High-performance Liquid Chromatography Fingerprint Research Articles

Spectrum-effect relationship between HPLC fingerprint and hypoglycemic of litchi leaves (Litchi chinensis Sonn) in vitro.

Litchi chinensis Sonn (Litchi) has been listed in the Chinese Pharmacopeia, and is an economically and medicinally valuable species within the family Sapindaceae. However, the material basis of its pharmacological action and the pharmacodynamic substances associated with its hypoglycemic effect are still unclear. The predominant objective of this study was to establish the fingerprint profile of litchi leaves and to evaluate the relationship between the components of the high-performance liquid chromatography (HPLC) fingerprint of litchi leaves, assess its hypoglycemic effect by measuring α-glucosidase and α-amylase inhibition, and find the spectrum-effect relationship of litchi leaves by bivariate correlation analysis, Grey relational analysis and partial least squares regression analysis. In this study, the fingerprint of litchi leaves was established by HPLC, and a total of 15 common peaks were identified that clearly calibrated eight components, with P1 being gallic acid, P2 being protocatechuic acid, P3 being catechin, P6 being epicatechin, P12 being rutin, P13 being astragalin, P14 being quercetin and P15 being kaempferol. The similarities between the fingerprints of 11 batches of litchi leaves were 0.766-0.979. Simultaneously, the results of the spectrum-effect relationship showed that the chemical constituents represented by peaks P8, P3, P12, P14, P2, P13, and P11 were relevant to the hypoglycemic effect.

Chemical constituent characterization and determination of Quisqualis fructus based on UPLC-Q-TOF-MS and HPLC combined with fingerprint and chemometric analysis.

Quisqualis fructus (QF) is a traditional Chinese medicine (TCM) that it has a long history in the therapeutic field of killing parasites, eliminating accumulation, and stopping diarrhea. However, the therapeutic material basis of QF is remaining ambiguous nowadays. The geographical origin differences of QF are also usually ignored in the process of medication. In this study, the alcohol-aqueous soluble constituents in QF from different origins were systematically characterized and accurately measured by ultra-high performance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and high-performance liquid chromatography (HPLC) respectively. Chemometric analysis was performed for origin differentiation and screening of potential quality marker (Q-marker). Finally, A total of 106 constituents were tentatively characterized in positive and negative ion modes, including 29 fatty acids, 26 organic acids, 11 amino acids and derivatives, 10 glycosides, 9 alkaloids and derivatives, and 21 other constituents. QF from different origins were effectively distinguished and 16 constituents were selected as the potential Q-markers subsequently. Four representative components (trigonelline, adenosine, ellagic acid, and 3,3'-di-O-methylellagic acid) in QF samples were simultaneously determined. HPLC fingerprint analysis indicated that the similarity between 16 batches of QF was in the range of 0.870-0.999. The above results provide some insights for the research on the pharmacodynamic constituents, quality control, and geographical discrimination of QF.

Notoginseng leaf triterpenes promotes angiogenesis by activating the Nrf2 pathway and AMPK/SIRT1-mediated PGC-1/ERα axis in ischemic stroke

Notoginseng leaf triterpenes (PNGL), derived from the dried stems and leaves of P. notoginseng, is a phytoestrogen that exerts many neuroprotective effects in vivo and in vitro of ischemic stroke. However, its impact on neurological restoration specifically in relation to angiogenesis following ischemic stroke remains unexplored. The aim of this study was to assess the effects of PNGL on angiogenesis subsequent to ischemic stroke. Male Sprague-Dawley rats were utilized in this study and were subjected to middle cerebral artery occlusion/reperfusion (MCAO/R). Post-ischemia, PNGL were administered through intraperitoneal (i.p.) injection. The high-performance liquid chromatography (HPLC) fingerprinting, triphenyltetrazolium chloride (TTC) staining, immunofluorescent staining, network pharmacology and western blot analyses were assessed to determine the therapeutical effect and molecular mechanisms of PNGL on cerebral ischemia/reperfusion injury. Our findings demonstrate that PNGL effectively reduced infarct volume, enhanced cerebral blood flow, and induced angiogenesis in rats subjected to MCAO/R. Notably, PNGL also facilitated neuronal proliferation and migration in HUMECs in vitro. The proangiogenic effects of PNGL were found to be linked to the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway and the AMPK/SIRT1-mediated PGC-1/ERα axis, as well as the activation of neurological function. Our study provides evidence that PNGL hold promise as an active ingredient of inducing proangiogenic effects, potentially through the activation of the Nrf2 pathway and the AMPK/SIRT1-mediated PGC-1/ERα axis. These findings contribute to the understanding of novel mechanisms involved in the restoration of neurological function following PNGL treatment for ischemic stroke.

