstName":"Heera","middleName":"","lastName":"Ram","suffix":""},{"id":288229847,"identity":"15507d63-44d9-4a44-876e-46f3137e3177","order_by":6,"name":"Bhim Pratap Singh","email":"","orcid":"","institution":"NIFTEM: National Institute of Food Technology Entrepreneurship and Management","correspondingAuthor":false,"prefix":"","firstName":"Bhim","middleName":"Pratap","lastName":"Singh","suffix":""}],"badges":[],"createdAt":"2024-03-31 10:40:28","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4195244/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4195244/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12010-024-05005-2","type":"published","date":"2024-08-08T15:57:29+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":54490254,"identity":"c4119058-262a-4912-8445-656aaade0bae","added_by":"auto","created_at":"2024-04-11 10:12:51","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":130003,"visible":true,"origin":"","legend":"\u003cp\u003eBase peak chromatograms of a mixture of purified standards and extracts of \u003cem\u003eA. huostonianum \u003c/em\u003e(MNP: Location 1; DTR: Location 2).\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4195244/v1/f4da656b70548edb08d3f715.jpg"},{"id":54490241,"identity":"7a4e5569-bc50-4376-b874-bf28a1c2b5a7","added_by":"auto","created_at":"2024-04-11 10:12:50","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":126621,"visible":true,"origin":"","legend":"\u003cp\u003eBase peak chromatograms of a mixture of purified standards and extracts of \u003cem\u003eM. micrantha \u003c/em\u003e(MNP: Location 1; DTR: Location 2).\u003c/p\u003e","description":"","filename":"Picture2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4195244/v1/5d23dfed2d73946102936032.jpg"},{"id":62298345,"identity":"bbcf86d2-7be1-42eb-ba21-b32fb17c8dda","added_by":"auto","created_at":"2024-08-12 16:12:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1457934,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4195244/v1/a8aac2e2-3ee2-41d9-b277-bc06b52ecdf8.pdf"},{"id":54490249,"identity":"787e2c35-a5d9-4ce9-9eb8-30cc35a3b9d5","added_by":"auto","created_at":"2024-04-11 10:12:50","extension":"docx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":136322,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFiles.docx","url":"https://assets-eu.researchsquare.com/files/rs-4195244/v1/96588c11ddeeafc5546e6dcb.docx"},{"id":54490259,"identity":"7136700c-d4a2-48f0-a47e-8925dda52635","added_by":"auto","created_at":"2024-04-11 10:12:51","extension":"docx","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":19769,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryTablesS1S4.docx","url":"https://assets-eu.researchsquare.com/files/rs-4195244/v1/87f58e4d48f15ddf1a756445.docx"}],"financialInterests":"","formattedTitle":"Determination of UPLC-ESI MS/MS- and GC-MS-based altitudinal variations in the bioactive potential of traditional medicinal plants","fulltext":[{"header":"Introduction","content":"\u003cp\u003ePlant species produce a tremendous range of primary and secondary metabolites such as alkaloids, terpenes, and polyphenols. Primary metabolites are essential for growth, while secondary metabolites are non-essential and mainly involved in functional roles such as the defense and protection of plants against herbivores, pathogens, and biotic and abiotic stress [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Plants contain diverse chemical entities that may exhibit novel and beneficial pharmacological activity. Medicinal plants with bioactive compounds can be beneficial to human health. For instance, they can function as antibiotics, antioxidants, mycotoxins, insecticides, herbicides, etc. [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Medicinal plants have been exploited for their phytochemicals and used for treating various prevalent diseases and common ailments such as malaria, tuberculosis, diarrhea, and asthma [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Some researchers have previously suggested that environmental factors such as light intensity, photoperiod, rainfall, soil properties, and temperature can affect the biosynthesis of secondary metabolites in plants [\u003cspan additionalcitationids=\"CR5\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Other factors, including plant age, season, nutritional status, grazing, or microbial attack, may also impact the composition and quantity of phytocompounds found in plants.\u003c/p\u003e \u003cp\u003eElevated levels of UV-B radiation, which occur at high altitudes, have been reported to induce the biosynthesis of secondary metabolites (phytochemicals) to mitigate radiation damage in plants [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. In 2015, Nascimento et al. reported that UV-B radiation stimulates the biosynthesis of secondary metabolites [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Phenolic compounds are generally known to enhance the bioactivity of plants. Thus, we hypothesized that variations in altitude could affect the chemical composition and bioactivity of extracts derived from plants. Using the HPLC and GC-MS methods, we characterized the variations in the chemical composition of \u003cem\u003eMikania micrantha\u003c/em\u003e and \u003cem\u003eAgeratum houstonianum\u003c/em\u003e collected at different altitudes. We also determined the antioxidant and anticancer properties of the extracts from both plants. \u003cem\u003eM. micrantha\u003c/em\u003e and \u003cem\u003eA. huostonianum\u003c/em\u003e, which are members of the Asteraceae family, are considered to be invasive weeds. Both plant species have a long history of being widely used in traditional medicine in many regions, specifically by the Mizo tribe in Mizoram, Northeast India [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. The antitumor activity of \u003cem\u003eM. micrantha\u003c/em\u003e and its ability to inhibit the growth of cancer cell lines have previously been reported [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. \u003cem\u003eM. micrantha\u003c/em\u003e is also supposed to have antifungal, antibacterial [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], and anti-inflammatory properties [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. \u003cem\u003eA. houstonianum\u003c/em\u003e has been examined for antimicrobial, mosquitocidal, and other types of bioactivity [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In the present study, the variation in the pharmacological properties of both species was evaluated in plants collected from sites at two different altitudes. One site was the Dampa Tiger Reserve (DTR), which is a 500 km\u003csup\u003e2\u003c/sup\u003e protected area of forest located at 23\u0026deg;32'42\"- 23\u0026deg;41\u0026rsquo;36\u0026rdquo; N and 92\u0026deg;13\u0026rsquo;12\u0026rdquo;- 92\u0026deg;27\u0026rsquo;27\u0026rdquo; E. The second site, Murlen National Park (MNP), is also a protected area of forest that has an area of approximately 100 km\u003csup\u003e2\u003c/sup\u003e and is located at 23\u003csup\u003eo\u003c/sup\u003e32\u0026rsquo; -23 \u003csup\u003eo\u003c/sup\u003e 42\u0026rsquo; N and 92\u003csup\u003eo\u003c/sup\u003e13\u0026rsquo; -92\u003csup\u003eo\u003c/sup\u003e 27\u0026rsquo; E. The climate in Mizoram is moderate, and temperatures vary from 10 \u003csup\u003eo\u003c/sup\u003eC to 30 \u003csup\u003eo\u003c/sup\u003eC throughout the year.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant material\u003c/h2\u003e \u003cp\u003eSamples of \u003cem\u003eMikania micrantha\u003c/em\u003e and \u003cem\u003eAgeratum houstonianum\u003c/em\u003e were collected from two different locations in June 2016 from the Dampa Tiger Reserve and Murlen National Park in Mizoram, India. The authenticity of the plants was confirmed by Dr. Kalidas Upadhyay from the Department of Forestry in Mizoram University, and voucher specimens were deposited in the Mizoram University herbarium. Genomic DNA was isolated from both the plant samples using PureLink Genomic Plant DNA Purification Kit (Invitrogen-K183001) according to manufacturer\u0026rsquo;s protocol and amplified maturase K (matK) gene using universal primer (matK Forward 5'-CGATCTATTCATTCAATATTTC-3\u0026lsquo;and matK Reverse 5'-TCTAGCACACGAAAGTCGAAGT-3') a per Cuenoud et al. (2002). PCR reaction conditions for matK gene was initial denaturation at 95\u0026deg;C for 5 min, followed by 40 cycles of denaturation at 95\u0026deg;C for 30 sec, annealing at 49\u0026deg;C for 35 sec and extension at 72\u0026deg;C for 1 min with a final extension step at 72\u0026deg;C for 8 min. The obtained PCR products were analyzed using 1.2% of agarose gel and visualized under Bio-Rad Gel Doc XR\u0026thinsp;+\u0026thinsp;system (Hercules, CA, USA). Finally, the PCR products was purified using the In-vitrogen Pure Link kit (Catalogue No: K310001), and were sequenced at DBT-State Biotech Hub, Department of Biotechnology, Mizoram University, Aizawl, India and sequence deposited in NCBI GenBank with the accession number of \u003cem\u003eAgeratum houstonianum\u003c/em\u003e (MF770205) and \u003cem\u003eMikania micrantha\u003c/em\u003e (MF770206) respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003ePreparation of plant extracts\u003c/h2\u003e \u003cp\u003eCollected plant samples was washed and plant parts were shade dried at room temperature (30 \u003csup\u003e0\u003c/sup\u003eC \u0026plusmn; 2 \u003csup\u003e0\u003c/sup\u003eC). The dried plant material was grinded to powder by using domestic mixture grinder. Around 100 g of dried powder was extracted thrice in 1000 ml of methanol for 48 h with occasional stirring. The extracts were concentrated using rotary evaporator (Buchi, India) at 40 \u003csup\u003e0\u003c/sup\u003eC under reduced pressure and the obtained crude extract was stored at -20 \u003csup\u003e0\u003c/sup\u003eC until further use [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of total phenolics and flavonoids\u003c/h2\u003e \u003cp\u003eThe level of total phenolics in the plant extracts was determined using Folin\u0026ndash;Ciocalteu reagent, as previously described by Singh et al. [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. An aliquot of 10\u0026micro;l of extract was mixed with 90 \u0026micro;l folin reagent (1:10 v/v in water) and 100 \u0026micro;l of 15% Na2CO3 to make the 200 \u0026micro;l volume in a 96 well microplate and recorded using a UV/VIS microplate spectrophotometer (MultiscanTM GO, Thermo Scientific, MA, USA). Gallic acid was used as a standard, and total phenolic content was expressed in gallic acid equivalents/mg of plant extract. Similarly, the level of total flavonoids was evaluated using a modified version of the aluminum colorimetric method, as previously described in [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. 150 \u0026micro;l of methanol extract is mixed with 150 \u0026micro;l of 2% ethanolic AlCl3 and allowed to incubate in dark for 1 h and the absorbance was recorded at 420 nm. The level of total flavonoids in the samples of plant extracts was expressed in \u0026micro;g quercetin equivalents (QE)/mg of plant dry weight, quercetin was used as a standard.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of free radical scavenging potential\u003c/h2\u003e \u003cp\u003eFree radical scavenging capability of methanolic leaves extract of both plants were determined by DPPH assay as described by previous studies [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Briefly, 200 \u0026micro;l of freshly prepared DPPH methanolic solution was added along with different concentrations of plant extracts and incubated for 30 minutes to observe the absorbance at 517nm. Likewise, the level of free radical scavenging potential present in the methanolic plant extracts was determined using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical assays [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Each experiment was repeated three times. The IC\u003csub\u003e50\u003c/sub\u003e concentration of each sample of plant extract was calculated as the amount that could scavenge 50% of the DPPH and ABTS free radicals. Ascorbic acid was used as a positive control.