An Ultra-rapid and Green Method for Simultaneous Determination of Honokiol and Magnolol in Magnoliae Officinalis Cortex with One Standard by HPLC-UV at Equal Absorption Wavelength

An Ultra-rapid and Green Method for Simultaneous Determination of Honokiol and Magnolol in Magnoliae Officinalis Cortex with One Standard by HPLC-UV at Equal Absorption Wavelength | 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 An Ultra-rapid and Green Method for Simultaneous Determination of Honokiol and Magnolol in Magnoliae Officinalis Cortex with One Standard by HPLC-UV at Equal Absorption Wavelength Zhengming Qian, Jing Chen, Qinggui Lei, Guoying Tan, Yuansheng Zou, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2381002/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 An ultra-rapid and green assay method for simultaneous determination of honokiol and magnolol in Magnoliae Officinalis Cortex with one standard was developed by HPLC-UV at equal absorption wavelength. The sample was prepared by ultrasonic-assisted matrix solid-phase dispersion. The HPLC separation was performed on a Poroshell C18 column with an eco-friendly mobile phase. The detection wavelength was set at the equal absolution wavelength of honokiol and magnolol (247 nm). The contents of honokiol and magnolol in six batches of samples, obtained by developed method with one marker and external standard method with two markers, were comparable. In addition, the developed HPLC method only took 2.5 min and 4.55 mL green organic solution (ethanol), which including the sample extraction and separation. The developed method was rapid, green and standard saving, which would be helpful to improve the quality evaluation of Magnoliae Officinalis Cortex. rapid green matrix solid-phase dispersion HPLC equal absorption wavelength Figures Figure 1 Figure 2 Introduction Magnoliae Officinalis Cortex (MOC), the stem or root bark of Magnoliae Officinalis , is a traditional used food addictive (Greenberg et al., 2007 ; Luo et al., 2019 ). For example, it had been used in biscuits, beverages and chewing gum. More than 200 components were found in MOC, such as lignans, alkaloids, steroids (Luo et al., 2019 ). Honokiol and magnolol were believed as two major bioactive components in MOC, which had a variety of biological activities, such as anti-cancer (Mottaghi and Abbaszadeh, 2022 ), cardiovascular modulating (Yuan et al., 2022), inhibiting pathogenic bacteria and inflammation (Chiu et al., 2021 ; Kim et al., 2021). As the excellent activities and high content, honokiol and magnolol were selected as the quality markers of MOC in Chinese pharmacopoeia (Committee of the Chinese Pharmacopoeia, 2020 ). Several high-performance liquid chromatography (HPLC) methods for determination of honokiol and magnolol in MOC had been reported, such as HPLC with ultraviolet detection (HPLC-UV) (Ma et al., 2022 ; Zhang et al., 2014 ; He et al., 2015 ), HPLC with fluorescence detector (HPLC-FLD) (Hu and Yin, 2015 ), HPLC with electrochemical detection (HPLC-ECD) (Yang et al., 2011 ) and HPLC with mass spectrometry (HPLC-MS) (Wu et al., 2006 ). However, the reported HPLC methods were time-consuming and took amounts of harmful organic solvents. For example, in Chinese pharmacopoeia, the maceration extraction (ME) combined with HPLC-UV, which took more than 24 hours and consumed 60.5 mL methanol (Ma et al., 2022 ). The reported ultrasonic extraction (UE) and HPLC-UV method took 60 min and consumed 24 mL methanol (He et al., 2015 ). In addition, the reported methods used external standard method (ESM) for determination and two reference compounds were required. In order to reduce the reference compounds consumption, quantitative analysis of multi-components by single marker (QAMS) method was developed for determination of honokiol and magnolol in MOC by using a single reference compound with relative calibration factors (RCF) (Wu and Yan, 2016 ). However, detection of RCF in QAMS method increased the operational complexity (Chen et al., 2022 ; Liu et al, 2022 ). Therefore, it was necessary to develop a simple, rapid, green and standards-saving method to improve assay of honokiol and magnolol in MOC. Ultrasonic-assisted matrix solid-phase dispersion (UAMSPD) was proved as a fast extraction method, which had the advantage of saving solvent, time and material (Tu and Chen, 2018 ; Dos Santos et al., 2019 ). The poroshell column was a type of rapid column, which could provide rapid separation of compounds with low back pressure (Qian et al., 2020 ; Yıldırım and Yaşar, 2018 ; Li et al., 2023 ). Ethanol was considered as an eco-friendly regent in HPLC analysis (Capello et al., 2007 ; Shimamoto and Aricetti, 2016; Okcu et al., 2016 ; Raspe et al., 2021 ). Therefore, the combination of UAMSPD extraction and poroshell column HPLC method with ethanol as green extraction solvent and mobile phase would provide a rapid and eco-friendly method for herbal medicine analysis. In HPLC-UV quantitative analysis, the content of analyte was determined by comparing peak area with a known concentration reference compound at detection wavelength (Rodrigues et al., 2007 ). The maximum absorption wavelength of analyte was usually selected as the detection wavelength because of the good sensitivity. For example, the maximum absorption wavelength 294nm was always selected as detection wavelength for HPLC analysis of honokiol and magnolol (Ma et al., 2022 ; Zhang et al., 2014 ; He et al., 2015 ). The UV responses of honokiol and magnolol at 294 nm were different, so the quantification of honokiol and magnolol needed two reference compounds. When honokiol and magnolol had the same UV response at someone UV detection wavelength, this UV detection wavelength was called as equal absorption wavelength (EAW). Therefore, at the EAW, the determination of honokiol and magnolol with one reference compound and without RCF could be achieved. In present experiment, an ultra-fast and eco-friendly analytical method for determination of honokiol and magnolol with one reference compound was developed by UAMSPD and HPLC-UV EAW. The developed method was applied in six batches of MOC samples. Materials And Methods Chemicals and materials HPLC-grade formic acid was obtained from Aladdin (Shanghai, China), HPLC-grade ethaanol was obtained from Krude company (Losangeles, USA), analytical grade ethanol was purchased from Xilong Scientific Co., Ltd (Sichuan, China). The deionized water was purified by Milli-Q purification system (Millipore, USA). Diatomaceous earth was purchased from Sigma (Merck, Germany). Honokiol (98%) and magnolol (98%) were purchased from Shanghai Acmec Biochemical Co., Ltd. (Shanghai, China). Six batches of MOC samples were collected from the Guangdong markets (Table 1 ). The authenticity of all samples was identified by Dr. Zhengming Qian. Voucher specimens were deposited at Dongguan HEC Cordyceps R&D Co., Ltd, Dongguan, Guangdong. All crude materials were pulverized with a mill and the powders were then obtained by passing through a 50 mesh sieve. Table 1 The contents of honokiol and magnolol in MOC by HPLC-UV ESM and HPLC-UV EAW (%) Sample ESM EAW Honokiol Magnolol Honokiol Magnolol (determined by honokiol) S1 2.22 2.17 2.22 2.15 S2 3.19 3.05 3.19 3.02 S3 2.21 2.58 2.21 2.56 S4 2.60 2.74 2.60 2.72 S5 2.50 2.65 2.50 2.63 S6 0.75 1.76 0.75 1.75 Preparation Of Reference Compound Solution The mixed reference compounds solution containing honokiol (4.453 mg mL − 1 ) and magnolol (4.449 mg mL − 1 ) was prepared with 70% (v/v) ethanol and then diluted with 70% ethanol to obtain the intended concentrations. All solutions were kept under 4 ℃. Preparation Of Sample Solution The sample powder (200 mg) and dispersant (200 mg) were precisely weighed, putted into an agate mortar and homogeneously ground for 0.5 min. Then 50 mg of the mixture was transferred into a 10 mL glass tube and 5 mL 70% ethanol was added. The sample was extracted for 1.0 min by ultrasonic extraction (33 kHz, 250W). Small amount of 70% ethanol solution was added to compensate the lost weight. The final sample solution was filtered by 0.45 µm membrane before HPLC injection. Hplc Condition The HPLC analysis was operated on Agilent 1260 HPLC system (Agilent, USA) consisting of a binary pump, a thermostatic column compartment and an auto-sampler. The sample solution was separated on Poroshell 120 EC-C18 column (Agilent, Germany) (50 ×4.6 mm, 2.7 µm) by 70% ethanol aqueous solution with 0.1% formic acid at a flow rate of 1.5 mL min − 1 . The column temperature was maintained at 35°C, the injection volume was 1uL, and the wavelength was set at 247 nm (bandwidth 4 nm). Method Validation The method was validated for linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy and stability. Linearity was evaluated by the calibration curve, which was obtained by plotting peak areas versus the concentrations of reference compound solutions (0.018–0.445 mg mL − 1 ). The limits of detection (LODs) and quantification (LOQs) were obtained by analyzing the reference compound solution based on the signal-to-noise ratios (S/N) of approximately 3 and 10, respectively. The intra-day precision was determined by analyzing the mixed reference compounds solution in six replicates within one day, and the inter-day precision was determined by analyzing the mixed reference compounds solution twice per day for three days. The repeatability was tested by analyzing sample S3 in six replicates. The relative standard deviation (RSD, %) was used to evaluate the precision and repeatability. Accuracy was evaluated by recovery. Known amount of two analytes were added into sample S3. The mixture was extracted and analyzed. Six replicates were performed. The recovery was calculated as the ratio of detected amount to added amount. The stability was tested by analyzing sample solution 6 times in 24 h at room temperature. The variation was expressed as RSD. The robustness tests were carried out by the developed HPLC method, which included flow rates (1.5 ± 0.1 mL min − 1 ), column temperatures (35 ± 5℃), and different columns (Agilent Poroshell 120 EC-C18, Waters CORTECS C18 and ACE UltraCore SuperC18). The relative retention time (RRT), relative peak area (RPA) and resolution of honokiol to magnolol were recorded for evaluating robustness. Results And Discussion Optimization of extraction condition The UAMSPD extraction parameters including sample grinding and ultrasonic extraction, the grinding time, extraction solvent and ultrasonic time were optimized to obtain the satisfactory extraction yield. The extraction yields of current method were evaluated by comparing with Pharmacopeia of China 2020 method of MOC. Diatomaceous earth was employed as the dispersant based on previous research (Zhang et al., 2012 ). Two ratios of sample to diatomaceous earth (1:1 and 1:2) were compared. There was no difference between the two ratios, thus 1:1 was used. Four different grinding times (10, 30, 60 and 120 s) were also compared. The results indicated that grinded time of 30 s could provide a satisfied extraction yield of target compounds (Fig. S1 A ). Ethanol as an eco-friendly solvent was selected as extraction solvent. Three concentrations of ethanol (50%, 70%, 90%) were tested. The results showed that 70% ethanol aqueous solution was more suitable (Fig. S1 B ). The different ratios of sample to extraction solvent (1: 100, 1: 200, 1: 400) were tested. The results (Fig. S1 C ) of the three tests were similar. Considering the suitable HPLC peak areas, 1: 200 was chosen. In addition, ultrasonic extraction time (30, 60, 120 and 240 s) were evaluated. According to Fig. S1 D , when extraction time was more than 60 s, the extraction yields were similar. Thus, 60 s was used as extraction time. To confirm the extraction efficiency of the developed UAMSPD, sample S3 was extracted with USMAPD and Chinese Pharmacopeia extraction method. Three replicates were performed. The contents of honokiol and magnolol extracted by UAMSPD were 2.27 ± 0.029% and 2.63 ± 0.042%, and by Chinese Pharmacopeia method were 2.32 ± 0.021% and 2.73 ± 0.027%. These results showed that the extraction efficiency of the developed method was similar with Chinese Pharmacopeia method. Thus, the developed UAMSPD could be used as a good substitute method for the Chinese Pharmacopeia method. Optimization Of Hplc Condition Honokiol and magnolol were usually separated on traditional C18 columns and more than 10 min was consumed (Ma et al., 2022 ; Zhang et al., 2014 ; He et al., 2015 ; Wu et al., 2006 ). In order to reduce the HPLC separation time, poroshell column was employed in experiment. To reduce the harmful solvent consumption, ethanol was selected as the organic mobile phase. The 25%, 30% and 35% ethanol aqueous solution with 0.1% formic acid were compared as the mobile phase. The 30% ethanol aqueous solution with 0.1% formic acid was chosen because of the good peak shape and resolution. The flow rates of 1.0 and 1.5 mL min − 1 were compared and 1.5 mL min − 1 was used based on the shorter time. The column temperatures (30, 35 and 40 ℃) were also compared, the separations of two analytes were similar. The 35 ℃ was selected, because it was easy to control. To ensure the specificity of the method, the peak purities of target compounds were checked. The results showed that the purities of honokiol and magnolol in samples were higher than 99.99%. The selection of EAW was important in this experiment, which included two steps. Firstly, screening the EAW of honokiol and magnolol by UV spectrophotometer. The honokiol solution (0.0222 mg mL − 1 ) and magnolol solution (0.0222 mg mL − 1 ) were prepared with the mobile phase, and then the two reference compound solutions were scanned from 200 nm to 350 nm with Agilent Cary 60 ultraviolet spectrophotometer (Agilent, USA). The UV spectra of the two compounds were shown in Fig. 1 . There were three EAWs (247 nm, 281 nm and 298 nm) for honokiol and magnolol. The 247 nm was chosen for further experiment due to the highest UV response. Secondly, confirming the EAW of honokiol and magnolol by HPLC-UV. The mixture reference solution of honokiol (0.0890 mg mL − 1 ) and magnolol (0.0890 mg mL − 1 ) was separated on Poroshell 120 EC-C18 column (50 ×4.6 mm, 2.7 µm) with 70% ethanol aqueous solution with 0.1% formic acid, and detected at 246 nm, 247 nm and 248 nm. The results (Table S1) showed that the peak areas of honokiol and magnolol were same at 247 nm. Therefore, 247 nm was used as EAW. In addition, different bandwidths (1 nm, 2 nm, 4 nm and 8 nm) were tested. The 4 nm showed the best precision of peak areas, and was used in developed HPLC method. Method Validation The regression equation for honokiol was y = 1410.2523 x + 0.8718 (r = 0.9999), for magnolol was y = 1404.0385 x + 1.3179 (r = 0.9999) in the test range of 0.018–0.445 mg mL − 1 , which indicated a good linearity of the two compounds. The LODs for honokiol and magnolol were 0.3559 and 0.3562 µg mL − 1 . The LOQs for honokiol and magnolol were 0.8898 and 0.8906 µg mL − 1 . The RSDs of intra- and inter-day precision (n = 6) for honokiol were 0.38% and 0.45%; the RSDs of intra- and inter-day precision (n = 6) for magnolol were 0.36% and 0.49%. The RSDs of repeatability (n = 6) for honokiol and magnolol were 1.9% and 1.8%. The recoveries (Table S2) of honokiol and magnolol (n = 6) were 99.28% (RSD = 2.3%) and 98.08% (RSD = 1.8%), respectively. The RSDs of stability (24 h) for honokiol and magnolol were 0.86% and 0.47%. The RRTs, RPAs, and resolution of honokiol to magnolol in the robustness tests were listed in Table S3. The RSD for RRTs were 1.3%. The RPA values were all close to 1.00. The resolutions were all larger than 1.5. These results indicated a good robustness of the developed method within the test ranges. Above all, the developed HPLC-UV EAW was accurate and stable for quantitative analysis of honokiol and magnolol in MOC. Analysis Of Moc Sample The reference compounds and MOC sample chromatograms were shown in Fig. 2 . To evaluate the feasibility of the developed HPLC-UV EAW, six MOC samples were determined by HPLC-UV EAW and ESM, respectively. In HPLC-UV EAW, honokiol and magnolol were determined by one reference compound (honokiol). While in HPLC-UV ESM, honokiol and magnolol were determined by two reference compounds. The results were listed in Table 1 . The contents of magnolol in six MOC samples obtained by the two different methods were compared by t-test (SPSS 19.0 software). The result (p > 0.05) indicated that there was no significant difference between HPLC-UV EAW (only use honokiol as reference compound) and HPLC-UV ESM (use two reference compounds). This indicated that HPLC-UV EAW could be a good alternative method of HPLC-UV ESM for determination of honokiol and magnolol in MOC. The contents of honokiol and magnolol obtained by HPLC-UV EAW were 0.75 ~ 3.19% and1.75 ~ 3.02%, respectively, which were agreed with the literatures (Jiang et al., 2011 ). The total contents of honokiol and magnolol were 2.50 ~ 6.21%, which indicated that the six batch MOC samples were all complied with the Chinese Pharmacopeia (≥ 2.0%). Comparison Of The Developed Method And Reported Method Comparisons of the current method with the reported HPLC methods (Method 1, 2, 3 and 4) (Ma et al., 2022 ;Zhang et al., 2014 ; He et al., 2015 ; Wu et al., 2006 ) were summarized in Table 2 . The developed method had advantages in less reference compounds, shorter analytical time and solvent eco-friendly. Table 2 The comparison of the developed and reported HPLC methods for determination of honokiol and magnolol in MOC. * The mobile phase additives (1.5 min HPLC Methanol 7.8mL 10 min 3 UE Polyethylene glycol 20mL 30 min HPLC Methanol 24mL 30 min 4 BE Ethanol 47.5mL 13 min LC-MS/MS Acetonitrile 3mL 5 min This work VAMSPD Ethanol 3.5ml 1.5 min HPLC Ethanol 1.05mL 1 min Firstly, the developed method only need one reference compound for determining two analytes, while in other literatures two reference compounds were required. Compared with the QAMS method, the current method was simpler without RCF used. Secondly, the developed method was the fastest. The total analytical time was 2.5 min including sample extraction time (1.5 min) and HPLC separation time (1 min). The reported methods took more than 11 min. For example, method 1 took 1440 min in sample extraction and 20 min in HPLC separation. Method 3 took 30 min in the ultrasonic extraction and 30 min in HPLC separation. Although a few rapid analytical methods (method 2, 4) were achieved by MSPD extraction or the LC-MS/MS system, more than 11 min were still needed. Thirdly, the developed method was the greenest. “Analytical Eco-Scale” approach (Sereshti et al.,2022; Gałuszka et al., 2012 ) was carried out to evaluate the greenness of developed method and reported ones. The results (Table 3 ) showed that developed HPLC method got the highest Eco-Scale score of 89. In the current method, eco-friendly solvent ethanol was employed to replace those harmful solvents, such as acetonitrile and methanol. The rapid sample extraction and HPLC separation also made less solvent and energy consumed. Methanol or acetonitrile was the main weak points for other methods (method 1, 2, 3, 4). Although polyethylene glycol as an eco-friendly solvent was used in sample extraction of method 3, methanol was still used in the HPLC separation. The sample extraction (30 min) and HPLC separation (30 min) also consumed more energy and produced more waste. Thus, it got the lowest score of 75. Table 3 Greenness assessment of the developed and reported HPLC methods for determination of honokiol and magnolol in MOC Penalty points (PPs) Method 1 Method 2 Method 3 Method 4 This work Reagents Methanol 12 12 12 / / Ethanol / / / 4 2 Acetonitrile / / / 4 / Water 0 0 0 0 0 Formic acid / / / / 2 Acetic acid / / 4 / / Multiwalled carbon nanotubes(60mg) / 2 / / / Diatomaceous earth (200mg) / / / / 4 Polyethylene glycol / / 2 / / Instrument Energy HPLC 1 1 1 / 0 HPLC-MS / / / 1 / Sonicator / / 1 / 0 Centrifuge / / / 1 / Occupational hazard 0 0 0 3 0 Waste 