Study on the material basis of Zhujing pill in treating fundus lesions through component analysis and network pharmacology.

Zhujing pill (ZP) is a famous Chinese herbal formula that has been widely used to treat diabetic retinopathy, macular degeneration, retinitis pigmentosa and other fundus lesions. In this study, the material basis and mechanism of ZP in the treatment of fundus lesions were evaluated via the high-performance liquid chromatography fingerprint, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry, network pharmacology and molecular docking. A total of 32 common components were found and 31 components were identified in 15 batches of ZP samples. Moreover, 134 common key targets and 17 putative active components that are connected to fundus lesions were identified. Molecular docking revealed that quercetin, kaempferol, isorhamnetin, 5-O-feruloylquinic acid, plantagoside and 2'-acetylacteoside have the ability to interact with the core targets such as AKT1, TP53, TNF, IL-6 and Jun. Our findings revealed that the therapeutic effects of ZP on fundus lesions are mediated by multiple components, targets and pathways, including at least six active ingredients and 11 targets. The study provides new ideas for further research on the material basis and mechanisms of traditional Chinese medicine prescriptions.

Experimental design approach for the quantitative analysis of multicomponents by single marker and HPLC fingerprinting of Thunbergia laurifolia aqueous extract.

Thunbergia laurifolia is used in traditional Thai medicine to reduce fever and treat mouth ulcers. However, the quantitative analysis of chemical markers has not yet been officially defined. The objective of this study is to develop a high-performance liquid chromatography (HPLC) method using a design of experiment (DoE) for the quantitative analysis of multicomponents by single marker (QAMS) and fingerprinting of the T. laurifolia aqueous extract. Critical variables were screened using a two-level fractional factorial design, followed by the optimization of the selected variables using a central composite design. The validated method was applied for quality assessment based on QAMS and fingerprinting of the extract. Optimum conditions of DoE for the analysis of caffeic acid, vicenin-2, and rosmarinic acid were determined. The relative correction factors for caffeic acid and vicenin-2 were calculated using rosmarinic acid as an internal reference standard, and their contents in 30 samples were determined. The differences between the external standard method (ESM) and QAMS were compared. No significant difference was observed in the quantitative determination, proving the consistency QAMS and ESM. HPLC fingerprints of T. laurifolia were established with 8 of 12 characteristic peaks that were structurally characterized using HPLC-diode array detection-electrospray ionization/tandem mass spectrometry. The similarity of the fingerprints in all samples was ≥0.74, and the pattern recognition of the characteristic peaks was satisfied. The proposed method efficiently detected multiple components of the T. laurifolia extract. Thus, the method is beneficial in providing references for enhancing the quality control of other herbal medicines.

The Enhanced Anti-anxiety Potentiality and Relevant Material Basis for the Extracts of Bile Processed Coptidis Rhizoma

Objective Bile-processed Coptidis Rhizoma (CR) exhibits stronger cold properties and central effect. However, the commonly used bile includes pig/cattle/sheep bile. It is currently unclear which kind of bile performs the best synergistic effect, and the enhanced efficacy is mainly contributed by what components. In this work, a comparative study on pig/cattle/sheep bile processed CR (pCR/cCR/sCR) was carried out. Materials and Methods Firstly, the anxiety model of sthenic heat type was established. Then, pharmacological research involving general state observation, biochemical examination (succinate dehydrogenase, glucokinase, corticosterone, adrenocor ticotropic hormore), pathological evaluation (liver and hippocampus tissue), and behavioral tests (elevated plus maze, open field test) was implemented. Finally, chemical assessment involving high-performance liquid chromatography (HPLC) fingerprints of pCR/cCR/sCR extracts and mass spectrograms of pig/cattle/sheep bile were performed in combination. Results cCR performed the most prominent enhancement in improving hyperactive energy metabolism in liver and abnormal hyperactivity of hypothalamic–pituitary–adrenal axis axis. As for chemical assessment, HPLC fingerprints of pCR/cCR/sCR extracts indicated that alkaloid components detected in cCR extracts were relatively low, mass spectrograms of pig/cattle/sheep bile suggested that bile acids in cattle bile were relatively abundant, and the contents of taurocholic acid, glycocholic acid, and glycodeoxycholic acid were particularly high. Conclusion Cattle bile could be specified as the processing excipient for bile-processed CR, and the high levels of taurocholic acid, glycocholic acid, and glycodeoxycholic acid mainly contributed to the enhanced efficacy of cCR.