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of antiproliferative\u003c/h2\u003e \u003cp\u003eThe antiproliferative potential of the plant extracts was calculated using an MTT assay. To evaluate cell viability, four cell lines (HepG2: liver hepatocellular carcinoma cell line; MCF7: human breast cancer cell line; AGS: human gastric adenocarcinoma cell line; HeLa: human cervical cancer cell line) were procured from the National Centre for Cell Science (NCCS) in Pune, India. The cells were cultured in DMEM medium supplemented with 10% inactivated fetal bovine serum (FBS), 100 \u0026micro;g/mL penicillin, 100 \u0026micro;g/mL streptomycin, and 5\u0026micro;g/mL amphotericin B, in a humidified atmosphere of 5% CO\u003csub\u003e2\u003c/sub\u003e at 37◦C.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eMTT Assay\u003c/h2\u003e \u003cp\u003eThe viability of the cell lines treated with different concentrations of the plant extracts was first evaluated using the MTT assay, as previously described in [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. All four selected cell lines were trypsinized and seeded in 96-well microtiter plates (approx. 1 \u0026times; 10\u003csup\u003e4\u003c/sup\u003e cells/well) and incubated for 24h at 37\u0026deg;C. The cells were then treated with different concentrations of the plant extracts (10, 25, 50, 75, 100, 125, 150, 175, 200, 225, or 250 \u0026micro;g/mL) and incubated for 72 h. An appropriate blank and control were used in each assay. The culture medium was removed by aspiration, and 20\u0026micro;L of MTT was added to each well and incubated for 4 h. DMSO was added to each well to dissolve the purple formazan of MTT. Absorbance values were recorded at 570nm, and cell viability was calculated based on a standard curve. All MTT assays were carried out in triplicate.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eHPLC determination of phenolic compounds\u003c/h2\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003eInstrumentation\u003c/h2\u003e \u003cdiv id=\"Sec11\" class=\"Section4\"\u003e \u003ch2\u003eHPLC/QTOF-MS conditions\u003c/h2\u003e \u003cp\u003eThe instrumentation conditions were the same as those reported by Singh et al. (Singh et al., 2020), with all parameters set identically for the standards and samples.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eChromatographic acquisitions\u003c/h2\u003e \u003cp\u003eChromatographic separation was carried out using an Xbridge\u0026reg; C18 column (100 mm \u0026times; 4.6 mm, 3.5\u0026micro;m) operated at 25\u0026deg;C. The mobile phase consisted of a 0.1% formic acid aqueous solution (A) and methanol (B); it had a flow rate of 0.5 mL/min with an isocratic elution of 90% (B) and a duration of 15\u0026ndash;25 minutes. The sample injection volume was 1\u0026micro;L. The UV spectra were obtained by scanning the samples in the range 200\u0026ndash;600 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eMass spectrometry\u003c/h2\u003e \u003cp\u003eMass spectrometric analysis was performed on an Agilent 6520 QTOF mass spectrometer in positive ESI mode, as reported by Bajpai et al. [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Appropriate measurements were taken using the auto mass calibration method with an external mass calibration solution, as reported by Kumar et al. [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eGC/MS analysis of volatile compounds\u003c/h2\u003e \u003cp\u003eVolatile organic compounds (VOCs) in the plant extracts were identified using GC/MS spectrometry [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. A Clarus 680 GC utilizing a fused silica column and packed with Elite-5MS (5% biphenyl 95% dimethylpolysiloxane, 30 m \u0026times; 0.25 mm ID \u0026times; 250\u0026micro;m df) was employed, and the components were separated using helium as a carrier gas at a constant flow of 1 mL/min. The injector temperature was set at 260\u0026deg;C during the chromatographic run. A 1\u0026micro;L sample of the plant extract was injected into the instrument, and the oven temperature was adjusted as follows: 60\u0026deg;C for 2 min; followed by 300\u0026deg;C at the rate of 10\u0026deg;C min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; and 300\u0026deg;C, where the sample was held for 6 min. The mass detector conditions were as follows: transfer line temperature, 240\u0026deg;C; ion source temperature, 240\u0026deg;C; ionization mode electron impact, 70 eV; scan time, 0.2 sec; scan interval, 0.1 sec. The fragments ranged from 40 to 600 Da, and the spectra of the components were compared with a database containing the spectra of known components stored in the GC-MS NIST (2008) library.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eAll of the experiments were performed in triplicate, and the obtained results were statistically analyzed via one-way ANOVA using SPSS 20.0 software [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. The presented data represent the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. P-values less than 0.05 were considered significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eTotal phenolic content (TPC)\u003c/h2\u003e \u003cp\u003eThe total phenolic content in the plant extracts was calculated based on the use of a standard curve (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.988). Both \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e plant samples collected from the MNP location had higher total phenolic contents than the corresponding plant samples collected from the DTR location. The obtained data are summarized and presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTotal phenolic content (TPC) and Total flavonoids content (TFC) of methanolic extract of selected plants collected from two locations and their EC\u003csub\u003e50\u003c/sub\u003e values using DPPH and ABTS assay (n\u0026thinsp;=\u0026thinsp;3)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eName of the plant (Location)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTPC, GAE (\u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTFC, QE (\u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eEC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDPPH assay\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eABTS assay\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eAgeratum huostonianum\u003c/em\u003e (MNP)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e453.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e120.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e388.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e241.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eAgeratum huostonianum\u003c/em\u003e (DTR)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e414.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e104.86\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e265.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e371.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eMikania micrantha\u003c/em\u003e (MNP)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e615.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e135.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e109.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e164.80\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eMikania micrantha\u003c/em\u003e(DTR)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e440.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e123.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e194.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e201.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eValues were taken as mean of three replicates (\u0026plusmn;\u0026thinsp;SD). QE: Querctin; GAE: Gallic acid equivalent; MNP: Murlen national park; DTR: Dampa tiger reserve.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eTotal flavonoids\u003c/h2\u003e \u003cp\u003eSamples of the plant extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e collected from the MNP site exhibited a higher concentration of total flavonoids than those collected from the DTR site (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Extracts of \u003cem\u003eM. Micrantha\u003c/em\u003e collected from the MNP site had a higher total flavonoid content (135.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46) than \u003cem\u003eM. Micrantha\u003c/em\u003e extracts from the DTR site (123.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28). Similarly, extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e plants collected from the MNP site exhibited a higher amount of total flavonoids (120.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.93) than what was present in \u003cem\u003eA. houstonianum\u003c/em\u003e collected from the DTR site (104.86\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of free radical scavenging potential\u003c/h2\u003e \u003cp\u003eThe antioxidant potential of the plant extracts was determined by calculating the IC\u003csub\u003e50\u003c/sub\u003e, which represents the concentration of the plant extract needed to scavenge 50% of the free radicals present in the test solution. Extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e collected from the DTR site exhibited good antioxidant potential against DPPH radicals (IC\u003csub\u003e50\u003c/sub\u003e 265.10\u0026micro;g/mL) as compared to extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e plants collected from the MNP site (IC\u003csub\u003e50\u003c/sub\u003e 388.00 \u0026micro;g/mL). The extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e plants collected from the MNP site exhibited better scavenging of ABTS free radicals (IC\u003csub\u003e50\u003c/sub\u003e 241.60 \u0026micro;g/mL) than the extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e collected from the DTR site (IC\u003csub\u003e50\u003c/sub\u003e 371.20 \u0026micro;g/mL) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eAntiproliferative potential of plant extracts\u003c/h2\u003e \u003cp\u003eThe antiproliferative of the plant extracts was evaluated with the use of four cell lines (HepG2: liver hepatocellular carcinoma cell line; MCF7: human breast cancer cell line; AGS: human gastric adenocarcinoma cell line; HeLa: human cervical cancer cell line) and using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium] assay. Extracts obtained from both \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. Micrantha\u003c/em\u003e plants collected from the MNP site were found to be more cytotoxic to the HepG2, MCF7, and HeLa cell lines than the corresponding extracts of both plant species collected from the DTR site (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In contrast, extracts of the \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e plants collected from the DTR site were found to be more cytotoxic to AGS cell lines than the corresponding extracts collected from the MNP site (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCytotoxicity screening of selected plants collected from two locations against cancer cell lines.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eName of the plant (Location)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eCell lines (IC\u003csub\u003e50\u003c/sub\u003e in \u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHepG2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHeLa\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMCF-7\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAGS\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eA. huostonianum\u003c/em\u003e (MNP)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e34.