Simultaneous Determination of Saponins in Dripping Pills Made from Astragali Radix and Panax notoginseng by UPLC-ELSD

Objective To develop a simple and fast method for removing polyethylene glycol(PEG) and simultaneous determination of fives saponins, i.e. astragaloside IV, notoginsenoside R_1, ginsenoside Rg_1, ginsenoside Rb_1, and ginsenoside Rd in dripping pills made from Astragali Radix and Panax notoginseng. Methods The extraction method was based on a liquid-liquid extraction using water-saturated n-butanol and the quantitative determination was based on ultra-performance liquid chromatography coupled with evaporative light scattering detection(UPLC-ELSD). The chromatographic analysis was performed on an Acquity UPLC HSS T3 column(100 mm × 2.1 mm, 1.8 μm) with a gradient elution of acetonitrile-0.1% formic acid aqueous solution within a runtime of 15 min. Results Compared to different methods, the proposed method could remove the interference of PEG in formulation. And the calibration curves showed good linearity during the test ranges. The method was validated for limits of detection and quantification, precision, and reproducibility. The recoveries were within the range of 96.87%-99.97%. In addition, the verified method was firstly applied to determination of the five active ingredients in Qishen Yiqi Dripping Pills(QYDP) simultaneously.Conclusion The contents of five active ingredients are stable and homogeneous in QYDP, which indicates that the method could be readily utilized as a quality evaluation method for this traditional Chinese medicine dripping pills made from Astragali Radix and Panax notoginseng.     

Determination of drug release in vitro of atractylenolide Ⅰ liposome by RP-HPLC

Objective To establish a RP-HPLC method for the determination of drug release in vitro of atractylenolide Ⅰ liposomes. Methods The release behavior of the drug from liposomes was studied by the third method for dissolution. ZORBAX C18 column (4.6 mm × 250 mm, 5 μm) was used with a mobile phase of Methanol-Acetonitrile-0.2% from liposomes in vitro fitted the log-normal distribution equation and had a property of sustained release. Conclusion The method is simple, fast and selective. It is suitable for the determination of release profile in vitro of atractylenolide Ⅰ liposomes.     

Comparative chemical characters of Pseudostellaria heterophylla from geographical origins of China

Objective: Pseudostellaria heterophylla has been paid more attention in recent years, mainly as a medicine food homology plant. The content determination of P. heterophylla is not specified in the Chinese Pharmacopoeia (version 2020). The environmental conditions in different production areas could exert an influence on the quality of P. heterophylla. The purpose of this study is to discriminate P. heterophylla collected from different geographical origins of China. Methods: In this study, the content of polysaccharide in 28 batches of P. heterophylla was determined using phenol-sulfuric acid. HPLC fingerprints were established under optimised HPLC-PDA methods. Subsequently, the similarity analysis (SA) and the quantification of heterophyllin B were analyzed. The metabolites of P. heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and orthogonal PLS-DA (OPLS-DA) were performed based on all peak areas. Results: The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas, which varied significant from different origins. While the content of heterophyllin B in Anhui and Jiangsu was high. The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990, and the characteristic map can be used to identify and evaluate the quality of P. heterophylla. The samples from Fujian, Guizhou, Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA, PLS-DA, HCA, OPLS-DA, indicating that their metabolic compositions were significantly different. Ultimately, a total of 15 metabolites which were filtrated by a VIP-value > 1 and a P-value ＜ 0.05 associated with the separation of different origins were identified. Conclusion: HPLC fingerprint was established to evaluate the quality and authenticity of P. heterophylla. The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions. A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P. heterophylla from different origins in China for the first time. Overall, this study provides insights to metabolomics of P. heterophylla and supplies important reference values for the development of functional foods.     

