High-performance Liquid Chromatography Fingerprint Research Articles

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.

Chromatographic Fingerprinting Based on Column Switching Technology for Quality Evaluation of Tianmeng Oral Liquid.

Separation power was limited when the conventional high-performance liquid chromatography (HPLC) fingerprinting method based on a single column was used to analyze very complex traditional Chinese medicine (TCM) preparations. In this research, a novel HPLC fingerprinting method based on column switching technology by using a single pump was established for evaluating the quality of Tianmeng oral liquid (TMOL). Twelve batches of TMOL samples were used for constructing HPLC fingerprints. Compared with the 16 common peaks in fingerprinting with a single column, 25 common peaks were achieved with two columns connected through a six-way valve. The similarity analysis combined with bootstrap method was applied to determine the similarity threshold, which was 0.992 to distinguish expired samples and unexpired samples. Principal component analysis (PCA) and hierarchical clustering analysis (HCA) were also applied to classify the TMOL samples, and results revealed that expired and unexpired samples are classified into two categories. The HPLC fingerprinting based on column switching technology with better separation power and higher peak capacity could characterize chemical composition information more comprehensively, providing an effective and alternative method to control and evaluate the quality of TMOL, which would offer a valuable reference for other TCM preparations.

Comprehensive multicomponent characterization and fingerprinting analysis of Lanqin Oral Liquid by ultra-high-performance liquid chromatography coupled with ion mobility-quadrupole time-of-flight mass spectrometry.

Comprehensive characterization of the chemical constituents of Chinese patent medicine poses a great challenge due to the frustrating complexity resulting from superposition of multiple drugs. Lanqin Oral Liquid is a five-component Chinese patent medicine widely applied to treat pharyngeal inflammation in clinic. Here, we streamline a universal three-dimensional separation approach to efficiently identify the multicomponents from Lanqin Oral Liquid by ultra-high-performance liquid chromatography/ion mobility quadrupole time-of-flight mass spectrometry and UNIFI/in-house library-driven automatic peak annotation. Because of the systematic optimization, the use of an HSS T3 column enabled good separation of the multiple components within 42min, while high-definition MSE in both the negative and positive modes could characterize more classes of herbal components, thus providing the retention, collision cross-section, and MS information for each component. Benefiting from the ion mobility separation, cleaner MS1 and MS2 spectra were acquired. Aided by comparison and analysis of the fragmentation pathways of 49 reference compounds, we could characterize 175 compounds from Lanqin Oral Liquid. A validated high-performance liquid chromatography fingerprinting approach unveiled good similarity (0.985-1.000) among 22 batches of commercial samples. Conclusively, we demonstrated a practical solution to elucidating the chemical composition of Chinese patent medicines, with the potential of popularization.

Quality evaluation of Aralia taibaiensis based on spectrum-activity relationship

A spectrum-activity relationship is established with high performance liquid chromatography(HPLC) fingerprints and the in vitro antioxidant activity to improve the quality evaluation system of Aralia taibaiensis. The HPLC profiles of 12 batches of samples were collected, and the similarity evaluation, heat map analysis and principal component analysis were conducted for the chemometric study of the fingerprint data. Combined with grey correlation analysis, the contributions of the common peaks in the fingerprints to the antioxidant activity were clarified, and the important peaks reflecting the efficacy were identified. The results showed that 17 common peaks were found in 12 batches of A. taibaiensis samples, and 6 of them were identified as saponins. Similarity evaluation, heat map analysis and principal component analysis roughly classified the A. taibaiensis herbs into two categories, i.e.,(1) S1-S10, S12 and(2) S11. Twelve batches of samples showed different antioxidant activities in a dose-dependent manner. In particular, S9 had the strongest antioxidant activity, while S11 was the weakest in antioxidant capacity, which was basically consistent with the overall score results. The results of grey correlation analysis demonstrated that the 17 common peaks scavenged DPPH radicals in the following order: X_3&gt;X_(17)&gt;X_4&gt;X_8&gt;X_7&gt;X_(13)&gt;X_2&gt;X_6&gt;X_(11)&gt;X_(10)&gt;X_(16)&gt;X_(12)&gt;X_9&gt;X_5&gt;X_(14)&gt;X_1&gt;X_(15), and scavenged ABTS radicals in the order of X_4&gt;X_3&gt;X_7&gt;X_8&gt;X_2&gt;X_(17)&gt;X_(13)&gt;X_6&gt;X_(16)&gt;X_(11)&gt;X_5&gt;X_(12)&gt;X_(10)&gt;X_9&gt;X_(14)&gt;X_1&gt;X_(15). Among them, X_3, X_4, X_7(araloside C), X_8 and X_(17) were the important peaks reflecting the efficacy of A. taibaiensis, which were basically consistent with those contained in the principal component 1. In this study, the correlation between the HPLC fingerprints of 12 batches of A. taibaiensis and its antioxidant activity provides a reference for the Q-marker screening and quality control of A. taibaiensis.

