The role of secondary metabolites of Pontederia parviflora Alexander and Salvinia auriculata Aubl. in phytoremediation of cadmium contaminated water

The role of secondary metabolites of Pontederia parviflora Alexander and Salvinia auriculata Aubl. in phytoremediation of cadmium contaminated water | 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 The role of secondary metabolites of Pontederia parviflora Alexander and Salvinia auriculata Aubl. in phytoremediation of cadmium contaminated water Augusto César Rodrigues, Samara Requena Nocchi, Jorge Raposo, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4356239/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Heavy metals represent a serious source of water and soil contamination, mainly due to their non-biodegradable nature. Their bioaccumulation occurs in plants and other trophic levels, including animals, consequently affecting humans and causing various side effects. Phytoremediation offers a biological, cost-effective, and environmentally friendly cleaning solution, utilizing plants to remove contaminants from soil and water. This study aims to establish the chemical composition of Pontederia parviflora and Salvinia auriculata using liquid chromatography and evaluate the role of their secondary metabolites in passive phytoremediation by these plants. Although the results are not aligned with the initial hypothesis of metal removal from the liquid medium (sequestration and chelation of metals), the amount of metal removed still represents a positive outlook for technique enhancement. Furthermore, this research challenges the notion presented in the literature, as secondary metabolites were considered contaminant sequesters. The removal of these metabolites from the biomass of these macrophytes did not significantly impact their phytoremediation performance. Aquatic Macrophytes Heavy Metals Metabolites Phytoremediation Potential Figures Figure 1 Figure 2 Introduction In the last decades, rapid industrialization, urbanization, and population growth have led to significant increases in pollutant levels in water and soil resources (CPBC, 2008; SOOD et al., 2012 ). This poses a serious problem, affecting aquatic ecosystems, agriculture, and human health (GUPTA et al., 2010), with heavy metals being a major source of water and soil contamination (SELLAL et al., 2019 ). These heavy metals cannot be degraded through microbial or chemical processes, and their accumulation in the bodies of animals and humans along the food chain can cause DNA damage, carcinogenic effects, and the induction of mutations (EBRAHEM et al., 2019 ). Several technologies have been employed for the removal of toxic metals from contaminated environments, but they tend to be expensive, ineffective at low concentration levels, and may generate a large amount of waste (SELLAL et al., 2019 ). Phytoremediation, on the other hand, represents a biological, cost-effective, and eco-friendly clean-up approach that utilizes plants, their rhizospheres, and associated micro-organisms to degrade, remove, or remediate contaminants from soil and water while promoting vegetation on the land. This method is considered a green alternative since it has identified over 400 plant species as potential phytoremediators (ENSLEY, 2000 ; EBRAHEM et al., 2019 ). The use of aquatic macrophytes as a tool for phytoremediation is well-documented (NEWETE & BYRNE, 2016). Macrophytes have been demonstrated to efficiently sorb and desorb metals, exhibit high selectivity, reusability, and can be easily separated from solutions, making them excellent biosorbents for effluents (VIEIRA et al., 2019 ). Pontederia is a genus of emergent aquatic weeds from the Pontederiaceae family. While Eichornia crassipes (Water hyacinth), which belongs to this family, has been extensively studied for the removal of contaminants from wastewater (MALIK et al., 2007; REZANIA et al., 2015; NEWETE & BYRNE, 2016; FENG et al., 2017; MISHRA & MAITI, 2017), few studies are available regarding the phytoremediation capacity of other Pontederiaceae species. In 2000, Wilson et al. observed that phytoremediation with Pontederia cordata reduced the activity of simazine, a commercial herbicidal active ingredient, by 34% in just seven days. In 2010, Balassa et al. showed that Pontederia parviflora Alexander, the species studied in this work, exhibits a high capacity for extraction and storage of copper (Cu), reducing 96% of this metal in solution. Salvinia auriculata , another macrophyte studied in this work, is a free-floating weed belonging to the Salvinaceae family that has demonstrated positive results in cadmium bioaccumulation (WOLFF et al., 2012), lead (ESPINOZA-QUINONES et al., 2009 ; VESELY et al., 2011 ), chrome, and other metals, putting this species in the spotlight as one of the most efficient species in the accumulation and removal of "multielementary" samples (SOARES et al., 2008 ). Phytoremediation research associates some chemical functions, such as hydroxyl and carboxylic groups, as agents that contribute to the potential of contaminant uptake in solution. These structures are present in secondary metabolites found in the tested plants, suggesting that the plants could detoxify the water or soil through their metabolic activities (DHIR et al., 2010; ALI et al., 2013 ). In the literature, there are few studies available concerning the chemical composition of aquatic macrophytes. Although some studies have indicated the presence of phenolic compounds, alkaloid, and terpenoid derivatives in Eichornia crassipes (YI et al., 2006 ; ABOUL-ENEIN et al., 2014), which belongs to the same family as Pontederia parviflora , no studies could be found for the latter. Regarding S. auriculata , Lima et al. (2013) isolated two steroids (stigmasterone and stigmasterol) and one triterpene (friedelinol) and observed that only stigmasterone showed activity against Staphylococcus aureus strains. Secondary metabolites are synthesized with the purpose of not only attracting insects for pollination but also defending the plants from herbivory, UV light, and other mechanisms that may cause damage, such as in response to the presence of heavy metals in the environment. Bizzo et al. ( 2014 ) reported an increased production of phenolic compounds, especially anthocyanin, in S. auriculata in the presence of high copper concentrations, with a reduction in flavonoid concentrations after a short exposure period. Another study reported the reduction of antioxidant activity in samples of S. auriculata and E. crassipes under cadmium stress (VESTENA et al., 2011 ), possibly due to the decrease in secondary metabolites responsible for this activity. Recently, Eichhornia crassipes and Salvinia molesta were compared in terms of the quantity of some secondary metabolites, and the highest levels of tannins, flavonoids, and terpenoids were found in E. crassipes, while the content of alkaloid groups, saponins, and phenols was higher in S. molesta (GAYA et al., 2017 ). Since macrophytes are known to be powerful phytoremediators, and the mechanisms responsible for trapping heavy metals are not yet very clear, this study aims to establish a relationship between the chemical profile of two aquatic macrophytes and their performance in phytoremediation. Additionally, considering that secondary metabolites play an important role in maintaining plants in the environment, the study seeks to understand the role of secondary metabolites in this process. So, in this study, the chemical composition of P. parviflora and S. auriculata was established based on liquid chromatography-diode array detector-mass spectrometry (LC-DAD-MS) and MS/MS. The phenol and tannin content in each species was determined, and the concentration of arsenic (As), cadmium (Cd), chromium (Cr), magnesium (Mg), manganese (Mn), molybdenum (Mo), nickel (Ni), and lead (Pb) in the biomass of macrophyte samples was evaluated before and after the extraction process. We also assessed the influence of secondary metabolites on the fixation of metals in the biomass of samples, besides evaluating the potential for Cd removal in the ex-situ phytoremediation system. Materials and Methods Collection and Extraction of Plant Material We collected Pontederia parviflora and Salvinia auriculata in the Lagoa Comprida Municipal Natural Park, in the hydrographic basin of the River Aquidauana, Aquidauana, Mato Grosso do Sul, Brazil, located between the coordinates 20°23’36.8” S and 20°23’28.8” S, and 55°48’51.5” W and 55°47’04.9” W. This location is situated between the ecotone Cerrado and Pantanal, characterized by a large biological diversity combining features from both ecosystems. We separated the submerged petiole and the emerged leaf blade of P. parviflora into roots (PPA-R), petioles/stems (PPA-S), and leaves (PPA-L). For S. auriculata (SAL), the whole plant was used since the floating leaves are also in contact with the water. The plants were dried in an oven with air circulation at 50 ºC for 72 hours until a constant weight was achieved. The dry biomass was ground in a knife mill and sieved through a 60-mesh screen to obtain a uniform granulometry. A portion of the pulverized material was extracted using a pressurized fluid extractor (Dionex, ASE 150®), equipped with a 100 ml extraction cartridge. The extraction was performed with ethanol: water (7:3 v/v) following parameters repeated three times during each extraction: temperature of 125°C, static extraction time of 15 min, washing of the cell with 100% of the volume of its capacity, and purging for 60 seconds. The solvents were evaporated in a rotavapor (Büchi R-3, Switzerland) under reduced pressure and then lyophilized. The P. parviflora leaves extract yielded 29.4%, while the petioles/stems and roots extracts yielded 27.4% and 2.6%, respectively, and the S. auriculata extract yielded 6.9%. After the extraction process, the plant material was dried again and reserved for heavy metal removal analyses, named as P. parviflora roots (PPA-R II), petioles/stems (PPA-S II), leaves (PPA-L II), and S. auriculata (SAL II) because they are used as biomass without secondary metabolites. Total Phenols Content To evaluate the content of total phenols and tannins (addressed in the next section) in P. parviflora and S. auriculata extracts, we followed the methodology proposed by Herald et al. (2012). In a 96-well plate, 75 µL of the sample or standard (gallic acid) at various concentrations was added to each well. Then, 75 µL of Folin-Ciocalteu reagent (1:1 v/v, deionized water) was added to each well, and the plate was homogenized. After 6 minutes of mixing, 100 µL of Na2CO3 at 75 g/L was added to the wells, remixed, and the mixture was left for 90 minutes in darkness. The measurement was taken at 765 nm using a spectrophotometric microplate reader. The analyses were conducted in triplicate. The total phenolic content was expressed as mg of gallic acid equivalent (GAE) per gram of each extract. Total Tannins Content To determine the total tannin content, the extracts were solubilized in water (4 mg/mL), brought into contact with skin powder (20 mg, Hide Powder - protease substrate purchased from Sigma-Aldrich), and stirred for 60 minutes. After centrifugation, the supernatant was utilized to undergo the same process as described above for phenols. The concentration of total tannins was calculated as the difference in the concentration between total phenolics and tannin phenols. The total tannin content is expressed as mg of GAE per gram of each extract. Identification of chemical constituents in P. parviflora and S. auriculata extracts by LC-DAD-MS/MS Each extract was prepared at a concentration of 1 mg/mL in methanol: water (7:3, v/v), then filtered through a PTFE filter (Millex 0.22 mm x 13 mm, Millipore®). Subsequently, 3 µL of the filtered solution was injected into a Shimadzu Prominence UFLC system coupled with a diode array detector (DAD) and a MicrOTOF-Q III mass spectrometer (Bruker Daltonics). The chromatographic column used was a Kinetex C18 column (2.6 µm, 150 × 2.1 mm, Phenomenex). The injection volume, flow rate, and oven temperature were set at 1 µL, 0.3 mL/min, and 50 ºC, respectively. The mobile phase comprised 0.1% formic acid (v/v) in both water (solvent A) and acetonitrile (solvent B), utilizing a gradient elution profile as follows: 0–2 min: 3% B; 2–25 min: 3–25% B; 25–40 min: 25–80% B; 40–43 min: 80% B. The analyses were carried out in both negative and positive ion modes. Nitrogen gas was used as both a nebulizer gas (at 4 Bar) and a dry gas (at 9 L/min). The capillary voltage was set to 2.5 kV. Quantification of heavy metals (As, Cd, Cr, Mg, Mn, Mo, Ni, Pb) in P. parviflora and S. auriculata biomass To quantify heavy metals in macrophyte biomass, the two studied plant species were divided into two groups: one labeled "non-extracted," where biomass underwent no secondary metabolite extraction, indicated by adding "I" after each sample's acronyms; and another group labeled "extracted," where biomass was subjected to extraction, marked by adding "II" after each sample's acronyms, as described in the "Collection and Extraction of Plant Material" section. Calibration curves for each metal (As, Cd, Cr, Mg, Mn, Mo, Ni, Pb) were prepared in triplicate using seven standard concentrations (0.05; 0.1; 0.25; 0.5; 1.0; 1.5; and 2.0 µg/mL). For Cd, the calibration curve was generated with 12 different concentrations (0.05; 0.1; 0.25; 0.5; 1.0; 1.5; 2.0; 4.0; 6.0; 8.0; and 10.0 µg/mL). These solutions were derived from aqueous mono-elemental standards acidified with 1% HNO3. The limit of detection (LOD) and limit of quantification (LOQ) were calculated from the three standard curves using the standard deviation (SD𝑎) of the intercept with the 𝑦-axis and the calibration curve slope (S), according to: LOD = SD𝑎 x 3/ S LOQ = SD𝑎 x 10/ S Before analysis, the PRE and POS samples underwent digestion to remove organic residues. For this, 400 mg of dry biomass from each sample (PRE and POS of P. parviflora roots, petioles/stems, and leaves, as well as PRE and POS of S. auriculata ) was placed in a digester tube within a Berghof® Speedwave four microwave digester. To this, 3 mL of nitric acid, 2 mL of ultrapure water, and 1 mL of hydrogen peroxide were added, and the protocol specified for aquatic macrophytes in the equipment manual (Speedwave) was followed. The microwave digester was set with the following parameters: Step (1) utilized 80% power at 150 ºC, with a 5-minute ramp time, 10-minute retention time, and 80 Bar pressure. In Step (2), power was set at 90%, temperature at 190 ºC, with a 2-minute ramp time, 20-minute retention time, and 80 Bar pressure. Finally, in Step (3), power was set to 0%, temperature to 50 ºC, with a 1-minute ramp time, 10-minute retention time, and 60 Bar pressure. Aqueous residual solutions from each digester tube were brought to a volume of 15 mL with water and analyzed using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) (iCAP 6000, Thermo®). The analysis conditions were as follows: power at 1250 W, sample flow at 0.35 L/min, plasma gas flow at 12 L/min, integration time at 5 s, stabilization time at 20 s, nebulization time at 30 psi, plasma view axial, and air gas (99.999%). Emission lines (nm) for each analyte were as follows: As 189.042, Cd 228.802, Cr 283.563, Mg 279.553, Mn 257.610, Mo 202.030, Ni 221.647, and Pb 220.353. The quantification calculation for metals present in the macrophyte biomass was performed using the following formula: Total metal in the sample (µg/g) = Metal mass (µg) found / Sample mass used (g) Cadmium Removal trials Removal trials were conducted to establish the relationship between secondary metabolite presence in plant biomass and metal removal through passive phytoremediation. The experimental design is depicted in Fig. 1 . In plastic tubes, we prepared 20 mL of Cd mono-elemental solutions at concentrations of 0.2 and 3.5 µg/mL, acidified with water. To these solutions, 0.2 g of biomass from each sample (I and II of P. parviflora roots, petioles/stems, and