Simultaneous Determination of Eight Constituents in Fruits of Rubus chingii by UPLC

Objective To develop a simple,efficient,and reliable method for routine quantitative analysis of main constituents presented in the fruits of Rubus chingii,which is widely used in Chinese materia medica(CMM),known as Fupenzi(FPZ) in Chinese.Methods An ultra performance liquid chromatography-photo diode array(UPLC-PDA) system was employed for simultaneous quantification of eight compounds,i.e.adenosine,gallic acid,brevifolin carboxylic acid,ethyl gallate,ellagic acid,kaempferol-3-O-rutinoside,kaempferol-3-O-β-D-glucopyranoside,and tiliroside.The chromatographic analysis was performed on a C_(18) column using a gradient elution of acetonitrile-0.1%formic acid aqueous solution within a runtime of 25 min.Results All calibration curves were linear(R~2 0.9997) over the tested ranges.The intra- and inter-day precisions as determined from sample solutions were both less than 2.45%and 2.78%,respectively.The average recoveries for the eight constituents ranged from 94.77%to 101.35%with RSD ≤ 4.41%.The newly-developed method was applied to the quality assessment of various R.chingii samples,including both ripe and unripe fruits of R.chingii from different habitats.Conclusion The relative levels of the investigated compounds vary remarkably in the fruits of R.chingii collected from different habitats.As only two of the eight compounds,adenosine and ellagic acid,are determined in the ripe fruits of R.chingii,the results may explain the reason why only the unripe fruits can be used in CMM.     

Simultaneous Determination of 10 Anthraquinones in Rhubarb Based on HPLC-Q-HR/MS

Objective To develop a simple, sensitive, and precise method for simultaneous determination of 10 anthraquinones in Rhubarb. Methods HPLC-Q-HR/MS was employed for simultaneous quantification of free anthraquinones(aloe-emodin, emodin, chrysophanol, physcion, and rhein) and their glycosides. Chromatographic analysis was performed on an XBridge~(TM) C_(18) column(2.1 mm × 150 mm, 5 μm) with mobile phases consisting of 3 mmol/L ammonium acetate(A) and methanol(B) at a flow rate of 0.3 mL/min. Results All calibration curves exhibited good linear relationship(R~2 0.999). The limits of detection(LOD) and quantification(LOQ) were in the range of0.39-2.97 ng/mL and 0.56-8.90 ng/mL, respectively. The overall intra-and inter-day precisions of analytes presented as relative standard deviations(RSDs) were less than 2.79%. Relative recoveries varied between 97.83% and 104.28%. The validated method was applied to assess the quality of Rhubarb collected from different regions of China. Results showed that chrysophanol and rhein-8-O-β-D-glucoside was the largest portion of free anthraquinones and anthraquinone glycosides in Rhubarb, respectively. The total content of anthraquinones was higher in Rhubarbs from Sichuan, Qinghai, Yunnan, and Gansu provinces than that in those from Shandong and Henan provinces, while no significant variability existed in different regions of the same province.Conclusion HPLC-Q-HR/MS method is accurate and reliable for simultaneous quantification of above free anthraquinones and their glycosides in Rhubarb and can be applied to standardize the quality of Rhubarb and its quality control in different regions.     

Simultaneous quantitative determination of five alkaloids in Catharanthus roseus by HPLC-ESI-MS/MS

AIM： To establish a method to simultaneously determine the main five alkaloids of Catharanthus roseus for trace samples, a high-performance liquid chromatography–electrospray ionization-tandem mass spectrometry（HPLC-ESI-MS/MS） analysis method was developed. METHOD： The five Catharanthus alkaloids, vinblastine, vincristine, vinleurosine, vindoline, and catharanthine were chromatographically separated on a C18 HPLC column. The mobile phase was methanol-15 nmol？L–1 ammonium acetate containing 0.02% formic acid（65 ： 35, V/V）. The quantification of these alkaloids was based on the Multiple Reaction Monitoring（MRM） mode. RESULTS： This method was validated, and the results achieved the aims of the study. The intra- and inter-day precision and accuracy of the five alkaloids were within 1.2%-11.5%（RSD%） and-10.9%-10.5%（RE%）. The recovery rates of the five alkaloids of samples were from 79.9% to 91.5%. The five analytes were stable at room temperature for 2 h, at 4 °C for 12 h, and at-20 °C for two weeks. The developed method was applied successfully to determine the content of the five alkaloids in three plant parts of three batches of C. roseus with a minute amount collected from three regions of China. CONCLUSION： The HPLC-ESI-MS/MS method can be used for the simultaneous determination of five important alkaloids in trace C. roseus samples.     