Identification of Codonopsis Radix from different sources by integrating high‐performance liquid chromatography fingerprinting with chemometric approaches and Lobetyolin determination

AbstractCodonopsis Radix (CR) is often used as both a medicine and a food ingredient. However, identifying CR can be difficult due to various factors such as variety, sulfur fumigation, and storage time. Consequently, a comprehensive strategy based on fingerprinting and chemometric approaches was developed to distinguish different CRs. First, the fingerprints of CR were acquired by high‐performance liquid chromatography (HPLC). Secondly, various chemical pattern recognition methods were employed to identify CR. Additionally, 14 and 16 major compounds were identified as marker compounds to differentiate between different varieties and sulfur‐fumigated samples, respectively. Furthermore, the amount of Lobetyolin present in different samples was determined, showing that the content of Lobetyolin varied significantly across the samples. Codonopsis pilosula Nannf. var. modesta (Nannf.) L.T.Shen had the highest content of Lobetyolin, followed by Codonopsis tangshen Oliv., and Codonopsis pilosula (Franch.) Nannf. In addition, the content of Lobetyolin was highest when it was non‐sulfur‐fumigated and stored for 0 years. Conclusively, HPLC fingerprint in conjunction with chemical pattern recognition and component content determination, can be employed to differentiate different varieties and sulfur‐fumigated CR. Additionally, it is a reliable, comprehensive, simple, and rapid method for the identification of CR.

Distinguishing the intrinsic differences of Cynomorium songaricum from different origins using chemical fingerprinting combined with chemometric methods

Cynomorium songaricum is a commonly used medicinal and culinary plant renowned for its effects in tonifying kidney yang, as well as nourishing essence and blood. Consequently, it has garnered considerable attention within the fields of pharmacology and chemistry. However, given the distinct geographical distribution of C. songaricum resources, establishing an accurate analytical method to differentiate its origin becomes indispensable in ensuring its quality and efficacy. In this study, high-performance liquid chromatography (HPLC) was utilized to obtain comprehensive information on the chemical composition of C. songaricum samples from various regions and establish HPLC fingerprints. Through chemometrics analysis, essential chemical markers for classifying them were successfully identified. Additionally, the Kruskal–Wallis rank sum test was employed to evaluate differences in the content of intrinsic constituents of C. songaricum from different origins. The results demonstrated that C. songaricum samples from Neimenggu, Gansu, and Xinjiang could be accurately classified, highlighting the significance of chemical markers in assessing the quality of C. songaricum. This study provides a scientific basis for systematic quality control of Chinese herbal medicines, thereby contributing to advancements in the Chinese medicine industry.

Quality Evaluation and Identification of Phyllanthi fructus (Yuganzi) Based on the Spectrum-Effect Relationship

