High-performance Liquid Chromatography Fingerprint Analysis Research Articles

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.

HPLC Fingerprint Analysis of Rana chensinensis Eggs from Different Habitats and Their Antitussive Effect.

In this paper, a novel fingerprint method was established for the quality control of Rana chensinensis eggs (RE) by high-performance liquid chromatography (HPLC). Cluster analysis and principal component analysis were performed. Besides, the antitussive effect of RE was explored. The analysis was achieved on a Kromasil 100-5C18 (4.6 mm × 250 mm, 5 μm) column by gradient elution using methanol-0.1% phosphoric acid solution as the mobile phase. The influence of RE on cough latent periods and cough times of mice was investigated via an ammonia cough-inducing experiment. The validated HPLC method was precise, reproducible, and stable. The HPLC fingerprints of 10 batches of RE samples displayed 31 well-resolved common peaks in the chromatogram. Three of these peaks were identified and assigned to 1-methyl hydantoin, estradiol, and 4-cholestene-3-one. The similarities of 10 batches of samples were more than 0.95. RE from different origins could be classified into three groups via SPSS 23.0 software, suggesting RE samples from various provinces (Jilin, Liaoning, and Heilongjiang) can be well distinguished via the established method. High dose and middle dose of the RE group can significantly prolong the cough latent periods of mice (P < 0.05 or P < 0.01) and inhibit the cough times of mice (P < 0.01), indicating RE had a good antitussive effect. HPLC fingerprint combined with multicomponent determination can be an efficient and useful method for monitoring the quality of RE. This study also provided a more comprehensive strategy for the quality evaluation of RE.

Quality Evaluation of Market Acacia catechu by Fingerprint-Chemical Pattern Recognition

Acacia catechu (L.f.) Willd, a leguminous plant, is included in the 2020 edition of the Chinese Pharmacopoeia and is mainly used to treat eczema, mouth ulcers, diarrhea, bruising, and traumatic hemorrhage. However, there are imported and domestic Acacia catechu samples available in China, and their quality and price are very different, which seriously affects the safety and stability of their clinical application. Importantly, there is no simple and effective method for identifying or classifying grades of Acacia catechu. In this study, 47 batches of commercial Acacia catechu were used for identifying or classifying grades of Acacia catechu using high performance liquid chromatography (HPLC) combined with chemometric analysis. Firstly, gradient elution was adopted with 0.05% phosphoric acid water (A)-methanol (B) as the mobile phase to establish chromatographic conditions. The HPLC chromatograms of 47 batches of Acacia catechu samples were analyzed by the “Similarity Evaluation System for Chromatographic Fingerprint of TCM” software (version 2012A). The common peaks of Acacia catechu were identified to evaluate the similarity. Based on the determination results of fingerprint chromatographic peak area, the quality of the collected Acacia catechu was evaluated by chemometric methods such as CA, PCA, and OPLS-DA. The results showed that the collected Acacia catechu samples were significantly divided into three categories. The first-class samples were all imported Acacia catechu except S9 sample, which was domestic Acacia catechu; the second-class samples were partly domestic Acacia catechu and partly imported Acacia catechu; and the third-class samples were all domestic Acacia catechu. Moreover, OPLS-DA of 47 batches of samples showed that the contents of catechin and the total contents of catechin and epicatechin could be used as key indicators for assessing the quality of Acacia catechu. The developed HPLC fingerprint and quantitative analysis method of multi-indicator components can be used for classification and quality evaluation of market Acacia catechu, which has a significant reference value for developing Acacia catechu grade quality standards.

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.

Genotoxicity, nitric oxide level modulation and cardio-protective potential of Kalanchoe Integra Var. Crenata (Andr.) Cuf Leaves in murine models