leaves, as well as I and II of S. auriculata ) was added. Subsequently, the solutions were stirred for 15 minutes and then filtered. The resulting aqueous residuals underwent microwave digestion, as detailed in the preceding section, followed by analysis using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) (iCAP 6000, Thermo®). The amount of Cd removed from the solution by the dry biomass of the macrophytes was calculated as follows: % Cadmium removed = (ΔC / Standard Concentration) x 100 ΔC = Control concentration – Sample concentration Statistical analysis To quantify heavy metals (As, Cd, Cr, Mg, Mn, Mo, Ni, Pb) in P. parviflora and S. auriculata biomass, as well as for Cd removal tests, uniform data underwent analysis using one-way analysis of variance (ANOVA), and means were compared using the Tukey test ( p < 0.05 ). Results are presented as mean ± standard deviation (SD). Results and Discussion Total phenolic and tannin contents in macrophyte extracts are shown in Table 1 . S. auriculata and P. parviflora leaves contained the highest phenol and tannin levels, with no statistical difference ( p < 0.05). Table 1 Total phenolic and total tannin content in extracts of macrophytes samples. Sample Phenol content (mg GAE g − 1 ) Tannin content (mg GAE g − 1 ) P. parviflora leaves 61.12 c (1.26) 5.32 b,c (1.29) P. parviflora petiole/stem 16.36 a (0.35) 0.56 a (0.13) P. parviflora roots 29.23 b (1.24) 4.24 b (0.12) S. auriculata 62.56 c (1.24) 6.21 c (0.46) The results are expressed in mg GAE (gallic acid equivalent) per g of extract. Different letters represent significantly different means (p < 0.05). The variation among P. parviflora samples could be linked to the roles of analyzed structures. Leaves, which are photosynthetic, displayed the highest phenol content. Phenolic compounds, linked to UV absorption, might be more abundant in leaves due to their photoprotective effect. The primary compounds in macrophyte samples were identified using UV, MS, and MS/MS data, compared with published information (Table 2 – 5 ). Predominantly, they comprised flavonoids and phenylpropanoids. Table 2 Identification of the constituents from hydroethanolic extract of Salvinia auriculata by LC-DAD-MS/MS. Peak RT (min) Compound Molecular Formula Error Ppm UV (nm) Negative mode (m/z) Positive mode (m/z) MS MS/MS MS MS/MS 1 5.3 Unknown C 14 H 16 O 9 -2,4 260–297 327.0729 191.0635;173.0485 329.0874 - 2 6.4 4- O-E -caffeoylquinic C 16 H 18 O 9 -2,8 297–325 353.0888 191.0548;179.0277 173.0345; 161.0196 163.0386 - 3 10.7 4 -O-E -caffeoylquinic C 16 H 18 O 9 -2,1 300–325 353.0885 191.0571; 179.0375; 173.0456; 161.0161 355.1037 163.0390; 145.0279 4 14.8 Unknown C 10 H 10 O 3 2,8 304–334 177.0548 161.0244 - - 5 16.6 Unknown C 18 H 24 O 10 3,3 285 399.1284 153.0273 - - 6 18.0 Unknown C 31 H 30 O 15 -0,7 283–329 641.1516 449.1000; 405.0954; 327.0566; 297.0419; 271.0588; 191.0570; 173.0441; 161.0223 643.1681 311.0581; 299.0575; 271.0578; 231.0693; 191.0350; 163.0403 7 19.0 Kaempferol- O -hexoside C 21 H 18 O 12 -1,2 266–346 461.0731 285.0388 463.0880 287.0569 8 20.0 3,4-dicafeoylquinic acid C 25 H 24 O 12 -2,1 297–325 515.1206 335.0795; 191.0562; 179.0344; 173.0477; 161.0250; 155.0380 517.1361 163.0399 9 20.2 Unknown C 31 H 30 O 14 -0,1 283–329 625.1564 433.0786; 281.0457; 191.0571; 161.0251 627.1737 283.0591; 163.0395 10 20.6 3,5-dicafeoylquinic acid C 25 H 24 O 12 -1,4 298–329 515.1202 191.0570; 179.0340; 161.0270 - - 11 21.9 Salviniside II derivative C 22 H 18 O 12 -1,3 330 473.0732 413.0480; 285.0481; 241.0490 475.0890 313.0342; 295.0262; 267,0279 12 22.1 4,5-dicafeoylquinic acid C 25 H 24 O 12 -0,3 298–329 515.1197 325.0149; 191.0596; 179.0330; 173.0470; 161.0213 499.1243 - 13 23.1 Salviniside II derivative C 22 H 18 O 12 -1,0 330 473.0730 413.0548; 327.0514; 267.0216; 241.0516 475.0894 313.0362; 267.0297; 239.0362 14 25.3 Unknown C 27 H 23 NO 13 -1,9 284–329 568.1108 413.0473; 241.0603 570.1233 313.0331; 295.0226 “Unknown” are the non identified compounds. In the S. auriculata extract, we observed 14 peaks (Table 2 ). Among these, peaks 2 and 3 had the ion at m/z 353 as the base peak in the negative ionization spectrum, along with two bands near 299 and 325 nm in the UV spectrum, characteristic of caffeoylquinic acids. Peaks 2 and 3 exhibited product ions at m/z 191 due to the loss of the caffeoyl unit, m/z 179 related to the quinic unit's loss, and m/z 173 for the dehydrated quinic acid unit. As only the caffeoylquinic acid esterified at position 4 produces the product ion at m/z 173, these peaks were identified as 4-O-E-caffeoylquinic acid (GOBBO-NETO, 2007). Compounds 11 and 13 displayed intense ions at m/z 473.0732 and 473.0730, respectively, in alignment with the molecular formula C22H18O12. These peaks were recognized as derivatives of salviniside II (Li et al., 2013). Peak 7 exhibited bands at 265 and 345 nm in the UV spectrum, characteristic of flavonol structures. This compound bore the molecular formula C21H18O12, with ions m/z 461.0731 [M-H]- and 463.0880 [M + H]-, and a key fragment at m/z 285, harmonizing with flavonoid aglycone kaempferol. Hence, this compound was identified as kaempferol-O-hexoside (SADEER et al., 2019). Peaks 8, 10, and 12, with retention times of 20.0, 20.6, and 22.1 min, displayed bands at 285 and 314 nm in the UV spectrum, along with m/z 515 [M + H] + and 513 [M-H]-, encompassing molecular formula C25H24O12, compatible with dicaffeoylquinic acids (CLIFFORD et al., 2003, 2005). Peaks 10 and 12 were validated as 3,5 and 4,5-dicaffeoylquinic acids, respectively, using genuine standard injection. The fragment ions of peak 8 were juxtaposed with identification cues suggested by Clifford et al. (2003, 2005) for dicaffeoylquinic acids. In these guidelines, the fragment ion at m/z 173 functions as a diagnostic indicator of a caffeic unit esterified at the fourth position of quinic acid. This data, coupled with the absence of characteristic ions of 1,4-dicaffeoylquinic acid ( m/z 299 and m/z 203), substantiated the identification of this compound as 3,4-dicaffeoylquinic acid (GOBBO-NETO 2007). In the extract of P. parviflora leaves (Table 3 ), 19 peaks were detected. Peaks 1, 2, 4, and 7, displaying the base ion at m/z 209, were suggested based on their molecular formulas and UV spectra as derivatives of caffeoyl glucarate. Peaks 3 and 6, exhibiting two bands in the UV spectra within the range of 300 to 324 nm and 299 to 325 nm, respectively, displayed the base ion at m/z 371. Both peaks 3 and 6 also showed product ions at m/z 191, with peak 6 additionally featuring an ion at m/z 209, characterizing them as isomers of caffeoyl glucarates (LORENZ et al. 2014). Table 3 Identification of the constituents from hydroethanolic extract of Pontederia parviflora (leaves) by LC-DAD-MS/MS. Peak RT (min) Compound Molecular Formula Error Ppm UV (nm) Negative mode (m/z) Positive mode (m/z) MS MS/MS MS MS/MS 1 3.2 Caffeoyl glucarate derivatives C 6 H 10 O 8 -3.9 300–324 209.0311 - - - 2 4.2 Caffeoyl glucarate derivatives C 6 H 10 O 8 0.1 300–324 209.0303 - - - 3 4.5 Isomers of caffeoyl glucarates C 15 H 16 O 11 -1.1 300–326 371.0624 191.0234 - - 4 4.8 Caffeoyl glucarate derivatives C 6 H 10 O 8 -0.5 300–324 209.0304 - - - 5 5.7 Tryptophan derivative C 11 H 11 N 2 O 2 1.2 280 203.0824 - - - 6 6.2 Isomers of caffeoyl glucarates C 15 H 16 O 11 -2.2 299–325 371.0628 209.0254; 191.0182 - - 7 6.5 Caffeoyl glucarate derivative C 6 H 10 O 8 -0.9 300–324 209.0303 - - - 8 9.9 Unit of caffeic acid C 9 H 8 O 4 0.6 299–320 179.0349 - 163.0371 - 9 10.6 Isomer of caffeoylquinic acid C 16 H 18 O 9 1.9 285–325 353.0871 191.0572; 161.0197 163.0375 - 10 12.1 Caffeic acid derivatives C 14 H 15 O 8 3.6 299–324 311.0786 271.0942; 255.0767; 179.0354; 161.0316; 149.0488 625.1748 163.0377 11 12.9 Unknown C 13 H 12 O 8 -4.1 300–325 295.0471 179.0384 - - 12 13.3 Caffeic acid derivatives C 14 H 15 O 8 1.0 300–324 311.0787 311.0823; 271.0922; 243.0617; 179.0356; 161.0252; 149.0456 313.0898 163.0379 13 14.1 Unknown C 16 H 16 O 8 -4.8 285–320 335.0789 179.0434; 161.0292 - - 14 16.8 Unknown C 14 H 14 O 7 -3.0 298–324 293.0676 179.0378; 161.0228 - - 15 20.0 Kaempferol-O-rutinoside C 27 H 30 O 15 -2.2 269–346 593.1525 477.0882; 285.0415 595.1640 2870549 16 20.0 3,4 dicafeoylquinic acid C 25 H 24 O 12 -0.9 269–346 515.1200 191.0603; 179.0335; 173.0407; 161.0183 - - 17 20.6 3,5 dicafeoylquinic acid C 25 H 24 O 12 -1.5 280–320 515.1162 - - - 18 22.2 4,5 dicafeoylquinic acid C 25 H 24 O 12 -1.5 280–320 515.1202 - - - 19 23.1 Kaempferol-O-ramnoside-O-malonil-O-hexoside C 30 H 32 O 18 0.2 267–347 679.1515 635.1623; 489.1055; 285.0380 681.1630 287.0554 20 26.4 Kaempferol derivative C 29 H 32 O 16 -0.6 273–335 635.1622 - - - 21 30.4 Unknown C 18 H 32 O 5 -3.4 276–311 327.2188 171.1031 - - 22 31.4 Unknown C 18 H 34 O 5 -3.5 276–311 329.2345 229.1431; 211.1313; 171.0995 - - 23 31.6 Unknown C 19 H 12 O 4 2.9 276–308 303.0654 274.0646 305.0817 277.0761; 259.0711; 249.0938; 231.0808; 213.0638; 203.0872 24 33.3 Unknown C 19 H 12 O 3 -4.3 276 287.0726 287.0773 289.0848 271.0725; 261.0924; 243.0786; 233.0945; 215.0837 25 35.0 Unknown C 18 H 30 O 4 -1.0 273–311 309.2075 183.0064 - - “Unknown” are the non identified compounds. Peak 5 demonstrated a base ion at m/z 203 in negative ionization mode, along with two ions at m/z 188 and m/z 205 in positive ionization mode. Furthermore, a product ion at m/z 170 was observed. This compound was identified as a derivative of tryptophan (LIU et al. 2015). Peak 8 displayed an ion at m/z 179 and lacked product ions in the fragmentation spectrum. However, relying on its UV spectrum and the molecular formula obtained from the software, this peak was suggested to represent a unit of caffeic acid, a product ion resulting from the fragmentation of caffeoylquinic acid (GOBBO-NETO, 2007). Peak 9, akin to peaks 2 and 3 of S. auriculata , exhibited the base ion at m/z 353 and two bands in the UV spectrum ranging from 285 to 325 nm, along with product ions at m/z 191 and m/z 161. This peak was identified as an isomer of caffeoylquinic acid (GOBBO-NETO, 2007). Peaks 10 and 12 displayed ions at m/z 311 [M-H]-, consistent with the molecular formula C14H15O8. However, peak 10 featured a main ion at m/z 625 [M + H]+, potentially indicating a dimer, while peak 12 exhibited m/z 313 [M + H]+. These compounds were recognized as derivatives of caffeic acid connected to a unit of D-erythrono-1,4-lactone or D-threono-1,4-lactone (CCANA-CCAPATINA et al., 2017). Peaks 15 and 19 exhibited two bands in the UV spectrum near wavelengths of 265 and 345 nm (typical of flavonols), along with ions at m/z 593.1525 [M-H]- and 679.1515 [M-H]-, respectively. The compounds possessed the molecular formulas C27H29O15 and C30H31O18, sharing the main fragment at m/z 285, akin to compound 7 of S. auriculata . Compound 15 was identified as Kaempferol-O-rutinoside (HERRANZ-LÓPEZ et al., 2012), and compound 19 as Kaempferol-3-O-rutinoside-7-O-rhamnoside (JU et al., 2018). Peak 20 featured a base ion at m/z 635, with the identical molecular formula as peak 19, albeit without any fragmentation. Hence, this peak was identified as a derivative of kaempferol (SADEER et al., 2019). Compounds 16, 17, and 18 were designated as 3,4, 3,5, and 4,5-dicaffeoylquinic acids, congruent with their presence in S. auriculata peaks 8, 10, and 12 (CCANA-CCAPATINA et al., 2017). In P. parviflora petiole/stem (Table 4 ), 7 peaks were detected. Peaks 2 and 3 exhibited the base ion at m/z 311 [M-H]-, with two bands in the wavelength range of 294 to 329 nm and 300 to 322 nm in the UV spectra, respectively, along with the molecular formula C14H15O8. Although peak 2 lacked fragmentation in the MS/MS spectrum, based on UV data and the molecular formula, this compound was tentatively identified as a derivative of chlorogenic acid. Peak 3 displayed a product ion at m/z 179 [M-H]-, linked to the caffeoyl unit. Furthermore, it demonstrated the ion at m/z 313 [M + H] + with a product ion at m/z 163 [M + H]+. Considering the calculated molecular formulae, these resemblances were noted with peaks 10 and 12 of P. parviflora leaves. Consequently, this compound was identified as a caffeic acid derivative connected to a unit of D-erythrono-1,4-lactone or D-threono-1,4-lactone (CCANA-CCAPATINA et al., 2017). Table 4 Identification of the constituents from hydroethanolic extract of Pontederia parviflora (petiole/stem) by LC-DAD-MS/MS. Peak RT (min) Compound Molecular Formula Error Ppm UV (nm) Negative mode (m/z) Positive mode (m/z) MS MS/MS MS MS/MS 01 5.7 Unknown C 11 H 12 N 2 O 2 0.0 283 203.0826 - - - 02 12.1 Chlorogenic acid derivative C 14 H 16 O 8 -5.0 294–329 311.0788 - - - 03 13.4 Caffeic acid derivative C 14 H 16 O 8 -1.0 300–322 311.0775 179.0401 313.0921 163.0386 04 20.0 Unknown C 27 H 30 O 15 0.2 285–330 593.1511 - 595.1664 287.0555 05 23.3 Unknown C 30 H 32 O 18 1.2 290–345 - - 681.1653 287.0550 06 30.5 Unknown C 18 H 31 O 5 -1.8 285 327.2197 211.1362 - - 07 32.5 Unknown C 14 H 17 O 4 1.8 283 249.1128 - - - “Unknown” are the non identified compounds. For P. parviflora roots (Table 5 ), 9 peaks were discerned. Peak 3 exhibited two bands within the wavelengths of 280 to 310 nm in the UV spectra and the ion at m/z 163 [M-H]-, consistent with the molecular formula C9H7O3, implying a coumaric acid molecule (DUGO et al., 2008). Table 5 Identification of the constituents from hydroethanolic extract of Pontederia parviflora (roots) by LC-DAD-MS/MS. Peak RT (min) Compound Molecular Formula Error Ppm UV (nm) Negative mode (m/z) Positive mode (m/z) MS MS/MS MS MS/MS 01 1.2 Unknown C 12 H 22 O 11 -7.7 269 341.1116 191.0588 365.1092 203.0522 02 1.2 Unknown C 18 H 17 O 9 -4.1 269 377.0893 - 03 13.3 Coumaric acid C 9 H 9 O 3 -0.1 280–310 163.0401 - - - 04 19.1 Unknown C 9 H 10 O 4 -1.5 275–308 181.0509 - - - 05 19.9 Unknown C 9 H 16 O 4 -12.7 276–310 187.1000 - - - 06 30.1 Unknown C 17 H 14 O 7 -4.3 276–311 329.0681 299.0270; 271.0236; 227.0467; 199.0431; 161.0193 331.0822 315.0521; 302.0464; 287.0549; 270.0489; 258.0514; 242.0549 07 31.5 Unknown C 18 H 34 O 5 -2.1 276–312 329.2340 211.1341; 171.1042 667.3233 433.2096; 389.1835; 261.1318; 217.1060; 173.0796; 155.0716 08 31.7 Unknown C 13 H 18 O 3 -3.1 276–310 221.0663 - - - 09 33.1 Unknown C 18 H 10 O 3 -1.7 276–312 273.0562 245.0636 - - “Unknown” are the non identified compounds. The results obtained from the heavy metal quantification tests in the biomass of the studied macrophytes (Table 6 ) demonstrated that, barring Ni and Mo in P. parviflora , the metal concentration escalated after the extraction of secondary metabolites. Employing ICP-OES, it was apparent that the heavy metals existing in the macrophytes' biomass were not predominantly complexed with the plants' metabolites, or at least, a majority of these metals were not. This is inferred from the elevation of metal levels in samples where metabolites were removed. Although secondary metabolites encompass clusters such as –OH- and –COO-, known to form bonds with metals (SARASWAT and RAI, 2010; ALI et al., 2013 ), our results imply that metals are chiefly associated with plant structures that persist throughout the process of secondary metabolite extraction, such as cell walls and vacuoles. Additionally, no correlation was established between the content of total phenols and tannins in the samples and the metal accumulation capacity, further affirming that the secondary metabolites in these plants are not primarily responsible for phytoremediation. The calibration parameters acquired for the ICP-OES are accessible in the Supplementary Material (Table S1 ). Table 6 Concentration (mean ± deviation, n = 3) in µg g − 1 of As, Cd, Cr, Mg, Mn, Mo, Ni and Pb found in P. parviflora -leaf (PPA-F), P. parviflora - petioles/stems (PPA-C), P. parviflora -roots (PPA-R) and S. auriculata . Sample Means (standard deviation) As Cd Cr Mg Mn Mo Ni Pb PPA-F – I 0.331 a (0.013) LPC >LPC 0.419 c (0.001) 0.1254 a (0.0003) 0.256 a (0.010) PPA-F – II 0.371 b (0.016) LPC >LPC 0.464 d (0.003) 0.2108 c (0.0003) 0.318 c (0.005) PPA-C – I 0.361 ab (0.006) LPC >LPC 0.267 b (0.004) 0.2142 c (0.001) 0.287 b (0.003) PPA-C – II 0.425 c (0.013) LPC >LPC 0.192 a (0.001) 0.1805 b (0.0008) 0.385 d (0.006) PPA-R – I 0.835 d (0.005) 0.0232 a (0.0002) 1.026 d (0.004) >LPC >LPC 0.186 a (0.001) 0.2497 d (0.001) 0.924 e (0.0005) PPA-R – II 1.134 f (0.015) 0.0318 b (0.0004) 1.264 e (0.007) >LPC >LPC 0.266 b (0.005) 0.3034 e (0.0007) 1.077 g (0.006) SAL – I 1.081 e (0.012) 0.0402 c (0.0004) 1.254 e (0.006) >LPC >LPC 0.541 e (0.003) 0.3660 f (0.002) 1.054 f (0.008) SAL – II 1.471 g (0.016) 0.0459 d (0.0002) 1.516 f (0.009) >LPC >LPC 0.623 f (0.004) 0.4506 g (0.0005) 1.235 h (0.007) <DL: samples with concentrations below the detection limit of the equipment. Means followed by the same letter in the column do not differ from each other by the Tukey test, considering the nominal value of 5% of significance. > UPC: sample with concentration above the highest concentration point of the calibration curve (2µg g − 1 ). Samples with “I” are samples of dried plant material that have not gone through the extraction process and “II” samples are samples that have had their secondary metabolites removed in the extraction process. We also observed that the roots of P. parviflora exhibit higher metal accumulation compared to the petiole/stem and leaves. This disparity could be attributed to the roots' closer interaction with the contaminated environment. P. parviflora , as an emerging macrophyte, has its roots firmly affixed to the substrate, while the leaves and a section of the petiole/stem remain above the water surface. To assess the plant biomass's capacity (pre- and post-extraction procedure), Cadmium Removal trials were conducted using three different Cd solution concentrations (0.2 and 3.5 µg/mL of Cd in acidic water). The outcomes (Table 7 ) revealed that at the 0.2 µg/mL solution, all samples demonstrated a decrease in Cd concentration, signifying the macrophytes' ability to capture this metal. However, in the 3.5 µg/mL solutions, the samples exhibited minimal reduction in Cd content. Notably, when in contact with sample P. parviflora - roots I, there was an elevation in Cd concentration. These findings imply a saturation of the trapping system in the plant material and suggest that the presence of metabolites in the biomass (samples before the extraction process) does not influence the metal trapping system. This conclusion is drawn from the lack of correlation between the presence of these metabolites and the enhanced removal of metals. Table 7 Mean values (± standard deviation, n = 3) of the residual concentration evaluated after the removal tests, the values presented are the residual cadmium concentration after the contact time of the control solution with the biomass. Sample Assay 0.2 µg mL -1 Assay 3.5 µg mL -1 Control 0.200 (± 0.00010) 3.350 (± 0.0024) PPA-F – I 0.026 (± 0.00008) e 2.940 (± 0.0110) b PPA-F – II 0.058 (± 0.00007) c 3.260 (± 0.0020) c PPA-C – I 0.019 (± 0.00008) g 3.260 (± 0.0020) c PPA-C – II 0.012 (± 0.00014) a 3.200 (± 0.0010) c PPA-R – I 0.014 (± 0.00017) d 3.430 (± 0.0010) cd PPA-R – II 0.008 (± 0.00006) f 3.300 (± 0.0010) e SAL – I 0.030 (± 0.00001) bc 3.380 (± 0.0020) e SAL – II 0.011 (± 0.000005) a 2.530 (± 0.0008) a Means followed by the same letter in the column do not differ from each other by the Tukey test, considering the nominal value of 5% of significance. The concentration of Cd in (µg mL − 1 ). Samples with “I” are samples of dried plant material that have not gone through the extraction process and “II” samples are samples that have had their secondary metabolites removed in the extraction process. Despite the absence of extraction enhancement, the attained results regarding extract yield and the percentage of phenolic compounds present promising potential. Tannins and phenolic compounds commonly possess well-recognized antioxidant and antiradical properties, constituting the primary biological activities linked to phytoremediation processes, involving metal sequestration and chelation (ZHENG and WANG, 2001; DAI and MUMPER, 2010). Regarding the disparity in metal content between biomass samples with and without secondary metabolites, it's noteworthy that the removal of metabolites led to elevated metal levels within the structures of the samples. Although existing literature links metal complexation with secondary metabolites, it's important to consider that plant tissues contain catechol subunits within lignins and lignans, which provide potential binding sites for metal atoms within the plant structure (SARASWAT and RAI, 2010; ALI et al., 2013 ). As demonstrated in Table 6 , the metal content escalates upon the removal of plant biomass metabolites. Samples with removed metabolites comprise a higher proportion of structures that contribute to plant tissue formation, such as lignins, which remain intact during the extraction process. Consequently, we hypothesize that the metals detected in these samples could be complexed within these structures. The exception to this trend is observed in the case of Mo and Ni in PPA-C, where this logic is reversed. Among the assessed metals, certain ones are integral to plant metabolism, constituting essential micronutrients. However, in most instances, these metals are present at levels designating them as contaminants within the collection areas of these plants. Generally, they possess high toxic potential. Furthermore, their presence in aquatic ecosystems fuels extensive debate, interest, and concern. This is particularly significant since their concentrations in natural waters are also subject to stringent limitations. The results presented for Cd removal assays in a contaminated aqueous medium (Fig. 1 , Table 7 ) indicate a low saturation point for the phytoremediation system in relation to this metal. At Cd concentrations of 0.2 µg mL-1, there was a reduction in all residual aqueous samples, ranging from 71% (PPA-F II) to 96% (PPA-R II). At concentrations of 3.5 µg mL-1 Cd, certain samples exhibited an increase in residual aqueous concentrations, with higher Cd contents than the control (PPA-R I and SAL I, both containing metabolites). In assays with a concentration of 3.5 µg mL-1, the most favorable removal result was achieved in SAL II, with a 24.5% reduction. The presence of other ions can impact the potential for metal removal due to competition for binding sites. It is essential to observe the ionic balance in control solutions and the dried biomass to enhance contaminant removal potential (Saraswat and RAI 2010). The elevated Cd contents observed after biomass intervention (cadmium removal assay) at higher metal concentrations can be explained by the scheme depicted in Fig. 2 . This scheme illustrates situations where biomass binding sites become saturated and complexed with other ions from the environment. This fact potentially compromises the ability of these vegetables to remove ions. Nevertheless, as the dry biomass persists, water absorption continues, resulting in the residual (aqueous) solution retaining Cd mass in the reduced volume of the medium (water), consequently elevating the Cd concentration in this residual solution. Conclusion The study demonstrated the effects of biosorption system saturation using biomass of P. parviflora and S. auriculata. In some cases, the concentration of cadmium ions did not decrease; instead, it increased due to a reduction in the volume of the dilution medium (water) without effective reduction of cadmium ions. This suggests low bioaccumulation in a phytotreatment system, or even bioaccumulation of metals in the biomass of these plants from the environment during vegetal development. Our study indicates the absence of interference in the cadmium removal processes, as well as in the bioaccumulation of metals, by metabolite contents, as samples without metabolites outperformed samples with metabolites in metal removal assays. Thus, there is no correlation between the metabolite content and the accumulated or removed metal content. Declarations Conflict of Interest There is no conflict of interest between the primary author and the co-authors. Funding This study was funded by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Author Contribution All authors have participated in the research and/or preparation of the article:Augusto César Rodrigues: conceptualization, methodology, validation, formalanalysis, investigation, writing original draft.Samara Requena Nocchi: methodology, validation, formal analysis, investigation,writing original draft.Jorge Raposo: conceptualization, resources, data curation, writing – review.Valter Aragão do Nascimento: conceptualization, resources, data curation,writing – review.Carlos Alexandre Carollo: conceptualization, resources, data curation, writing –review and editing, visualization, supervision, project administration, fundingacquisition. Acknowledgments We are grateful to Instituto Nacional de Áreas Úmidas (INAU), Laboratório de Produtos Naturais e Espectrometria de Massas (LaPNEM) and Programa de Pós-graduação em Biologia Vegetal da Universidade Federal de Mato Grosso do Sul (PPGBV-UFMS). Data availability statements The authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information files. Should any raw data files be needed in another format they are available from the corresponding author upon reasonable request. References ALI, H.; KHAN, E.; SAJAD, M. A. Phytoremediation of heavy metals–concepts and applications. Chemosphere, v. 91, n. 7, p. 869–881. 2013. BALASSA, G. C.; DE SOUZA, D. C.; BARBOSA DE LIMA, S. Evaluation of the potential of Pontederia parviflora Alexander in the absorption of copper (Cu) and its effects on tissues. Acta Scientiarum, Biological Sciences, v. 32, n. 3, p. 311–316, 2010. BIZZO, A. L. T. et al. Short-term physiological responses to copper stress in Salvinia auriculata Aubl. Acta Limnologica Brasiliensia, v. 26, n. 3, p. 268–277. 2014. DHIR, B.; SRIVASTAVA, S. Heavy metal removal from a multi-metal solution and wastewater by Salvinia natans . Ecological Engineering, v. 37, n. 6, p. 893–896. 2011. EBRAHEM, M.E; KAMAL, H.S; FARAHT, S.M; MOHAMED, S.G.Y; EITAR, E; SOLIMAN, A.H. Bioaccumulation and translocation of nine heavy metals by Eichhornia crassipes in Nile Delta, Egypt:perspectives for phytoremediation. International Journal of Phytoremediation, vol.21, p.821–830. 2019. ENSLEY, B.D. Rationale for use of phytoremediation. In Phytoremediation of Toxic Metals: Using Plants to Clean up the Environment; Raskin, I., Ensley, B.D., Eds.; Wiley: New York, NY, USA, 2000; pp. 3–11. ESPINOZA-QUINONES, F. R. et al. Study of the bioaccumulation kinetic of lead by living aquatic macrophyte Salvinia auriculata . Chemical Engineering Journal (Amsterdam, Netherlands), v. 150, n. 2–3, p. 316–322, 2009. GAYA, K. S.; MATHEW, L.; RAMESH, B. M. G. A preliminary quantitative phytochemical screening of five macrophytes, N. Paravur, Ernakulam, Kerala, India. CIBTech Journal of Pharmaceutical Sciences, v. 6, n. 1, p. 22–27. 2017. HERALD, T. J.; GADGIL, P.; TILLEY, M. High-throughput micro plate assays for screening flavonoid content and DPPH-scavenging activity in sorghum bran and flour. Journal of the Science of Food and Agriculture, v. 92, n. 11, p. 2326–2331, 2012/08/30 2012. ISSN 0022–5142. LIMA, S.; DIAZ, G.; DIAZ, M.A.N. Antibacterial Chemical Constituent and Antiseptic Herbal Soap from Salvinia auriculata Aubl. Evidence-Based Complementary and Alternative Medicine Volume 2013, Article ID 480509, 5 pages http://dx.doi.org/10.1155/2013/480509 SELLAL, A; BELATTAR, R; BOUZIDI, A. Trace elements removal ability and antioxidant activity of Phragmites australis (from Algeria). International Journal of Phytoremediation, vol.21, p.456–460. 2019. SOARES, D. C. F. et al. Salvinia auriculata : Aquatic bioindicator studied by instrumental neutron activation analysis (INAA). Applied Radiation and Isotopes, v. 66, n. 5, p. 561–564, 2008. SOLOMON W. NEWETE & MARCUS J. BYRNE. The capacity of aquatic macrophytes for phytoremediation and their disposal with specific reference to water hyacinth. Environ Sci Pollut Res (2016) 23:10630–10643 DOI 10.1007/s11356-016-6329-6 SOOD, A; UNIYAL, P.L; PRASANNA, R; AHLUWALIA, A.S. Phytoremediation Potencial of Aquatic Macrophyte, Azolla . AMBIO vol.41, p.122–137. 2012. VESELY, T.; TLUSTOS, P.; SZAKOVA, J. The Use of Water Lettuce ( Pistia Stratiotes L.) for Rhizofiltration of a Highly Polluted Solution by Cadmium and Lead. International Journal of Phytoremediation, v. 13, n. 9, p. 859–872, 2011. VESTENA, S. et al. Cadmium-induced oxidative stress and antioxidative enzyme response in water Hyacinth and Salvinia. Brazilian Journal of Plant Physiology, v. 23, n. 2, p. 131–139. 2011. VIEIRA, L.C; ARAUJO, L.G; FERREIRA, R.V.P; SILVA, E.A; CANEVESI, R.L.S; MARUMO, J.T. Uranium biosorption by Lemna sp. and Pistia stratiotes , vol.203, p.179–186. 2019. WILSON, P.C; WHITWELL, T; KLAINE, S.J. Phytotoxicity, uptake, and distribution of 14C-simazine in Acorus gramenius and Pontederia cordata. Weed Science Society of America, vol.48, p.701–709. 2000. YI, Z.; ZHANG, M.; LING, B.; XU, D.; YE, J. Inhibitory effects of Lantana camera and its contained phenolic compounds in Eichhornia crassipes growth. Ying Yong Sheng Tai Xue Bao. 2006;17(9):1637–40. Additional Declarations No competing interests reported. Supplementary Files SuplementarInformation.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-4356239","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":307956082,"identity":"6fefe1fe-66ac-409f-ac28-530536034c60","order_by":0,"name":"Augusto César Rodrigues","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDElEQVRIiWNgGAWjYFADCcYGhgdAmh/ESSggpDwBqgVESzaABAyI0gKlDQ6ARPBo4Z+Rfk3y5w8befPZzY0PEipq5YzPr0788MCAQZ5f7AB2H9zIKZPmSUgznHPnYLNBwpnjxmY33m6WADrMcObsBOzW3MhJk2ZIOJwgIZHYBkTHErfdOLsBpCXB4DZ2LfJALZI/kLVsnnF28w98WgxupB+T4EFoqUncwN+7Da8thmfeMFvzpKUZzpAB++WAscQN3m0WCQYSOP0idzz94c0fNjbyEtLtDx98qKiT4+8/u/nmjwobeX5pHN5n4EGJgsPAQASrlMChHATYHyDz6oBRdQCP6lEwCkbBKBiJAABRymRl6ZUmcAAAAABJRU5ErkJggg==","orcid":"","institution":"Universidade Federal de Mato Grosso do Sul","correspondingAuthor":true,"prefix":"","firstName":"Augusto","middleName":"César","lastName":"Rodrigues","suffix":""},{"id":307956083,"identity":"29ada3cf-bdae-434f-8c56-39a6b31d62a3","order_by":1,"name":"Samara Requena Nocchi","email":"","orcid":"","institution":"Universidade Federal de Mato Grosso do Sul","correspondingAuthor":false,"prefix":"","firstName":"Samara","middleName":"Requena","lastName":"Nocchi","suffix":""},{"id":307956084,"identity":"505e2576-0f58-498e-9872-9952f05e105e","order_by":2,"name":"Jorge Raposo","email":"","orcid":"","institution":"Universidade Federal de Mato Grosso do Sul","correspondingAuthor":false,"prefix":"","firstName":"Jorge","middleName":"","lastName":"Raposo","suffix":""},{"id":307956085,"identity":"6238e6c3-9be6-4e41-a897-28c0d99b71e6","order_by":3,"name":"Valter Aragão Nascimento","email":"","orcid":"","institution":"Universidade Federal de Mato Grosso do Sul","correspondingAuthor":false,"prefix":"","firstName":"Valter","middleName":"Aragão","lastName":"Nascimento","suffix":""},{"id":307956086,"identity":"9c56b928-c7a6-4af9-af59-7d1e5436f809","order_by":4,"name":"Carlos Alexandre Carollo","email":"","orcid":"","institution":"Universidade Federal de Mato Grosso do Sul","correspondingAuthor":false,"prefix":"","firstName":"Carlos","middleName":"Alexandre","lastName":"Carollo","suffix":""}],"badges":[],"createdAt":"2024-05-02 00:53:23","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4356239/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4356239/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":58055329,"identity":"63714314-0535-405a-b38a-f509df36ea0f","added_by":"auto","created_at":"2024-06-10 14:00:02","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":58967,"visible":true,"origin":"","legend":"\u003cp\u003eExperimental design of the assays of cadmium removal, where samples of dry biomass \u003cem\u003ePontederia parviflora\u003c/em\u003e leaves (PPA-F), petiole/stem (PPA-C), roots (APP-R) and \u003cem\u003eSalvinia auriculata\u003c/em\u003e (SAL) were classified into I (samples in which there is the presence of metabolites secondary) and II (samples that had their metabolites removed). The data of removal where obtained with the difference between the first concentration and the final concentration (in the residual).