To establish a quality evaluation and identification method for Phyllanthi fructus (Yuganzi), the spectrum-effect relationship was explored. A high-performance liquid chromatography (HPLC) fingerprint was established using ultraviolet spectrophotometry, and the in vitro antioxidant activity was determined using a total antioxidant capacity assay kit. Similarity analysis, hierarchical cluster analysis (HCA), and partial least squares regression (PLSR) were performed to establish the spectrum-effect relationships. Thirteen batches of Yuganzi were collected for testing. The results revealed that the optimal chromatographic conditions for the HPLC fingerprint were as follows: the mobile phase consisted of 0.1% phosphoric acid solution (A) and acetonitrile (B), the detection wavelength was 214 nm, the column temperature was 30 °C, and the flow rate was 0.8 mL/min. Among the batches of samples, the similarity values of 10 samples (S1–S10) from Yunnan were larger than 0.995; the similarity values of 3 samples (S11–S13) from India, Gaoligong Mountain, and Fujian were less than or equal to 0.986. Furthermore, nineteen characteristic peaks of Yuganzi were calibrated using fingerprint evaluation software. The study on the spectrum-effect relationship further revealed that compounds corresponding to peaks 5 and 8 were potentially key ingredients for the quality evaluation and identification of Yuganzi, closely related to the stable antioxidant activities of Yuganzi. The spectrum-effect relationship is an agile and efficient approach that can ensure the intra-assay stability of Yuganzi from same region and identify Yuganzi from different regions. Compounds with antioxidant activity can be identified as quality markers for Yuganzi.

Recognition of antitussive components in Farfarae Flos based on grey relational analysis and partial least squares regression.

Farfarae Flos has the effect of cough suppression and phlegm elimination, with cough suppression as the main function. Studies have revealed that certain components of Farfarae Flos may be related to its cough suppressant effect, and some components have been confirmed to have cough suppressant activity. However, the antitussive material basis of Farfarae Flos has not been systematically elucidated. This study aims to elucidate the group of active ingredients in Farfarae Flos with cough suppressant activity by correlating the high performance liquid chromatography (HPLC) fingerprint of Farfarae Flos extract with its cough suppressant activity. HPLC was used to establish the fingerprint profiles of 10 batches of Farfarae Flos extract and obtain their chemical composition data. Guinea pigs were selected as experimental animals and the citric acid-induced cough model was used to evaluate the antitussive efficacy data of 10 batches of Farfarae Flos extract. SPF-grade healthy male Hartley guinea pigs were randomly divided into the S1 to S10 groups, a positive control group, and a blank control group (12 groups in total), with 10 guinea pigs in each group. The S1 to S10 groups were respectively administered Farfarae Flos extract S1 to S10 (4 g/kg), the positive control group was administered pentoverine citrate (10 mg/kg), and the blank control group was administered purified water. Each group received continuous oral administration for 5 days. The guinea pigs were placed in 5 L closed wide-mouth bottles, and 17.5% citric acid was sprayed into the bottle with an ultrasonic atomizer at the maximum spray intensity for 0.5 minutes. The cough latency period and cough frequency in 5 minutes were recorded for each guinea pig. Grey relational analysis (GRA) and partial least squares regression (PLSR) were used to conduct spectral-effect correlation analysis of the chemical composition data of Farfarae Flos extract and the antitussive efficacy data, and predict the group of active ingredients in Farfarae Flos with antitussive activity. The bioequivalence verification was conducted to verify the predicted group of active ingredients in Farfarae Flos with antitussive activity: SPF-grade healthy male Hartley guinea pigs were randomly divided into a S9 group, an active ingredient group, a positive control group, and a blank control group (4 groups in total), with 10 guinea pigs in each group. The S9 group was administered Farfarae Flos extract S9 (4 g/kg), the active ingredient group was administered the predicted combination of antitussive active ingredients (dose equivalent to 4 g/kg of Farfarae Flos extract S9), the positive control group was administered pentoverine citrate (10 mg/kg), and the blank control group was administered purified water. Each group received continuous oral administration for 5 days, and animal modeling and observation of efficacy indicators were the same as above. The HPLC fingerprint of 10 batches of Farfarae Flos extract was established, and the peak area data of 14 main common peaks were obtained. The antitussive effect data of 10 batches of Farfarae Flos extract were obtained. Compared with the blank control group, the cough latence in the positive control group and S1, S2, S3, S4, S6, S7, S8, S9, S10 groups was prolonged (all P<0.01), while the cough frequency in 5 minutes in the positive control group and S1, S2, S4, S6, S8, S9, S10 groups was decreased (all P<0.05). The analysis of spectrum-effect relationship revealed that isochlorogenic acid C, isochlorogenic acid A, chlorogenic acid, isochlorogenic acid B, isoquercitrin, and rutin had high contribution to the antitussive effect of Farfarae Flos, and the 6 components were predicted to be the antitussive component group of Farfarae Flos. The verification of bioequivalence showed that there were no statistically significant differences in the antitussive effect between the S9 group and the antitussive component composition group(all P>0.05), which confirmed that isochlorogenic acid C, isochlorogenic acid A, chlorogenic acid, isochlorogenic acid B, isoquercetin, and rutin were the antitussive component group of Farfarae Flos. The analysis of spectrum-effect relationship combined with the verification of bioequivalence could be used to study the antitussive material basis of Farfarae Flos. The antitussive effect of Farfarae Flos is the result of the joint action of many components.