中国不同地区三棱的化学特征比较

Objective: To compare the chemical characters of Sparganii Rhizoma from different areas via chromatographic analysis and to establish a sensitive LC/MS method for quality assessment of Sparganii Rhizoma.Methods: Under the optimised HPLC-PDA chromatographic conditions,twenty batches of Sparganii Rhizoma were analyzed by HPLC fingerprints.Principal component analysis(PCA),orthogonal projections to latent structures discriminant analysis(OPLS-DA)and hierarchical cluster analysis(HCA)were performed based on all peak areas of Sparganii Rhizoma fingerprints.Meanwhile,part of common peaks were subsequently quantified by UFLC-QTRAP-MS.Results: The similarity values of HPLC fingerprints fluctuated in a wide range of 0.511–0.973,which showed variable differences of chemical characters among Sparganii Rhizoma from twenty habitats.PCA,OPLS-DA and HCA indicated that samples could be divided into five groups with different chemical characters,which generally corresponded with their geographical distributions.A total of 31 peaks in HPLC fingerprints were marked,and eight of them were identified and quantified.The quantitative result was generally in agreement with the classifications based on HPLC fingerprints,which indicated that Sparganii Rhizoma samples from eastern China mostly contained more contents including phenolic acids and flavonoids.Conclusion: This study not only proved that there were relationships between geographic distributions and internal chemical compositions of plants,which could provide evidence to the traditional Chinese medicine concept "geo-authentic",but also supplied a sensitive and rapid simultaneous quantitive method for the quality estimation of Sparganii Rhizoma.     

Fingerprint analysis of Zhimu-Huangbai herb pair and simul- taneous determination of its alkaloids,xanthone glycosides and steroidal saponins by HPLC-DAD-ELSD

AIM： To develop and validate a high performance liquid chromatography （HPLC） coupled with diode array and evaporative light scattering detectors （DAD-ELSD） method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes （namely, xanthone glycosides, steroidal saponins, and alkaloids） in Zhimu-Huangbai herb pair （ZB）. METHOD： Chromatographic separation was performed on a Diamonsil C18 column （4.6 mm× 250 mm, 5 μm, Dikma） by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL.min-1 at 260 nm. The drift tube temperature of ELSD was set to 60 ℃ and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy. RESULTS： The HPLC-DAD-ELSD method allowed the quantification of ten compounds （phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-Ⅱ, timosaponin B, and timosaponin A-Ⅲ）, and was successfully applied to fingerprint analysis for ten batches of ZB samples. CONCLUSION： This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for as sessment of complex TCM formulas.     