Ethnopharmacological relevanceAdvancement in cancer therapy has improved survival among patients. However, use of anticancer drugs like anthracyclines (e.g., doxorubicin) is not without adverse effects. Notable among adverse effects of doxorubicin (DOX) is cardiotoxicity, which ranges from mild transient blood pressure changes to potentially serious heart failure. Anecdotal reports suggest that Kalanchoe integra (KI) may have cardio-protective potential. Aims of the studyThis study sought to determine the cardio-protective potential of KI against doxorubicin-induced cardiotoxicity and also examined any possible genotoxic potential of KI in selected organs. Additionally, the nitric oxide modulatory potential of KI was assessed. Materials and methodsThe leaves of KI were collected, air-dried, pulverised and extracted using 70% ethanol. High-performance liquid chromatography (HPLC) fingerprinting was done for KI. Also, the single-cell gel electrophoresis assay (Comet assay) was employed to ascertain the genotoxic potential of KI. In assessment of cardio-protective potential of KI against doxorubicin-induced cardiotoxicity, a total of 42 female Sprague-Dawley rats were put into 7 groups (n = 6). Group I: vehicle control, received normal saline (1 mL/kg p.o) for 30 days. Group II: toxic control, received DOX (20 mg/kg i.p.) once on the 29th day. Group III: KI control, received KI (300 mg/kg p.o) for 30 days. Group IV: vitamin E control, received vitamin E (100 mg/kg p.o) for 30 days. Group V: KI treated-1, received KI (300 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Group VI: KI treated-2, received KI (600 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Group VII: vitamin E treated, received vitamin E (100 mg/kg p.o) for 30 days and DOX (20 mg/kg i.p) on the 29th day. Thirty-six (36) hours after last administration, rats were sacrificed. Blood samples were taken via cardiac puncture to determine levels of aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), lactate dehydrogenase (LDH), enzymatic antioxidants such as glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT). Nitric oxide level was also determined. Hearts of rats in each group were excised and taken through histopathological examination. ResultsIn the HPLC fingerprint analysis, 13 peaks were identified, and peak with retention time of 24.0 min had the highest peak area (3.223 x104 mAU). Comet assay showed that the KI extract was non-genotoxic. Pretreatment with KI protected rats against doxorubicin-induced cardiotoxicity as evidenced by the low levels of AST, ALT, ALP, CK and LDH compared with the controls (p < 0.05). SOD, CAT and GPX levels were also high for rats administered KI extracts, further showing that KI protected rats against doxorubicin-induced cardiotoxicity. KI also inhibited nitric oxide levels at 300 mg/kg and 600 mg/kg effective doses. Histological examination revealed that rats pretreated with KI showed no signs of abnormal myocardial fibres (shape, size and configuration). ConclusionEthanolic (70%) leaf extract of KI showed no genotoxic potential and possessed cardioprotective effects against doxorubicin-induced cardiotoxicity in Sprague-Dawley rats. KI also inhibited nitric oxide production, thus, a potential nitric oxide scavenger.

Carica papaya leaf water extract promotes innate immune response via MAPK signaling pathways

The emergence and rapid spread of the potentially fatal coronavirus disease 2019, caused due to infection by severe acute respiratory syndrome coronavirus-2, has led to worldwide interest in developing functional bioactive ingredients that act as immunomodulatory agents. In this study, we aimed to characterize Carica papaya extract and explore its potential as an immuno-modulator by performing in vitro cell screening. Papaya leaf water extract (PLW) was found to significantly increase the levels of nitric oxide (NO) and prostaglandin E2 (PGE2) by upregulating inducible nitric oxide synthase and cyclo-oxygenase-2 activity, respectively. Additionally, PLW increased the production of tumor necrosis factor-α and interleukin 1β in RAW 264.7 cells. Furthermore, PLW activated the expression of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) but not that of p38 mitogen-activated protein kinase. These results indicate that PLW increased the production of NO, PGE2, and pro-inflammatory cytokines by activating the JNK and ERK pathways in macrophages, thus demonstrating immunomodulatory properties. Finally, high-performance liquid chromatography fingerprint analysis indicated the presence of rutin, narirutin, and ρ-coumaric acid in PLW (6.30, 119.76, and 47.25 ppm, respectively). Treating cells with these compounds at non-toxic concentrations had no effect on NO production. Taken together, these results suggest that PLW may have potential as an immunity-enhancing supplement. © The Korean Society for Applied Biological Chemistry 2021.

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 ( R2 ≥ 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.

Chemical Fingerprint Analysis and Ultra-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry-Based Metabolomics Study of the Protective Effect of Buxue Yimu Granule in Medical-Induced Incomplete Abortion Rats.