Establishment of multivariate digital high performance liquid chromatography fingerprints of fruit juice essence and its flavor quality evaluation

Quality control problem of essence and spices has always been the focus of food chemists in recent years. In this study, a multi-variate digital fingerprint profile of fruit juice essence sample was established using the combination of HPLC-DAD spectra and UPLC-MS spectra, which were collected to obtain the information about organic acids, polyphenols, sugars and sugar alcohols of the samples. The stability, reliability and repeatability of the combined fingerprinting method were verified in the work. A total of 36 markers in the established fingerprint were common (20 compounds were confirmed) belonging to different classes: organic acids, polyphenols, sugars, sugar alcohols, and other miscellaneous compounds. Those chromatographic peaks that significantly affect the quality of apple juice flavor were screened from principal component analysis (PCA). Moreover, PCA and cluster analysis of the proposed combined fingerprint gave the better evaluation of the sample quality stability and distinguish sample groups than the fingerprint from single data, which demonstrated its great significance to the quality control of fruit juice samples.

Discovery of Potential Anti-Microbial Molecules and Spectrum Correlation Effect of Ardisia crenata Sims via High-Performance Liquid Chromatography Fingerprints and Molecular Docking.

Ardisia crenata Sims, an important ethnic medicine, is recorded in the Chinese Pharmacopoeia for treating laryngeal diseases and upper respiratory tract infections. This study aimed to evaluate the antimicrobial effect of extracts and potential antimicrobial compounds of A. crenata Sims. It was found that the roots of A. crenata Sims have a potential inhibitory effect on Candida albicans and Aspergillus flavus, with MICs of 1.56 mg/mL and 0.39 mg/mL, and the leaves of A. crenata Sims have a potential inhibitory effect on Pseudomonas aeruginosa and Staphylococcus aureus, with MICs of 3.12 mg/mL and 6.77 mg/mL, respectively. Meanwhile, five compounds including one catechin and four bergenins were obtained from roots. These components were identified on the fingerprint spectrum, representing chromatographic peaks 16, 21, 22, 23, and 25, respectively. Among these, 11-β-d-glucopyranosyl-bergenin and (-)-gallocatechin showed potential inhibition for Staphylococcus aureus and Pseudomonas aeruginosa with MIC of 0.26 and 0.33 mg/mL, respectively. The roots, stems, and leaves of A. crenata Sims are very similar in chemical composition, with large differences in content. Principal component analysis (PCA) and Hierarchical cluster analysis (HCA) showed that 16 batches of A. crenata Sims could be divided into four main production areas: Guizhou, Jiangsu, Guangxi, and Jiangxi. Furthermore, molecular docking results showed that 11-β-d-glucopyranosyl-bergenin had a better affinity for Casein lytic proteinase P (ClpP), and (-)-gallocatechin possessed a strong affinity for LasA hydrolysis protease and LasB elastase. These findings suggest catechin and bergenins from A. crenata Sims can be used as antimicrobial activity molecules.

Application of polyphenolic compound-based HPLC fingerprint in Chinese golden camellias (Camellia sect. Chrysantha)