\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4356239/v1/51a78cab63ad23e4ded30b06.jpeg"},{"id":58055804,"identity":"c820679c-10fa-4484-bbea-9d59fd678903","added_by":"auto","created_at":"2024-06-10 14:08:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":108430,"visible":true,"origin":"","legend":"\u003cp\u003eScheme to elucidate efficiency dynamics in samples featuring both complexed and non-complexed binding sites.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-4356239/v1/c505bab8e4f41a27b0ed1ba7.png"},{"id":61026574,"identity":"50792b41-1fe7-490a-9185-15f5157d5b2a","added_by":"auto","created_at":"2024-07-24 17:49:42","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1405984,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4356239/v1/3c94d904-aa75-4cfd-b123-9a0389dfd235.pdf"},{"id":58055331,"identity":"ce401447-1c5a-4fbe-ab17-80cb7d4a3bc5","added_by":"auto","created_at":"2024-06-10 14:00:02","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":101579,"visible":true,"origin":"","legend":"","description":"","filename":"SuplementarInformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-4356239/v1/e1d677e6042fa887881dd6a4.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"The role of secondary metabolites of Pontederia parviflora Alexander and Salvinia auriculata Aubl. in phytoremediation of cadmium contaminated water","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIn the last decades, rapid industrialization, urbanization, and population growth have led to significant increases in pollutant levels in water and soil resources (CPBC, 2008; SOOD et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). This poses a serious problem, affecting aquatic ecosystems, agriculture, and human health (GUPTA et al., 2010), with heavy metals being a major source of water and soil contamination (SELLAL et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). These heavy metals cannot be degraded through microbial or chemical processes, and their accumulation in the bodies of animals and humans along the food chain can cause DNA damage, carcinogenic effects, and the induction of mutations (EBRAHEM et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSeveral technologies have been employed for the removal of toxic metals from contaminated environments, but they tend to be expensive, ineffective at low concentration levels, and may generate a large amount of waste (SELLAL et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Phytoremediation, on the other hand, represents a biological, cost-effective, and eco-friendly clean-up approach that utilizes plants, their rhizospheres, and associated micro-organisms to degrade, remove, or remediate contaminants from soil and water while promoting vegetation on the land. This method is considered a green alternative since it has identified over 400 plant species as potential phytoremediators (ENSLEY, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; EBRAHEM et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe use of aquatic macrophytes as a tool for phytoremediation is well-documented (NEWETE \u0026amp; BYRNE, 2016). Macrophytes have been demonstrated to efficiently sorb and desorb metals, exhibit high selectivity, reusability, and can be easily separated from solutions, making them excellent biosorbents for effluents (VIEIRA et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003ePontederia\u003c/em\u003e is a genus of emergent aquatic weeds from the Pontederiaceae family. While \u003cem\u003eEichornia crassipes\u003c/em\u003e (Water hyacinth), which belongs to this family, has been extensively studied for the removal of contaminants from wastewater (MALIK et al., 2007; REZANIA et al., 2015; NEWETE \u0026amp; BYRNE, 2016; FENG et al., 2017; MISHRA \u0026amp; MAITI, 2017), few studies are available regarding the phytoremediation capacity of other Pontederiaceae species. In 2000, Wilson et al. observed that phytoremediation with Pontederia cordata reduced the activity of simazine, a commercial herbicidal active ingredient, by 34% in just seven days. In 2010, Balassa et al. showed that Pontederia parviflora Alexander, the species studied in this work, exhibits a high capacity for extraction and storage of copper (Cu), reducing 96% of this metal in solution.\u003c/p\u003e \u003cp\u003e \u003cem\u003eSalvinia auriculata\u003c/em\u003e, another macrophyte studied in this work, is a free-floating weed belonging to the Salvinaceae family that has demonstrated positive results in cadmium bioaccumulation (WOLFF et al., 2012), lead (ESPINOZA-QUINONES et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; VESELY et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2011\u003c/span\u003e), chrome, and other metals, putting this species in the spotlight as one of the most efficient species in the accumulation and removal of \"multielementary\" samples (SOARES et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePhytoremediation research associates some chemical functions, such as hydroxyl and carboxylic groups, as agents that contribute to the potential of contaminant uptake in solution. These structures are present in secondary metabolites found in the tested plants, suggesting that the plants could detoxify the water or soil through their metabolic activities (DHIR et al., 2010; ALI et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the literature, there are few studies available concerning the chemical composition of aquatic macrophytes. Although some studies have indicated the presence of phenolic compounds, alkaloid, and terpenoid derivatives in \u003cem\u003eEichornia crassipes\u003c/em\u003e (YI et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; ABOUL-ENEIN et al., 2014), which belongs to the same family as \u003cem\u003ePontederia parviflora\u003c/em\u003e, no studies could be found for the latter. Regarding \u003cem\u003eS. auriculata\u003c/em\u003e, Lima et al. (2013) isolated two steroids (stigmasterone and stigmasterol) and one triterpene (friedelinol) and observed that only stigmasterone showed activity against Staphylococcus aureus strains.\u003c/p\u003e \u003cp\u003eSecondary metabolites are synthesized with the purpose of not only attracting insects for pollination but also defending the plants from herbivory, UV light, and other mechanisms that may cause damage, such as in response to the presence of heavy metals in the environment. Bizzo et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) reported an increased production of phenolic compounds, especially anthocyanin, in \u003cem\u003eS. auriculata\u003c/em\u003e in the presence of high copper concentrations, with a reduction in flavonoid concentrations after a short exposure period. Another study reported the reduction of antioxidant activity in samples of \u003cem\u003eS. auriculata\u003c/em\u003e and \u003cem\u003eE. crassipes\u003c/em\u003e under cadmium stress (VESTENA et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2011\u003c/span\u003e), possibly due to the decrease in secondary metabolites responsible for this activity. Recently, \u003cem\u003eEichhornia crassipes\u003c/em\u003e and \u003cem\u003eSalvinia molesta\u003c/em\u003e were compared in terms of the quantity of some secondary metabolites, and the highest levels of tannins, flavonoids, and terpenoids were found in E. crassipes, while the content of alkaloid groups, saponins, and phenols was higher in S. molesta (GAYA et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSince macrophytes are known to be powerful phytoremediators, and the mechanisms responsible for trapping heavy metals are not yet very clear, this study aims to establish a relationship between the chemical profile of two aquatic macrophytes and their performance in phytoremediation. Additionally, considering that secondary metabolites play an important role in maintaining plants in the environment, the study seeks to understand the role of secondary metabolites in this process.\u003c/p\u003e \u003cp\u003eSo, in this study, the chemical composition of \u003cem\u003eP. parviflora\u003c/em\u003e and \u003cem\u003eS. auriculata\u003c/em\u003e was established based on liquid chromatography-diode array detector-mass spectrometry (LC-DAD-MS) and MS/MS. The phenol and tannin content in each species was determined, and the concentration of arsenic (As), cadmium (Cd), chromium (Cr), magnesium (Mg), manganese (Mn), molybdenum (Mo), nickel (Ni), and lead (Pb) in the biomass of macrophyte samples was evaluated before and after the extraction process. We also assessed the influence of secondary metabolites on the fixation of metals in the biomass of samples, besides evaluating the potential for Cd removal in the ex-situ phytoremediation system.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eCollection and Extraction of Plant Material\u003c/h2\u003e \u003cp\u003eWe collected \u003cem\u003ePontederia parviflora\u003c/em\u003e and \u003cem\u003eSalvinia auriculata\u003c/em\u003e in the Lagoa Comprida Municipal Natural Park, in the hydrographic basin of the River Aquidauana, Aquidauana, Mato Grosso do Sul, Brazil, located between the coordinates 20\u0026deg;23\u0026rsquo;36.8\u0026rdquo; S and 20\u0026deg;23\u0026rsquo;28.8\u0026rdquo; S, and 55\u0026deg;48\u0026rsquo;51.5\u0026rdquo; W and 55\u0026deg;47\u0026rsquo;04.9\u0026rdquo; W. This location is situated between the ecotone Cerrado and Pantanal, characterized by a large biological diversity combining features from both ecosystems.\u003c/p\u003e \u003cp\u003eWe separated the submerged petiole and the emerged leaf blade of \u003cem\u003eP. parviflora\u003c/em\u003e into roots (PPA-R), petioles/stems (PPA-S), and leaves (PPA-L). For \u003cem\u003eS. auriculata\u003c/em\u003e (SAL), the whole plant was used since the floating leaves are also in contact with the water. The plants were dried in an oven with air circulation at 50 \u0026ordm;C for 72 hours until a constant weight was achieved. The dry biomass was ground in a knife mill and sieved through a 60-mesh screen to obtain a uniform granulometry.\u003c/p\u003e \u003cp\u003eA portion of the pulverized material was extracted using a pressurized fluid extractor (Dionex, ASE 150\u0026reg;), equipped with a 100 ml extraction cartridge. The extraction was performed with ethanol: water (7:3 v/v) following parameters repeated three times during each extraction: temperature of 125\u0026deg;C, static extraction time of 15 min, washing of the cell with 100% of the volume of its capacity, and purging for 60 seconds. The solvents were evaporated in a rotavapor (B\u0026uuml;chi R-3, Switzerland) under reduced pressure and then lyophilized. The \u003cem\u003eP. parviflora\u003c/em\u003e leaves extract yielded 29.4%, while the petioles/stems and roots extracts yielded 27.4% and 2.6%, respectively, and the \u003cem\u003eS. auriculata\u003c/em\u003e extract yielded 6.9%. After the extraction process, the plant material was dried again and reserved for heavy metal removal analyses, named as \u003cem\u003eP. parviflora\u003c/em\u003e roots (PPA-R II), petioles/stems (PPA-S II), leaves (PPA-L II), and \u003cem\u003eS. auriculata\u003c/em\u003e (SAL II) because they are used as biomass without secondary metabolites.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eTotal Phenols Content\u003c/h2\u003e \u003cp\u003eTo evaluate the content of total phenols and tannins (addressed in the next section) in \u003cem\u003eP. parviflora\u003c/em\u003e and \u003cem\u003eS. auriculata\u003c/em\u003e extracts, we followed the methodology proposed by Herald et al. (2012). In a 96-well plate, 75 \u0026micro;L of the sample or standard (gallic acid) at various concentrations was added to each well. Then, 75 \u0026micro;L of Folin-Ciocalteu reagent (1:1 v/v, deionized water) was added to each well, and the plate was homogenized. After 6 minutes of mixing, 100 \u0026micro;L of Na2CO3 at 75 g/L was added to the wells, remixed, and the mixture was left for 90 minutes in darkness. The measurement was taken at 765 nm using a spectrophotometric microplate reader. The analyses were conducted in triplicate. The total phenolic content was expressed as mg of gallic acid equivalent (GAE) per gram of each extract.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eTotal Tannins Content\u003c/h2\u003e \u003cp\u003eTo determine the total tannin content, the extracts were solubilized in water (4 mg/mL), brought into contact with skin powder (20 mg, Hide Powder - protease substrate purchased from Sigma-Aldrich), and stirred for 60 minutes. After centrifugation, the supernatant was utilized to undergo the same process as described above for phenols. The concentration of total tannins was calculated as the difference in the concentration between total phenolics and tannin phenols. The total tannin content is expressed as mg of GAE per gram of each extract.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eIdentification of chemical constituents in \u003cem\u003eP. parviflora\u003c/em\u003e and \u003cem\u003eS. auriculata\u003c/em\u003e extracts by LC-DAD-MS/MS\u003c/h2\u003e \u003cp\u003eEach extract was prepared at a concentration of 1 mg/mL in methanol: water (7:3, v/v), then filtered through a PTFE filter (Millex 0.22 mm x 13 mm, Millipore\u0026reg;). Subsequently, 3 \u0026micro;L of the filtered solution was injected into a Shimadzu Prominence UFLC system coupled with a diode array detector (DAD) and a MicrOTOF-Q III mass spectrometer (Bruker Daltonics). The chromatographic column used was a Kinetex C18 column (2.6 \u0026micro;m, 150 \u0026times; 2.1 mm, Phenomenex). The injection volume, flow rate, and oven temperature were set at 1 \u0026micro;L, 0.3 mL/min, and 50 \u0026ordm;C, respectively. The mobile phase comprised 0.1% formic acid (v/v) in both water (solvent A) and acetonitrile (solvent B), utilizing a gradient elution profile as follows: 0\u0026ndash;2 min: 3% B; 2\u0026ndash;25 min: 3\u0026ndash;25% B; 25\u0026ndash;40 min: 25\u0026ndash;80% B; 40\u0026ndash;43 min: 80% B. The analyses were carried out in both negative and positive ion modes. Nitrogen gas was used as both a nebulizer gas (at 4 Bar) and a dry gas (at 9 L/min). The capillary voltage was set to 2.5 kV.\u003c/p\u003e \u003cp\u003e \u003cb\u003eQuantification of heavy metals (As, Cd, Cr, Mg, Mn, Mo, Ni, Pb) in\u003c/b\u003e \u003cb\u003eP. parviflora\u003c/b\u003e \u003cb\u003eand\u003c/b\u003e \u003cb\u003eS. auriculata\u003c/b\u003e \u003cb\u003ebiomass\u003c/b\u003e\u003c/p\u003e \u003cp\u003eTo quantify heavy metals in macrophyte biomass, the two studied plant species were divided into two groups: one labeled \"non-extracted,\" where biomass underwent no secondary metabolite extraction, indicated by adding \"I\" after each sample's acronyms; and another group labeled \"extracted,\" where biomass was subjected to extraction, marked by adding \"II\" after each sample's acronyms, as described in the \"Collection and Extraction of Plant Material\" section.