Medical abortion is a common method to terminate an early pregnancy and often causes serious complications such as abnormal uterine bleeding and endometritis. Buxue Yimu granule (BYG) is a well-known traditional Chinese medicine prescription composed of five kinds of drugs and is widely used in gynecology and obstetrics. The aim of the present study was to establish the quality standard of BYG and investigate its protective effect on incomplete abortion. The chemical fingerprint of BYG was established by high performance liquid chromatography (HPLC). The major compounds of BYG were determined by ultra-performance liquid chromatography with triple quadrupole mass spectrometry. An incomplete abortion rat model was induced by intragastric administration of mifepristone (8.3 mg·kg−1) combined with misoprostol (100.0 μg·kg−1) during early pregnancy. The serum levels of human chorionic gonadotrophin (HCG), estradiol (E2), and progesterone (PG) were determined. The serum endogenous metabolites were analyzed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Multivariate analysis, including partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA), was employed to analyze the metabolic profiles, and MetaboAnalyst was used to investigate the metabolic pathways. Furthermore, hematoxylin-eosin staining (HE) was used to evaluate the histopathological changes in uterine tissue. The expression levels of VEGFA and NF-κB were detected by immunohistochemistry. The results indicated that HPLC fingerprint analysis can be successfully used to assess the quality of BYG. The medical-induced incomplete abortion rats were clearly separated from control rats, and the biochemical changes were gradually restored to normal after administration of BYG. Moreover, 19 potential biomarkers, including N-lactoylleucine, 2-piperidinone, isobutyryl-l-carnitine, eicosapentaenoylcholine, LysoPC(14:0), LysoPC(20:5), physagulin C, LysoPC(18:3), leukotriene D5, deoxycholic acid 3-glucuronide, glycine, pregnanediol 3-O-glucuronide, LysoPC(18:2), LysoPC(17:0/0:0), N-acetyl-leukotriene E4, LysoPC(18:0), platelet-activating factor, LysoPA(24:1), and LysoPC(18:1), which were mainly related to the amino acids metabolism, lipids metabolism, and bile acid biosynthesis, were identified. Consequently, BYG exerts a potential protective role in the intervention of incomplete abortion by anti-inflammatory, promote endometrial repair, and regulate the metabolic disorders.

The first assessment on antioxidant and antidiabetic activities of leaves and stems of Vaccinium secundiflorum Hook. (Ericaceae), an endemic plant of Madagascar

Vaccinium secundiflorum Hook. (Ericaceae family) is an endemic shrub of the Tapia forests of Madagascar widely used by local Malagasy population to treat diabetes mellitus. This study aims to preliminarily determine the antioxidant activity and anti-diabetic properties of this plant. Plant leaf and stem powders were macerated in methanol 95%. The methanol extract (MVS) was used for (i) phytochemical fingerprint by High Performance Liquid Chromatography (HPLC), (ii) total phenolic content (TPC) evaluation by the Folin–Ciocalteu colourimetric method, (iii) 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenger spectrophotometric assay, (iv) antioxidant capacity by Ferric Reducing Antioxidant Power (FRAP) assay, and (v) antidiabetic activity evaluation by oral glucose tolerance test and acute toxicity in mice. Leaves and stems of V. secundiflorum presented a high rate of TPC (4151.02 ± 745.21mgGAE/100 g of dry weight). The HPLC fingerprint analysis showed the presence of several secondary metabolite classes including organic acids (55.31%), phenolic compounds (29.66%), carotenoids (10.90%), vitamin C (3.96%), and monoterpenes (0.10%). The DPPH free radical scavenger spectrometric assay showed a MVS high potential scavenging activity (IC50 = 76.06 ± 1.08 µg/mL). This potential antioxidant capacity was confirmed by the FRAP assay results (69.31 ± 3.31 mmol Fe2+/Kg of dry weight). The oral glucose tolerance test in mice showed a highly significant blood glucose-lowering effect. The acute toxicity in mice revealed that the MVS did not show toxicity at the dose of 2000 mg/kg. Leaves and stems of V. secundiflorum showed significant antioxidant and antidiabetic properties supporting the plant traditional use, even if further studies on bioactive compounds and their action mechanisms should be carried out to confirm these preliminary results.