5 5 5 3 3 Total PPs ∑18 ∑20 ∑25 ∑16 ∑11 Analytical EcoScale 82 80 75 84 89 total score Conclusions In present study, an ultra-fast and eco-friendly method based on UAMSPD and HPLC-UV EAW was developed for determination of honokiol and magnolol in MOC. The developed assay method only used one reference compound for determination of honokiol and magnolol in MOC without RCF. Comparing with the reported methods, the developed procedure was ultra-fast, green and reference compound saving. It was an improved analytical method for determination of honokiol and magnolol in MOC and related products. Declarations Authorship contribution Zhengming Qian: Investigations, Methodology, Writing- original draft preparation . Jing Chen: Formal analysis, Investigations. Qinggui Lei: Investigations, Data curation, Methodology. Guoying Tan: writing- review & editing. Yuansheng Zou: Investigations. Gang Peng: Collecting sample. Wenqing Li: Conceptualization, Funding acquisition, Writing- review & editing. Juying Xie: Supervision, Project administration, Writing- review & editing. Funding This study was financially supported by the Key Laboratory of Guangdong Drug Administration (2021ZDB05). Data Availability The authors declare that all the data supporting the findings of this study are included in the manuscript or in the supplements Compliance with ethical standards Conflict of Interest The authors declare no potential conflicts of interest with respect to the research, authorship, and or publication of this article Ethical Approval Not applicable. Informed Consent Not applicable. References Capello C, Fischer U, Hungerbühler K (2007) What is a green solvent? A comprehensive framework for the environmental assessment of solvents. Green Chem 9:927-934. https://doi.org/10.1039/b617536h. Chen Q, Wang Z, Yang B, Yang Q, Kan J (2022) Determination of main alkylamides responsible for Zanthoxylum bungeanum pungency through quantitative analysis of multi-components by a single marker. Food Chem 396: 133645. https://doi.org/10.1016/j.foodchem.2022.133645. Chiu KC, Shih YH, Wang TH, Lan WC, Li PJ, Jhuang HS, Hsia SM, Shen YW, Chen MYC, Shieh TM (2021) In vitro antimicrobial and antiproinflammation potential of honokiol and magnolol against oral pathogens and macrophages. J Formosan Med Assoc 120: 827-837. https://doi.org/10.1016/j.jfma.2020.09.002. Committee of the Chinese Pharmacopoeia (2020). Pharmacopoeia of the People’s Republic of China, Vol. 4. China Medical Science Press, Bejing, pp 236. Dos Santos EO, Gonzales O, Ores JC, Marube LC, Caldas SS, Furlong EB, Primel EG (2019) Sand as a solid support in ultrasound-assisted MSPD: A simple, green and low-cost method for multiresidue pesticide determination in fruits and vegetables. Food Chem 297:124926. https://doi.org/10.1016/j.foodchem.2019.05.200. Gałuszka A, Konieczka PM, Migaszewski Z, Namieśnik J (2012) Analytical eco-scale for assessing the greenness of analytical procedures. TrAC, Trends Anal Chem 37:61-72. http://dx.doi.org/10.1016/j.trac.2012.03.013. Greenberg M, Urnezis P, Tian M (2007) Compressed mints and chewing gum containing magnolia bark extract are effective against bacteria responsible for oral malodor. J Agric Food Chem 55: 9465-9469. https://doi.org/10.1021/jf072122h. He L, Fan T, Hu J, Zhang L (2015) Polyethylene glycol-based ultrasound-assisted extraction of magnolol and honokiol from cortex magnoliae officinalis . Nat Prod Res 29:31-36. http://dx.doi.org/10.1080/14786419.2014.955800. Hu L, Yin C (2015) Multi-way calibration coupling with fluorescence spectroscopy to determine magnolol and honokiol in herb and plasma samples. Anal Methods 7:5913-5923. http://dx.doi.org/10.1039/c5ay01285f. Jiang Y, Vaysse J, Gilard V, Balayssac S, Déjean S, Malet-Martino M, David B, Fiorini C, Barbin Y (2011) Quality assessment of commercial magnoliae officinalis cortex by 1H-NMR-based metabolomics and HPLC methods. Phytochem Anal 23:387-395. http://dx.doi.org/10.1002/pca.1369. Kim H, Lim CY, Chung MS (2020) Magnolia officinalis and its honokiol and magnolol constituents inhibit human norovirus surrogates. Foodborne Pathog Dis 18:24-30. https://doi.org/10.1089/fpd.2020.2805. Li W, Qian Z, Lei Q, Lian Y, Zou Y, Wang Y, Lan D (2023) An ultra-rapid and eco-friendly method for determination of loganic acid and gentiopicroside from Gentianae Macrophyllae Radix by vortex-assisted matrix solid-phase dispersion extraction and LC-MS. J Pharm Biomed Anal 222:115085. https://doi.org/10.1016/j.jpba.2022.115085. Liu M, Yao J, Zhang C, Hao R, Zhang Z, Cao W (2022) Quantitative analysis of multi-components by single-marker: An effective method for the chemical characterization of wood vinegar. Ind Crops Prod 182:114862. https://doi.org/10.1016/j.indcrop.2022.114862. Luo H, Wu H, Yu X, Zhang X, Lu Y, Fan J, Tang L, Wang Z (2019) A review of the phytochemistry and pharmacological activities of magnoliae officinalis cortex. J Ethnopharmacol 236:412-442. https://doi.org/10.1016/j.jep.2019.02.041. Ma DX, Liu JF, Hu MJ, Li BZ, Chen J, Liu RL (2022) Analysis on the contents of medicinal components in the back of houpoea officinalis at different heights. J Hebei Norm Univ, Nat Sci Ed 46:304-309. http://dx.doi.org/10.13763/j.cnki.jhebnu.nse.202204007. Mottaghi S, Abbaszadeh H (2022) Natural lignans honokiol and magnolol as potential anticarcinogenic and anticancer agents. A comprehensive mechanistic review. Nutr Cancer 74:761-778, https://doi.org/10.1080/01635581.2021.1931364. Okcu G, Ayhan K, Altuntas EG, Vural N, Poyrazogiu ES (2016) Determination of phenolic acid decarboxylase produced by lactic acid bacteria isolated from shalgam, salgam juice using green analytical chemistry method. LWT-Food Sci Technol 66:615-621. http://dx.doi.org/10.1016/j.lwt.2015.10.072. Qian Z, Wu Z, Li C, Tan G, Hu H, Li W (2020) A green liquid chromatography method for rapid determination of ergosterol in edible fungi based on matrix solid-phase dispersion extraction and a core–shell column. Anal Methods 12:3337-3343. https://doi.org/10.1039/d0ay00714e. Raspe DT, Ciotta SR, Zorzenon MRT, Dacome AS, Silva CD, Milani PG, Costa SC (2021) Ultrasound-assisted extraction of compounds from Stevia leaf pretreated with ethanol. Ind Crops Prod 172:114035. https://doi.org/10.1016/j.indcrop.2021.114035. Rodrigues CI, Marta L, Maia R, Miranda M, Ribeirinho M, Máguas C (2007) Application of solid-phase extraction to brewed coffee caffeine and organic acid determination by UV/HPLC. J Food Compos Anal 20:440-448. https://doi.org/10.1016/j.jfca.2006.08.005. Sereshti H, Zarei-Hosseinabadi M, Soltani S, Taghizadeh M (2022). Green vortex-assisted emulsification microextraction using a ternary deep eutectic solvent for extraction of tetracyclines in infant formulas. Food Chem 396:133743. https://doi.org/10.1016/j.foodchem.2022.133743. Shimamoto GG, Aricetti JA, Tubino M (2016) A Simple, Fast, and Green Titrimetric Method for the Determination of the Iodine Value of Vegetable Oils Without Wijs Solution (ICl). Food Anal Methods 9:2479–2483. https://doi.org/10.1007/s12161-016-0401-1. Tu X, Chen W (2018). A review on the recent progress in matrix solid phase dispersion. Molecules 23:2767. http://dx.doi.org/10.3390/molecules23112767. Wu W, Yan Q (2016) Content determination of magnolol and honokiol in magnoliae officmalis cortex by quantitative analysis of multi-components by single-marker. China Licensed Pharm 13:23-25. http://dx.doi.org/10.3969/j.issn.1672-5433.2016.01.005. Wu YT, Lin LC, Tsai TH (2006) Simultaneous determination of honokiol and magnolol in Magnolia officinalis by liquid chromatography with tandem mass spectrometric detection. Biomed Chromatogr 20:1076-1081. http://dx.doi.org/10.1002/bmc.644. Yang X, Gao M, Hu H, Zhang H (2011) Electrochemical detection of honokiol and magnolol in traditional Chinese medicines using acetylene black nanoparticle-modified electrode. Phytochem Anal 22:291-295. http://dx.doi.org/10.1002/pca.1279. Yıldırım S, Yaşar A (2018) A core-shell column approach to fast determination of synthetic dyes in foodstuffs by high-performance liquid chromatography. Food Anal Methods 11:1581–1590. https://doi.org/10.1007/s12161-017-1138-1 Yuan Y, Zhou X, Wang Y, Wang Y, Teng X, Wang S (2020) Cardiovascular modulating effects of magnolol and honokiol, two polyphenolic compounds from traditional Chinese medicine- magnolia officinalis . Curr Drug Targets 21:559-72. https://doi.org/10.2174/1389450120666191024175727. Zhang Q, Hong B, Liu J, Mu G, Cong H, Li G, Cai DF (2014). Multiwalled-carbon-nanotubes-based matrix solid-phase dispersion extraction coupled with high performance liquid chromatography for the determination of honokiol and mangolol in magnoliae cortex. J Sep Sci 37:1330-1336. http://dx.doi.org/10.1002/jssc.201301046. Zhang Y, Xu X, Qi X, Gao W, Sun S, Li X, Jiang C, Yu A, Zhang H, Yu Y (2012). Determination of sulfonamides in livers using matrix solid-phase dispersion extraction high-performance liquid chromatography. J Sep Sci 35: 45-52. http://dx.doi.org/10.1002/jssc.201100600. Additional Declarations No competing interests reported. Supplementary Files 20221215Supplementarydata.