\u003c/p\u003e \u003cp\u003eCalibration curves for each metal (As, Cd, Cr, Mg, Mn, Mo, Ni, Pb) were prepared in triplicate using seven standard concentrations (0.05; 0.1; 0.25; 0.5; 1.0; 1.5; and 2.0 \u0026micro;g/mL). For Cd, the calibration curve was generated with 12 different concentrations (0.05; 0.1; 0.25; 0.5; 1.0; 1.5; 2.0; 4.0; 6.0; 8.0; and 10.0 \u0026micro;g/mL). These solutions were derived from aqueous mono-elemental standards acidified with 1% HNO3. The limit of detection (LOD) and limit of quantification (LOQ) were calculated from the three standard curves using the standard deviation (SD\u0026#119886;) of the intercept with the \u0026#119910;-axis and the calibration curve slope (S), according to:\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e \u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eLOD\u0026thinsp;=\u003c/span\u003e\u0026thinsp;SD\u0026#119886; \u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003ex 3/\u003c/span\u003e\u003cspan type=\"ItalicSmallCaps\" class=\"ItalicSmallCaps\" name=\"Emphasis\"\u003eS\u003c/span\u003e\u003c/p\u003e\u003cp\u003e \u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eLOQ\u0026thinsp;=\u003c/span\u003e\u0026thinsp;SD\u0026#119886; \u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003ex 10/\u003c/span\u003e\u003cspan type=\"ItalicSmallCaps\" class=\"ItalicSmallCaps\" name=\"Emphasis\"\u003eS\u003c/span\u003e\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eBefore analysis, the PRE and POS samples underwent digestion to remove organic residues. For this, 400 mg of dry biomass from each sample (PRE and POS of \u003cem\u003eP. parviflora\u003c/em\u003e roots, petioles/stems, and leaves, as well as PRE and POS of \u003cem\u003eS. auriculata\u003c/em\u003e) was placed in a digester tube within a Berghof\u0026reg; Speedwave four microwave digester. To this, 3 mL of nitric acid, 2 mL of ultrapure water, and 1 mL of hydrogen peroxide were added, and the protocol specified for aquatic macrophytes in the equipment manual (Speedwave) was followed.\u003c/p\u003e \u003cp\u003eThe microwave digester was set with the following parameters: Step (1) utilized 80% power at 150 \u0026ordm;C, with a 5-minute ramp time, 10-minute retention time, and 80 Bar pressure. In Step (2), power was set at 90%, temperature at 190 \u0026ordm;C, with a 2-minute ramp time, 20-minute retention time, and 80 Bar pressure. Finally, in Step (3), power was set to 0%, temperature to 50 \u0026ordm;C, with a 1-minute ramp time, 10-minute retention time, and 60 Bar pressure.\u003c/p\u003e \u003cp\u003eAqueous residual solutions from each digester tube were brought to a volume of 15 mL with water and analyzed using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) (iCAP 6000, Thermo\u0026reg;). The analysis conditions were as follows: power at 1250 W, sample flow at 0.35 L/min, plasma gas flow at 12 L/min, integration time at 5 s, stabilization time at 20 s, nebulization time at 30 psi, plasma view axial, and air gas (99.999%).\u003c/p\u003e \u003cp\u003eEmission lines (nm) for each analyte were as follows: As 189.042, Cd 228.802, Cr 283.563, Mg 279.553, Mn 257.610, Mo 202.030, Ni 221.647, and Pb 220.353.\u003c/p\u003e \u003cp\u003eThe quantification calculation for metals present in the macrophyte biomass was performed using the following formula:\u003c/p\u003e \u003cp\u003eTotal metal in the sample (\u0026micro;g/g)\u0026thinsp;=\u0026thinsp;Metal mass (\u0026micro;g) found / Sample mass used (g)\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eCadmium Removal trials\u003c/h2\u003e \u003cp\u003eRemoval trials were conducted to establish the relationship between secondary metabolite presence in plant biomass and metal removal through passive phytoremediation. The experimental design is depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn plastic tubes, we prepared 20 mL of Cd mono-elemental solutions at concentrations of 0.2 and 3.5 \u0026micro;g/mL, acidified with water. To these solutions, 0.2 g of biomass from each sample (I and II of \u003cem\u003eP. parviflora\u003c/em\u003e roots, petioles/stems, and leaves, as well as I and II of \u003cem\u003eS. auriculata\u003c/em\u003e) was added. Subsequently, the solutions were stirred for 15 minutes and then filtered. The resulting aqueous residuals underwent microwave digestion, as detailed in the preceding section, followed by analysis using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) (iCAP 6000, Thermo\u0026reg;).\u003c/p\u003e \u003cp\u003eThe amount of Cd removed from the solution by the dry biomass of the macrophytes was calculated as follows:\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e% Cadmium removed = (ΔC / Standard Concentration) x 100\u003c/h2\u003e \u003cdiv id=\"Sec9\" class=\"Section4\"\u003e \u003ch2\u003eΔC\u0026thinsp;=\u0026thinsp;Control concentration \u0026ndash; Sample concentration\u003c/h2\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eTo quantify heavy metals (As, Cd, Cr, Mg, Mn, Mo, Ni, Pb) in \u003cem\u003eP. parviflora\u003c/em\u003e and \u003cem\u003eS. auriculata\u003c/em\u003e biomass, as well as for Cd removal tests, uniform data underwent analysis using one-way analysis of variance (ANOVA), and means were compared using the Tukey test (\u003cem\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/em\u003e). Results are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eTotal phenolic and tannin contents in macrophyte extracts are shown in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. \u003cem\u003eS. auriculata\u003c/em\u003e and \u003cem\u003eP. parviflora\u003c/em\u003e leaves contained the highest phenol and tannin levels, with no statistical difference (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eTotal phenolic and total tannin content in extracts of macrophytes samples.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSample\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePhenol content\u003c/p\u003e\n\u003cp\u003e(mg GAE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTannin content\u003c/p\u003e\n\u003cp\u003e(mg GAE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP. parviflora\u003c/em\u003e leaves\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e61.12\u003csup\u003ec\u003c/sup\u003e (1.26)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.32\u003csup\u003eb,c\u003c/sup\u003e (1.29)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP. parviflora\u003c/em\u003e petiole/stem\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16.36\u003csup\u003ea\u003c/sup\u003e (0.35)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.56\u003csup\u003ea\u003c/sup\u003e (0.13)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP. parviflora\u003c/em\u003e roots\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e29.23\u003csup\u003eb\u003c/sup\u003e (1.24)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.24\u003csup\u003eb\u003c/sup\u003e (0.12)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eS. auriculata\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e62.56\u003csup\u003ec\u003c/sup\u003e (1.24)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.21\u003csup\u003ec\u003c/sup\u003e (0.46)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"3\"\u003eThe results are expressed in mg GAE (gallic acid equivalent) per g of extract.\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"3\"\u003eDifferent letters represent significantly different means (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe variation among \u003cem\u003eP. parviflora\u003c/em\u003e samples could be linked to the roles of analyzed structures. Leaves, which are photosynthetic, displayed the highest phenol content. Phenolic compounds, linked to UV absorption, might be more abundant in leaves due to their photoprotective effect.\u003c/p\u003e\n\u003cp\u003eThe primary compounds in macrophyte samples were identified using UV, MS, and MS/MS data, compared with published information (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). Predominantly, they comprised flavonoids and phenylpropanoids.\u0026nbsp;\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eIdentification of the constituents from hydroethanolic extract of \u003cem\u003eSalvinia auriculata\u003c/em\u003e by LC-DAD-MS/MS.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePeak\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eRT\u003c/p\u003e\n\u003cp\u003e(min)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eCompound\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eMolecular\u003c/p\u003e\n\u003cp\u003eFormula\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eError\u003c/p\u003e\n\u003cp\u003ePpm\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eUV\u003c/p\u003e\n\u003cp\u003e(nm)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eNegative mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePositive mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-2,4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e260\u0026ndash;297\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e327.0729\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e191.0635;173.0485\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e329.0874\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4-\u003cem\u003eO-E\u003c/em\u003e-caffeoylquinic\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-2,8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e297\u0026ndash;325\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e353.0888\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e191.0548;179.0277\u003c/p\u003e\n\u003cp\u003e173.0345; 161.0196\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0386\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e10.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4\u003cem\u003e-O-E\u003c/em\u003e-caffeoylquinic\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-2,1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;325\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e353.0885\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e191.0571; 179.0375; 173.0456; 161.0161\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e355.1037\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0390; 145.0279\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e14.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e10\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2,8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e304\u0026ndash;334\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e177.0548\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e161.0244\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e16.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3,3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e285\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e399.1284\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e153.0273\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e18.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e15\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0,7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e283\u0026ndash;329\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e641.1516\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e449.1000; 405.0954; 327.0566; 297.0419; 271.0588; 191.0570; 173.0441; 161.0223\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e643.1681\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e311.0581; 299.0575; 271.0578; 231.0693; 191.0350; 163.0403\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e19.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eKaempferol-\u003cem\u003eO\u003c/em\u003e-hexoside\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-1,2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e266\u0026ndash;346\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e461.0731\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e285.0388\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e463.0880\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e287.0569\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3,4-dicafeoylquinic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-2,1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e297\u0026ndash;325\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e515.1206\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e335.0795; 191.0562; 179.0344; 173.0477; 161.0250; 155.0380\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e517.1361\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0399\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e31\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e14\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0,1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e283\u0026ndash;329\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e625.1564\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e433.0786; 281.0457; 191.0571; 161.0251\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e627.1737\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e283.0591; 163.0395\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3,5-dicafeoylquinic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-1,4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e298\u0026ndash;329\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e515.1202\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e191.0570; 179.0340; 161.0270\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e21.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSalviniside II derivative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-1,3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e330\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e473.0732\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e413.0480; 285.0481; 241.0490\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e475.0890\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e313.0342; 295.0262; 267,0279\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e22.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4,5-dicafeoylquinic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0,3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e298\u0026ndash;329\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e515.1197\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e325.0149; 191.0596; 179.0330; 173.0470; 161.0213\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e499.1243\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e23.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSalviniside II derivative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-1,0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e330\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e473.0730\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e413.0548; 327.0514; 267.0216; 241.0516\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e475.0894\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e313.0362; 267.0297; 239.0362\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e25.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e23\u003c/sub\u003eNO\u003csub\u003e13\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-1,9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e284\u0026ndash;329\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e568.1108\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e413.0473; 241.0603\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e570.1233\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e313.0331; 295.0226\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"10\"\u003e\u0026ldquo;Unknown\u0026rdquo; are the non identified compounds.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"char\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"char\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003eIn the \u003cem\u003eS. auriculata\u003c/em\u003e extract, we observed 14 peaks (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). Among these, peaks 2 and 3 had the ion at \u003cem\u003em/z\u003c/em\u003e 353 as the base peak in the negative ionization spectrum, along with two bands near 299 and 325 nm in the UV spectrum, characteristic of caffeoylquinic acids. Peaks 2 and 3 exhibited product ions at \u003cem\u003em/z\u003c/em\u003e 191 due to the loss of the caffeoyl unit, \u003cem\u003em/z\u003c/em\u003e 179 related to the quinic unit's loss, and \u003cem\u003em/z\u003c/em\u003e 173 for the dehydrated quinic acid unit. As only the caffeoylquinic acid esterified at position 4 produces the product ion at \u003cem\u003em/z\u003c/em\u003e 173, these peaks were identified as 4-O-E-caffeoylquinic acid (GOBBO-NETO, 2007).