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e23.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e83.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e35.83\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eA. huostonianum\u003c/em\u003e (DTR)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e54.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e46.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e111.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e31.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eM. micrantha\u003c/em\u003e (MNP)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e22.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e124.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e162.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e144.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eM .micrantha\u003c/em\u003e (DTR)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e110.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e211.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e487.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e79.97\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eHepG2: liver hepatocellular carcinoma cell line; MCF7: human breast cancer cell line; AGS: human gastric adenocarcinoma cell line; HeLa: human cervical cancer cell line; MNP: Murlen national park; DTR: Dampa tiger reserve.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003ePhenolic compounds identified via HPLC/QTOF-MS\u003c/h2\u003e \u003cp\u003eThe methanolic extracts of \u003cem\u003eA. huostonianum\u003c/em\u003e (from MNP and DTR) and \u003cem\u003eM. micrantha\u003c/em\u003e (from MNP and DTR) were analyzed with the use of mobile phase consisting of aqueous formic acid (0.1% formic acid) and methanol. Parameters such as column type, column temperature, mobile phase, elution conditions, flow rate, and MS conditions were optimized. The base peak chromatograms (BPCs) of the methanolic extract of \u003cem\u003eA. huostonianum\u003c/em\u003e (from MNP and DTR) are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, whereas the base peak chromatograms of the methanolic extract of \u003cem\u003eM. micrantha\u003c/em\u003e (from MNP and DTR) are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The retention time (calculated as \u003cem\u003em/z\u003c/em\u003e [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e), molecular formula, error (Δppm), and distribution, along with a compound assignment, are presented in Tables\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. All of the compounds were identified based on their exact mass, molecular formula calculation, and database search results. Twenty-four compounds were identified in the \u003cem\u003eA. huostonianum\u003c/em\u003e (MNP) and \u003cem\u003eA. huostonianum\u003c/em\u003e (DTR) plant extracts. The presence of catechin, chlorogenic acid, p-coumaric acid, vanillic acid, ferulic acid, and rutin was also confirmed through a comparison with purified standards. All of the compounds were identified in the extract derived from \u003cem\u003eA. huostonianum\u003c/em\u003e (MNP), with the exception of catechin \u003cb\u003e(1)\u003c/b\u003e, heliohoustine \u003cb\u003e(2)\u003c/b\u003e, agecorynin D \u003cb\u003e(6)\u003c/b\u003e, and precocene 2 \u003cb\u003e(21)\u003c/b\u003e. In comparison, extracts of \u003cem\u003eA. huostonianum\u003c/em\u003e (DTR) contained all of the compounds, except for vanillic acid \u003cb\u003e(5)\u003c/b\u003e, cinnamic acid \u003cb\u003e(7)\u003c/b\u003e, ferulic acid \u003cb\u003e(9)\u003c/b\u003e, agehoustin D \u003cb\u003e(10)\u003c/b\u003e, benzoic acid \u003cb\u003e(12)\u003c/b\u003e, ononin \u003cb\u003e(13)\u003c/b\u003e, and penduletin \u003cb\u003e(16)\u003c/b\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDetermination of bioactive compounds in methanolic extract of \u003cem\u003eA. huostonianum\u003c/em\u003e in positive ionization mode by using HPLC-QTOF-MS/MS collected from two different locations\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePeak No.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003et\u003csub\u003eR\u003c/sub\u003e\u003c/p\u003e \u003cp\u003e(min)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eM. formula\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCal. mass [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eObs. Mass [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eError (Δppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIdentification\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003eDistribution\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003eA. huostonianum\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003e(MNP)\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003eA. huostonianum (DTR)\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e291.0863\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e291.0859\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCatechin*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e23\u003c/sub\u003eNO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e270.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e270.1700\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eHeliohoustine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e355.1024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e355.1026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eChlorogenic acid*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e165.0546\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e165.0546\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e-Coumaric acid*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e169.0495\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e169.0495\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eVanillic acid*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e391.1024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e391.1019\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAgecorynin D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e149.0597\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e149.0587\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCinnamic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e165.0546\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e165.0546\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eo\u003c/em\u003e-Coumaric acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e10\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e195.0652\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e195.0653\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eFerulic acid*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e435.1286\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e435.1281\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAgehoustin D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e16\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e611.1607\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e611.1607\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eRutin*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e7\u003c/sub\u003eH\u003csub\u003e6\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e123.0441\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e123.0442\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eBenzoic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e431.1337\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e431.1327\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOnonin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e389.1231\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e389.1229\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAgehoustin G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e419.1337\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e419.1340\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAgehoustin E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e345.0969\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e345.0962\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePenduletin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e373.1282\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e373.1279\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSinensetin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e403.1387\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e403.1387\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNobiletin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e387.1074\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e387.1074\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-1.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eLinderoflavone B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e403.1387\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e403.1384\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5,6,8,3',4',5'-Hexamethoxyflavone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e13\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e221.1172\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e221.1172\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePrecocene 2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e433.1493\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e433.1494\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAgecorynin C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e417.118\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e417.1180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eEupalestin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e26\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e295.1904\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e295.1904\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-1.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1-O-Acetylageratriol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003e*Detected as compared with standard\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDetermination of bioactive compounds in methanolic extract of \u003cem\u003eM. micrantha\u003c/em\u003e in negative ionization mode by using HPLC-QTOF-MS/MS collected from two different locations\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePeak No.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003et\u003csub\u003eR\u003c/sub\u003e\u003c/p\u003e \u003cp\u003e(min)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eM. formula\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCal. mass [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eObs. Mass [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eError (Δppm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eIdentification\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003eDistribution\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003eM. micrantha\u003c/em\u003e (MNP)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003eM. micrantha\u003c/em\u003e (DTR)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e291.0863\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e291.0859\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCatechin*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e355.1024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e355.1026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eChlorogenic acid*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e199.0601\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e199.0597\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSyringic acid*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e293.1020\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e293.1021\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eDihydromikanolide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e275.0914\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e275.0912\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMikacynancholide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e335.1125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e335.1126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eScandenolide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e16\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e611.1607\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e611.1607\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eRutin*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e329.102\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e329.1039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-1.