A Strategy for Quality Control of Vespa magnifica (Smith) Venom Based on HPLC Fingerprint Analysis and Multi-Component Separation Combined with Quantitative Analysis

As a folk medicine of the Jingpo minority in Yunnan province, the venom of Vespa magnifica has been commonly used for the treatment of rheumatoid arthritis. Quality standardization of the wasp venom is a necessary step for its pharmaceutical research and development. To control the quality of the wasp venom, a method based on high-performance liquid chromatography (HPLC) was developed for chemical fingerprint analysis. In the chromatographic fingerprinting, chemometrics procedures, including similarity analysis (SA), hierarchical clustering analysis (HCA), and principal component analysis (PCA), were applied to classify 134 batches (S1–S134) of wasp venom from different origins. The HPLC fingerprint method displayed good precision (Relative standard deviation, RSD < 0.27%), stability (in 16 h, RSD < 0.34%), and repeatability (RSD < 1.00%). Simultaneously, four compounds (VMS1, VMS2, VMS3, and VMS4) in the wasp venom were purified and identified. VMS1 was 5-hydroxytryptamine, and the other compounds were three peptides that were sequenced as follows: Gly–Arg–Pro–Hyp–Gly–Phe–Ser–Pro–Phe–Arg–Ile–Asp–NH2 (VMS2), Ile–Asn–Leu–Lys–Ala–Ile–Ala–Ala–Leu–Ala–Lys–Lys–Leu–Leu–NH2 (VMS3), and Phe–Leu–Pro–Ile–Ile–Gly–Lys–Leu–Leu–Ser–Gly–Leu–Leu–NH2 (VMS4). The quantifications for these components were 110.2 mg/g, 26.9 mg/g, 216.3 mg/g, and 58.0 mg/g, respectively. The results of this work indicated that the combination of the chemical fingerprint and quantitative analysis offers a reasonable way to evaluate the quality of wasp venom.

ZhiShiXiaoPi tang inhibits autophagy induced by corticosterone and functional dyspepsia through blockade of the mTOR pathway

Ethnopharmacological relevanceZhiShiXiaoPi Tang (ZSXPT) is a Chinese traditional medicine formula that contains 10 Chinese traditional medicine substances. It has been widely used to treat patients with functional dyspepsia (FD). However, the protective effect of ZSXPT and its molecular mechanisms in FD still remain elusive. Aim of the studyTo investigate the protective effect of ZSXPT on autophagy induced by Corticosterone (Cort) in PC12 cells which have typical neuron characteristics and have been widely used as a model system for depression studies and FD rats, and explore its underlying mechanisms. Materials and methodsIn this study, high-performance liquid chromatography fingerprint analysis was performed to characterize the chemical composition of ZSXPT. Depression-induced autophagy, ROS generation, and changes in mitochondrial membrane potential (MMP) were investigated in Cort-induced PC12 cells and in FD rats to evaluate the protective effects of ZSXPT. ResultsOur results show that ZSXPT treatment protects neurons against Cort-induced damage and apoptosis by increasing cell viability and reducing the release of lactate dehydrogenase. ZSXPT decreased Cort-induced ROS generation, increased MMP, and accelerated autophagy through the blockade of the mammalian target of rapamycin (mTOR) pathway. Moreover, we observed similar findings when we studied ZSXPT in a rat model of FD. ConclusionsOur in vitro and in vivo results indicate that the neuroprotective effect of ZSXPT against autophagy-induced damage and apoptosis occurs mainly by blocking the mTOR pathway in Cort-induced PC12 cells and in FD rats. Taken together, these data provide reliable experimental evidence and explain the molecular mechanism by which ZSXPT ameliorates FD.

DiDang Tang Inhibits Endoplasmic Reticulum Stress-Mediated Apoptosis Induced by Oxygen Glucose Deprivation and Intracerebral Hemorrhage Through Blockade of the GRP78-IRE1/PERK Pathways.