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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-2381002","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":160593205,"identity":"53a9e2c8-74bc-478b-945b-9833cf8d0e34","order_by":0,"name":"Zhengming Qian","email":"","orcid":"","institution":"Xiangnan University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zhengming","middleName":"","lastName":"Qian","suffix":""},{"id":160593208,"identity":"d3990391-e29d-4146-ba6a-c3a7d46cd0f6","order_by":1,"name":"Jing Chen","email":"","orcid":"","institution":"Xiangnan University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jing","middleName":"","lastName":"Chen","suffix":""},{"id":160593211,"identity":"eefa6ed9-102d-4595-85a0-73b084bc8c2c","order_by":2,"name":"Qinggui Lei","email":"","orcid":"","institution":"Dongguan HEC Cordyceps R\u0026D Co., Ltd","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Qinggui","middleName":"","lastName":"Lei","suffix":""},{"id":160593213,"identity":"5c9b911d-7b7c-49d4-8059-c387448dc5f9","order_by":3,"name":"Guoying Tan","email":"","orcid":"","institution":"Dongguan HEC Cordyceps R\u0026D Co., Ltd","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Guoying","middleName":"","lastName":"Tan","suffix":""},{"id":160593216,"identity":"360799ee-d0ec-42bc-af7f-4ab03918b705","order_by":4,"name":"Yuansheng Zou","email":"","orcid":"","institution":"Dongguan HEC Cordyceps R\u0026D Co., Ltd","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yuansheng","middleName":"","lastName":"Zou","suffix":""},{"id":160593217,"identity":"25eb9d9a-c99d-4bc5-a6a6-bf56c9cd8d98","order_by":5,"name":"Gang Peng","email":"","orcid":"","institution":"Lingnan Traditional Chinese Medicine Tablets Co., Ltd","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gang","middleName":"","lastName":"Peng","suffix":""},{"id":160593218,"identity":"59e438b5-67d8-47b7-b3f7-c8be5d94dbcb","order_by":6,"name":"Wenqing Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2klEQVRIiWNgGAWjYBACPmYwJcHAwN7Y+CChooawFja4Fp7DzQYPzhwjQgucJeHeJvmwhZkILew8ho8Lyizy5CMY2yoSG9gY+Nu7Ewg4jMfYeMY5iWLD241tNxJ3yDBInDm7gYAW3m3SvG0SiRvnHARqOcPGYCCRS1DL9t9gLTMS2woS25iJ0rKNGaRlvkRiGwORWvg/S/Ock0jcwHOwWSLhzDEegn7h5z+W+JmnrC5xfnv7w48/Kmrk+Nt78WuB2sXAYHAAwuQhQjlUi3wDkWpHwSgYBaNg5AEAvPVEeizNSYsAAAAASUVORK5CYII=","orcid":"","institution":"Dongguan HEC Cordyceps R\u0026D Co., Ltd","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Wenqing","middleName":"","lastName":"Li","suffix":""},{"id":160593219,"identity":"45bfb7c2-20d4-42f9-b6ff-279fa6347ea0","order_by":7,"name":"Juying Xie","email":"","orcid":"","institution":"Xiangnan University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Juying","middleName":"","lastName":"Xie","suffix":""}],"badges":[],"createdAt":"2022-12-15 09:44:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2381002/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2381002/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":30548268,"identity":"b522f47d-efd2-4fe1-bf85-a8e128d0fd63","added_by":"auto","created_at":"2022-12-20 05:22:30","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":26389,"visible":true,"origin":"","legend":"\u003cp\u003eThe UV spectra of honokiol and magnolol.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-2381002/v1/03198ce7da9943b58109ce5d.png"},{"id":30548269,"identity":"78a08be3-f9c8-4c1b-9e78-f694c77750d9","added_by":"auto","created_at":"2022-12-20 05:22:30","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":46235,"visible":true,"origin":"","legend":"\u003cp\u003eChromatograms of the blank solution, reference compound and MOC sample.\u003c/p\u003e\n\u003cp\u003e(1) honokiol, (2) magnolol.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-2381002/v1/adbe72e4a789b7cf0c9cc07f.png"},{"id":31527819,"identity":"16b375d1-65b0-4aaa-afff-bd64973a04eb","added_by":"auto","created_at":"2023-01-13 10:29:42","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":446634,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2381002/v1/fa2d4498-4bf3-4aee-89b1-01f773e08b63.pdf"},{"id":30548708,"identity":"7140600d-2512-45df-b6b8-d4e334f1dc73","added_by":"auto","created_at":"2022-12-20 05:30:30","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":53614,"visible":true,"origin":"","legend":"","description":"","filename":"20221215Supplementarydata.docx","url":"https://assets-eu.researchsquare.com/files/rs-2381002/v1/3afee168147d15522e8ae50f.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"An Ultra-rapid and Green Method for Simultaneous Determination of Honokiol and Magnolol in Magnoliae Officinalis Cortex with One Standard by HPLC-UV at Equal Absorption Wavelength","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMagnoliae Officinalis Cortex (MOC), the stem or root bark of \u003cem\u003eMagnoliae Officinalis\u003c/em\u003e, is a traditional used food addictive (Greenberg et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Luo et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). For example, it had been used in biscuits, beverages and chewing gum. More than 200 components were found in MOC, such as lignans, alkaloids, steroids (Luo et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Honokiol and magnolol were believed as two major bioactive components in MOC, which had a variety of biological activities, such as anti-cancer (Mottaghi and Abbaszadeh, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), cardiovascular modulating (Yuan et al., 2022), inhibiting pathogenic bacteria and inflammation (Chiu et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Kim et al., 2021). As the excellent activities and high content, honokiol and magnolol were selected as the quality markers of MOC in Chinese pharmacopoeia (Committee of the Chinese Pharmacopoeia, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSeveral high-performance liquid chromatography (HPLC) methods for determination of honokiol and magnolol in MOC had been reported, such as HPLC with ultraviolet detection (HPLC-UV) (Ma et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; He et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), HPLC with fluorescence detector (HPLC-FLD) (Hu and Yin, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), HPLC with electrochemical detection (HPLC-ECD) (Yang et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) and HPLC with mass spectrometry (HPLC-MS) (Wu et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). However, the reported HPLC methods were time-consuming and took amounts of harmful organic solvents. For example, in Chinese pharmacopoeia, the maceration extraction (ME) combined with HPLC-UV, which took more than 24 hours and consumed 60.5 mL methanol (Ma et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The reported ultrasonic extraction (UE) and HPLC-UV method took 60 min and consumed 24 mL methanol (He et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In addition, the reported methods used external standard method (ESM) for determination and two reference compounds were required. In order to reduce the reference compounds consumption, quantitative analysis of multi-components by single marker (QAMS) method was developed for determination of honokiol and magnolol in MOC by using a single reference compound with relative calibration factors (RCF) (Wu and Yan, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). However, detection of RCF in QAMS method increased the operational complexity (Chen et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Liu et al, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Therefore, it was necessary to develop a simple, rapid, green and standards-saving method to improve assay of honokiol and magnolol in MOC.\u003c/p\u003e \u003cp\u003eUltrasonic-assisted matrix solid-phase dispersion (UAMSPD) was proved as a fast extraction method, which had the advantage of saving solvent, time and material (Tu and Chen, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Dos Santos et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The poroshell column was a type of rapid column, which could provide rapid separation of compounds with low back pressure (Qian et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Yıldırım and Yaşar, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Li et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Ethanol was considered as an eco-friendly regent in HPLC analysis (Capello et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Shimamoto and Aricetti, 2016; Okcu et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Raspe et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Therefore, the combination of UAMSPD extraction and poroshell column HPLC method with ethanol as green extraction solvent and mobile phase would provide a rapid and eco-friendly method for herbal medicine analysis.\u003c/p\u003e \u003cp\u003eIn HPLC-UV quantitative analysis, the content of analyte was determined by comparing peak area with a known concentration reference compound at detection wavelength (Rodrigues et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The maximum absorption wavelength of analyte was usually selected as the detection wavelength because of the good sensitivity. For example, the maximum absorption wavelength 294nm was always selected as detection wavelength for HPLC analysis of honokiol and magnolol (Ma et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; He et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). The UV responses of honokiol and magnolol at 294 nm were different, so the quantification of honokiol and magnolol needed two reference compounds. When honokiol and magnolol had the same UV response at someone UV detection wavelength, this UV detection wavelength was called as equal absorption wavelength (EAW). Therefore, at the EAW, the determination of honokiol and magnolol with one reference compound and without RCF could be achieved.\u003c/p\u003e \u003cp\u003eIn present experiment, an ultra-fast and eco-friendly analytical method for determination of honokiol and magnolol with one reference compound was developed by UAMSPD and HPLC-UV EAW. The developed method was applied in six batches of MOC samples.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cdiv class=\"Section2\" id=\"Sec3\"\u003e\n \u003ch2\u003eChemicals and materials\u003c/h2\u003e\n \u003cp\u003eHPLC-grade formic acid was obtained from Aladdin (Shanghai, China), HPLC-grade ethaanol was obtained from Krude company (Losangeles, USA), analytical grade ethanol was purchased from Xilong Scientific Co., Ltd (Sichuan, China). The deionized water was purified by Milli-Q purification system (Millipore, USA). Diatomaceous earth was purchased from Sigma (Merck, Germany). Honokiol (98%) and magnolol (98%) were purchased from Shanghai Acmec Biochemical Co., Ltd. (Shanghai, China).