Optimization of high pressure machine decocting process for Dachengqi Tang using HPLC fingerprints combined with the Box–Behnken experimental design

Using Dachengqi Tang（DCQT） as a model, high performance liquid chromatography（HPLC）fingerprints were applied to optimize machine extracting process with the Box–Behnken experimental design. HPLC fingerprints were carried out to investigate the chemical ingredients of DCQT; synthetic weighing method based on analytic hierarchy process（AHP） and criteria importance through intercriteria correlation（CRITIC） was performed to calculate synthetic scores of fingerprints; using the mark ingredients contents and synthetic scores as indicators, the Box–Behnken design was carried out to optimize the process parameters of machine decocting process under high pressure for DCQT. Results of optimal process showed that the herb materials were soaked for 45 min and extracted with 9 folds volume of water in the decocting machine under the temperature of 140 1C till the pressure arrived at 0.25 MPa;then hot decoction was excreted to soak Dahuang and Mangxiao for 5 min. Finally, obtained solutions were mixed, filtrated and packed. It concluded that HPLC fingerprints combined with the Box–Behnken experimental design could be used to optimize extracting process of traditional Chinese medicine（TCM）.

HPLC fingerprint of Nauclea officinalis stems in different harvesting period

Objective:To establish HPLC fingerprint of Nauclea officinalis stems in different harvesting periods,and analyze the effect of harvest time on the quality of the medicinal materials by combining with chemical pattern recognition.Methods:The analysis was performed on Sun Fire-C18(150 mm×4.6 mm,5μm)column.The mobile phase consisted of acetonitrile-0.1%phosphoric acid solution(gradient elution)at a flow rate of 1.0 mL/min.Results:The HPLC fingerprint of Nauclea officinalis stems was established and 12 common peaks were determined,and 3 chromatographic peaks were identified by comparison with the mixed references.There were some differences in the quality of Nauclea officinalis in different harvesting periods.The OPLS-DA analysis successfully predicted four main markers of quality difference.Conclusion:The established HPLC fingerprint could reflect the composition characteristics of Nauclea officinalis stems in different harvesting period,and the main markers that influence the composition difference of the stems could be used as key indicators for the quality control.

Authentication of Cassia seeds on the basis of two-wavelength HPLC fingerprinting with the use of chemometrics

High performance liquid chromatographic(HPLC) fingerprints of Cassia seed,a traditional Chinese medicine(TCM),were developed by means of the chromatograms at two wavelengths of 238 and 282 nm.Then,the two data sets were combined into one matrix.The application of principal component analysis(PCA) for this data matrix showed that the samples were clustered into four groups in accordance with the plant sources and preparation procedures.Furthermore,partial least squares(PLS),back propagation artificial neural...

HPLC Fingerprint with Multi-components Analysis for Quality Consistency Evaluation of Traditional Chinese Medicine Si-Mo-Tang Oral Liquid Preparation

Si-Mo-Tang（SMT） oral liquid preparation, a traditional Chinese medicine, was prepared from four crude herbal drugs, Fructus Aurantii Submaturus, Radix Aucklandiae, Semen Arecae and Radix Linderae Aggregatae. A combinative method using HPLC fingerprint and quantitative analysis was developed and validated for quality consistency evaluation of SMT. Individual HPLC chromatograms were evaluated against the mean chromatogram generated via a similarity evaluation computer program. Data from chromatographic fingerprints were also processed with principal component analysis（PCA） and hierarchical cluster analysis（HCA）. Additionally, six components （naringin, isonaringin, hesperidin, neohesperidin, norisoboldine and potassium sorbate） in SMT were simultaneously determined to interpret the quality consistency. For fingerprint analysis, 20 peaks were selected as the characteristic peaks to evaluate the similarities of 26 SMT collected from different manufacturers. Among the 20 characteristic peaks, 10 peaks were assigned to be naringin, hesperidin, neohesperidin, isonaringin, neoeriocitrin, tangeretin, nobiletin, norisoboldine, 5-（ethoxymethyl）furan-2-carbaldehyde and potassium sorbate, respectively. The results of similarity analysis, PCA and HCA, indicate that the samples from different manufacturers were consistent with each other in composition. The results from the quantitative data show that the contents of six compounds were significantly different in SMT oral liquid preparations from different manufacturers. The combinative method of chromatographic fingerprint with quantitative analysis developed here offered an efficient way for the quality consistency evaluation of the traditional Chinese medicine SMT.