DiDang Tang (DDT), a Chinese traditional medicine formula, contains 4 Chinese traditional medicine substances, has been widely used to treat intracerebral hemorrhage (ICH) patients. However, the molecular mechanisms of DDT for protecting neurons from oxygen and glucose deprivation (OGD)-induced endoplasmic reticulum (ER) stress and apoptosis after ICH still remains elusive. In this study, high-performance liquid chromatography fingerprint analysis was performed to learn the features of the chemical compositions of DDT. OGD-induced ER stress, Ca2+ overload, and mitochondrial apoptosis were investigated in nerve growth factor -induced PC12, primary neuronal cells, and ICH rats to evaluate the protective effect of DDT. We found that DDT treatment protected neurons against OGD-induced damage and apoptosis by increasing cell viability and reducing the release of lactate dehydrogenase. DDT decreased OGD-induced Ca2+ overload and ER stress through the blockade of the glucose-regulated protein 78 (GRP78)- inositol-requiring protein 1α (IRE1)/ protein kinase RNA-like ER kinase (PERK) pathways and also inhibited apoptosis by decreasing mitochondrial damage. Moreover, we observed similar findings when we studied DDT for inhibition of ER stress in a rat model of ICH. In addition, our experiments further confirmed the neuroprotective potential of DDT against tunicamycin (TM)-induced neural damage. Our in vitro and in vivo results indicated that the neuroprotective effect of DDT against ER stress damage and apoptosis occurred mainly by blocking the GPR78-IRE1/PERK pathways. Taken together, it provides reliable experimental evidence and explains the molecular mechanism of DDT for the treatment of patients with ICH.

Survey of pyrrolizidine alkaloids in the tribe Lithospermeae (Boraginaceae) from Pan-Himalaya and their chemotaxonomic significance

Family Boraginaceae has drawn the attention of many botanists for many years because of its systematic position and the presence of toxic pyrrolizidine alkaloids (PAs). Some researcher used these alkaloids as chemotaxonomic markers at the family and subfamily level in several botanical families. Therefore, this explorative study carried out with an aim to determine the distribution of selected PAs and their chemotaxonomic significance in the tribe Lithospermeae. For this, Lithospermeae species along with other Boraginaceae members from different parts of Pan-Himalayan regions in Pakistan and China were brought to Beijing Normal University in Beijing, China. For HPLC fingerprint analysis, dried silica gel aerial parts (500 mg) of twenty-four mature flowering species of Lithospermeae and the available PAs: supinine, heliotrine, lycopsamine, europine, and echimidine were used as chemotaxonomic markers. In the laboratory, the plants of Lithospermeae were investigated for pyrrolizidine alkaloids by using high performance liquid chromatography (HPLC). Five pyrrolizidine alkaloids were identified qualitatively in 24 species of tribe Lithospermeae from Pan-Himalaya. The results of the study support the previous research that PAs can be used as a taxonomic marker. The PAs lycopsamine and echimidine were common in all Onosma species except O. sinicum Diels, O. waddellii Duthie, and O. griffithii Vatke. So we proposed that lycopsamine and echimidine can be used as taxonomic markers for Onosma species from Pan-Himalayan regions. In the case of the genus Maharanga, M. emodi (Wall.) A. DC. and M. bicolor (Wall. ex G. Don) A. DC. show the same composition of PAs lycopsamine and echimidine. But these two species could be easily distinguished from each other; heliotrine is present in M. emodi but absent in M. bicolor. Arnebia guttata Bunge was the only species in the genus Arnebia which was detected for only europine. A. benthamii (Wall. ex) G. Don. I.M. Johnst. and A. euchroma (Royle) I.M. Johnst. were similar in PA composition for heliotrine, lycopsamine, and echimidine. These two species can easily be distinguished from each other: europine is present in the former genus but absent in the latter one. Moreover, the result of HPLC fingerprint analysis shows that supinine is distributed throughout all investigated species from the Pan-Himalayan regions except Lithospermum arvense L. So we proposed that supinine could serve as a chemotaxonomic marker in the tribe Lithospermeae.

Comparison of the chromatographic fingerprint, multicomponent quantitation and antioxidant activity of Salvia miltiorrhiza Bge. between sweating and nonsweating.

Salvia miltiorrhiza Bge. is a traditional Chinese medicine applied in the treatment of various diseases in clinical practice. In the course of its processing, S. miltiorrhiza Bge. is usually processed by sweating. This study employed 10-component contents determination coupled with high-performance liquid chromatography (HPLC) fingerprint and antioxidant activity to investigate the effect of sweating on S. miltiorrhiza Bge. so as to evaluate the quality of S. miltiorrhiza Bge. The HPLC method was performed using C18 and 0.05% phosphoric acid aqueous solution-acetonitrile with a gradient elution system. It was validated for linearity, precision, repeatability, stability and recovery. Similarity analysis, principal components analysis and antioxidant activity assays were used to compare sweated S. miltiorrhiza Bge. (SSM) and nonsweated S. miltiorrhiza Bge. (NSSM). SSM and NSSM showed good similarities in HPLC fingerprint (>0.9), but principal components analysis could classify the HPLC fingerprint and 10-component quantitation analysis. Meanwhile, the antioxidant activity of SSM was significantly higher than that of NSSM (p < 0.01). The results of this study indicated that sweating could alter the content of chemical constituents in S. miltiorrhiza Bge., and could also improve its antioxidant activity. In addition, the method not only affords a viable strategy for comparing SSM and NSSM and assessing the quality of S. miltiorrhiza Bge., but also provides a reference for other herbal medicine that suffers from sweating.