\u003c/p\u003e\n \u003cp\u003eSix batches of MOC samples were collected from the Guangdong markets (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The authenticity of all samples was identified by Dr. Zhengming Qian. Voucher specimens were deposited at Dongguan HEC Cordyceps R\u0026amp;D Co., Ltd, Dongguan, Guangdong. All crude materials were pulverized with a mill and the powders were then obtained by passing through a 50 mesh sieve.\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eThe contents of honokiol and magnolol in MOC by HPLC-UV ESM and HPLC-UV EAW (%)\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\" style=\"width: 11.119%;\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\" style=\"width: 29.3993%;\"\u003e\n \u003cp\u003eESM\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\" style=\"width: 49.7527%;\"\u003e\n \u003cp\u003eEAW\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" style=\"width: 15.0766%;\"\u003e\n \u003cp\u003eHonokiol\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 14.1343%;\"\u003e\n \u003cp\u003eMagnolol\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 13.5689%;\"\u003e\n \u003cp\u003eHonokiol\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 36.1837%;\"\u003e\n \u003cp\u003eMagnolol\u003c/p\u003e\n \u003cp\u003e(determined by honokiol)\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\" style=\"width: 11.119%;\"\u003e\n \u003cp\u003eS1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.0766%;\"\u003e\n \u003cp\u003e2.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 14.1343%;\"\u003e\n \u003cp\u003e2.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 13.5689%;\"\u003e\n \u003cp\u003e2.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 36.1837%;\"\u003e\n \u003cp\u003e2.15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 11.119%;\"\u003e\n \u003cp\u003eS2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.0766%;\"\u003e\n \u003cp\u003e3.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 14.1343%;\"\u003e\n \u003cp\u003e3.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 13.5689%;\"\u003e\n \u003cp\u003e3.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 36.1837%;\"\u003e\n \u003cp\u003e3.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 11.119%;\"\u003e\n \u003cp\u003eS3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.0766%;\"\u003e\n \u003cp\u003e2.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 14.1343%;\"\u003e\n \u003cp\u003e2.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 13.5689%;\"\u003e\n \u003cp\u003e2.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 36.1837%;\"\u003e\n \u003cp\u003e2.56\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 11.119%;\"\u003e\n \u003cp\u003eS4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.0766%;\"\u003e\n \u003cp\u003e2.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 14.1343%;\"\u003e\n \u003cp\u003e2.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 13.5689%;\"\u003e\n \u003cp\u003e2.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 36.1837%;\"\u003e\n \u003cp\u003e2.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 11.119%;\"\u003e\n \u003cp\u003eS5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.0766%;\"\u003e\n \u003cp\u003e2.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 14.1343%;\"\u003e\n \u003cp\u003e2.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 13.5689%;\"\u003e\n \u003cp\u003e2.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 36.1837%;\"\u003e\n \u003cp\u003e2.63\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 11.119%;\"\u003e\n \u003cp\u003eS6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.0766%;\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 14.1343%;\"\u003e\n \u003cp\u003e1.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 13.5689%;\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 36.1837%;\"\u003e\n \u003cp\u003e1.75\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003c/p\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003ePreparation Of Reference Compound Solution\u003c/h3\u003e\n\u003cp\u003eThe mixed reference compounds solution containing honokiol (4.453 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and magnolol (4.449 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) was prepared with 70% (v/v) ethanol and then diluted with 70% ethanol to obtain the intended concentrations. All solutions were kept under 4 ℃.\u003c/p\u003e\n\u003ch3\u003ePreparation Of Sample Solution\u003c/h3\u003e\n\u003cp\u003eThe sample powder (200 mg) and dispersant (200 mg) were precisely weighed, putted into an agate mortar and homogeneously ground for 0.5 min. Then 50 mg of the mixture was transferred into a 10 mL glass tube and 5 mL 70% ethanol was added. The sample was extracted for 1.0 min by ultrasonic extraction (33 kHz, 250W). Small amount of 70% ethanol solution was added to compensate the lost weight. The final sample solution was filtered by 0.45 \u0026micro;m membrane before HPLC injection.\u003c/p\u003e\n\u003ch3\u003eHplc Condition\u003c/h3\u003e\n\u003cp\u003eThe HPLC analysis was operated on Agilent 1260 HPLC system (Agilent, USA) consisting of a binary pump, a thermostatic column compartment and an auto-sampler. The sample solution was separated on Poroshell 120 EC-C18 column (Agilent, Germany) (50 \u0026times;4.6 mm, 2.7 \u0026micro;m) by 70% ethanol aqueous solution with 0.1% formic acid at a flow rate of 1.5 mL min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The column temperature was maintained at 35\u0026deg;C, the injection volume was 1uL, and the wavelength was set at 247 nm (bandwidth 4 nm).\u003c/p\u003e\n\u003ch3\u003eMethod Validation\u003c/h3\u003e\n\u003cp\u003eThe method was validated for linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy and stability.\u003c/p\u003e\n\u003cp\u003eLinearity was evaluated by the calibration curve, which was obtained by plotting peak areas versus the concentrations of reference compound solutions (0.018\u0026ndash;0.445 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). The limits of detection (LODs) and quantification (LOQs) were obtained by analyzing the reference compound solution based on the signal-to-noise ratios (S/N) of approximately 3 and 10, respectively.\u003c/p\u003e\n\u003cp\u003eThe intra-day precision was determined by analyzing the mixed reference compounds solution in six replicates within one day, and the inter-day precision was determined by analyzing the mixed reference compounds solution twice per day for three days. The repeatability was tested by analyzing sample S3 in six replicates. The relative standard deviation (RSD, %) was used to evaluate the precision and repeatability.\u003c/p\u003e\n\u003cp\u003eAccuracy was evaluated by recovery. Known amount of two analytes were added into sample S3. The mixture was extracted and analyzed. Six replicates were performed. The recovery was calculated as the ratio of detected amount to added amount.\u003c/p\u003e\n\u003cp\u003eThe stability was tested by analyzing sample solution 6 times in 24 h at room temperature. The variation was expressed as RSD. The robustness tests were carried out by the developed HPLC method, which included flow rates (1.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 mL min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), column temperatures (35\u0026thinsp;\u0026plusmn;\u0026thinsp;5℃), and different columns (Agilent Poroshell 120 EC-C18, Waters CORTECS C18 and ACE UltraCore SuperC18). The relative retention time (RRT), relative peak area (RPA) and resolution of honokiol to magnolol were recorded for evaluating robustness.\u003c/p\u003e"},{"header":"Results And Discussion","content":"\u003cdiv class=\"Section2\" id=\"Sec9\"\u003e\n \u003ch2\u003eOptimization of extraction condition\u003c/h2\u003e\n \u003cp\u003eThe UAMSPD extraction parameters including sample grinding and ultrasonic extraction, the grinding time, extraction solvent and ultrasonic time were optimized to obtain the satisfactory extraction yield. The extraction yields of current method were evaluated by comparing with Pharmacopeia of China 2020 method of MOC.\u003c/p\u003e\n \u003cp\u003eDiatomaceous earth was employed as the dispersant based on previous research (Zhang et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e). Two ratios of sample to diatomaceous earth (1:1 and 1:2) were compared. There was no difference between the two ratios, thus 1:1 was used. Four different grinding times (10, 30, 60 and 120 s) were also compared. The results indicated that grinded time of 30 s could provide a satisfied extraction yield of target compounds (Fig. S1\u003cstrong\u003eA\u003c/strong\u003e). Ethanol as an eco-friendly solvent was selected as extraction solvent. Three concentrations of ethanol (50%, 70%, 90%) were tested. The results showed that 70% ethanol aqueous solution was more suitable (Fig. S1\u003cstrong\u003eB\u003c/strong\u003e). The different ratios of sample to extraction solvent (1: 100, 1: 200, 1: 400) were tested. The results (Fig. S1\u003cstrong\u003eC\u003c/strong\u003e) of the three tests were similar. Considering the suitable HPLC peak areas, 1: 200 was chosen. In addition, ultrasonic extraction time (30, 60, 120 and 240 s) were evaluated. According to Fig. S1\u003cstrong\u003eD\u003c/strong\u003e, when extraction time was more than 60 s, the extraction yields were similar. Thus, 60 s was used as extraction time.\u003c/p\u003e\n \u003cp\u003eTo confirm the extraction efficiency of the developed UAMSPD, sample S3 was extracted with USMAPD and Chinese Pharmacopeia extraction method. Three replicates were performed. The contents of honokiol and magnolol extracted by UAMSPD were 2.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.029% and 2.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042%, and by Chinese Pharmacopeia method were 2.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.021% and 2.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.027%. These results showed that the extraction efficiency of the developed method was similar with Chinese Pharmacopeia method. Thus, the developed UAMSPD could be used as a good substitute method for the Chinese Pharmacopeia method.