HPLC/MS Fingerprint Analysis of Tangshenosides

Radix codonopsis(党参)，which is derived fromCodonopsis pilosula, C.pilosula subsp. Modesta andC. Tangshen(ChP,2000),has been used as a remedy fora decrease of appetite, psychoneurosis, fatigue, dyspepsiaetc.or as a substitute for Panax ginseng in traditionalChinese medicine. Relating to the constituents of theseherbs, triterpenes, phytosterols, furaldehyde, sesquitesand some phenolic glycosides have been reported, butnone of these compounds seems to be responsible for thebiological activities of this drug. As regards the quality evaluation of Radix Codonopsis, a HPLC analysis of atractylenolide Ⅲ has been described, but this sesquiterpene lactone exists in a very low content and is difficultfor conventional analysis. The determination of polysaccharide also met limitation because of its non-specialization. In this paper, we report the HPLC/MS fingerprintsanalysis of phelolic glycosides from Radix Codonopsis including three species.

An HPLC Fingerprint Identification of Dried Barks of Ilex rotunda and Ilex godajam

[Objectives] A simple and reliable HPLC fingerprint method was developed for the identification of dried barks of Ilex rotunda and I. godajam. [Methods] Nine batches of dried barks of I. rotunda,and seven batches of dried barks of I. godajam collected from different pharmacies and arboretums in different regions of China were used to establish fingerprints. The software Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine( 2004 A Edition) was used to evaluate the fingerprints. [Results]The fingerprints of dried barks of I. rotunda and I. godajam were established. Methodological study met the technical requirements of fingerprints. The similarities of the fingerprints of dried barks of I. rotunda and I. godajam were all more than 0. 8 and 0. 9 respectively. There were 31 and 28 common peaks in I. rotunda and I. godajam,which could be classified into two clusters by principal component analysis( PCA) and hierarchical cluster analysis. [Conclusions] The feasibility and advantages of used HPLC fingerprints were verified,and the results indicated that the HPLC fingerprint as a characteristic distinguishing method combining similarity evaluation,principal component analysis and hierarchical cluster analysis can be successfully used to identify the authenticity of dried barks of I. rotunda and I. godajam.

The fingerprint and flavonoids contents by HPLC and the UPLC-DAD-Q-TOF-MS analysis of Sedum aizoon L.

Fingerprint analysis of Sedum aizoon L.was carried by High-performance liquid chromatography.There were 27 common peaks in the fingerprint of 10 samples.Eight of these common peaks,peaks 1(arbutin),4(gallic acid),10(myricetin-3-O-β-D-glucopyranoside),14(myricetin-3-O-α-L-rhamnopyranoside),19(quercetin-3-O-α-L-rhamnopyranoside),22(kaempferol-3-O-α-L-rhamnopyranoside),24(quercetin)and 26(kaempferol)were identified by reference substances.Moreover,six flavonoid compounds were simultaneously determined by the above method.By comparing with the constituents in the aerial parts,roots and flowers,it’s easy found that the aerial parts of S.aizoon had abundant flavonoids and the flavonoid aglycones mostly exist in the roots,while the flavonoid glycosides largely existing in the flowers.In addition,an UPLC-DAD-Q-TOF-MS method was developed for qualitative analysis of the aerial parts of S.aizoon.This study may provide a preliminary reference for the quality control of S.aizoon.