HPLC Fingerprint Analysis Combined with Chemometrics for Authentication of &lt;i&gt;Kaempferia galanga&lt;/i&gt; from Related Species

Fingerprint analysis using high performance liquid chromatography (HPLC) has been developed for authentication of Kaempferia galanga from related species, such as Kaempferia pandurata and K. rotunda. By comparing the fingerprint chromatograms of K. galanga, K. pandurata and K. rotunda, we could identify K. galanga samples and detect adulteration of K. galanga from K. pandurata and K. rotunda by using their marker peaks. We also combined HPLC fingerprint with chemometrics for discrimination the three species and also for authentication of K. galanga. All the three species and K. galanga adulterated with K. pandurata and K. rotunda were discriminated successfully by using principal component analysis (PCA) and discriminant analysis (DA). This result indicates that HPLC fingerprint analysis in combination with PCA (PC1 = 30.06% and PC2 = 34.74%) and DA (DF1 = 94.59% and DF2 = 3.32%) could be used for authentication of K. galanga samples from the related species.

Quality evaluation of raw and processed Crataegi Fructus by color measurement and fingerprint analysis.

Crataegi Fructus and its processed products have been used as a traditional medicine for a long time, and numerous active components are responsible for their curative effects. However, a comprehensive and fast method for the quality control of its processed products is still lacking. In this study, two analytical methods based on color measurements and fingerprint analysis are established. In the color measurements, the color values of the peel and flesh of Crataegi Fructus were evaluated spectrophotometrically. Based on the results, a color reference range was established using percentiles, and the standard color difference value was established using the median color values. Then, the color values of Crataegi Fructus and its processed products were analyzed using Bayes linear discriminant analysis and mathematical functions were built in order to predict the degree of processing. Moreover, high-performance liquid chromatography fingerprint analysis was performed on a Hibar C18 column, and a high-performance liquid chromatography fingerprint pattern was obtained, from which nine peaks were identified. Chemometric methods were successfully applied to differentiate raw and processed Crataegi Fructus.

Development of Quality Control Method for Glucofarmaka Antidiabetic Jamu by HPLC Fingerprint Analysis

Herbal medicines become increasingly popular all over the world for preventive and therapeutic purposes. Quality control of herbal medicines is important to make sure their safety and efficacy. Chromatographic fingerprinting has been accepted by the World Health Organization as one reliable strategy for quality control method in herbal medicines. In this study, high-performance liquid chromatography fingerprint analysis was developed as a quality control method for glucofarmaka antidiabetic jamu. The optimum fingerprint chromatogram were obtained using C18 as the stationary phase and linear gradient elution using 10–95% acetonitrile:water as the mobile phase within 60 minutes of elution and detection at 210 nm. About 20 peaks were detected and could be used as fingerprint of glucofarmaka jamu. To evaluate the analytical performance of the method, we determined the precision, reproducibility, and stability. The result of the analytical performance showed reliable results. The proposed method could be used as a quality control method for glucofarmaka antidiabetic jamu and also for its raw materials.

High perfomance liquid chromatography fingerprint analysis for quality control of brotowali (Tinospora crispa)

Brotowali (Tinospora crispa) is widely used in Indonesia as ingredient of herbal medicine formulation. To ensure the quality, safety, and efficacy of herbal medicine products, its chemical constituents should be continuously evaluated. High performance liquid chromatography (HPLC) fingerprint is one of powerful technique for this quality control process. In this study, HPLC fingerprint analysis method was developed for quality control of brotowali. HPLC analysis was performed in C18 column and detection was performed using photodiode array detector. The optimum mobile phase for brotowali fingerprint was acetonitrile (ACN) and 0.1% formic acid in gradient elution mode at a flow rate of 1 mL/min. The number of peaks detected in HPLC fingerprint of brotowali was 32 peaks and 23 peaks for stems and leaves, respectively. Berberine as marker compound was detected at retention time of 20.525 minutes. Evaluation of analytical performance including precision, reproducibility, and stability prove that this HPLC fingerprint analysis was reliable and could be applied for quality control of brotowali.