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eSalvigenin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e13.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e10\u003c/sub\u003eS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e425.0537\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e425.0536\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMikanin 3-sulfate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e13.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e317.0656\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e317.0656\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNodifloretin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH1\u003csub\u003e6\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e345.0969\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e345.0967\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eEupatorin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e249.1485\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e249.1482\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eQuadrangolide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e287.2369\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e287.2366\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMikanifuran\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e345.0969\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e345.0966\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMikanin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e235.1693\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e235.1693\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5-[3-(3,3-Dimethyloxiranyl)-1-methylenepropyl]-2-methyl-2-cyclohexen-1-one\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e305.2475\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e305.2466\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eent-8(17),12E,14-Labdatriene-3beta,19-diol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003e*Detected as compared with standard\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eSimilarly, sixteen compounds were identified in the methanolic extracts of \u003cem\u003eM. micrantha\u003c/em\u003e (MNP) and \u003cem\u003eM. micrantha\u003c/em\u003e (DTR). All of the compounds were identified in the extracts of \u003cem\u003eM. micrantha\u003c/em\u003e (MNP), except for catechin \u003cb\u003e(1)\u003c/b\u003e, dihydromikanolide \u003cb\u003e(4)\u003c/b\u003e, scandenolide \u003cb\u003e(6)\u003c/b\u003e, and mikanifuran \u003cb\u003e(13)\u003c/b\u003e. In comparison, all of the compounds were identified in the extracts of \u003cem\u003eM. micrantha\u003c/em\u003e (DTR), except for rutin \u003cb\u003e(7)\u003c/b\u003e and salvigenin \u003cb\u003e(8)\u003c/b\u003e. Catechin \u003cb\u003e(1)\u003c/b\u003e, chlorogenic acid \u003cb\u003e(2)\u003c/b\u003e, syringic acid \u003cb\u003e(3)\u003c/b\u003e, and rutin \u003cb\u003e(7)\u003c/b\u003e were also identified through a cross-evaluation with the spectra of purified standards.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eVolatile compounds identified in the extracts via GC-MS\u003c/h2\u003e \u003cp\u003eThe volatile compounds found in the methanol extracts of \u003cem\u003eA. huostonianum\u003c/em\u003e (MNP and DTR) and \u003cem\u003eM. micrantha\u003c/em\u003e (MNP and DTR) are shown in \u003cb\u003eTables S1-S4.\u003c/b\u003e A total of nine volatile compounds were tentatively detected in the \u003cem\u003eA. huostonianum\u003c/em\u003e (MNP and DTR) plant extracts. Notably, seven volatile compounds\u0026mdash;Benzenamine, 2,4,6-tribromo; methyl trans-9-(2-butylcyclopentyl)nonanoate; 7-hexadecenoic acid, methyl ester, (Z)-; 1-hexyl-2-nitrocyclohexane; 1,2,3,4-butanetetrol, 1,4-diacetate 2,3-dibenzoate, [S-(R*,R*)]; Cyclotrisiloxane, hexamethyl; and trimethyl[4-(1,1,3,3,-tetramethylbutyl) phenoxy]silane\u0026mdash;were found in the \u003cem\u003eA. houstonianum\u003c/em\u003e (MNP) plant extracts (\u003cb\u003eTable \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e\u003c/b\u003e), while only two volatile compounds\u0026mdash;2H-1-Benzopyran, 6,7-dimethoxy-2,2-dimethyl and 2,6-DI-T-butyl-4-methylphenol acetate\u0026mdash;were found in the extracts of \u003cem\u003eA. houstonianum (\u003c/em\u003eDTR) (\u003cb\u003eTable \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e\u003c/b\u003e). A total of seven volatile compounds were detected in the \u003cem\u003eM. micrantha\u003c/em\u003e (MNP and DTR) plant extracts. Notably, six volatile compounds\u0026mdash;Decanal; Cyclotrisiloxane hexamethyl; 2,4,6-Cycloheptatrien-1-one, 3,5-bis-trimethylsilyl; Trimethyl[4-(2-methyl-4-oxo-2-pentyl) phenoxy]silane; 1,2-BIS(Trimethylsilyl)benzene; and Trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy] silane\u0026mdash;were present in the \u003cem\u003eM. micrantha\u003c/em\u003e MNP plant extracts (\u003cb\u003eTable S3\u003c/b\u003e), while four compounds\u0026mdash;11-Tridecen-1-ol; Cyclotrisiloxane, hexamethyl; Trimethyl[4-(1,1,3,3,-tetramethylbutyl) phenoxy]silane; and 1,2-BIS(trimethylsilyl) benzene\u0026mdash;were identified in the \u003cem\u003eM. micrantha\u003c/em\u003e DTR plant extracts (\u003cb\u003eTable S4\u003c/b\u003e). The GC-MS peaks were identified by comparing the measured mass spectra with the known mass spectra of the compounds in the National Institute of Standards and Technology (NIST, USA) database.\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn the present study, the bioactive potential of methanolic plant extracts from \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. Micrantha\u003c/em\u003e was determined using samples that had been collected from two locations at different altitudes. The total phenolic content, total flavonoid content, antioxidant potential, and cytotoxicity of the \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. Micrantha\u003c/em\u003e plant extracts were evaluated. The presence of phenolic compounds in the plant extracts was determined using HPLC/QTOF-MS, and the VOCs were determined through the GC-MS analysis of the collected plant extracts. The results of this study clearly show variations in the bioactivity potential and composition of the phytochemical constituents of the plant extracts collected from the Dampa Tiger Reserve (23\u0026deg;34\u0026prime; N, 92\u0026deg;22\u0026prime; E) and Murlen National Park (23\u0026deg;37\u0026prime;01\u0026Prime; N, 93\u0026deg;18\u0026prime;00\u0026Prime; E) in Mizoram, India. Phenolics comprise a large group of secondary metabolites that are reported to provide a protective effect against UV-B radiation. Spitaler et al. [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e] reported that the biosynthesis of phenolic compounds is enhanced in plants growing at high altitudes. Jin et al. [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] reported that the levels of total phenolics and antioxidant activity in red wine significantly increase with the altitude at which the grape plants are grown. This may occur in response to the reactive oxygen species (ROS) that are induced by the elevated UV-B radiation at high altitudes. Similarly, Rieger et al. [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] reported greater levels of flavonoids in \u003cem\u003eC. vulgaris\u003c/em\u003e and \u003cem\u003eS. nigra\u003c/em\u003e with increasing altitude. In the present study, both species of medicinal plants growing in the MNP location, which had a higher elevation than the DTR site, exhibited a high level of both total phenolics and total flavonoids. Phenolic compounds are excellent reducing agents that effectively scavenge free radicals. The ortho-dihydroxylated structure of flavonoids traps ultraviolet radiation and scavenges free radicals [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. This may be the reason why the plant extracts with high total phenolics and flavonoids exhibited good antioxidant activity. Copaja et al. [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] reported elevated phytochemical production in plants collected from sites with adverse environmental conditions. The primary reason behind the higher production of phytochemicals may be the extreme fluctuations in temperature and the lack of nutrients.\u003c/p\u003e \u003cp\u003eAntioxidants are the primary source of protection that organisms use against free radical molecules that induce oxidative stress [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Free radicals are an essential aspect of cellular biochemistry [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. High levels of free radicals, however, can oxidize and damage cellular lipids, proteins, and DNA and cause a metabolic imbalance that induces aging [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. In comparison to humans, who have an antioxidant system to protect them, plants contain a wide array of phenolic compounds that possess antioxidant properties [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Interestingly, the number of antioxidants produced by plants can vary between low and high altitudes. In our study, the plant extracts derived from \u003cem\u003eA. houstonianum\u003c/em\u003e (MNP) exhibited significantly greater antioxidant activity against ABTS (IC\u003csub\u003e50\u003c/sub\u003e 241.6 \u0026micro;g/mL) radicals as compared to the extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e (DTR) (IC\u003csub\u003e50\u003c/sub\u003e 371.2 \u0026micro;g/mL). A higher level of DPPH (IC\u003csub\u003e50\u003c/sub\u003e 109.8 \u0026micro;g/mL) and ABTS (IC\u003csub\u003e50\u003c/sub\u003e 194.1 \u0026micro;g/mL) radical scavenging activity was exhibited by the extracts of \u003cem\u003eMikania micrantha\u003c/em\u003e from the MNP site than those from the DTR site. We found that the methanolic extracts of the \u003cem\u003eM. micrantha\u003c/em\u003e and \u003cem\u003eA. houstonianum\u003c/em\u003e plants growing at the higher altitude exhibited a greater level of DPPH and ABTS radical scavenging activity than the extracts of the same plants collected from the site at the lower altitude (DTR). Overall, the extracts of the \u003cem\u003eM. micrantha\u003c/em\u003e plants collected from the MNP site exhibited higher scavenging activity against DPPH and ABTS free radicals as compared to the extracts of the \u003cem\u003eM. micrantha\u003c/em\u003e plants collected from the DTR site. Kishore et al. [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e] reported that the maximum amount of DPPH and beta-carotene radical scavenging potential had been detected in methanolic extracts of tartar buckwheat seed samples obtained from plants growing at high altitudes. In our study, however, the extracts of the \u003cem\u003eA. houstonianum\u003c/em\u003e (DTR) plants growing at the lower altitude had significantly greater antioxidant activity against DPPH (IC\u003csub\u003e50\u003c/sub\u003e 265.1 \u0026micro;g/mL) radicals than the extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e (MNP) obtained from plants growing at the higher altitude (IC\u003csub\u003e50\u003c/sub\u003e 388.0 \u0026micro;g/mL).