黄芩不同部位中15种黄酮类成分的含量测定及化学计量学研究

Objective: The aerial parts of Scutellaria baicalensis were used as Huangqin Tea for thousands of years and mainly contain flavonoids which contribute to its bioactivities. However, there is no appropriate quality evaluation method of Huangqin Tea, and three flavanones of isocarthamidin-7-O-β-D-glucuronide,carthamidin-7-O-β-D-glucuronide, and isoscutellarein-8-O-β-D-glucuronide with high contents in the aerial parts have never been defined quantitatively. Here, an HPLC-DAD method for simultaneous determination of 15 flavonoids and systematically compared their contents and distribution in the roots,stems, leaves, and flowers of S. baicalensis was established.Methods: Under the HPLC-DAD chromatographic conditions, 77 batches of samples of S. baicalensis were analyzed. Meanwhile, the chromatographic fingerprint of different parts of S. baicalensis was established.Subsequently, principal component analysis(PCA), orthogonal projections to latent structures discriminant analysis(OPLS-DA), and clustering heat map were performed based on the contents of 15 flavonoids in different parts of S. baicalensis.Results: The results showed significant differences in the contents and distributions of 15 flavonoids among the different parts of S. baicalensis. The chemical composition of stems showed some similarities to leaves, and their contents were all lower than leaves. The contents of isocarthamidin-7-O-β–glucuronide [(106.66 66 ± 22.68) mg/g], carthamidin-7-O-β-D-glucuronide [(19.82 ± 11.17) mg/g],and isoscutellarein-8-O-β-D-glucuronide [(3.10 ± 1.73) mg/g] were the highest in leaves. The content of apigenin-7-O-β-D-glucopyranoside and chrysin-7-O-β-D-glucuronide were the highest in flowers. The contents of baicalin, baicalein, wogonoside, wogonin, alpinetin, and oroxylin A were higher in roots than in other parts.Conclusion: The method was fully validated and could be effectively used to characterize the contents and distributions of main flavonoids in the different parts of S. baicalensis. It may lay a foundation to establish the quality evaluation system for Huangqin Tea.     

Simultaneous Determination of Seven Components in Gamboge and Its Processed Products Using a Single Reference Standard

Objective To establish a quality control method for simultaneous determination of multiple components in gamboge. Methods A single reference standard for the determination of multiple components(SSDMC) with HPLC was proposed. Seven major components of gamboge including gambogenic acid(S), β-morellic acid(C1), 2R-30-hydroxygambogic acid(C2), isogambogenic acid(C3), gambogellic acid(C4), 2R-gambogic acid(C5), and 2S-gambogic acid(C6) were simultaneously analyzed using gambogenic acid as reference standard. The credibility and feasibility of SSDMC method were validated with respect to linearity, limits of detection and quantification, precision, stability, repeatability, accuracy, ruggedness, and robustness. The relative conversion factors(RCFs) of S and C1-6 were calculated. Twelve batches of gamboge including crude and processed products were successfully analyzed by applying the SSDMC and traditional external standard(ES) methods. Results The SSDMC method was credible and feasible. The RCFs of S and C1-6 were 1.000, 0.913, 0.864, 1.064, 0.777, 0.921, and 0.919, respectively. No significant difference was observed in the contents of the seven components between SSDMC and ES methods. The heat-processing technique caused a reduction in the seven components. Conclusion SSDMC is a simple, reliable, and effective method for the analysis of the complex multiple components in gamboge, and it is also a practical and economical approach.     

Comprehensive Quality Evaluation of ShuXueNing Injection Employing Quantitative High-Performance Liquid Chromatography Fingerprint and Chemometrics

Objective: In this study, a comprehensive and effective quality method for evaluating the efficacy of ShuXueNing injection (SXNI) was developed. Materials and Methods: Quantitative high?performance liquid chromatography fingerprint, the quantitative analysis of multicomponents by a single marker (QAMS) method, hierarchical cluster analysis(HCA)?, and orthogonal partial leastsquares discrimination analysis (OPLS?DA) were used to distinguish 53 batches of SXNI samples from 7 manufacturers. Results: A total of 53 batches of samples were analyzed to establish antithesis fingerprint of SXNI, and 12 peaks of the common model were collected and used for the similarity analysis. Meanwhile, six index flavonoid components were determined by the QAMS method, using rutin as internal reference substance. The accuracy of the QAMS method was confirmed by investigating the relative deviation between the QAMS method and the traditional external standard method. The results demonstrated that there was no significant difference (RE < 1%), suggesting that QAMS was a reliable and convenient method for the content determination of multiple components. The HCA and OPLS?DA methods drew a similar conclusion. The 53 batches of SXNI samples from 7 manufacturers were categorized into five groups, indicating that chemometrics could reveal the quality differences of SXNI between the manufacturers. Conclusions: The method established herein was efficient and successful in assessing the quality of SXNI, and that it may be potentially employed in the quality control of related products composed of Ginkgo biloba extract.     