Chinese golden camellias (Camellia sect. Chrysantha) are traditional substitute tea with medicinal and food properties. The development of fingerprinting techniques specific to Chinese golden camellias may provide a new means of reliably assessing the quality of tea and traditional medicine. Herein, 32 total specimens were collected in Southern China and used to establish a polyphenolic compound-based high-performance liquid chromatography (HPLC) signature profile. The established chromatographic fingerprints were subjected to similarity, hierarchical clustering analysis (HCA), together with principal component analysis (PCA) evaluations, and the uncertainty and reliability for such HPLC fingerprints were also evaluated. Similarity values for C. sect. Chrysantha samples ranged from 0.721 to 0.969 relative to the reference chromatographic fingerprint (RFP). HCA and PCA approaches were performed to classify 13 specimens of C. sect. Chrysantha from different species and regions into four categories that respectively contained 7, 1, 2, and 3 specimens. In this HPLC fingerprint, peak 11 was authenticated as rutin, while peaks 2, 4, 6, 7, 9, 12 and 13 were tentatively characterized via liquid chromatography-high-resolution mass spectrometry (LC-HRMS) as corresponding with other polyphenolic agents. Credibility analyses indicated that the macroscopic qualitative and quantitative reliability values for C. fascicularis RFP were greater than 0.9500. Taken together, statistical analyses thus confirmed that this HPLC fingerprinting strategy can be reliably used to classify and assess the quality of C. sect. Chrysantha specimens. And the qualitative and quantitative reliability analysis and systematic quantitative fingerprint methods are promising tools for future efforts to ensure the quality of C. fascicularis.

HPLC Fingerprint Analysis of Cibotii rhizoma from Different Regions and Identification of Common Peaks by LC-MS.

To establish the fingerprint of Cibotii rhizoma using high-performance liquid chromatography (HPLC) and evaluate the quality of Cibotii rhizoma from different regions using chemometrics to identify the potential quality markers, thirteen batches of Cibotii rhizoma samples were analyzed. the similarity evaluation system of TCM chromatographic fingerprint similarity evaluation was used to confirm common peaks. The SPSS 27 software was used for hierarchical cluster analysis (HCA), and SIMCA 14.1 software was used for principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Moreover, a batch of Cibotii rhizoma was selected for LC-MS analysis and speculated on 15 common components. HPLC fingerprint were established, 15 common peaks were matched, two chromatographic peaks were identified using standard substances (protocatechuic acid and protocatechuic aldehyde), and 13 common components were inferred through liquid chromatograph-mass spectrometer (LC-MS). The 13 batches of the samples showed good similarities (>0.910). The results of HCA, PCA and OPLS-DA showed that 13 batches of samples were divided into three groups, and different markers were selected. The method is simple, rapid and reproducible, and can provide a reference for the overall quality evaluation of Cibotii rhizoma.

Study on the Fingerprints of Acanthopanax trifoliatus Fractions with Varying Polarities and the Relationship Between Anti-inflammatory and Antioxidant Activity Using Multivariate Analysis

Objectives: To evaluate the Spectrum-Effect relationships between the High-Performance Liquid Chromatography (HPLC) Fingerprints of fractions with different polarities and the anti-inflammatory and antioxidant effects of A. trifoliatus and to identify its pharmacodynamic material basis. Materials and Methods: Chemical Fingerprints of A. trifoliatus fractions with different polarities (all fractions and ethyl acetate, n-butanol and water fractions) were obtained by HPLC, and Principal Component Analysis (PCA) was performed. The foot swelling and prostaglandin E2 (PGE2) content in mice induced by carrageenan were used as anti-inflammatory markers, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods were used as antioxidant indicators. Moreover, the Spectrum-Effect relationship between common peaks and efficacy value was conducted by Grey Relational Analysis (GRA) and Partial Least Squares analysis (PLS). Results: PCA identified three principal components, and all fractions of A. trifoliatus had good anti-inflammatory and antioxidant effects. Based on GRA and PLS analysis, peaks 1 and 2 (neochlorogenic acid), 4 and 5 (chlorogenic acid), 8, 9, 10, 11 (isochlorogenic acid B), and 12 were involved in anti-inflammatory efficacy, and peaks 1 and 2 (neochlorogenic acid), 3, 4, and 5 (chlorogenic acid), and 6, 7, 9, 10, and 13 (isochlorogenic acid A) were involved in antioxidant efficacy. Conclusion: This study on the Spectrum-Effect relationship of different polar fractions of A. trifoliatus provides data to support the discovery of the pharmacodynamic material basis of the anti-inflammatory and antioxidant properties of A. trifoliatus.

Holistic quality evaluation method of Epimedii Folium based on NIR spectroscopy and chemometrics.