\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eOptimization Of Hplc Condition\u003c/h3\u003e\n\u003cp\u003eHonokiol and magnolol were usually separated on traditional C18 columns and more than 10 min was consumed (Ma et al., \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhang et al., \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e; He et al., \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e; Wu et al., \u003cspan class=\"CitationRef\"\u003e2006\u003c/span\u003e). In order to reduce the HPLC separation time, poroshell column was employed in experiment. To reduce the harmful solvent consumption, ethanol was selected as the organic mobile phase. The 25%, 30% and 35% ethanol aqueous solution with 0.1% formic acid were compared as the mobile phase. The 30% ethanol aqueous solution with 0.1% formic acid was chosen because of the good peak shape and resolution. The flow rates of 1.0 and 1.5 mL min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e were compared and 1.5 mL min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e was used based on the shorter time. The column temperatures (30, 35 and 40 ℃) were also compared, the separations of two analytes were similar. The 35 ℃ was selected, because it was easy to control. To ensure the specificity of the method, the peak purities of target compounds were checked. The results showed that the purities of honokiol and magnolol in samples were higher than 99.99%.\u003c/p\u003e\n\u003cp\u003eThe selection of EAW was important in this experiment, which included two steps. Firstly, screening the EAW of honokiol and magnolol by UV spectrophotometer. The honokiol solution (0.0222 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and magnolol solution (0.0222 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) were prepared with the mobile phase, and then the two reference compound solutions were scanned from 200 nm to 350 nm with Agilent Cary 60 ultraviolet spectrophotometer (Agilent, USA). The UV spectra of the two compounds were shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. There were three EAWs (247 nm, 281 nm and 298 nm) for honokiol and magnolol. The 247 nm was chosen for further experiment due to the highest UV response. Secondly, confirming the EAW of honokiol and magnolol by HPLC-UV. The mixture reference solution of honokiol (0.0890 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and magnolol (0.0890 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) was separated on Poroshell 120 EC-C18 column (50 \u0026times;4.6 mm, 2.7 \u0026micro;m) with 70% ethanol aqueous solution with 0.1% formic acid, and detected at 246 nm, 247 nm and 248 nm. The results (Table S1) showed that the peak areas of honokiol and magnolol were same at 247 nm. Therefore, 247 nm was used as EAW. In addition, different bandwidths (1 nm, 2 nm, 4 nm and 8 nm) were tested. The 4 nm showed the best precision of peak areas, and was used in developed HPLC method.\u003c/p\u003e\n\u003ch3\u003eMethod Validation\u003c/h3\u003e\n\u003cp\u003eThe regression equation for honokiol was y\u0026thinsp;=\u0026thinsp;1410.2523 \u003cem\u003ex\u003c/em\u003e\u0026thinsp;+\u0026thinsp;0.8718 (r\u0026thinsp;=\u0026thinsp;0.9999), for magnolol was \u003cem\u003ey\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1404.0385 \u003cem\u003ex\u003c/em\u003e\u0026thinsp;+\u0026thinsp;1.3179 (r\u0026thinsp;=\u0026thinsp;0.9999) in the test range of 0.018\u0026ndash;0.445 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, which indicated a good linearity of the two compounds. The LODs for honokiol and magnolol were 0.3559 and 0.3562 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The LOQs for honokiol and magnolol were 0.8898 and 0.8906 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eThe RSDs of intra- and inter-day precision (n\u0026thinsp;=\u0026thinsp;6) for honokiol were 0.38% and 0.45%; the RSDs of intra- and inter-day precision (n\u0026thinsp;=\u0026thinsp;6) for magnolol were 0.36% and 0.49%. The RSDs of repeatability (n\u0026thinsp;=\u0026thinsp;6) for honokiol and magnolol were 1.9% and 1.8%. The recoveries (Table S2) of honokiol and magnolol (n\u0026thinsp;=\u0026thinsp;6) were 99.28% (RSD\u0026thinsp;=\u0026thinsp;2.3%) and 98.08% (RSD\u0026thinsp;=\u0026thinsp;1.8%), respectively. The RSDs of stability (24 h) for honokiol and magnolol were 0.86% and 0.47%.\u003c/p\u003e\n\u003cp\u003eThe RRTs, RPAs, and resolution of honokiol to magnolol in the robustness tests were listed in Table S3. The RSD for RRTs were 1.3%. The RPA values were all close to 1.00. The resolutions were all larger than 1.5. These results indicated a good robustness of the developed method within the test ranges.\u003c/p\u003e\n\u003cp\u003eAbove all, the developed HPLC-UV EAW was accurate and stable for quantitative analysis of honokiol and magnolol in MOC.\u003c/p\u003e\n\u003ch3\u003eAnalysis Of Moc Sample\u003c/h3\u003e\n\u003cp\u003eThe reference compounds and MOC sample chromatograms were shown in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. To evaluate the feasibility of the developed HPLC-UV EAW, six MOC samples were determined by HPLC-UV EAW and ESM, respectively. In HPLC-UV EAW, honokiol and magnolol were determined by one reference compound (honokiol). While in HPLC-UV ESM, honokiol and magnolol were determined by two reference compounds. The results were listed in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. The contents of magnolol in six MOC samples obtained by the two different methods were compared by t-test (SPSS 19.0 software). The result (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) indicated that there was no significant difference between HPLC-UV EAW (only use honokiol as reference compound) and HPLC-UV ESM (use two reference compounds). This indicated that HPLC-UV EAW could be a good alternative method of HPLC-UV ESM for determination of honokiol and magnolol in MOC.\u003c/p\u003e\n\u003cp\u003eThe contents of honokiol and magnolol obtained by HPLC-UV EAW were 0.75\u0026thinsp;~\u0026thinsp;3.19% and1.75\u0026thinsp;~\u0026thinsp;3.02%, respectively, which were agreed with the literatures (Jiang et al., \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e). The total contents of honokiol and magnolol were 2.50\u0026thinsp;~\u0026thinsp;6.21%, which indicated that the six batch MOC samples were all complied with the Chinese Pharmacopeia (\u0026ge;\u0026thinsp;2.0%).\u003c/p\u003e\n\u003ch3\u003eComparison Of The Developed Method And Reported Method\u003c/h3\u003e\n\u003cp\u003eComparisons of the current method with the reported HPLC methods (Method 1, 2, 3 and 4) (Ma et al., \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e;Zhang et al., \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e; He et al., \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e; Wu et al., \u003cspan class=\"CitationRef\"\u003e2006\u003c/span\u003e) were summarized in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The developed method had advantages in less reference compounds, shorter analytical time and solvent eco-friendly.\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab2\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eThe comparison of the developed and reported HPLC methods for determination of honokiol and magnolol in MOC. \u003cstrong\u003e*\u003c/strong\u003e The mobile phase additives (\u0026lt;\u0026thinsp;0.1 mL) were not reflected\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\" style=\"width: 8.8316%;\"\u003e\n \u003cp\u003eNO.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\" style=\"width: 39.2942%;\"\u003e\n \u003cp\u003eSample preparation\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\" style=\"width: 31.6146%;\"\u003e\n \u003cp\u003eHPLC separation\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" style=\"width: 8.7036%;\"\u003e\n \u003cp\u003eMethod\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 22.271%;\"\u003e\n \u003cp\u003eSolvent\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 8.3196%;\"\u003e\n \u003cp\u003eTime\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 10.1115%;\"\u003e\n \u003cp\u003eMethod\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 15.3593%;\"\u003e\n \u003cp\u003eSolvent*\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 6.1437%;\"\u003e\n \u003cp\u003eTime\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\" style=\"width: 8.8316%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.7036%;\"\u003e\n \u003cp\u003eME\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 22.271%;\"\u003e\n \u003cp\u003eMethanol 45mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.3196%;\"\u003e\n \u003cp\u003e1440 min\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.1115%;\"\u003e\n \u003cp\u003eHPLC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 15.3593%;\"\u003e\n \u003cp\u003eMethanol 15.6mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.1437%;\"\u003e\n \u003cp\u003e20 min\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 8.8316%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.7036%;\"\u003e\n \u003cp\u003eMSPD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 22.271%;\"\u003e\n \u003cp\u003eMethanol 10mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.3196%;\"\u003e\n \u003cp\u003e\u0026gt;1.5 min\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.1115%;\"\u003e\n \u003cp\u003eHPLC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 15.3593%;\"\u003e\n \u003cp\u003eMethanol 7.8mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.1437%;\"\u003e\n \u003cp\u003e10 min\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 8.8316%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.7036%;\"\u003e\n \u003cp\u003eUE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 22.271%;\"\u003e\n \u003cp\u003ePolyethylene glycol 20mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.3196%;\"\u003e\n \u003cp\u003e30 min\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.1115%;\"\u003e\n \u003cp\u003eHPLC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 15.3593%;\"\u003e\n \u003cp\u003eMethanol 24mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.1437%;\"\u003e\n \u003cp\u003e30 min\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 8.8316%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.7036%;\"\u003e\n \u003cp\u003eBE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 22.271%;\"\u003e\n \u003cp\u003eEthanol 47.5mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.3196%;\"\u003e\n \u003cp\u003e13 min\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.1115%;\"\u003e\n \u003cp\u003eLC-MS/MS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 15.3593%;\"\u003e\n \u003cp\u003eAcetonitrile 3mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.1437%;\"\u003e\n \u003cp\u003e5 min\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 8.8316%;\"\u003e\n \u003cp\u003eThis work\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.7036%;\"\u003e\n \u003cp\u003eVAMSPD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 22.271%;\"\u003e\n \u003cp\u003eEthanol 3.5ml\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 8.3196%;\"\u003e\n \u003cp\u003e1.5 min\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 10.1115%;\"\u003e\n \u003cp\u003eHPLC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 15.3593%;\"\u003e\n \u003cp\u003eEthanol 1.05mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 6.1437%;\"\u003e\n \u003cp\u003e1 min\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cp\u003eFirstly, the developed method only need one reference compound for determining two analytes, while in other literatures two reference compounds were required. Compared with the QAMS method, the current method was simpler without RCF used.\u003c/p\u003e\n\u003cp\u003eSecondly, the developed method was the fastest. The total analytical time was 2.5 min including sample extraction time (1.5 min) and HPLC separation time (1 min). The reported methods took more than 11 min. For example, method 1 took 1440 min in sample extraction and 20 min in HPLC separation. Method 3 took 30 min in the ultrasonic extraction and 30 min in HPLC separation. Although a few rapid analytical methods (method 2, 4) were achieved by MSPD extraction or the LC-MS/MS system, more than 11 min were still needed.\u003c/p\u003e\n\u003cp\u003eThirdly, the developed method was the greenest. \u0026ldquo;Analytical Eco-Scale\u0026rdquo; approach (Sereshti et al.,2022; Gałuszka et al., \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e) was carried out to evaluate the greenness of developed method and reported ones. The results (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e) showed that developed HPLC method got the highest Eco-Scale score of 89. In the current method, eco-friendly solvent ethanol was employed to replace those harmful solvents, such as acetonitrile and methanol. The rapid sample extraction and HPLC separation also made less solvent and energy consumed. Methanol or acetonitrile was the main weak points for other methods (method 1, 2, 3, 4). Although polyethylene glycol as an eco-friendly solvent was used in sample extraction of method 3, methanol was still used in the HPLC separation. The sample extraction (30 min) and HPLC separation (30 min) also consumed more energy and produced more waste. Thus, it got the lowest score of 75.\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab3\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eGreenness assessment of the developed and reported HPLC methods for determination of honokiol and magnolol in MOC\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003ePenalty points (PPs)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMethod 1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMethod 2\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMethod 3\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMethod 4\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eThis work\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eReagents\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMethanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\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\u003eEthanol\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 \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAcetonitrile\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 \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\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\u003eWater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFormic acid\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 \u003ctd align=\"left\"\u003e\n \u003cp\u003e/\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAcetic acid\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\u003e4\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\u003eMultiwalled carbon nanotubes(60mg)\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\u003e2\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\u003eDiatomaceous earth (200mg)\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 \u003ctd align=\"left\"\u003e\n \u003cp\u003e/\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePolyethylene glycol\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\u003e2\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\u003e\u003cstrong\u003eInstrument Energy\u003c/strong\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 \u003ctd align=\"left\"\u003e\u0026nbsp;\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\u003eHPLC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\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\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHPLC-MS\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 \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\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\u003eSonicator\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\u003e1\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\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCentrifuge\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 \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\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\u003e\u003cstrong\u003eOccupational hazard\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eWaste\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal PPs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026sum;18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026sum;20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026sum;25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026sum;16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026sum;11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAnalytical EcoScale\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e82\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e80\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e75\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e84\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e89\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003etotal score\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Conclusions","content":"\u003cp\u003eIn present study, an ultra-fast and eco-friendly method based on UAMSPD and HPLC-UV EAW was developed for determination of honokiol and magnolol in MOC. The developed assay method only used one reference compound for determination of honokiol and magnolol in MOC without RCF. Comparing with the reported methods, the developed procedure was ultra-fast, green and reference compound saving. It was an improved analytical method for determination of honokiol and magnolol in MOC and related products.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthorship contribution\u0026nbsp;\u003c/strong\u003eZhengming Qian: Investigations, Methodology, Writing-\u0026nbsp;original draft preparation\u003cstrong\u003e.\u003c/strong\u003e Jing Chen: Formal analysis, Investigations.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eQinggui Lei: Investigations, Data curation, Methodology. Guoying Tan: writing- review \u0026amp; editing. Yuansheng Zou: Investigations. Gang Peng: Collecting sample. Wenqing Li: Conceptualization, Funding acquisition, Writing- review \u0026amp; editing. Juying Xie: Supervision, Project administration, Writing- review \u0026amp; editing.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThis study was financially supported by the Key Laboratory of Guangdong Drug Administration (2021ZDB05).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e The authors declare that all the data supporting the findings of this study are included in the manuscript or in the supplements\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompliance with ethical standards\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest\u0026nbsp;\u003c/strong\u003eThe authors declare no potential conflicts of interest with respect to the research, authorship, and or publication of this article\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval\u003c/strong\u003e Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent\u0026nbsp;\u003c/strong\u003eNot applicable.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eCapello C, Fischer U, Hungerb\u0026uuml;hler K (2007) What is a green solvent? A comprehensive framework for the environmental assessment of solvents. Green Chem 9:927-934. https://doi.org/10.1039/b617536h.\u003c/li\u003e\n\u003cli\u003eChen Q, Wang Z, Yang B, Yang Q, Kan J (2022) Determination of main alkylamides responsible for Zanthoxylum bungeanum pungency through quantitative analysis of multi-components by a single marker. Food Chem\u003cem\u003e \u003c/em\u003e396: 133645. https://doi.org/10.1016/j.foodchem.2022.133645.