A Simple High-Performance Liquid Chromatography Method for the Assay of Flavonoids in Ginkgo biloba Leaves

Objective: Ginkgo biloba leaves, as an herbal medicine or dietary supplement, have been widely used worldwide. In this study, an integrated analytical method was established for the comprehensive analysis of flavonoids in G. biloba leaves. Materials and Methods: A practical chromatographic method combining high-performance liquid chromatography fingerprint analysis and quantitation was used to simultaneously determine 11 flavonoids(6 flavonol glycosides and 5 biflavones) in G. biloba leaves from different regions. Results: A total of 11 characteristic peaks were identified accurately, and the similarity of fingerprints ranged from 0.944 to 0.996. Methodology validation revealed appropriate linearity(R~2 ≥0.9997), precision, repeatability, stability, and recovery. The total contents of the six flavonol glycosides and five biflavones were within the range of 2.142-8.378 mg/g and 3.759-5.675 mg/g in 19 batches of samples, respectively. Among them, two coumaroyl flavonol glycosides were the predominant components. Conclusions: In this study, a convenient and reliable approach was successfully employed for the comprehensive evaluation of flavonoids in G. biloba leaves, which also provided a reference for its quality standard.     

Quality Evaluation of Astragali Radix based on DPPH Radical Scavenging Activity and Chemical Analysis

Objective To assess the quality of Astragali Radix from different areas based on the biological evaluation and chemical analysis. Methods The bioassay method of 1,1-diphenyl-2-picryl-hydrazyl(DPPH) radical scavenging activity for Astragali Radix was established. The parameters of DPPH assay including sample extraction time, reaction time, repeatability, and stability were detected. Furthermore, a method of HPLC-MS was developed to simultaneously determine calycosin-7-O-glucoside, ononin, formononetin, and astragaloside IV in Astragali Radix samples. And the total flavonoids and total saponins were detected by spectrophotometry. The relationship between DPPH evaluation and chemical analysis was studied by Pearson correlation analysis. Results Twelvebatches of Astragali Radix from different origins showed a wide range of DPPH radical scavenging activities(IC50 = 1.395-9.894 μg/mL). Based on DPPH assay, Sample 10 derived from Inner Mongolia Autonomous Region(IC50 = 1.395 μg/mL) showed the best quality of all samples. Chemical analysis showed that different compounds selected as indices would cause different results for quality evaluation. Pearson correlation analysis revealed that the contents of total flavonoids(P = 0.032), calycosin-7-glucoside(P =0.035), and astragaloside IV(P = 0.010) were positively correlated with DPPH radical scavenging activity. Conclusion Except for chemical analysis, DPPH radical scavenging activity can be used as a good alternative to assess and control the quality of Astragali Radix.     

Simultaneous Determination of Multiple Bioactive Constituents in Total Alkaloid of Sophora alopecuroides by HPLC-DAD

Objective To develop a qualitative and quantitative simultaneous determination of multiple bioactive constituents in total alkaloid in Sophora alopecuroides(TASA).Methods In the experiment,a new and simple HPLC-DAD method for the simultaneous determination of multiple constituents in TASA was developed.The separation was performed on a Kromasil C18 column(250 mm×4.6 mm,5.0μm)eluted with 0.02 mol/L potassium dihydrogen phosphate(adjusted pH 4.3 using 1%glacial acetic acid)and acetonitrile(75:25)at a flow-rate of 0.7 mL/min.The detection wavelength was set at 210 nm.Results Five constituents(sophoridine,matrine,oxymatrine,aloperine,and lehmannine)were simultaneously analyzed in this study.Four of them were identified and determinated by the developed method.The calibration curves exhibited linear regressions(r 2 >0.9995).The injection precision,the intra-day precision,and the analysis repeatability were validated with the RSD values less than 5.0%.The mean recoveries of the four constituents were ranged from 98.62%to 100.20%,and the RSD values were all less than 3.37%.Conclusion This method is convenient,fast,accurate,and is applicable to analyze the multi-constituents in TASA.     