\u003c/p\u003e \u003cp\u003eIn the present study, the antiproliferative of the plant extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e was evaluated using an MTT assay. The MTT assay shows the number of viable cells based on the change of the yellow tetrazolium MTT into purple formazan that results from the enzymatic activity of a mitochondrial dehydrogenase enzyme, which is present only in viable cells. Thus, the amount of formazan is what shows the amount of metabolically active viable cells [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The methanolic extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e obtained from the MNP site exhibited strong antiproliferative against the HepG2 (liver hepatocarcinoma cells), MCF7 (human breast cancer cell), and HeLa (human cervical cancer cell) cell lines as compared to extracts obtained from plants growing at the DTR location. In contrast, the methanolic extracts of the \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e plants collected from the DTR location exhibited greater antiproliferative against AGS (human adenoma gastric cancer) cell lines than the plant extracts collected from the MNP site. Bhagat et al. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] investigated the anticancer activity of different extracts of \u003cem\u003eAnisochilus carnosus\u003c/em\u003e (L.f.) (Lamiaceae) leaves collected from a high altitude. They reported that petroleum ether and ethanolic extracts of \u003cem\u003eA. carnosus\u003c/em\u003e had greater antiproliferative against the BT-549 cancer cell line than plant extracts collected at a lower altitude. In another study, ethyl acetate and butanol extracts of the rhizomes of \u003cem\u003eParis vietnamensis\u003c/em\u003e obtained from a high altitude (1579.6 min) in Sapa, Laocai province, Vietnam, exhibited antiproliferative against three cancer cell lines, including the SK-LU-1 cell line (human lung carcinoma), HeLa cell line (human cervical carcinoma), and MKN7 cell line (human gastric carcinoma), with IC50 values ranging from 1.07 to 4.37 \u0026micro;M [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe methanolic extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. Micrantha\u003c/em\u003e growing at two different locations showed the effect of altitude on the composition of phenolic compounds present in the plants, which was based on their mass, molecular weight, and retention time, along with the m/z ratio. A total of 24 phenolic compounds were identified in \u003cem\u003eA. huostonianum\u003c/em\u003e extracts using HPLC-QTOF-MS/MS. Among them, 20 compounds were detected in extracts obtained from plants collected from the MNP location, and 17 compounds were present in extracts obtained from plants growing at the DTR location. A total of 16 phenolic compounds were identified in \u003cem\u003eM. micrantha\u003c/em\u003e plant extracts, which included 12 phenolic compounds in extracts obtained from plants at the MNP site and 2 phenolic compounds in the extracts of plants growing at the DTR site. Notably, three compounds (catechin, chlorogenic acid, and rutin) were identified in the extracts of both \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha.\u003c/em\u003e The compounds identified in the \u003cem\u003eA. huostonianum\u003c/em\u003e extracts were catechin (\u003cb\u003e1\u003c/b\u003e) [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e], heliohoustine (\u003cb\u003e2\u003c/b\u003e) [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e], chlorogenic acid (\u003cb\u003e3\u003c/b\u003e) [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e], \u003cem\u003ep\u003c/em\u003e-coumaric acid (\u003cb\u003e4\u003c/b\u003e) [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e], vanillic acid (\u003cb\u003e5\u003c/b\u003e) [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e], agecorynin D (\u003cb\u003e6\u003c/b\u003e) [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e], cinnamic acid (\u003cb\u003e7\u003c/b\u003e) [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e], \u003cem\u003eo\u003c/em\u003e-coumaric acid (\u003cb\u003e8\u003c/b\u003e) [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], ferulic acid (\u003cb\u003e9\u003c/b\u003e) [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e], agehoustin D (\u003cb\u003e10\u003c/b\u003e) [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e], rutin (\u003cb\u003e11\u003c/b\u003e) [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e], benzoic acid (\u003cb\u003e12\u003c/b\u003e) [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e], ononin (\u003cb\u003e13\u003c/b\u003e), agehoustin G (\u003cb\u003e14\u003c/b\u003e), agehoustin E (\u003cb\u003e15\u003c/b\u003e), penduletin (\u003cb\u003e16\u003c/b\u003e), sinensetin (\u003cb\u003e17\u003c/b\u003e) [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], nobiletin (\u003cb\u003e18\u003c/b\u003e) [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], linderoflavone B (\u003cb\u003e19\u003c/b\u003e) [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], 5,6,8,3',4',5'-hexamethoxyflavone (\u003cb\u003e20\u003c/b\u003e) [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], precocene 2 (2\u003cb\u003e1\u003c/b\u003e) [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e], agecorynin C (\u003cb\u003e22\u003c/b\u003e) [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e], eupalestin (\u003cb\u003e23\u003c/b\u003e) [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], and 1-O-acetylageratriol (\u003cb\u003e24\u003c/b\u003e). The thephenolic compounds identified in the \u003cem\u003eM. micrantha\u003c/em\u003e extracts were catechin (\u003cb\u003e1\u003c/b\u003e), chlorogenic acid (\u003cb\u003e2\u003c/b\u003e) [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e], syringic acid (\u003cb\u003e3\u003c/b\u003e), dihydromikanolide (\u003cb\u003e4\u003c/b\u003e) [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e], mikacynancholide (\u003cb\u003e5\u003c/b\u003e), scandenolide (\u003cb\u003e6\u003c/b\u003e) [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e], rutin (\u003cb\u003e7\u003c/b\u003e) [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e], salvigenin (\u003cb\u003e8\u003c/b\u003e), mikanin 3-sulfate (\u003cb\u003e9\u003c/b\u003e) [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e], nodifloretin (\u003cb\u003e10\u003c/b\u003e), eupatorin (\u003cb\u003e11\u003c/b\u003e), quadrangolide (\u003cb\u003e12\u003c/b\u003e), mikanifuran (\u003cb\u003e13\u003c/b\u003e), mikanin (\u003cb\u003e14\u003c/b\u003e) [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e], 5-[3-(3,3-Dimethyloxiranyl)-1-methylenepropyl]-2-methyl-2-cyclohexen-1-one (\u003cb\u003e15\u003c/b\u003e), and ent-8(17),12E,14-Labdatriene-3-beta,19-diol (\u003cb\u003e16\u003c/b\u003e). Zhao and Hu [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e] reported that catechin has both antioxidant and anticancer properties. The phenolic compound rutin has antihypertensive, antiviral, and antiplatelet properties that can increase free radical scavenging activity and overall antioxidant capacity [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]. Chlorogenic acid has a wide array of biological properties, including antibacterial, antioxidant, and anti-carcinogenic activity [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo the best of our knowledge, this is the first study to report the five compounds in \u003cem\u003eA. huostonianum\u003c/em\u003e plant extracts, namely ononin (\u003cb\u003e13\u003c/b\u003e), agehoustin G (\u003cb\u003e14\u003c/b\u003e), agehoustin E (\u003cb\u003e15\u003c/b\u003e), penduletin (\u003cb\u003e16\u003c/b\u003e), and 1-O-acetylageratriol (\u003cb\u003e24\u003c/b\u003e). Similarly, ten compounds in the plant extracts of \u003cem\u003eMikania micrantha\u003c/em\u003e are reported for the first time, namely catechin (\u003cb\u003e1\u003c/b\u003e), syringic acid (\u003cb\u003e3\u003c/b\u003e), mikacynancholide (\u003cb\u003e5\u003c/b\u003e), salvigenin (\u003cb\u003e8\u003c/b\u003e), nodifloretin (\u003cb\u003e10\u003c/b\u003e), eupatorin (\u003cb\u003e11\u003c/b\u003e), quadrangolide (\u003cb\u003e12\u003c/b\u003e), mikanifuran (\u003cb\u003e13\u003c/b\u003e), 5-[3-(3,3-Dimethyloxiranyl)-1-methylenepropyl]-2-methyl-2-cyclohexen-1-one (\u003cb\u003e15\u003c/b\u003e), and ent-8(17),12E,14-Labdatriene-3-beta,19-diol (\u003cb\u003e16\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eKurade et al. [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] and Zeeshan et al. [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e] previously identified the presence of volatile organic compounds (VOCs) in \u003cem\u003eA. huostonianum\u003c/em\u003e, while Ishak et al. [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e] and Perez-Amador et al. [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] identified the presence of volatile compounds in \u003cem\u003eM. Micrantha\u003c/em\u003e extracts. In the present study, we identified volatile compounds in these two plant species growing at two different altitudes. A total of nine VOCs were found in the extracts of \u003cem\u003eA. houstonianum\u003c/em\u003e plants growing at the MNP and DTR sites. Seven volatile compounds\u0026mdash;Benzenamine, 2,4,6-tribromo [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]; methyl trans-9-(2-butylcyclopentyl)nonanoate [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]; 7-hexadecenoic acid, methyl ester, (Z)- [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]; 1-hexyl-2-nitrocyclohexane [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]; 1,2,3,4-butanetetrol, 1,4-diacetate 2,3-dibenzoate, [S-(R*,R*)] [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]; Cyclotrisiloxane, hexamethyl; and trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy]silane [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u0026mdash;were found in the \u003cem\u003eA. houstonianum\u003c/em\u003e (MNP) plant extracts, while only two VOCs\u0026mdash;2H-1-Benzopyran, 6,7-dimethoxy-2,2-dimethyl and 2,6-DI-T-butyl-4-methylphenol acetate\u0026mdash;were detected in the \u003cem\u003eA. houstonianum\u003c/em\u003e (DTR) plant extracts. A total of seven VOCs were identified in \u003cem\u003eM. micrantha\u003c/em\u003e (MNP) and \u003cem\u003eM. micrantha\u003c/em\u003e (DTR) plant extracts. Six VOCs\u0026mdash;Decanal [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]; Cyclotrisiloxane, hexamethyl [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]; 2,4,6-Cycloheptatrien-1-one, 3,5-bis-trimethylsilyl [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]; Trimethyl[4-(2-methyl-4-oxo-2-pentyl)phenoxy]silane [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]; 1,2-BIS(Trimethylsilyl)benzene [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]; and Trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy]silane [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u0026mdash;were detected in the \u003cem\u003eM. micrantha\u003c/em\u003e (MNP) plant extracts, while four VOCs\u0026mdash;11-Tridecen-1-ol [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]; Cyclotrisiloxane, hexamethyl [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]; Trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy]silane [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]; and 1,2-BIS(trimethylsilyl)benzene [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]\u0026mdash;were detected in the \u003cem\u003eM. micrantha\u003c/em\u003e (DTR) plant extracts.