Inulin-type fructan and infusion of Artemisia vulgaris protect the liver against carbon tetrachloride-induced liver injury

BackgroundInfusions of aerial parts of Artemisia vulgaris L. (Asteraceae) are used in herbal medicine to treat several disorders, including hepatosis. PurposeEvaluation of in vivo hepatoprotective effects of A. vulgaris infusion (VI) and inulin (VPI; i.e., the major polysaccharide of VI). Study designThe hepatoprotective effect of A. vulgaris extracts on carbon tetrachloride (CCl4)-induced hepatotoxicity and the probable mechanism involved in this protection were investigated in mice. MethodsA. vulgaris infusion (VI) was prepared according to folk medicine using the aerial parts of the plant. Carbohydrate, protein, and total phenolic content was determined in VI, and its phenolic profile was determined by high-performance liquid chromatography (HPLC). Male Swiss mice were orally pretreated for 7 days with VI or VPI (once per day). On days 6 and 7 of treatment, the mice were intraperitoneally challenged with CCl4. Liver and blood were collected and markers of hepatic damage in plasma and oxidative stress in the liver were analyzed. Hepatic histology and inflammatory parameters were also studied in the liver. The scavenging activity of VI and VPI were evaluated in vitro using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. ResultsVI contained 40% carbohydrates, 2.9% proteins and 9.8% phenolic compounds. The HPLC fingerprint analysis of VI revealed chlorogenic, caffeic and dicaffeoylquinic acids as major low-molar-mass constituents. Oral pretreatment with VI and VPI significantly attenuated CCl4-induced liver damage, reduced the activity of alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) in plasma, and prevented reactive oxygen species accumulation and lipid peroxidation in the liver. Comparisons with the CCl4-treated group showed that VI and VPI completely prevented necrosis, increased the levels of reduced glutathione (GSH), and reduced tumor necrosis factor alpha (TNF-α) level in the liver. VI and VPI also exhibited high radical scavenging activity in vitro. ConclusionVI and VPI had remarkable hepatoprotective effects in vivo, which were likely attributable to antioxidant and immunomodulatory properties. The present findings support the traditional use of A. vulgaris infusion for the treatment of hepatic disorders.

Hydroethanolic extract of Baccharis trimera promotes gastroprotection and healing of acute and chronic gastric ulcers induced by ethanol and acetic acid.

Ethanol is a psychoactive substance highly consumed around the world whose health problems include gastric lesions. Baccharis trimera is used in folk medicine for the treatment of gastrointestinal disorders. However, few studies have evaluated its biological and toxic effects. To validate the popular use of B. trimera and elucidate its possible antiulcerogenic and cytotoxic mechanisms, a hydroethanolic extract of B. trimera (HEBT) was evaluated in models of gastric lesions. Rats and mice were used to evaluate the protective and antiulcerogenic effects of HEBT on gastric lesions induced by ethanol, acetic acid, and chronic ethanol consumption. The effects of HEBT were also evaluated in a pylorus ligature model and on gastrointestinal motility. The LD50 of HEBT in mice was additionally estimated. HEBT was analyzed by nuclear magnetic resonance, and a high-performance liquid chromatography fingerprint analysis was performed. Oral HEBT administration significantly reduced the lesion area and the oxidative stress induced by acute and chronic ethanol consumption. However, HEBT did not protect against gastric wall mucus depletion and did not alter gastric secretory volume, pH, or total acidity in the pylorus ligature model. Histologically, HEBT accelerated the healing of chronic gastric ulcers in rats, reflected by contractions of the ulcer base. Flavonoids and caffeoylquinic acids were detected in HEBT, which likely contributed to the therapeutic efficacy of HEBT, preventing or reversing ethanol- and acetic acid-induced ulcers, respectively. HEBT antiulcerogenic activity may be partially attributable to the inhibition of free radical generation and subsequent prevention of lipid peroxidation. Our results indicate that HEBT has both gastroprotective and curative activity in animal models, with no toxicity.