Rapid Analysis of Polar Components in Ophiocordyceps sinensis by Conventional Liquid Chromatography System

Objective To develop a rapid high-performance liquid chromatography coupled with diode array detection(HPLC-DAD) method for the simultaneous determination of sixpolar compounds in Ophiocordyceps sinensis. Methods A poroshell SB Aq column(50mm × 4.6 mm, 2.7 μm) and gradient elution were used; The detection wavelength of compounds was set at 260 nm. The chromatographic peaks of the six investigated compounds in sample were identified by comparing their retention times with reference compounds. Results All calibration curves showed good linearity(r 0.999) within the tested ranges. The intra- and inter-day precisions of the six analytes were less than 0.8% and 2.1%, respectively, and the recoveries of the six analytes were between 95% and 103%. The validated method was successfully applied to the determination of sixpolar compounds in O. sinensis samples. Conclusion The poroshell SB Aq column is suitable for the rapid analysis of polar components in Chinese materia medica on conventional HPLC system and the developed HPLC method is also helpful to the quality control of O. sinensis.     

Simultaneous Determination of Four Major Steroidal Saponins in Seven Species of Dioscorea L. by HPLC-ELSD

Objective To control the quality of the species in Dioscorea L.better.Methods An HPLC-ELSD method was developed for the first time to simultaneously determine four bioactive ingredients:dioscin,gracillin,protoneodioscin,and protoneogracillin in 31 samples belonging to seven species of Dioscorea L.from different areas.The column was an Inertsil HILIC(250 mm×4.6 mm,5μm).The separation was carried out with a gradient program.The mobile phase was acetonitrile-water at a flow rate of 0.8 mL/min.Results The standard curve was rectilinear in the range of 0.464-12.97μg(r=0.9969)for dioscin,0.310-7.09μg(r=0.9953)for gracillin,0.469- 11.66μg(r=0.9970)for protoneodioscin,and 0.276-6.87μg(r=0.9992)for protoneogracillin.The recoveries of the markers were 98.1%,100.1%,97.2%,and 96.4%,respectively.The contents of the four components were quite different among the seven species of Dioscorea L.Conclusion The proposed HPLC-ELSD method is convenient,fast,accurate,and applicable for simultaneous analysis of multiple bioactive components of species in Dioscorea L.for quality control,which could facilitate discovering new natural resources of steroidal saponin.     

Quantitative Analysis of Eight Ginsenosides in Red Ginseng Using Ginsenoside Rg1 as Single Reference Standard

Objective: To develop a reversed?phase high?performance liquid chromatography method for the quantification of major ginsenosides in red ginseng (RG, the steamed and dried root of the cultivar of Panax ginseng C. A. Mey). Methods: A feasible method was developed in strict accordance with chromatographic properties of eight ginsenosides. Their contents could be unveiled with conventional external standard method, or as an alternative, using ginsenoside Rg1 as the single reference standard by means of seven conversion factors. Those parameters had been validated on different chromatographic columns and instruments. Results: Twenty-one batches of RG samples were determined. In addition, the chromatograms of RG and confusing species, including Panax ginseng, Panax quinquefolium, and Panax notoginseng, were apparently different. Conclusions: The method was proved to be efficient for the quality control of RG.     