There are some problems in the quality control of Epimedii Folium (leaves of Epimedium brevicornum Maxim.), such as the mixed use of Epimedii Folium from different harvesting periods and regions, incomplete quality evaluation, and time-consuming analysis methods. Near-infrared (NIR) spectroscopy was conducted to establish a rapid overall quality evaluation method for Epimedii Folium. Quantitative models of the total solid, moisture, total flavonoid, and flavonol glycoside (Epimedin A, Epimedin B, Epimedin C, Icariin) contents of Epimedii Folium were established by partial least squares regression (PLSR). The root mean square error (RMSE) and correlation coefficient (R) were used to evaluate the performance of models. The qualitative models of Epimedii Folium from different geographic origins and harvest periods were established based on K-nearest neighbor (KNN), back-propagation neural network (BPNN), and random forest (RF). Accuracy and Kappa values were used to evaluate the performance of models. A new multivariable signal conversion strategy was proposed, which combines NIR spectroscopy with the PLSR model to predict the absorbance values of retention time points in the high-performance liquid chromatography (HPLC) fingerprint to obtain the predicted HPLC fingerprint. The Pearson correlation coefficient and cosine coefficient were used to evaluate the similarity between real and predicted HPLC fingerprints. Qualitative models, quantitative models, and the similarity between real and predicted HPLC fingerprints are satisfactory. The method serves as a fast and green analytical quality evaluation method of Epimedii Folium and can replace traditional methods to achieve the overall quality evaluation of Epimedii Folium.

Quality evaluation of Lonicerae Japonicae Flos from different origins based on high-performance liquid chromatography (HPLC) fingerprinting and multicomponent quantitative analysis combined with chemical pattern recognition.

Lonicerae Japonicae Flos (LJF) is widely used in food and traditional Chinese medicine. To meet demand, Lonicera japonica Thunb. is widely cultivated in many provinces of China. However, reported studies on the quality evaluation of LJF only used a single or a few active components as indicators, which could not fully reflect the quality of LJF. In the present study, we aimed to develop a methodology for comprehensively evaluating the quality of LJF from different origins based on high-performance liquid chromatography (HPLC) fingerprinting and multicomponent quantitative analysis combined with chemical pattern recognition. The HPLC method was developed for fingerprint analysis and was used to determine the contents of 19 components of LJF. To distinguish between samples and identify differential components, similarity analysis, hierarchical cluster analysis, principal component analysis, and orthogonal partial least squares discriminant analysis were performed. The HPLC fingerprint was established. Using the developed method, the contents of 19 components recognized in the fingerprint analysis were determined. Samples from different origins could be effectively distinguished. HPLC fingerprinting and multicomponent quantitative analysis combined with chemical pattern recognition is an efficient method for evaluating LJF.

In vivo toxicity, anti-hyperlipidaemic, antioxidant and anti-atherogenic activities of ‘LIPO A’ A traditional herbal product in rodents

Hyperlipidemia accounts for about 17 million deaths worldwide each year. High cost and side effects have limited the use of conventional anti-lipidaemic agents in some cases, majority of whom resort to traditional medicine. The current research focused on validating the safety and efficacy of a herbal product, ‘LIPO A’ used in the management of hyperlipidaemia. Induction of hyperlipidaemia was achieved by oral administration of 3 mL of cholesterol in coconut oil for 4 weeks in male Sprague Dawley rats with water available as 40 % sucrose. Subsequently, the animals were treated with 100, 200 and 400 mg/kg of the product ‘LIPO A’ for 4 additional weeks with atorvastatin as reference drug (at 2 mg/kg body weight). Blood samples were taken for serum biochemistry and atherogenic ratios were then calculated. 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) scavenging assay, total antioxidant capacity, physicochemical and phytochemical analysis were also carried out using standard methods. Treatment resulted in a dose-dependent reduction in total cholesterol with maximum reduction of 46.01 % at 400 mg/kg compared to atorvastatin with 49.30 %. There were significant changes in the low-density lipoprotein cholesterol and high-density lipoprotein cholesterol (LDL-c/HDL-c) and Total Cholesterol (TC/HDL-c) ratios which measures the atherogenic and coronary risk indices respectively. Acute and subacute toxicity studies did not reveal any signs of toxicity. High Performance Liquid Chromatography (HPLC) fingerprint revealed six well resolved peaks with two prominent compounds with retention times 24.88 and 23.95 min, which could serve as quality control markers for the product. The herbal product showed considerable antihyperlipidemic and antioxidant actions in rodent models and lend credence to its use in traditional medicine for hyperlipidaemia.