\u003c/li\u003e\n\u003cli\u003eChiu KC, Shih YH, Wang TH, Lan WC, Li PJ, Jhuang HS, Hsia SM, Shen YW, Chen MYC, Shieh TM (2021) In vitro antimicrobial and antiproinflammation potential of honokiol and magnolol against oral pathogens and macrophages. J Formosan Med Assoc 120: 827-837. https://doi.org/10.1016/j.jfma.2020.09.002.\u003c/li\u003e\n\u003cli\u003eCommittee of the Chinese Pharmacopoeia (2020). Pharmacopoeia of the People\u0026rsquo;s Republic of China, Vol. 4. China Medical Science Press, Bejing, pp 236.\u003c/li\u003e\n\u003cli\u003eDos Santos EO, Gonzales O, Ores JC, Marube LC, Caldas SS, Furlong EB, Primel EG (2019) Sand as a solid support in ultrasound-assisted MSPD: A simple, green and low-cost method for multiresidue pesticide determination in fruits and vegetables. Food Chem 297:124926. https://doi.org/10.1016/j.foodchem.2019.05.200.\u003c/li\u003e\n\u003cli\u003eGałuszka A, Konieczka PM, Migaszewski Z, Namieśnik J (2012) Analytical eco-scale for assessing the greenness of analytical procedures. TrAC, Trends Anal Chem\u003cem\u003e \u003c/em\u003e37:61-72. http://dx.doi.org/10.1016/j.trac.2012.03.013.\u003c/li\u003e\n\u003cli\u003eGreenberg M, Urnezis P, Tian M (2007) Compressed mints and chewing gum containing magnolia bark extract are effective against bacteria responsible for oral malodor. J Agric Food Chem\u003cem\u003e \u003c/em\u003e55: 9465-9469. https://doi.org/10.1021/jf072122h.\u003c/li\u003e\n\u003cli\u003eHe L, Fan T, Hu J, Zhang L (2015) Polyethylene glycol-based ultrasound-assisted extraction of magnolol and honokiol from\u003cem\u003e \u003c/em\u003ecortex\u003cem\u003e magnoliae officinalis\u003c/em\u003e. Nat Prod Res 29:31-36. http://dx.doi.org/10.1080/14786419.2014.955800.\u003c/li\u003e\n\u003cli\u003eHu L, Yin C (2015) Multi-way calibration coupling with fluorescence spectroscopy to determine magnolol and honokiol in herb and plasma samples. Anal Methods 7:5913-5923. http://dx.doi.org/10.1039/c5ay01285f. \u003c/li\u003e\n\u003cli\u003eJiang Y, Vaysse J, Gilard V, Balayssac S, D\u0026eacute;jean S, Malet-Martino M, David B, Fiorini C, Barbin Y (2011) Quality assessment of commercial magnoliae officinalis cortex by 1H-NMR-based metabolomics and HPLC methods. Phytochem Anal 23:387-395. http://dx.doi.org/10.1002/pca.1369.\u003c/li\u003e\n\u003cli\u003eKim H, Lim CY, Chung MS (2020) \u003cem\u003eMagnolia officinalis\u003c/em\u003e and its honokiol and magnolol constituents inhibit human norovirus surrogates. Foodborne Pathog Dis 18:24-30. https://doi.org/10.1089/fpd.2020.2805.\u003c/li\u003e\n\u003cli\u003eLi W, Qian Z, Lei Q, Lian Y, Zou Y, Wang Y, Lan D (2023) An ultra-rapid and eco-friendly method for determination of loganic acid and gentiopicroside from \u003cem\u003eGentianae Macrophyllae\u003c/em\u003e Radix by vortex-assisted matrix solid-phase dispersion extraction and LC-MS. J Pharm Biomed Anal 222:115085. https://doi.org/10.1016/j.jpba.2022.115085.\u003c/li\u003e\n\u003cli\u003eLiu M, Yao J, Zhang C, Hao R, Zhang Z, Cao W (2022) Quantitative analysis of multi-components by single-marker: An effective method for the chemical characterization of wood vinegar. Ind Crops Prod 182:114862. https://doi.org/10.1016/j.indcrop.2022.114862.\u003c/li\u003e\n\u003cli\u003eLuo H, Wu H, Yu X, Zhang X, Lu Y, Fan J, Tang L, Wang Z (2019) A review of the phytochemistry and pharmacological activities of \u003cem\u003emagnoliae officinalis\u003c/em\u003e cortex. J Ethnopharmacol 236:412-442. https://doi.org/10.1016/j.jep.2019.02.041. \u003c/li\u003e\n\u003cli\u003eMa DX, Liu JF, Hu MJ, Li BZ, Chen J, Liu RL (2022) Analysis on the contents of medicinal components in the back of\u003cem\u003e houpoea officinalis\u003c/em\u003e at different heights. J Hebei Norm Univ, Nat Sci Ed 46:304-309. http://dx.doi.org/10.13763/j.cnki.jhebnu.nse.202204007.\u003c/li\u003e\n\u003cli\u003eMottaghi S, Abbaszadeh H (2022) Natural lignans honokiol and magnolol as potential anticarcinogenic and anticancer agents. A comprehensive mechanistic review. Nutr Cancer 74:761-778, https://doi.org/10.1080/01635581.2021.1931364.\u003c/li\u003e\n\u003cli\u003eOkcu G, Ayhan K, Altuntas EG, Vural N, Poyrazogiu ES (2016) Determination of phenolic acid decarboxylase produced by lactic acid bacteria isolated from shalgam, salgam juice using green analytical chemistry method. LWT-Food Sci Technol 66:615-621. http://dx.doi.org/10.1016/j.lwt.2015.10.072.\u003c/li\u003e\n\u003cli\u003eQian Z, Wu Z, Li C, Tan G, Hu H, Li W (2020) A green liquid chromatography method for rapid determination of ergosterol in edible fungi based on matrix solid-phase dispersion extraction and a core\u0026ndash;shell column. Anal Methods 12:3337-3343. https://doi.org/10.1039/d0ay00714e.\u003c/li\u003e\n\u003cli\u003eRaspe DT, Ciotta SR, Zorzenon MRT, Dacome AS, Silva CD, Milani PG, Costa SC (2021) Ultrasound-assisted extraction of compounds from Stevia leaf pretreated with ethanol. Ind Crops Prod 172:114035. https://doi.org/10.1016/j.indcrop.2021.114035.\u003c/li\u003e\n\u003cli\u003eRodrigues CI, Marta L, Maia R, Miranda M, Ribeirinho M, M\u0026aacute;guas C (2007) Application of solid-phase extraction to brewed coffee caffeine and organic acid determination by UV/HPLC. J Food Compos Anal 20:440-448. https://doi.org/10.1016/j.jfca.2006.08.005.\u003c/li\u003e\n\u003cli\u003eSereshti H, Zarei-Hosseinabadi M, Soltani S, Taghizadeh M (2022). Green vortex-assisted emulsification microextraction using a ternary deep eutectic solvent for extraction of tetracyclines in infant formulas. Food Chem 396:133743. https://doi.org/10.1016/j.foodchem.2022.133743.\u003c/li\u003e\n\u003cli\u003eShimamoto GG, Aricetti JA, Tubino M (2016) A Simple, Fast, and Green Titrimetric Method for the Determination of the Iodine Value of Vegetable Oils Without Wijs Solution (ICl). Food Anal Methods 9:2479\u0026ndash;2483. https://doi.org/10.1007/s12161-016-0401-1.\u003c/li\u003e\n\u003cli\u003eTu X, Chen W (2018). A review on the recent progress in matrix solid phase dispersion. Molecules 23:2767. http://dx.doi.org/10.3390/molecules23112767.\u003c/li\u003e\n\u003cli\u003eWu W, Yan Q (2016) Content determination of magnolol and honokiol in magnoliae officmalis\u003cem\u003e \u003c/em\u003ecortex by quantitative analysis of multi-components by single-marker. China Licensed Pharm 13:23-25. http://dx.doi.org/10.3969/j.issn.1672-5433.2016.01.005. \u003c/li\u003e\n\u003cli\u003eWu YT, Lin LC, Tsai TH (2006) Simultaneous determination of honokiol and magnolol in \u003cem\u003eMagnolia officinalis\u003c/em\u003e by liquid chromatography with tandem mass spectrometric detection. Biomed Chromatogr 20:1076-1081. http://dx.doi.org/10.1002/bmc.644.\u003c/li\u003e\n\u003cli\u003eYang X, Gao M, Hu H, Zhang H (2011) Electrochemical detection of honokiol and magnolol in traditional Chinese medicines using acetylene black nanoparticle-modified electrode. Phytochem Anal 22:291-295. http://dx.doi.org/10.1002/pca.1279.\u003c/li\u003e\n\u003cli\u003eYıldırım S, Yaşar A (2018) A core-shell column approach to fast determination of synthetic dyes in foodstuffs by high-performance liquid chromatography. Food Anal Methods 11:1581\u0026ndash;1590. https://doi.org/10.1007/s12161-017-1138-1\u003c/li\u003e\n\u003cli\u003eYuan Y, Zhou X, Wang Y, Wang Y, Teng X, Wang S (2020) Cardiovascular modulating effects of magnolol and honokiol, two polyphenolic compounds from traditional Chinese medicine-\u003cem\u003emagnolia officinalis\u003c/em\u003e. Curr Drug Targets 21:559-72. https://doi.org/10.2174/1389450120666191024175727.\u003c/li\u003e\n\u003cli\u003eZhang Q, Hong B, Liu J, Mu G, Cong H, Li G, Cai DF (2014). Multiwalled-carbon-nanotubes-based matrix solid-phase dispersion extraction coupled with high performance liquid chromatography for the determination of honokiol and mangolol in magnoliae cortex. J Sep Sci 37:1330-1336. http://dx.doi.org/10.1002/jssc.201301046.\u003c/li\u003e\n\u003cli\u003eZhang Y, Xu X, Qi X, Gao W, Sun S, Li X, Jiang C, Yu A, Zhang H, Yu Y (2012). Determination of sulfonamides in livers using matrix solid-phase dispersion extraction high-performance liquid chromatography. J Sep Sci 35: 45-52. http://dx.doi.org/10.1002/jssc.201100600.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"rapid, green, matrix solid-phase dispersion, HPLC, equal absorption wavelength","lastPublishedDoi":"10.21203/rs.3.rs-2381002/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2381002/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eAn ultra-rapid and green assay method for simultaneous determination of honokiol and magnolol in Magnoliae Officinalis Cortex with one standard was developed by HPLC-UV at equal absorption wavelength. The sample was prepared by ultrasonic-assisted matrix solid-phase dispersion. The HPLC separation was performed on a Poroshell C18 column with an eco-friendly mobile phase. The detection wavelength was set at the equal absolution wavelength of honokiol and magnolol (247 nm). The contents of honokiol and magnolol in six batches of samples, obtained by developed method with one marker and external standard method with two markers, were comparable. In addition, the developed HPLC method only took 2.5 min and 4.55 mL green organic solution (ethanol), which including the sample extraction and separation. The developed method was rapid, green and standard saving, which would be helpful to improve the quality evaluation of Magnoliae Officinalis Cortex.\u003c/p\u003e","manuscriptTitle":"An Ultra-rapid and Green Method for Simultaneous Determination of Honokiol and Magnolol in Magnoliae Officinalis Cortex with One Standard by HPLC-UV at Equal Absorption Wavelength","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-12-20 05:22:25","doi":"10.21203/rs.3.rs-2381002/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":"0802792c-e485-4f86-8a35-da1d9c31fdbe","owner":[],"postedDate":"December 20th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-01-13T10:29:35+00:00","versionOfRecord":[],"versionCreatedAt":"2022-12-20 05:22:25","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2381002","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2381002","identity":"rs-2381002","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}