\u003c/div\u003e\n\u003c/div\u003e\n\u003cp\u003eCompounds 11 and 13 displayed intense ions at \u003cem\u003em/z\u003c/em\u003e 473.0732 and 473.0730, respectively, in alignment with the molecular formula C22H18O12. These peaks were recognized as derivatives of salviniside II (Li et al., 2013). Peak 7 exhibited bands at 265 and 345 nm in the UV spectrum, characteristic of flavonol structures. This compound bore the molecular formula C21H18O12, with ions \u003cem\u003em/z\u003c/em\u003e 461.0731 [M-H]- and 463.0880 [M\u0026thinsp;+\u0026thinsp;H]-, and a key fragment at \u003cem\u003em/z\u003c/em\u003e 285, harmonizing with flavonoid aglycone kaempferol. Hence, this compound was identified as kaempferol-O-hexoside (SADEER et al., 2019).\u003c/p\u003e\n\u003cp\u003ePeaks 8, 10, and 12, with retention times of 20.0, 20.6, and 22.1 min, displayed bands at 285 and 314 nm in the UV spectrum, along with \u003cem\u003em/z\u003c/em\u003e 515 [M\u0026thinsp;+\u0026thinsp;H]\u0026thinsp;+\u0026thinsp;and 513 [M-H]-, encompassing molecular formula C25H24O12, compatible with dicaffeoylquinic acids (CLIFFORD et al., 2003, 2005). Peaks 10 and 12 were validated as 3,5 and 4,5-dicaffeoylquinic acids, respectively, using genuine standard injection. The fragment ions of peak 8 were juxtaposed with identification cues suggested by Clifford et al. (2003, 2005) for dicaffeoylquinic acids. In these guidelines, the fragment ion at \u003cem\u003em/z\u003c/em\u003e 173 functions as a diagnostic indicator of a caffeic unit esterified at the fourth position of quinic acid. This data, coupled with the absence of characteristic ions of 1,4-dicaffeoylquinic acid (\u003cem\u003em/z\u003c/em\u003e 299 and \u003cem\u003em/z\u003c/em\u003e 203), substantiated the identification of this compound as 3,4-dicaffeoylquinic acid (GOBBO-NETO 2007).\u003c/p\u003e\n\u003cp\u003eIn the extract of \u003cem\u003eP. parviflora\u003c/em\u003e leaves (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e), 19 peaks were detected. Peaks 1, 2, 4, and 7, displaying the base ion at \u003cem\u003em/z\u003c/em\u003e 209, were suggested based on their molecular formulas and UV spectra as derivatives of caffeoyl glucarate. Peaks 3 and 6, exhibiting two bands in the UV spectra within the range of 300 to 324 nm and 299 to 325 nm, respectively, displayed the base ion at \u003cem\u003em/z\u003c/em\u003e 371. Both peaks 3 and 6 also showed product ions at \u003cem\u003em/z\u003c/em\u003e 191, with peak 6 additionally featuring an ion at \u003cem\u003em/z\u003c/em\u003e 209, characterizing them as isomers of caffeoyl glucarates (LORENZ et al. 2014).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab4\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eIdentification of the constituents from hydroethanolic extract of \u003cem\u003ePontederia parviflora\u003c/em\u003e (leaves) by LC-DAD-MS/MS.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePeak\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eRT\u003c/p\u003e\n\u003cp\u003e(min)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eCompound\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eMolecular\u003c/p\u003e\n\u003cp\u003eFormula\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eError\u003c/p\u003e\n\u003cp\u003ePpm\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eUV\u003c/p\u003e\n\u003cp\u003e(nm)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eNegative mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePositive mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaffeoyl glucarate derivatives\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;324\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e209.0311\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaffeoyl glucarate derivatives\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;324\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e209.0303\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIsomers of caffeoyl glucarates\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e11\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;326\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e371.0624\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e191.0234\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaffeoyl glucarate derivatives\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;324\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e209.0304\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTryptophan derivative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e11\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e280\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e203.0824\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIsomers of caffeoyl glucarates\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e11\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-2.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e299\u0026ndash;325\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e371.0628\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e209.0254; 191.0182\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaffeoyl glucarate derivative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e6\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;324\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e209.0303\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnit of caffeic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e299\u0026ndash;320\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e179.0349\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0371\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e10.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIsomer of caffeoylquinic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e285\u0026ndash;325\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e353.0871\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e191.0572; 161.0197\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0375\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e12.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaffeic acid derivatives\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e15\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e299\u0026ndash;324\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e311.0786\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e271.0942; 255.0767; 179.0354; 161.0316; 149.0488\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e625.1748\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0377\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e12.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e13\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-4.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;325\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e295.0471\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e179.0384\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e13.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaffeic acid derivatives\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e15\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;324\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e311.0787\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e311.0823; 271.0922; 243.0617; 179.0356; 161.0252; 149.0456\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e313.0898\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0379\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e14.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-4.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e285\u0026ndash;320\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e335.0789\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e179.0434; 161.0292\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e16.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e298\u0026ndash;324\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e293.0676\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e179.0378; 161.0228\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eKaempferol-O-rutinoside\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e15\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-2.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e269\u0026ndash;346\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e593.1525\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e477.0882; 285.0415\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e595.1640\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2870549\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3,4 dicafeoylquinic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e269\u0026ndash;346\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e515.1200\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e191.0603; 179.0335; 173.0407; 161.0183\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3,5 dicafeoylquinic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e280\u0026ndash;320\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e515.1162\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e18\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e22.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4,5 dicafeoylquinic acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e25\u003c/sub\u003eH\u003csub\u003e24\u003c/sub\u003eO\u003csub\u003e12\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e280\u0026ndash;320\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e515.1202\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e23.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eKaempferol-O-ramnoside-O-malonil-O-hexoside\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e30\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e18\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e267\u0026ndash;347\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e679.1515\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e635.1623; 489.1055; 285.0380\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e681.1630\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e287.0554\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e26.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eKaempferol derivative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e16\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e273\u0026ndash;335\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e635.1622\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e30.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;311\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e327.2188\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e171.1031\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e22\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e31.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;311\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e329.2345\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e229.1431; 211.1313; 171.0995\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e31.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;308\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e303.0654\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e274.0646\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e305.0817\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e277.0761; 259.0711; 249.0938; 231.0808; 213.0638; 203.0872\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e33.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-4.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e287.0726\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e287.0773\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e289.0848\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e271.0725; 261.0924; 243.0786; 233.0945; 215.0837\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e35.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e273\u0026ndash;311\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e309.2075\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e183.0064\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"10\"\u003e\u0026ldquo;Unknown\u0026rdquo; are the non identified compounds.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003ePeak 5 demonstrated a base ion at \u003cem\u003em/z\u003c/em\u003e 203 in negative ionization mode, along with two ions at \u003cem\u003em/z\u003c/em\u003e 188 and \u003cem\u003em/z\u003c/em\u003e 205 in positive ionization mode. Furthermore, a product ion at \u003cem\u003em/z\u003c/em\u003e 170 was observed. This compound was identified as a derivative of tryptophan (LIU et al. 2015).\u003c/p\u003e\n\u003cp\u003ePeak 8 displayed an ion at \u003cem\u003em/z\u003c/em\u003e 179 and lacked product ions in the fragmentation spectrum. However, relying on its UV spectrum and the molecular formula obtained from the software, this peak was suggested to represent a unit of caffeic acid, a product ion resulting from the fragmentation of caffeoylquinic acid (GOBBO-NETO, 2007). Peak 9, akin to peaks 2 and 3 of \u003cem\u003eS. auriculata\u003c/em\u003e, exhibited the base ion at \u003cem\u003em/z\u003c/em\u003e 353 and two bands in the UV spectrum ranging from 285 to 325 nm, along with product ions at \u003cem\u003em/z\u003c/em\u003e 191 and \u003cem\u003em/z\u003c/em\u003e 161. This peak was identified as an isomer of caffeoylquinic acid (GOBBO-NETO, 2007).\u003c/p\u003e\n\u003cp\u003ePeaks 10 and 12 displayed ions at \u003cem\u003em/z\u003c/em\u003e 311 [M-H]-, consistent with the molecular formula C14H15O8. However, peak 10 featured a main ion at \u003cem\u003em/z\u003c/em\u003e 625 [M\u0026thinsp;+\u0026thinsp;H]+, potentially indicating a dimer, while peak 12 exhibited \u003cem\u003em/z\u003c/em\u003e 313 [M\u0026thinsp;+\u0026thinsp;H]+. These compounds were recognized as derivatives of caffeic acid connected to a unit of D-erythrono-1,4-lactone or D-threono-1,4-lactone (CCANA-CCAPATINA et al., 2017).\u003c/p\u003e\n\u003cp\u003ePeaks 15 and 19 exhibited two bands in the UV spectrum near wavelengths of 265 and 345 nm (typical of flavonols), along with ions at \u003cem\u003em/z\u003c/em\u003e 593.1525 [M-H]- and 679.1515 [M-H]-, respectively. The compounds possessed the molecular formulas C27H29O15 and C30H31O18, sharing the main fragment at \u003cem\u003em/z\u003c/em\u003e 285, akin to compound 7 of \u003cem\u003eS. auriculata\u003c/em\u003e. Compound 15 was identified as Kaempferol-O-rutinoside (HERRANZ-L\u0026Oacute;PEZ et al., 2012), and compound 19 as Kaempferol-3-O-rutinoside-7-O-rhamnoside (JU et al., 2018).\u003c/p\u003e\n\u003cp\u003ePeak 20 featured a base ion at \u003cem\u003em/z\u003c/em\u003e 635, with the identical molecular formula as peak 19, albeit without any fragmentation. Hence, this peak was identified as a derivative of kaempferol (SADEER et al., 2019). Compounds 16, 17, and 18 were designated as 3,4, 3,5, and 4,5-dicaffeoylquinic acids, congruent with their presence in S. auriculata peaks 8, 10, and 12 (CCANA-CCAPATINA et al., 2017).\u003c/p\u003e\n\u003cp\u003eIn \u003cem\u003eP. parviflora\u003c/em\u003e petiole/stem (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e), 7 peaks were detected. Peaks 2 and 3 exhibited the base ion at \u003cem\u003em/z\u003c/em\u003e 311 [M-H]-, with two bands in the wavelength range of 294 to 329 nm and 300 to 322 nm in the UV spectra, respectively, along with the molecular formula C14H15O8. Although peak 2 lacked fragmentation in the MS/MS spectrum, based on UV data and the molecular formula, this compound was tentatively identified as a derivative of chlorogenic acid. Peak 3 displayed a product ion at \u003cem\u003em/z\u003c/em\u003e 179 [M-H]-, linked to the caffeoyl unit. Furthermore, it demonstrated the ion at \u003cem\u003em/z\u003c/em\u003e 313 [M\u0026thinsp;+\u0026thinsp;H]\u0026thinsp;+\u0026thinsp;with a product ion at \u003cem\u003em/z\u003c/em\u003e 163 [M\u0026thinsp;+\u0026thinsp;H]+. Considering the calculated molecular formulae, these resemblances were noted with peaks 10 and 12 of P. parviflora leaves. Consequently, this compound was identified as a caffeic acid derivative connected to a unit of D-erythrono-1,4-lactone or D-threono-1,4-lactone (CCANA-CCAPATINA et al., 2017).