Chemical Characteristics Combined with Bioactivity for Comprehensive Evaluation of Tumuxiang Based on HPLC-DAD and Multivariate Statistical Methods

Background: The dried roots of Inula helenium L.(IH) and Inula racemosa Hook f.(IR) are used commonly as folk medicine under the name of "tumuxiang(TMX)". Phenolic acid compounds and their derivatives, as main active constituents in IH and IR, exhibit prominent anti-inflammation effect.Objective: To develop a holistic method based on chemical characteristic and anti-inflammation effect for systematically evaluating the quality of twenty-seven TMX samples(including 18 IH samples and 9 IR samples) from different origins.Methods: HPLC fingerprints data of AL(Aucklandia lappa Decne.) whose dried root was similar with HR was added for classification analysis. The HPLC fingerprints of twenty-seven TMX samples and four AL samples were evaluated using hierarchical clustering analysis(HCA) and principle component analysis(PCA). The spectrum-efficacy model between HPLC fingerprints and anti-inflammatory activities was investigated by principal component regression(PCR) and partial least squares(PLS).Results: All samples were successfully divided into three main clusters and peaks 7, 9, 11, 22, 24 and 26 had a primary contribution to classify these medicinal herbs. The results were in accord with the appraisal results of herbs. The spectrum-efficacy relationship results indicated that citric acid, quinic acid, caffeic acid-β-D-glucopyranoside, chlorogenic acid, caffeic acid, 1,3-O-dicaffeoyl quinic acid, tianshic acid and 3β-Hydroxypterondontic acid had main contribution to anti-inflammatory activities.Conclusion: This comprehensive strategy was successfully used for identification of IH, IR and AL, which provided a reliable and adequate theoretical basis for the bioactivity relevant quality standards and studying the material basis of anti-inflammatory effect of TMX.

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 VIPvalue>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.

Relationship between DNA Barcoding and Chemical Classification of Salvia Medicinal Herbs

Objective To make the identification of medicinal herbs in Salvia L.quickly and accurately.Methods In this work,DNA barcoding and chemical fingerprint were compared for the identification of herbs in Salvia L.First,the nucleotide sequences of the internal transcribed spacer region two amplified from 48 medicinal plants in Salvia L.,and three other groups of medicinal plants in Lamiaceae were sequenced.A molecular phylogeny was constructed using the minimum evolution and maximum parsimony methods according to their sequence diversity.Second,the water-solution bioactive components and lipid soluble components were tested by HPLC.Then a chemical phylogeny was built using HPLC fingerprint data.Comparing the molecular and chemical phylogenetic trees revealed many similarities.Results DNA barcoding was sequencing based and could therefore provide more accurate results within a shorter time especially in large-scale studies.Conclusion The results show that ITS2 region is a novel DNA barcode for the authentication of the species in Salvia L.This is the first work to show the relationship between DNA barcoding and chemical components.

Comparison of Murraya exotica and Murraya paniculata by fingerprint analysis coupled with chemometrics and network pharmacology methods

There are two source plants for the traditional Chinese medicine Murrayae Folium et Cacumen(MFC)in Chinese Pharmacopoeia,i.e.Murraya exotica L.and M.paniculata(L.)Jack.Herein,a chemical comparison of M.exotica and M.paniculata by high performance liquid chromatography(HPLC)fingerprint analysis coupled with chemometrics and network pharmacology was performed.The main peaks in the fingerprints were identified by liquid chromatography coupled with ion trap/time-of-flight mass spectrometry(LC-IT-TOF-MS)and authenticated by references.The chemometrics results showed that the HPLC fingerprints of these two species were clearly divided into two categories using hierarchical cluster analysis(HCA)and principal component analysis(PCA),and a total of 13 significantly differentiated markers were screened out by orthogonal partial least squares-discriminant analysis(OPLS-DA).However,the following network pharmacology analysis showed that these discriminated markers were found to act via many common targets and metabolic pathways,indicating the possibly similar pharmacological effects and mechanisms for M.exotica and M.paniculata.The above results provide valuable evidence for the equivalent use of these two plants in clinical settings.Moreover,the chromatographic fingerprint analysis coupled with chemometrics and network pharmacology supplies an efficient approach for the comparative analysis of multi-source TCMs like MFC.