\u003c/p\u003e \u003cp\u003eThe effect of altitude on the phytochemical production of plants grown at a higher elevation has not been well-documented, especially for plants growing in their natural habitats. Therefore, there is a great need to investigate the differences in phytochemical composition, especially phenolics, in plants growing at low and high altitudes [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. Zidorn [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e] reported that the differences in plant secondary metabolites due to altitude are primarily dependent on ecological factors. Sampio et al. [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] reported that the seasonal variation in the production of secondary metabolites in \u003cem\u003eTithonia diversifolia\u003c/em\u003e was primarily due to changes in rainfall and temperature. The types of compounds detected in the present study and their respective bioactivities varied in both plant species. The observed variation may be due to several environmental factors, including differences in geographical location and soil fertility. Temperature and other environmental activities such as relative humidity, wind speed, water availability, and elevation are among the most critical factors that affect plant health. Notably, plant location and altitude can alter many eco-physiological factors as well as plant response to those factors [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe geographical distribution of plants is one of the most important factors that determine their degree of adaptation to environmental conditions. Their production of primary and secondary metabolites is a result of the interaction between their internal constitution and living environment. In our study, the extracts obtained from the \u003cem\u003eA. huostonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e plants growing at the MNP site (high elevation) exhibited a higher level of antioxidant activity and cytotoxicity against cancer cell lines relative to the extracts obtained from those growing at the DTR site (low elevation). The plants of both species obtained from the MNP site possessed a larger number of phenolic compounds and volatile organic compounds than the corresponding extracts obtained from the DTR site. The results of this study clearly indicate that plants growing at higher elevations have a greater ability to harness bioactive potential and synthesize more phytochemicals than plants growing at lower elevations. The present study also demonstrated the phytochemical potential of selected plants for the preparation of traditional formulations.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGS, BPS \u0026ndash; Conceptualization \u0026amp; editing; GS \u0026ndash; Investigation \u0026amp; Original writing; AKP, NSK, BK \u0026amp; CN \u0026ndash; Methodology; GDAQ-Editing.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Approval\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. \u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. \u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publication\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors have read and give the permission to publish the current version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data will be provided with reasonable request.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgement\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors acknowledged the support of the department of Zoology, Jai Narain Vyas University, Jodhpur for providing the essential facilities.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eWaterman, P. G. (1992). Roles for secondary metabolites in plants. \u003cem\u003eCiba Foundation symposium\u003c/em\u003e, \u003cem\u003e171\u003c/em\u003e, 255\u0026ndash;275. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/9780470514344.ch15\u003c/span\u003e\u003cspan address=\"10.1002/9780470514344.ch15\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVining, L. C. (1992). Secondary metabolism, inventive evolution and biochemical diversity - a review. \u003cem\u003eGene\u003c/em\u003e, \u003cem\u003e115\u003c/em\u003e(1\u0026ndash;2), 135\u0026ndash;140. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0378-1119(92)90551-Y\u003c/span\u003e\u003cspan address=\"10.1016/0378-1119(92)90551-Y\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVerpoorte, R. (1998). Exploration of nature\u0026rsquo;s chemodiversity: The role of secondary metabolites as leads in drug development. \u003cem\u003eDrug Discovery Today\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e(5), 232\u0026ndash;238. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S1359-6446(97)01167-7\u003c/span\u003e\u003cspan address=\"10.1016/S1359-6446(97)01167-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChapin, F. S., Bloom, A. J., Field, C. B., \u0026amp; Waring, R. H. (1987). Plant Responses to Multiple Environmental Factors. \u003cem\u003eBioScience\u003c/em\u003e, \u003cem\u003e37\u003c/em\u003e(1), 49\u0026ndash;57. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2307/1310177\u003c/span\u003e\u003cspan address=\"10.2307/1310177\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSampaio, B. L., Edrada-Ebel, R., \u0026amp; Da Costa, F. B. (2016). Effect of the environment on the secondary metabolic profile of Tithonia diversifolia: A model for environmental metabolomics of plants. \u003cem\u003eScientific Reports\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e(July), 1\u0026ndash;11. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/srep29265\u003c/span\u003e\u003cspan address=\"10.1038/srep29265\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDixon, R. A., \u0026amp; Dickinson, A. J. (2024). A century of studying plant secondary metabolism\u0026mdash;From what? to where, how, and why? \u003cem\u003ePlant Physiology\u003c/em\u003e, \u003cem\u003e00\u003c/em\u003e, 1\u0026ndash;19. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/plphys/kiad596\u003c/span\u003e\u003cspan address=\"10.1093/plphys/kiad596\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChaves, N., Escudero, J. C., \u0026amp; Gutierrez-Merino, C. (1997). Role of ecological variables in the seasonal variation of flavonoid content of Cistus ladanifer exudate. \u003cem\u003eJournal of Chemical Ecology\u003c/em\u003e, \u003cem\u003e23\u003c/em\u003e(3), 579\u0026ndash;603. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1023/B:JOEC.0000006398.79306.09\u003c/span\u003e\u003cspan address=\"10.1023/B:JOEC.0000006398.79306.09\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSpitaler, R., Schlorhaufer, P. D., Ellmerer, E. P., Merfort, I., Bortenschlager, S., Stuppner, H., \u0026amp; Zidorn, C. (2006). Altitudinal variation of secondary metabolite profiles in flowering heads of Arnica montana cv. \u003cem\u003eARBO Phytochemistry\u003c/em\u003e, \u003cem\u003e67\u003c/em\u003e(4), 409\u0026ndash;417. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.phytochem.2005.11.018\u003c/span\u003e\u003cspan address=\"10.1016/j.phytochem.2005.11.018\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNascimento, L. B. D. S., Leal-Costa, M. V., Menezes, E. A., Lopes, V. R., Muzitano, M. F., Costa, S. S., \u0026amp; Tavares, E. S. (2015). Ultraviolet-B radiation effects on phenolic profile and flavonoid content of Kalanchoe pinnata. \u003cem\u003eJournal of Photochemistry and Photobiology B: Biology\u003c/em\u003e, \u003cem\u003e148\u003c/em\u003e(3), 73\u0026ndash;81. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jphotobiol.2015.03.011\u003c/span\u003e\u003cspan address=\"10.1016/j.jphotobiol.2015.03.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma, H. K., Chhangte, L., \u0026amp; Dolui, A. K. (2001). Traditional medicinal plants in Mizoram, India. \u003cem\u003eFitoterapia\u003c/em\u003e, \u003cem\u003e72\u003c/em\u003e(2), 146\u0026ndash;161. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0367-326X(00)00278-1\u003c/span\u003e\u003cspan address=\"10.1016/S0367-326X(00)00278-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDou, X., Zhang, Y., Sun, N., Wu, Y., \u0026amp; Li, L. (2014). The anti-tumor activity of Mikania micrantha aqueous extract in vitro and in vivo. \u003cem\u003eCytotechnology\u003c/em\u003e, \u003cem\u003e66\u003c/em\u003e(1), 107\u0026ndash;117. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10616-013-9543-9\u003c/span\u003e\u003cspan address=\"10.1007/s10616-013-9543-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDebaprotim, D., Suvakanta, D., \u0026amp; Jashabir, C. (2014). Evaluation of Anticancer Activity of Mikania micrantha Kunth (Asteraceae) Against Ehrlich Ascites Carcinoma in Swiss Albino Mice. \u003cem\u003eInternational Journal of Pharmaceutical Research \u0026amp; Allied Sciences\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e(2), 9\u0026ndash;18.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhosh, A., Das, B. K., Roy, A., Mandal, B., \u0026amp; Chandra, G. (2008). Antibacterial activity of some medicinal plant extracts. \u003cem\u003eJournal of Natural Medicines\u003c/em\u003e, \u003cem\u003e62\u003c/em\u003e(2), 259\u0026ndash;262. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11418-007-0216-x\u003c/span\u003e\u003cspan address=\"10.1007/s11418-007-0216-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, Y., Li, J., Li, Y., Wang, X., \u0026amp; Cao, A. (2013). Antimicrobial Constituents of the Leaves of Mikania micrantha H. B. K. \u003cem\u003ePlos One\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(10 (e76725)), 1\u0026ndash;10. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0076725\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0076725\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eP\u0026eacute;rez-Amador, M. C., Mu\u0026ntilde;oz Ocotero, V., Balcazar, I., R., \u0026amp; Garc\u0026iacute;a Jim\u0026eacute;nez, F. (2010). Phytochemical and pharmacological studies on Mikania micrantha H.B.K. (Asteraceae). \u003cem\u003ePhyton\u003c/em\u003e, \u003cem\u003e79\u003c/em\u003e, 77\u0026ndash;80.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKurade, N. P., Jaitak, V., Kaul, V. K., \u0026amp; Sharma, O. P. (2010). Chemical composition and antibacterial activity of essential oils of Lantana camara, Ageratum houstonianum and Eupatorium adenophorum. \u003cem\u003ePharmaceutical Biology\u003c/em\u003e, \u003cem\u003e48\u003c/em\u003e(5), 539\u0026ndash;544. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3109/13880200903193336\u003c/span\u003e\u003cspan address=\"10.3109/13880200903193336\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar Passari, A., Vineeth Leo, V., Singh, G., Samanta, L., Ram, H., Siddaiah, N., \u0026amp; Pratap Singh, C., B (2020). In Vivo Studies of Inoculated Plants and in Vitro Studies Utilizing Methanolic Extracts of Endophytic Streptomyces sp. Strain DBT34 Obtained from Mirabilis Jalapa L. Exhibit ROS-Scavenging and Other Bioactive Properties. \u003cem\u003eInternational Journal Of Molecular Sciences\u003c/em\u003e, \u003cem\u003e21\u003c/em\u003e, 7364. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ijms21197364\u003c/span\u003e\u003cspan address=\"10.3390/ijms21197364\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh, G., Passsari, A. K., Singh, P., Leo, V. V., Subbarayan, S., Kumar, B., \u0026amp; Kumar, N. S. (2017). Pharmacological potential of Bidens pilosa L. and determination of bioactive compounds using UHPLC-QqQLIT-MS/MS and GC/MS. \u003cem\u003eBMC Complementary and Alternative Medicine\u003c/em\u003e, \u003cem\u003e17\u003c/em\u003e(1), 1\u0026ndash;16. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12906-017-2000-0\u003c/span\u003e\u003cspan address=\"10.1186/s12906-017-2000-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkpoveso, O. P., Ubah, E. E., \u0026amp; Obasanmi, G. (2023). Antioxidant Phytochemicals as Potential Therapy for Diabetic Complications. \u003cem\u003eAntioxidants\u003c/em\u003e, \u003cem\u003e12\u003c/em\u003e, 1\u0026ndash;30.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAl-dalahmeh, Y., Al-bataineh, N., Al-balawi, S. S., Lahham, J. N., Al-momani, I. F., Al-sheraideh, M. S., \u0026amp; Al-qudah, M. A. (2022). LC-MS/MS Screening, Total Phenolic, Flavonoid and Antioxidant Contents of Crude Extracts from Three Asclepiadaceae Species Growing in Jordan. \u003cem\u003eMolecules\u003c/em\u003e, \u003cem\u003e27\u003c/em\u003e, 1\u0026ndash;11.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh, P., Bajpai, V., Gond, V. K., Kumar, A., \u0026amp; Tadigoppula, N. (2020). and K. B. Determination of Bioactive Compounds of Fenugreek (Trigonella foenum-graecum) Seeds Using LC-MS Techniques. In \u003cem\u003eLLegume Genomics: Methods and Protocols. Methods in Molecular Biology. doi:10.1007/978-1\u003c/em\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e-0716-0235-5\u003c/span\u003e\u003cspan address=\"http://-0716-0235-5\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (pp. 377\u0026ndash;392). Retrieved from http://www.springer.com/series/7651.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBajpai, V., Singh, A., Arya, K. R., Srivastava, M., \u0026amp; Kumar, B. (2015). Rapid screening for the adulterants of Berberis aristata using direct analysis in real-time mass spectrometry and principal component analysis for discrimination. \u003cem\u003eFood Additives and Contaminants - Part A Chemistry, Analysis, Control\u003c/em\u003e. \u003cem\u003eExposure and Risk Assessment\u003c/em\u003e, \u003cem\u003e32\u003c/em\u003e(6), 799\u0026ndash;807. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/19440049.2015.1022885\u003c/span\u003e\u003cspan address=\"10.1080/19440049.2015.1022885\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, S., Singh, A., \u0026amp; Kumar, B. (2019). Screening of monoterpene indole alkaloids in six Rauwolfia species by ultra-high performance liquid chromatography orbitrap velos pro mass spectrometer. \u003cem\u003eSeparation Science Plus\u003c/em\u003e, \u003cem\u003e2\u003c/em\u003e(8), 300\u0026ndash;308. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/sscp.201900029\u003c/span\u003e\u003cspan address=\"10.1002/sscp.201900029\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShelke, V., \u0026amp; Bhot, M. (2019). GC-MS Analysis of Bio-active Compounds in Ethanolic Extract of Leaf and Stem of Asclepias curassavica L. \u003cem\u003eInt J Pharm Investigation\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(2), 67\u0026ndash;70. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5330/ijpi.2019.2.13\u003c/span\u003e\u003cspan address=\"10.5330/ijpi.2019.2.13\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAssaad, H. I., Zhou, L., Carroll, R. J., \u0026amp; Wu, G. (2014). Rapid publication-ready MS-Word tables for one-way ANOVA. \u003cem\u003eSpringerPlus\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e, 1\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/doi:10.1186/2193-1801-3-474\u003c/span\u003e\u003cspan address=\"doi:10.1186/2193-1801-3-474\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJin, X., Wu, X., \u0026amp; Liu, X. (2017). Phenolic Characteristics and Antioxidant Activity of Merlot and Cabernet Sauvignon Wines Increase with Vineyard Altitude in a High-altitude Region. \u003cem\u003eSouth African Journal of Enology \u0026amp; Viticulture\u003c/em\u003e, \u003cem\u003e38\u003c/em\u003e(2), 132\u0026ndash;143. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.21548/38-2-1068\u003c/span\u003e\u003cspan address=\"10.21548/38-2-1068\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHagaggi, N. S. A., Abdul-Raouf, U. M., \u0026amp; Radwan, T. A. A. (2024). Variation of antibacterial and antioxidant secondary metabolites and volatiles in leaf and callus extracts of Phulai (Acacia Modesta Wall). \u003cem\u003eBMC Plant Biology\u003c/em\u003e, \u003cem\u003e24\u003c/em\u003e(1), 1\u0026ndash;12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12870-024-04747-9\u003c/span\u003e\u003cspan address=\"10.1186/s12870-024-04747-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRieger, G., M\u0026uuml;ller, M., Guttenberger, H., \u0026amp; Bucar, F. (2008). Influence of altitudinal variation on the content of phenolic compounds in wild populations of Calluna vulgaris, Sambucus nigra, and Vaccinium myrtillus. \u003cem\u003eJournal of Agricultural and Food Chemistry\u003c/em\u003e, \u003cem\u003e56\u003c/em\u003e(19), 9080\u0026ndash;9086. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/jf801104e\u003c/span\u003e\u003cspan address=\"10.1021/jf801104e\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu, W., Yin, D., Li, N., Hou, X., Wang, D., Li, D., \u0026amp; Liu, J. (2016). Influence of environmental factors on the active substance production and antioxidant activity in Potentilla fruticosa L. and its quality assessment. \u003cem\u003eScientific Reports\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e(July), 1\u0026ndash;18. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/srep28591\u003c/span\u003e\u003cspan address=\"10.1038/srep28591\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCopaja, S. V., Blackburn, C., \u0026amp; Carmona, R. (2003). Variation of saponin contents in Quillaja saponica molina. \u003cem\u003eWood Science and Technology\u003c/em\u003e, \u003cem\u003e37\u003c/em\u003e(2), 103\u0026ndash;108. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00226-002-0150-8\u003c/span\u003e\u003cspan address=\"10.1007/s00226-002-0150-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHalliwell, B., \u0026amp; Gutteridge, J. M. C. (2015). \u003cem\u003eFree Radicals in Biology and Medicine\u003c/em\u003e. \u003cem\u003eFree Radicals in Biology and Medicine\u003c/em\u003e. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/acprof:oso/9780198717478.001.0001\u003c/span\u003e\u003cspan address=\"10.1093/acprof:oso/9780198717478.001.0001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh, R., Sharad, S., \u0026amp; Kapur, S. (2004). Free radicals and oxidative stress in neurodegenerative diseases: relevance of dietary antioxidants. \u003cem\u003eJ Indian Acad Clin Med\u003c/em\u003e, \u003cem\u003e5\u003c/em\u003e(3), 218\u0026ndash;225.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eValko, M., Leibfritz, D., Moncol, J., Cronin, M. T. D., Mazur, M., \u0026amp; Telser, J. (2007). Free radicals and antioxidants in normal physiological functions and human disease. \u003cem\u003eInternational Journal of Biochemistry and Cell Biology\u003c/em\u003e, \u003cem\u003e39\u003c/em\u003e(1), 44\u0026ndash;84. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.biocel.2006.07.001\u003c/span\u003e\u003cspan address=\"10.1016/j.biocel.2006.07.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGaikwad, P., Barik, A., Priyadarsini, K. I., \u0026amp; Rao, B. S. M. (2010). Antioxidant activities of phenols in different solvents using DPPH assay. \u003cem\u003eResearch on Chemical Intermediates\u003c/em\u003e, \u003cem\u003e36\u003c/em\u003e(9), 1065\u0026ndash;1072. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11164-010-0220-8\u003c/span\u003e\u003cspan address=\"10.1007/s11164-010-0220-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKishore, G., Ranjan, S., Pandey, A., \u0026amp; Gupta, S. (2010). Influence of altitudinal variation on the antioxidant potential of tartar buckwheat of Western Himalaya. \u003cem\u003eFood Science and Biotechnology\u003c/em\u003e, \u003cem\u003e19\u003c/em\u003e(5), 1355\u0026ndash;1363. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10068-010-0193-9\u003c/span\u003e\u003cspan address=\"10.1007/s10068-010-0193-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003evan de Loosdrecht, A. A., Beelen, R. H. J., Ossenkoppele, G. J., Broekhoven, M. G., \u0026amp; Langenhuijsen, M. M. A. C. (1994). A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. \u003cem\u003eJournal of Immunological Methods\u003c/em\u003e, \u003cem\u003e174\u003c/em\u003e(1\u0026ndash;2), 311\u0026ndash;320. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0022-1759(94)90034-5\u003c/span\u003e\u003cspan address=\"10.1016/0022-1759(94)90034-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBhagat, J., Lobo, R., Kumar, N., Mathew, J. E., lizabet, \u0026amp; Pai, A. (2014). Cytotoxic potential of Anisochilus carnosus (L.f.) wall and estimation of luteolin content by HPLC. \u003cem\u003eBMC complementary and alternative medicine\u003c/em\u003e, \u003cem\u003e14\u003c/em\u003e(October), 421. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1472-6882-14-421\u003c/span\u003e\u003cspan address=\"10.1186/1472-6882-14-421\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVu, T. T. T., Vu, L. T. K., Nguyen, Q. H., Pham, K., Van, Nguyen, D. T., Nguyen, L. T. N., \u0026amp; Chu, M. H. (2019). Cytotoxic effects of steroidal glycosides isolated from the Paris vietnamensis plant on cancer cell lines and against bacterial strains. \u003cem\u003eBiotechnology and Biotechnological Equipment\u003c/em\u003e, \u003cem\u003e33\u003c/em\u003e(1), 1516\u0026ndash;1524. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/13102818.2019.1676168\u003c/span\u003e\u003cspan address=\"10.1080/13102818.2019.1676168\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXuan, T. D., Shinkichi, T., Hong, N. H., Khanh, T. D., \u0026amp; Min, C. I. (2004). Assessment of phytotoxic action of Ageratum conyzoides L. (billy goat weed) on weeds. \u003cem\u003eCrop Protection\u003c/em\u003e, \u003cem\u003e23\u003c/em\u003e(10), 915\u0026ndash;922. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cropro.2004.02.005\u003c/span\u003e\u003cspan address=\"10.1016/j.cropro.2004.02.005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cem\u003ePharmacognosy Magazine\u003c/em\u003e, \u003cem\u003e10\u003c/em\u003e(37), 18\u0026ndash;26. https://doi.org/10.4103/0973-1296.126653.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOkereke, S., Chukwudoruo, C., \u0026amp; Nwaokezie, C. (2017). Phytochemical screening using GC-FID and sub- chronic assessment of Hydroethanolic leaf extract of Ageratum conyzoides Linn on albino rats. \u003cem\u003eJournal of Medicinal Plants Studies\u003c/em\u003e, \u003cem\u003e5\u003c/em\u003e(2), 282\u0026ndash;287.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQuijano, L., G\u0026oacute;mez-Garibay, F., Garcia-Perez, G., \u0026amp; Rios, T. (1992). Highly oxygenated flavonoids from Ageratum corymbosum. \u003cem\u003ePhytochemistry\u003c/em\u003e, \u003cem\u003e31\u003c/em\u003e(8), 2859\u0026ndash;2862. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0031-9422(92)83647-H\u003c/span\u003e\u003cspan address=\"10.1016/0031-9422(92)83647-H\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFaqueti, L. G., Sandjo, L. P., \u0026amp; Biavatti, M. W. (2017). Simultaneous identification and quantification of polymethoxyflavones, coumarin and phenolic acids in Ageratum conyzoides by UPLC-ESI-QToF-MS and UPLC-PDA. \u003cem\u003eJournal of Pharmaceutical and Biomedical Analysis\u003c/em\u003e, \u003cem\u003e145\u003c/em\u003e, 621\u0026ndash;628. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jpba.2017.07.034\u003c/span\u003e\u003cspan address=\"10.1016/j.jpba.2017.07.034\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkhtar, N., Ihsan-ul-Haq, \u0026amp; Mirza, B. (2018). Phytochemical analysis and comprehensive evaluation of antimicrobial and antioxidant properties of 61 medicinal plant species. \u003cem\u003eArabian Journal of Chemistry\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e(8), 1223\u0026ndash;1235. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.arabjc.2015.01.013\u003c/span\u003e\u003cspan address=\"10.1016/j.arabjc.2015.01.013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHerbert, N. N., \u0026amp; David, S. (1992). \u003cem\u003ePhytochemical Resources for medicine and agriculture\u003c/em\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWen, J., Kang, L., Liu, H., Xiao, Y., Zhang, X., \u0026amp; Chen, Y. (2012). A validated UV-HPLC method for determination of chlorogenic acid in Lepidogrammitis drymoglossoides (Baker) Ching, Polypodiaceae. \u003cem\u003ePharmacognosy Research\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(3), 148\u0026ndash;153. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4103/0974-8490.99076\u003c/span\u003e\u003cspan address=\"10.4103/0974-8490.99076\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cem\u003ePLoS Neglected Tropical Diseases\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e(9), 1\u0026ndash;16. https://doi.org/10.1371/journal.pntd.0005929.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, A., Hou, X., \u0026amp; Wei, Y. (2018). Fast screening of flavonoids from switchgrass and: Mikania micrantha by liquid chromatography hybrid-ion trap time-of-flight mass spectrometry. \u003cem\u003eAnalytical Methods\u003c/em\u003e, \u003cem\u003e10\u003c/em\u003e(1), 109\u0026ndash;122. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/c7ay02103h\u003c/span\u003e\u003cspan address=\"10.1039/c7ay02103h\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRufatto, L. C., Gower, A., Schwambach, J., \u0026amp; Moura, S. (2012). Genus Mikania: Chemical composition and phytotherapeutical activity. \u003cem\u003eBrazilian Journal of Pharmacognosy\u003c/em\u003e, \u003cem\u003e22\u003c/em\u003e(6), 1384\u0026ndash;1403. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1590/S0102-695X2012005000099\u003c/span\u003e\u003cspan address=\"10.1590/S0102-695X2012005000099\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKiang, A. K., Sim, K. Y., \u0026amp; Yoong, S. W. (1968). Constituents of Mikania cordata (burm. f.) B. L. Robinson (compositae)-II. \u003cem\u003ePhytochemistry\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(6), 1035\u0026ndash;1037. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0031-9422(00)82193-7\u003c/span\u003e\u003cspan address=\"10.1016/S0031-9422(00)82193-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhao, B., \u0026amp; Hu, M. (2013). Gallic acid reduces cell viability, proliferation, invasion and angiogenesis in human cervical cancer cells. \u003cem\u003eOncology Letters\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e(6), 1749\u0026ndash;1755. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3892/ol.2013.1632\u003c/span\u003e\u003cspan address=\"10.3892/ol.2013.1632\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYang, J., Guo, J., \u0026amp; Yuan, J. (2008). In vitro antioxidant properties of rutin. \u003cem\u003eLWT - Food Science and Technology\u003c/em\u003e, \u003cem\u003e41\u003c/em\u003e(6), 1060\u0026ndash;1066. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.lwt.2007.06.010\u003c/span\u003e\u003cspan address=\"10.1016/j.lwt.2007.06.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKasai, H., Fukada, S., Yamaizumi, Z., Sugie, S., \u0026amp; Mori, H. (2000). Action of chlorogenic acid in vegetables and fruits as an inhibitor of 8-hydroxydeoxyguanosine formation in vitro and in a rat carcinogenesis model. \u003cem\u003eFood and Chemical Toxicology\u003c/em\u003e, \u003cem\u003e38\u003c/em\u003e(5), 467\u0026ndash;471. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0278-6915(00)00014-4\u003c/span\u003e\u003cspan address=\"10.1016/S0278-6915(00)00014-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDos Santos, M. D., Almeida, M. C., Lopes, N. P., \u0026amp; De Souza, G. E. P. (2006). Evaluation of the anti-inflammatory, analgesic and antipyretic activities of the natural polyphenol chlorogenic acid. \u003cem\u003eBiological and Pharmaceutical Bulletin\u003c/em\u003e, \u003cem\u003e29\u003c/em\u003e(11), 2236\u0026ndash;2240. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1248/bpb.29.2236\u003c/span\u003e\u003cspan address=\"10.1248/bpb.29.2236\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZeeshan, M., Rizvi, S. M. D., Khan, M. S., \u0026amp; Kumar, A. (2012). Isolation, partial purification and evaluation of bioactive compounds from leaves of Ageratum houstonianum. \u003cem\u003eEXCLI Journal\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e, 78\u0026ndash;88. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.17877/DE290R-4911\u003c/span\u003e\u003cspan address=\"10.17877/DE290R-4911\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIshak, A. H., Shafie, N. H., Esa, N. M., Bahari, H., \u0026amp; Ismail, A. (2018). From weed to medicinal plant: Antioxidant capacities and phytochemicals of various extracts of Mikania micrantha. \u003cem\u003eInternational Journal of Agriculture and Biology\u003c/em\u003e, \u003cem\u003e20\u003c/em\u003e(3), 561\u0026ndash;568. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.17957/IJAB/15.0522\u003c/span\u003e\u003cspan address=\"10.17957/IJAB/15.0522\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGanzera, M., Guggenberger, M., Stuppner, H., \u0026amp; Zidorn, C. (2008). Altitudinal variation of secondary metabolite profiles in flowering heads of Matricaria chamomilla cv. BONA. \u003cem\u003ePlanta Medica\u003c/em\u003e, \u003cem\u003e74\u003c/em\u003e(4), 453\u0026ndash;457. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1055/s-2008-1034326\u003c/span\u003e\u003cspan address=\"10.1055/s-2008-1034326\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZidorn, C. (2010). Altitudinal variation of secondary metabolites in flowering heads of the Asteraceae: Trends and causes. \u003cem\u003ePhytochemistry Reviews\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(2), 197\u0026ndash;203. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11101-009-9143-7\u003c/span\u003e\u003cspan address=\"10.1007/s11101-009-9143-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWong, S. K., Lim, Y. Y., \u0026amp; Chan, E. W. C. (2009). Antioxidant properties of hibiscus: Species variation, altitudinal change, coastal influence and floral colour change. \u003cem\u003eJournal of Tropical Forest Science\u003c/em\u003e, \u003cem\u003e21\u003c/em\u003e(4), 307\u0026ndash;315.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"applied-biochemistry-and-biotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"abab","sideBox":"Learn more about [Applied Biochemistry and Biotechnology](https://www.springer.com/journal/12010)","snPcode":"12010","submissionUrl":"https://submission.nature.com/new-submission/12010/3","title":"Applied Biochemistry and Biotechnology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Ageratum huostonianum, Mikania micrantha, UPLC-ESI MS/MS, GC-MS, traditional medicinal plants, altitude variation","lastPublishedDoi":"10.21203/rs.3.rs-4195244/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4195244/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eTraditional medicinal plants have attracted scientific interest due to their bioactive compounds, and the levels of their constituents vary with location and altitude. The present study was designed to evaluate the pharmacological potential of two traditional medicinal plants, \u003cem\u003eMikania micrantha\u003c/em\u003e and \u003cem\u003eAgeratum huostonianum;\u003c/em\u003e these were collected from two sites, Murlen National Park (MNP) and Dampa Tiger Reserve (DTR), which are located at different altitudes. Both plant species are used by local traditional healers in Mizoram, Northeast India, for the treatment of various health problems. We hypothesized that altitudinal variation would affect these plants' chemical composition and bioactive potential. Plant extracts were evaluated for antioxidant, antimicrobial, and cytotoxic activities. The results show that the plants located at a higher altitude, i.e., MNP, showed higher TPC (615.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 and 453.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95 \u0026micro;g gallic acid equivalents/mg of plant extract dry weight (\u0026micro;g GAE/mg) for \u003cem\u003eM. micrantha\u003c/em\u003e and \u003cem\u003eA. huostonium\u003c/em\u003e, respectively) and TFC (135.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46 and 120.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.93 \u0026micro;g quercetin equivalents/mg of plant extract dry weight (\u0026micro;g GE/mg) for \u003cem\u003eM. micrantha\u003c/em\u003e and \u003cem\u003eA. huostonium\u003c/em\u003e, respectively). The extract of \u003cem\u003eA. houstonianum\u003c/em\u003e (MNP) exhibited significantly greater antioxidant activity against ABTS radicals (IC\u003csub\u003e50\u003c/sub\u003e 241.6 \u0026micro;g/mL) as compared to the extract of \u003cem\u003eA. houstonianum\u003c/em\u003e (DTR) (IC\u003csub\u003e50\u003c/sub\u003e 371.2 \u0026micro;g/mL). The composition of the bioactive compounds present in the plants was determined using UPLC-ESI MS/MS and GC/MS, which detected five and ten compounds in the \u003cem\u003eA. houstonianum\u003c/em\u003e and \u003cem\u003eM. micrantha\u003c/em\u003e extracts, respectively. Plant species collected from the Murlen National Park site had high bioactivity potential and contained several bioactive compounds. A distinct variation between the volatile and non-volatile compounds was revealed. The collective data in this study show the influence of altitude on the biological compound production of selected medicinal plants. The findings will be utilized in determining the plant material needed for the development of bioactive formulations.\u003c/p\u003e","manuscriptTitle":"Determination of UPLC-ESI MS/MS- and GC-MS-based altitudinal variations in the bioactive potential of traditional medicinal plants","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-11 10:10:20","doi":"10.21203/rs.3.rs-4195244/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Accept with revisions","date":"2024-05-11T15:05:30+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-04-16T06:10:45+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-04-06T13:10:50+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Applied Biochemistry and Biotechnology","date":"2024-04-05T04:42:54+00:00","index":"","fulltext":""},{"type":"submitted","content":"Applied Biochemistry and Biotechnology","date":"2024-04-04T06:18:58+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"applied-biochemistry-and-biotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"abab","sideBox":"Learn more about [Applied Biochemistry and Biotechnology](https://www.springer.com/journal/12010)","snPcode":"12010","submissionUrl":"https://submission.nature.com/new-submission/12010/3","title":"Applied Biochemistry and Biotechnology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"5fbb8fe1-a6fe-4893-9b6e-05cfe09ecf28","owner":[],"postedDate":"April 11th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-08-12T16:02:24+00:00","versionOfRecord":{"articleIdentity":"rs-4195244","link":"https://doi.org/10.1007/s12010-024-05005-2","journal":{"identity":"applied-biochemistry-and-biotechnology","isVorOnly":false,"title":"Applied Biochemistry and Biotechnology"},"publishedOn":"2024-08-08 15:57:29","publishedOnDateReadable":"August 8th, 2024"},"versionCreatedAt":"2024-04-11 10:10:20","video":"","vorDoi":"10.1007/s12010-024-05005-2","vorDoiUrl":"https://doi.org/10.1007/s12010-024-05005-2","workflowStages":[]},"version":"v1","identity":"rs-4195244","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4195244","identity":"rs-4195244","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}