Research on the quality markers of antioxidant activity of Kai-Xin-San based on the spectrum-effect relationship.

Background: Kai-Xin-San (KXS) is one of the classic famous traditional Chinese medicine prescriptions for amnesia, which has been applied for thousands of years. Modern pharmacological research has found that KXS has significant therapeutic efficacy on nervous system diseases, which is related to its antioxidant activity. However, the antioxidant material basis and quality markers (Q-makers) of KXS have not been studied. Objective: The objective of this study is to explore the Q-makers of antioxidant activity of KXS based on spectrum-effect relationship. Methods: Specifically, the metabolites in KXS extracts were identified by UPLC-Q-Exactive Orbitrap MS/MS. The fingerprint profile of KXS extracts were established by high-performance liquid chromatography (HPLC) and seven common peaks were identified. Meanwhile, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) test was used to evaluate the free radical scavenging ability of KXS. The spectrum-effect relationship between its HPLC fingerprint and DPPH free radical scavenging activity was preliminarily examined by the Pearson correlation analysis, grey relation analysis (GRA), and orthogonal partial least squares discrimination analysis (OPLS-DA). Further, the antioxidant effect of KXS and its Q-makers were validated through human neuroblastoma (SH-SY5Y) cells experiment. Results: The results showed that 103 metabolites were identified from KXS, and the similarity values between HPLC fingerprint of twelve batches of KXS were greater than 0.900. At the same time, the results of Pearson correlation analysis showed that the peaks 8, 1, 14, 17, 18, 24, 16, 21, 15, 13, 6, 5, and 3 from KXS were positively correlated with the scavenging activity values of DPPH. Combined with the results of GRA and OPLS-DA, peaks 1, 3, 5 (Sibiricose A6), 6, 13 (Ginsenoside Rg1), 15, and 24 in the fingerprints were screen out as the potential Q-makers of KXS for antioxidant effect. Besides, the results of CCK-8 assay showed that KXS and its Q-makers remarkably reduced the oxidative damage of SH-SY5Y cells caused by H2O2. However, the antioxidant activity of KXS was decreased significantly after Q-makers were knocked out. Conclusion: In conclusion, the metabolites in KXS were successfully identified by UPLC-Q-Exactive Orbitrap MS/MS, and the Q-makers of KXS for antioxidant effect was analyzed based on the spectrum-effect relationship. These results are beneficial to clarify the antioxidant material basis of KXS and provide the quality control standards for new KXS products development.

Elucidating the active ingredients and potential anti-inflammatory mechanism of Qingwei Huanglian Wan

Qingwei Huanglian Wan (QHW) is a traditional Chinese medicine used for treating inflammation. As the compositions of QHW preparations are complex and varied, it has proven difficult to identify the active ingredients in QHW and control the quality. In this study, high-performance liquid chromatography (HPLC) fingerprints, network pharmacology, molecular docking, and the Griess method were used to screen for and verify the potential active ingredients of QHW. First, sixteen batches of QHW were studied as two groups using multivariate statistical analysis. Thirteen peaks were detected in the HPLC results to establish a fingerprint similarity model, and six chemical constituents (phellodendrine hydrochloride, geniposide, berberine hydrochloride, baicalin, wogonoside, and glycyrrhizic acid) were identified. Among these six constituents, four components (glycyrrhizic acid, berberine hydrochloride, baicalin, and wogonoside) were considered as potential active ingredients. Second, network pharmacology, molecular docking, and the Griess method were used to further elucidate the potential active ingredients. There was no significant difference in the NO content of the active ingredients administration group (dose converted into the medium-dose group) and medium-dose QHW administration group. Then, the contents of berberine hydrochloride, baicalin, wogonoside, and glycyrrhizic acid were determined using the HPLC method. This study provides a comprehensive and reliable strategy for the quality control of QHW preparations and identifies potential active ingredients in QHW. Our methodology may also be useful for studying other traditional Chinese medicines.