Effects of drying process of Yuanzhi （Radix Palygalae） on its bioactire ingredients

OBJECTIVE： To study the effects of the drying processing in terms operational parameters on the bio- active constituents of six YuanzhJ （Radix Palygalae） samples across China. METHODS： Six Yuanzhi （Radix Palygalae） samples were investigated using thermogravimetry analysis. The heating courses were set in two ways： the temperature-programmed process from room temperature to 150℃ ,and the constant-temperature course at 50℃, 70℃ and 90℃. RESULTS： The peak temperature of six Yuanzhi （Radix Palygalae） samples ranged from 78℃ to 88℃. The mass loss rate of Yuanzhi （Radix Palygalae） alcohol-soluble extract was significantly increased when heated at 90℃. Four types of bioactive ingredients were detected in volatile oils of Yuanzhi （Radix Palygalae） sample from Shanxi province by Gas Chromatography-mass spectrometry analysis. Fourier Transform Infrared Spectroscopy results showed that the drying temperature exerted a great influence on types and amount of ingredients of Yuanzhi （Radix Palygalae）. The kinetic study showed that the constant-temperature drying process of Yuanzhi Radix Palygalae） samples could be well de- scribed by the Page Model, especially for the drying process at 50~C, in which R2 and SD values were more than 0.98 and less than 0.04, respectively.The drying constant k of three Yuanzhi （Radix Palygalae） samples from Shanxi, Gansu and Shaanxi provinces in China was corresponding to the Arrhenius equation, and their activation energies were 28.07, 2.5.38 and 21.48 kJ/mol, respectively. CONCLUSION： The drying process of Yuanzhi （Radix Palygalae） was very important for bioactive ingredients improvement in Yuanzhi （Radix Palygalae）. Temperature was a thermodynamic property significantly affecting the process.     

A Rapid and Effective HPLC Method for Identification of Bovis Calculus in Chinese Patent Medicines

Objective Due to the limited resource and the large demand,many kinds of Bovis Calculus(BC)including artificial Bovis Calculus(ABC),in vivo cultured Bovis Calculus(in vivo CBC),and in vitro cultured Bovis Calculus(in vitro CBC)were used in Chinese patent medicines(CPMs).Previous studies have shown that the chemical constituents of ABC and their properties were different from other BC.The two types of CBC with much higher price than ABC were approximately equivalent with natural Bovis Calculus in quality and clinical effect.The aim of the study is to establish a rapid and effective method for the identification of BC in CPMs.Methods An HPLC method with the higher specificity for analyzing bilirubin was established to distinguish ABC from other three kinds of BC by comparing the change of bilirubin content with the addition of EDTA-2Na as the extraction solvent and stabilizer.Results The bilirubin content in CPMs containing ABC was basically unchanged,while that in CPMs containing other kinds of BC showed significant difference.The proposed method was employed to analyze a variety of CPMs containing Bovis Calculus(CPMBCs)and proven to be universal.Conclusion An effective analytical method is established for the quality control of CPMBCs and further ensures the safety and efficacy of these drugs in clinical practice.     

Development and validation of a HPLC-UV-ESI-MS method for the simultaneous quantitation of ten bioactive compounds in Dahuang Fuzi Tang

AIM： To develop a high performance liquid chromatography （HPLC） coupled with electrospray ionization mass spectrometry （ESI-MS） and ultraviolet （UV） detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang （DFT）. METHOD： The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer （containing 0.15% formic acid, V/V） at 25 ℃ with a flow rate of 1.0 mL.min^-1 and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV.
RESULTS： All calibration curves showed good linear regression （r^2 ≥ 0.9990）. The limits of detection and limits of quantification were 0.021-0.155 μg.mL ^-1 and 0.076-0.520 μg.mL ^-1, respectively. Overall precision RSD （intra-day and inter-day） were less than 2.96%, and the average recoveries were 98.35% 101.45%, with RSD ranging from 1.54% to 3.01% for the analytes. CONCLUSION： The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.