Effects of Postharvest Processing and Geographical Source on Phytochemical Variation of Corydalis Rhizoma

Objective To assess the relative contributions of postharvest processing and geographical source to phytochemical variation of Corydalis Rhizoma, and rhizome of Corydalis yanhusuo, and to examine what phytochemical components are the most sensitive to the differences of each factor and how they change. Methods HPLC fingerprinting and LC-MS coupled with chemometric approaches were applied. Results The results of principal component analysis(PCA) and hierarchical cluster analysis(HCA) explicitly demonstrated the postharvest processing could produce a greater impact on the phytochemical profiles of Corydalis Rhizoma than geographical source. The contents of most compounds increased after water boiling while decreased after sulphur-fumigation.Protopine, coptisine, and palmatine were the most variable components in processing. Geographical sources also led to a remarkable phytochemical differentiation, in which the environmental variation of the three regions might play a role. Dehydrocorybulbine, coptisine, dehydrocorydaline, and protopine varied most among the three production regions and decreased sequentially in Zhejiang, Shaanxi, and Jiangsu provinces, China. Conclusion Both postharvest processing and geographical source should be enhanced with the priority for the former in the quality control of Corydalis Rhizoma. The application of boiling is supported but the consistency should be improved in practice. Sulphur-fumigation is strongly suggested to be abandoned.

Saponin Accumulation in Flower Buds of Panax notoginseng

Objective Panax notoginseng is an important Chinese medicinal plant. Saponin accumulation is higher in the flower buds than in the other parts of P. notoginseng. However, the flower bud compositions have not yet been quantified. The aim of this study is to investigate the formation and accumulation of saponins in the flower buds of P. notoginseng from different populations and at different growth years. Methods Fourteen types of P. notoginseng with different growing durations and from different areas of Wenshan County, Yunnan Province were collected. We separated P. notoginseng individually into the flower buds, stems, leaves, and roots at the places where it has the highest saponin content. An efficient high-performance liquid chromatography（HPLC） method was developed for simultaneously quantifying two active saponins, ginsenoside Rb1 and ginsenoside Rb3, in the flower buds of P. notoginseng. The total saponin content was determined by using a quantitative vanillin-sulfuric acid colorimetric method. HPLC method was used to establish the fingerprints of 13 saponins and then quantify the composition in the whole plant of P. notoginseng. Results The saponin contents of different parts differ significantly, and the total saponin content and those of Rb1 and Rb3 do not entirely correlated. The flower buds of P. notoginseng contain 27.79% of total saponins, which is the highest saponin content in the whole plant. Fingerprint result showed that different saponins were appeared in different parts of the plant, i.e. flower buds, stems, leaves, and roots.Conclusion The saponin contents from the flower buds of P. notoginseng vary depending on the growth area and duration. The fingerprints show that the saponin contents and compositions vary depending on the part of P. notoginseng. These results are useful for the pharmacological evaluation and quality control of P. notoginseng.

Standardization and Identification of Minor Components of Silymarin (MK-001)

Objective To develop a highly effective HPLC method for standardization of silymarin and for characterization of minor components, which has not been reported previously. Methods An HPLC fingerprinting method and 1 H-NMR spectroscopy were employed to provide assurance for a standardized silymarin product (MK-001). Results A total of 18 marker compounds were identified and characterized including ten flavonolignans and eight small molecules including adenine, adenosine, uridine, and 3,5,7-trihydroxychromone which were first isolated and characterized from silymarin. The HPLC fingerprinting method and 1 H-NMR had been established for complete assignments of HPLC peaks with intensity over 1.0%. Conclusion The successfully established HPLC fingerprinting method and 1 H-NMR could be applied for the standardization of commercial silymarin products. MK-001 represents the first standardized silymarin with the highest content of flavonolignans ( > 90%).