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab5\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eIdentification of the constituents from hydroethanolic extract of \u003cem\u003ePontederia parviflora\u003c/em\u003e (petiole/stem) by LC-DAD-MS/MS.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePeak\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eRT\u003c/p\u003e\n\u003cp\u003e(min)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eCompound\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eMolecular\u003c/p\u003e\n\u003cp\u003eFormula\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eError\u003c/p\u003e\n\u003cp\u003ePpm\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eUV\u003c/p\u003e\n\u003cp\u003e(nm)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eNegative mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePositive mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eN\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e283\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e203.0826\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e12.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eChlorogenic acid derivative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-5.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e294\u0026ndash;329\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e311.0788\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e13.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCaffeic acid derivative\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e8\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e300\u0026ndash;322\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e311.0775\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e179.0401\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e313.0921\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e163.0386\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e20.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e27\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003csub\u003e15\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e285\u0026ndash;330\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e593.1511\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e595.1664\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e287.0555\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e23.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e30\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e18\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e290\u0026ndash;345\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e681.1653\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e287.0550\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e30.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e31\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e285\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e327.2197\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e211.1362\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e32.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e14\u003c/sub\u003eH\u003csub\u003e17\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e283\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e249.1128\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"10\"\u003e\u0026ldquo;Unknown\u0026rdquo; are the non identified compounds.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eFor \u003cem\u003eP. parviflora\u003c/em\u003e roots (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e), 9 peaks were discerned. Peak 3 exhibited two bands within the wavelengths of 280 to 310 nm in the UV spectra and the ion at \u003cem\u003em/z\u003c/em\u003e 163 [M-H]-, consistent with the molecular formula C9H7O3, implying a coumaric acid molecule (DUGO et al., 2008).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eIdentification of the constituents from hydroethanolic extract of \u003cem\u003ePontederia parviflora\u003c/em\u003e (roots) by LC-DAD-MS/MS.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePeak\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eRT\u003c/p\u003e\n\u003cp\u003e(min)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eCompound\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eMolecular\u003c/p\u003e\n\u003cp\u003eFormula\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eError\u003c/p\u003e\n\u003cp\u003ePpm\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eUV\u003c/p\u003e\n\u003cp\u003e(nm)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eNegative mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePositive mode \u003cem\u003e(m/z)\u003c/em\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMS/MS\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e12\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e11\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-7.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e269\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e341.1116\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e191.0588\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e365.1092\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e203.0522\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e17\u003c/sub\u003eO\u003csub\u003e9\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-4.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e269\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e377.0893\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e13.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCoumaric acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e9\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e280\u0026ndash;310\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e163.0401\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e19.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e275\u0026ndash;308\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e181.0509\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e19.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-12.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;310\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e187.1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e06\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e30.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e17\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-4.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;311\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e329.0681\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e299.0270; 271.0236; 227.0467; 199.0431; 161.0193\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e331.0822\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e315.0521; 302.0464; 287.0549; 270.0489; 258.0514; 242.0549\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e07\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e31.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-2.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;312\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e329.2340\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e211.1341; 171.1042\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e667.3233\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e433.2096; 389.1835; 261.1318; 217.1060; 173.0796; 155.0716\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e31.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e13\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;310\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e221.0663\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e33.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eUnknown\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e276\u0026ndash;312\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e273.0562\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e245.0636\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"10\"\u003e\u0026ldquo;Unknown\u0026rdquo; are the non identified compounds.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe results obtained from the heavy metal quantification tests in the biomass of the studied macrophytes (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e) demonstrated that, barring Ni and Mo in \u003cem\u003eP. parviflora\u003c/em\u003e, the metal concentration escalated after the extraction of secondary metabolites. Employing ICP-OES, it was apparent that the heavy metals existing in the macrophytes' biomass were not predominantly complexed with the plants' metabolites, or at least, a majority of these metals were not. This is inferred from the elevation of metal levels in samples where metabolites were removed. Although secondary metabolites encompass clusters such as \u0026ndash;OH- and \u0026ndash;COO-, known to form bonds with metals (SARASWAT and RAI, 2010; ALI et al., \u003cspan class=\"CitationRef\"\u003e2013\u003c/span\u003e), our results imply that metals are chiefly associated with plant structures that persist throughout the process of secondary metabolite extraction, such as cell walls and vacuoles. Additionally, no correlation was established between the content of total phenols and tannins in the samples and the metal accumulation capacity, further affirming that the secondary metabolites in these plants are not primarily responsible for phytoremediation. The calibration parameters acquired for the ICP-OES are accessible in the Supplementary Material (Table \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab6\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eConcentration (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;deviation, n\u0026thinsp;=\u0026thinsp;3) in \u0026micro;g g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of As, Cd, Cr, Mg, Mn, Mo, Ni and Pb found in \u003cem\u003eP. parviflora\u003c/em\u003e-leaf (PPA-F), \u003cem\u003eP. parviflora\u003c/em\u003e - petioles/stems (PPA-C), \u003cem\u003eP. parviflora\u003c/em\u003e-roots (PPA-R) and \u003cem\u003eS. auriculata\u003c/em\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eSample\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"8\" align=\"left\"\u003e\n\u003cp\u003eMeans (standard deviation)\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAs\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eCd\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eCr\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMg\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMn\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMo\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eNi\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePb\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-F \u0026ndash; I\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.331\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.013)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;DL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.179\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.001)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.419\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.001)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.1254\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.0003)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.256\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.010)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-F \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.371\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.016)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;DL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.499\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.002)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.464\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.003)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.2108\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.0003)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.318\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.005)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-C \u0026ndash; I\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.361\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.006)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;DL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.434\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.008)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.267\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.004)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.2142\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.001)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.287\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.003)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-C \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.425\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.013)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;DL\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.519\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.012)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.192\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.001)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.1805\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.0008)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.385\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.006)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-R \u0026ndash; I\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.835\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.005)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.0232 a\u003c/p\u003e\n\u003cp\u003e(0.0002)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.026\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.004)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.186\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.001)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.2497\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.001)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.924\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.0005)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-R \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.134\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.015)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.0318 b\u003c/p\u003e\n\u003cp\u003e(0.0004)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.264\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.007)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.266\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.005)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.3034\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.0007)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.077\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.006)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSAL \u0026ndash; I\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.081\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.012)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.0402 c\u003c/p\u003e\n\u003cp\u003e(0.0004)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.254\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.006)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.541\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.003)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.3660\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.002)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.054\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.008)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSAL \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.471\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.016)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.0459 d\u003c/p\u003e\n\u003cp\u003e(0.0002)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.516\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.009)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;LPC\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.623\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.004)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.4506\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.0005)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.235\u003csup\u003eh\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e(0.007)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"9\"\u003e\u0026lt;DL: samples with concentrations below the detection limit of the equipment. Means followed by the same letter in the column do not differ from each other by the Tukey test, considering the nominal value of 5% of significance. \u0026gt; UPC: sample with concentration above the highest concentration point of the calibration curve (2\u0026micro;g g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e).