A mathematical model for quality evaluation of total saponins of Panax japonicas based on hypolipidemic activity

Objective: The chemical finger printing-based methods for evaluating TCMs quality can report partial of TCMs quality without linking to effective constituents. In this study, a mathematical model was established for the quality evaluation of total saponins of Panax japonicus(TSPJ), a folk medicine in China and Japan for treating diseases, through coupling the dynamic changes of chemical constitutions with corresponding activities.Methods: High-performance liquid chromatography(HPLC) fingerprints were applied to establish the chromatographic database of TSPJ. The associated hypolipidemic activity database was determined by TG assay using Hep G2 cell model. Correlation analyses of two databases were performed by partial least squares(PLS) for calculating regression coefficients, and the interval value of YZL value(the ratio of positive and negative peak-to-peak area coefficient) closely related to hypolipidemic activity was refined by the formula of Norminv function to value the quality of TSPJ.Results: In this study, the chromatographic data of 16 common peaks were obtained from 20 batches of TSPJ. After the estimate by this mathematical evaluation model, seven peaks were positively correlated with hypolipidemic activity, and nine peaks were negatively correlated with hypolipidemic activity.When the YZL value was less than 0.7861, the quality of sample was inferior, while YZL value was more than 6.6992, and the quality of samples was superior. The quality of another ten batches of TSPJ was further assessed to verify this method.Conclusion: These results indicated that the established model could be usefully applied to evaluate the quality of TSPJ in the hypolipidemic activity.

High-Performance Liquid Chromatography Fingerprint of Marine Traditional Chinese Medicine Haliotidis

Objective: Haliotidis, the dried muscle of abalone, has long been used as a marine traditional Chinese medicine(TCM) due to its high medicinal and nutritive values, whereas few reports contributed to its quality control. This study aimed to establish the high-performance liquid chromatography(HPLC) fingerprints of Haliotidis. Materials and Methods: The HPLC fingerprints of Haliotidis, the manufacturing products of dried muscle of abalone, were analyzed by reverse-phase HPLC, using a YMC HPLC C18 column(250 mm × 4.6 mm, 5 μm).The gradient elution mode was applied with mobile phase starting from 5% acetonitrile and 0.1% phosphoric acid aqueous solution to 100%acetonitrile in 70 min at a flow rate of 1.0 mL/min. The detection wavelength was set at 190 nm. Results: Through compared the HPLC fingerprint chromatograms of 14 batches of Haliotidis, 25 peaks were assigned as the characteristic common peaks, of which one index component L-phenylalanine was identified by comparison with its standard substance. The established HPLC fingerprint analysis method of Haliotidis was validated with good precision, repeatability, and stability. Based on the above study, a reference HPLC fingerprint chromatogram of Haliotidis was proposed. Conclusions: It is the first time to establish the HPLC fingerprint chromatogram of TCM Haliotidis, which could contribute to the establishment of its quality control system.

Reflecting the Steps of Modernization of Chinese Materia Medica from the Papers in Chinese Traditional and Herbal Drugs in 2009

The papers in the journal of Chinese Traditional and Herbal Drugs in Vol. 40, 2009 are briefly reviewed in the categories of chemical constituents, preparations and technologies, analysis and quality control, pharmacological and clinical studies, reviews, and finally healthy principles. Some comments, especially for hot topics have been personally provided.

New Progress in Modern Research on Chinese Herbal Medicines Published in Chinese Traditional and Herbal Drugs in 2010:Annual Highlights and Comments

As a series of and continuous publication,the papers published on Chinese Traditional and Herbal Drugs in 2010 were selectively and briefly highlighted to reflect the new progress on modern research of Chinese herbal medicines.Within 617 articles,chemical constituents(127),pharmaceutics and technology(149),pharmacological studies and clinical observation,and medicinal materials are still major categories.Some comments have also been personally provided.