\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"9\"\u003eSamples with \u0026ldquo;I\u0026rdquo; are samples of dried plant material that have not gone through the extraction process and \u0026ldquo;II\u0026rdquo; samples are samples that have had their secondary metabolites removed in the extraction process.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eWe also observed that the roots of \u003cem\u003eP. parviflora\u003c/em\u003e exhibit higher metal accumulation compared to the petiole/stem and leaves. This disparity could be attributed to the roots' closer interaction with the contaminated environment. \u003cem\u003eP. parviflora\u003c/em\u003e, as an emerging macrophyte, has its roots firmly affixed to the substrate, while the leaves and a section of the petiole/stem remain above the water surface.\u003c/p\u003e\n\u003cp\u003eTo assess the plant biomass's capacity (pre- and post-extraction procedure), Cadmium Removal trials were conducted using three different Cd solution concentrations (0.2 and 3.5 \u0026micro;g/mL of Cd in acidic water). The outcomes (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e) revealed that at the 0.2 \u0026micro;g/mL solution, all samples demonstrated a decrease in Cd concentration, signifying the macrophytes' ability to capture this metal. However, in the 3.5 \u0026micro;g/mL solutions, the samples exhibited minimal reduction in Cd content. Notably, when in contact with sample \u003cem\u003eP. parviflora\u003c/em\u003e - roots I, there was an elevation in Cd concentration. These findings imply a saturation of the trapping system in the plant material and suggest that the presence of metabolites in the biomass (samples before the extraction process) does not influence the metal trapping system. This conclusion is drawn from the lack of correlation between the presence of these metabolites and the enhanced removal of metals.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab8\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eMean values (\u0026plusmn;\u0026thinsp;standard deviation, n\u0026thinsp;=\u0026thinsp;3) of the residual concentration evaluated after the removal tests, the values presented are the residual cadmium concentration after the contact time of the control solution with the biomass.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSample\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAssay 0.2 \u0026micro;g mL\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAssay 3.5 \u0026micro;g mL\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.200 (\u0026plusmn;\u0026thinsp;0.00010)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.350 (\u0026plusmn;\u0026thinsp;0.0024)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePPA-F \u0026ndash; I\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e0.026 (\u0026plusmn;\u0026thinsp;0.00008)\u003csup\u003ee\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e2.940 (\u0026plusmn;\u0026thinsp;0.0110)\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-F \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.058 (\u0026plusmn;\u0026thinsp;0.00007)\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.260 (\u0026plusmn;\u0026thinsp;0.0020)\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-C \u0026ndash; I\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.019 (\u0026plusmn;\u0026thinsp;0.00008)\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.260 (\u0026plusmn;\u0026thinsp;0.0020)\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-C \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.012 (\u0026plusmn;\u0026thinsp;0.00014)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.200 (\u0026plusmn;\u0026thinsp;0.0010)\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-R \u0026ndash; I\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.014 (\u0026plusmn;\u0026thinsp;0.00017)\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.430 (\u0026plusmn;\u0026thinsp;0.0010)\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePPA-R \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.008 (\u0026plusmn;\u0026thinsp;0.00006)\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.300 (\u0026plusmn;\u0026thinsp;0.0010)\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSAL \u0026ndash; I\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.030 (\u0026plusmn;\u0026thinsp;0.00001)\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.380 (\u0026plusmn;\u0026thinsp;0.0020)\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSAL \u0026ndash; II\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.011 (\u0026plusmn;\u0026thinsp;0.000005)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.530 (\u0026plusmn;\u0026thinsp;0.0008)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"3\"\u003eMeans followed by the same letter in the column do not differ from each other by the Tukey test, considering the nominal value of 5% of significance.\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"3\"\u003eThe concentration of Cd in (\u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). Samples with \u0026ldquo;I\u0026rdquo; are samples of dried plant material that have not gone through the extraction process and \u0026ldquo;II\u0026rdquo; samples are samples that have had their secondary metabolites removed in the extraction process.\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eDespite the absence of extraction enhancement, the attained results regarding extract yield and the percentage of phenolic compounds present promising potential. Tannins and phenolic compounds commonly possess well-recognized antioxidant and antiradical properties, constituting the primary biological activities linked to phytoremediation processes, involving metal sequestration and chelation (ZHENG and WANG, 2001; DAI and MUMPER, 2010).\u003c/p\u003e\n\u003cp\u003eRegarding the disparity in metal content between biomass samples with and without secondary metabolites, it's noteworthy that the removal of metabolites led to elevated metal levels within the structures of the samples. Although existing literature links metal complexation with secondary metabolites, it's important to consider that plant tissues contain catechol subunits within lignins and lignans, which provide potential binding sites for metal atoms within the plant structure (SARASWAT and RAI, 2010; ALI et al., \u003cspan class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eAs demonstrated in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e, the metal content escalates upon the removal of plant biomass metabolites. Samples with removed metabolites comprise a higher proportion of structures that contribute to plant tissue formation, such as lignins, which remain intact during the extraction process. Consequently, we hypothesize that the metals detected in these samples could be complexed within these structures. The exception to this trend is observed in the case of Mo and Ni in PPA-C, where this logic is reversed.\u003c/p\u003e\n\u003cp\u003eAmong the assessed metals, certain ones are integral to plant metabolism, constituting essential micronutrients. However, in most instances, these metals are present at levels designating them as contaminants within the collection areas of these plants. Generally, they possess high toxic potential. Furthermore, their presence in aquatic ecosystems fuels extensive debate, interest, and concern. This is particularly significant since their concentrations in natural waters are also subject to stringent limitations.\u003c/p\u003e\n\u003cp\u003eThe results presented for Cd removal assays in a contaminated aqueous medium (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e) indicate a low saturation point for the phytoremediation system in relation to this metal. At Cd concentrations of 0.2 \u0026micro;g mL-1, there was a reduction in all residual aqueous samples, ranging from 71% (PPA-F II) to 96% (PPA-R II). At concentrations of 3.5 \u0026micro;g mL-1 Cd, certain samples exhibited an increase in residual aqueous concentrations, with higher Cd contents than the control (PPA-R I and SAL I, both containing metabolites). In assays with a concentration of 3.5 \u0026micro;g mL-1, the most favorable removal result was achieved in SAL II, with a 24.5% reduction.\u003c/p\u003e\n\u003cp\u003eThe presence of other ions can impact the potential for metal removal due to competition for binding sites. It is essential to observe the ionic balance in control solutions and the dried biomass to enhance contaminant removal potential (Saraswat and RAI 2010).\u003c/p\u003e\n\u003cp\u003eThe elevated Cd contents observed after biomass intervention (cadmium removal assay) at higher metal concentrations can be explained by the scheme depicted in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. This scheme illustrates situations where biomass binding sites become saturated and complexed with other ions from the environment. This fact potentially compromises the ability of these vegetables to remove ions. Nevertheless, as the dry biomass persists, water absorption continues, resulting in the residual (aqueous) solution retaining Cd mass in the reduced volume of the medium (water), consequently elevating the Cd concentration in this residual solution.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe study demonstrated the effects of biosorption system saturation using biomass of P. parviflora and S. auriculata. In some cases, the concentration of cadmium ions did not decrease; instead, it increased due to a reduction in the volume of the dilution medium (water) without effective reduction of cadmium ions. This suggests low bioaccumulation in a phytotreatment system, or even bioaccumulation of metals in the biomass of these plants from the environment during vegetal development.\u003c/p\u003e \u003cp\u003eOur study indicates the absence of interference in the cadmium removal processes, as well as in the bioaccumulation of metals, by metabolite contents, as samples without metabolites outperformed samples with metabolites in metal removal assays. Thus, there is no correlation between the metabolite content and the accumulated or removed metal content.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eConflict of Interest\u003c/h2\u003e \u003cp\u003eThere is no conflict of interest between the primary author and the co-authors.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis study was funded by the Coordena\u0026ccedil;\u0026atilde;o de Aperfei\u0026ccedil;oamento de Pessoal de N\u0026iacute;vel Superior (CAPES), Conselho Nacional de Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico (CNPq).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eAll authors have participated in the research and/or preparation of the article:Augusto C\u0026eacute;sar Rodrigues: conceptualization, methodology, validation, formalanalysis, investigation, writing original draft.Samara Requena Nocchi: methodology, validation, formal analysis, investigation,writing original draft.Jorge Raposo: conceptualization, resources, data curation, writing \u0026ndash; review.Valter Arag\u0026atilde;o do Nascimento: conceptualization, resources, data curation,writing \u0026ndash; review.Carlos Alexandre Carollo: conceptualization, resources, data curation, writing \u0026ndash;review and editing, visualization, supervision, project administration, fundingacquisition.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eWe are grateful to Instituto Nacional de \u0026Aacute;reas \u0026Uacute;midas (INAU), Laborat\u0026oacute;rio de Produtos Naturais e Espectrometria de Massas (LaPNEM) and Programa de P\u0026oacute;s-gradua\u0026ccedil;\u0026atilde;o em Biologia Vegetal da Universidade Federal de Mato Grosso do Sul (PPGBV-UFMS).\u003c/p\u003e\u003ch2\u003eData availability statements\u003c/h2\u003e \u003cp\u003eThe authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information files. Should any raw data files be needed in another format they are available from the corresponding author upon reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eALI, H.; KHAN, E.; SAJAD, M. A. Phytoremediation of heavy metals\u0026ndash;concepts and applications. Chemosphere, v. 91, n. 7, p. 869\u0026ndash;881. 2013.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBALASSA, G. C.; DE SOUZA, D. C.; BARBOSA DE LIMA, S. Evaluation of the potential of \u003cem\u003ePontederia parviflora\u003c/em\u003e Alexander in the absorption of copper (Cu) and its effects on tissues. Acta Scientiarum, Biological Sciences, v. 32, n. 3, p. 311\u0026ndash;316, 2010.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBIZZO, A. L. T. et al. Short-term physiological responses to copper stress in \u003cem\u003eSalvinia auriculata\u003c/em\u003e Aubl. 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Ying Yong Sheng Tai Xue Bao. 2006;17(9):1637\u0026ndash;40.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Aquatic Macrophytes, Heavy Metals, Metabolites, Phytoremediation Potential","lastPublishedDoi":"10.21203/rs.3.rs-4356239/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4356239/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHeavy metals represent a serious source of water and soil contamination, mainly due to their non-biodegradable nature. Their bioaccumulation occurs in plants and other trophic levels, including animals, consequently affecting humans and causing various side effects. Phytoremediation offers a biological, cost-effective, and environmentally friendly cleaning solution, utilizing plants to remove contaminants from soil and water. This study aims to establish the chemical composition of Pontederia parviflora and Salvinia auriculata using liquid chromatography and evaluate the role of their secondary metabolites in passive phytoremediation by these plants. Although the results are not aligned with the initial hypothesis of metal removal from the liquid medium (sequestration and chelation of metals), the amount of metal removed still represents a positive outlook for technique enhancement. Furthermore, this research challenges the notion presented in the literature, as secondary metabolites were considered contaminant sequesters. The removal of these metabolites from the biomass of these macrophytes did not significantly impact their phytoremediation performance.\u003c/p\u003e","manuscriptTitle":"The role of secondary metabolites of Pontederia parviflora Alexander and Salvinia auriculata Aubl. in phytoremediation of cadmium contaminated water","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-10 13:59:57","doi":"10.21203/rs.3.rs-4356239/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b7e7e6d4-b6f3-47f7-a161-4b8dfe63af2d","owner":[],"postedDate":"June 10th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-24T17:41:35+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-10 13:59:57","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4356239","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4356239","identity":"rs-4356239","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}