Spectrum-effect relationship of the blood-activating efficacy of Notoginseng Radix et Rhizoma

This study aims to establish the high-performance liquid chromatography(HPLC) fingerprints of different batches of Notoginseng Radix et Rhizoma, determine their pharmacodynamic indexes of promoting blood circulation, and explore the spectrum-effect relationship between the chemical components of Notoginseng Radix et Rhizoma and the efficacy of promoting blood circulation. Firstly, the HPLC fingerprints of different batches of Notoginseng Radix et Rhizoma were established. Then, the pharmacodynamic indexes were determined after the capillary coagulation experiment and the cerebral ischemia-reperfusion in rats, including capillary coagulation time, percentage of cerebral ischemic area, cerebral water loss rate, and brain-body index. Afterward, the partial least-squares method was used to explore the spectrum-effect relationship between the chemical components of Notoginseng Radix et Rhizoma and the pharmacodynamic indexes. The results showed that this study successfully established the HPLC fingerprints of different batches of Notoginseng Radix et Rhizoma, found 23 common peaks, and identified 12 of them, all of which were saponins. The method was proved stable and reliable. Both the capillary coagulation experiment and the middle cerebral artery occlusion(MCAO)-induced cerebral ischemia-reperfusion experiment on rats revealed that there were obvious differences in the pharmacodynamic indexes of different batches of Notoginseng Radix et Rhizoma. The relationships between 23 common components of Notoginseng Radix et Rhizoma in different batches and the pharmacodynamic indexes were discussed by means of spectrum-effect correlation analysis, of which 17 components had positive effects while 6 components had negative effects on the pharmacodynamic indexes. This study provides a certain reference basis for the clinical rational use and quality control of Notoginseng Radix et Rhizoma.

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.

Physicochemical Characterization, Phytochemical and HPTLC Fingerprinting Studies on Fruit of Couroupita Guianensis

Abstract Traditional knowledge and literature studies report that each part of a plant has tremendous medicinal values. Validation of these medicinal plants scientifically is an important criterion for the development of plant-based drugs. Couroupita guianensis (Family: Lecythidaceae) is a plant with immense medicinal properties. To authenticate its biological value, the present investigation aims to standardize the fruit of C. guianensis based on physicochemical characterization, phytochemical analysis both qualitatively and quantitatively, and high-performance thin-layer liquid chromatography (HPTLC) fingerprinting studies. Fruit pulp of C. guianensis was obtained, processed and extracted with solvents such as petroleum ether, chloroform, ethanol and hydroalcohol. Moisture content, total ash, water-soluble ash and acid-insoluble ash values were calculated. Preliminary phytochemical analysis revealed the existence of several secondary metabolites in the extracts. In addition, interpreting peaks obtained from HPTLC analysis revealed the presence of potential bioactive phytoconstituents in all the extracts. The quantitative determination proclaimed that fruit pulp was found to be rich in phenolics and flavonoids followed by tannin and saponin. Further, primary metabolites were quantified and they were found to be abundant in the fruit pulp. Henceforth, the outcome of these results provides information for assessing the quality of the sample that could help in ensuring its therapeutic efficacy. Keywords: Couroupita guianensis, HPTLC, Physicochemical characterization, Phytochemical, Fingerprinting

Comprehensive analysis of components in Chinese medicines derived from Apocynum venetum and Poacynum pictum in Xinjiang based on HPLC fingerprint and chemometrics

This study established high-performance liquid chromatography(HPLC) fingerprints of Chinese medicines derived from Apocynum venetum and Poacynum pictum in Xinjiang and explored their composition differences with the combination of content determination, similarity analysis, cluster analysis and principal component analysis. The HPLC conditions included Phenomenex Kinetex C_(18) column(4.6 mm ×100 mm, 2.6 μm), acetonitrile-0.01% trifluoroacetic acid aqueous solution as mobile phase, gradient elution, flow rate of 0.6 mL·min~(-1), detection wavelength of 281 nm and column temperature of 25 ℃. The content of chlorogenic acid, quercetin-3-O-sophoroside, rutin, hyperin, isoquercitrin, trifolin and astragalin was determined in 31 batches of medicinal materials, and fingerprint research and chemometric analysis were performed with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2004 A) and SPSS 21.0. In the Chinese Pharmacopoeia 2020, the quality of Apocyni Veneti Folium is controlled by character identification, microscopic identification, thin layer chromatography identification and quantitative determination of hyperin. There were 21 common peaks of A. venetum and P. pictum in the HPLC fingerprints, 5 of which were identified as chlorogenic acid, hyperin, isoquercitrin, trifolin and astragalin, with their content also determined. Except for 3 batches of medicinal materials, the similarity of other 28 batches was higher than 0.83, indicating good similarity. Two categories were formed in the cluster analysis based on content determination, which showed that some differences existed in similarities between different regions of Xinjiang. The medicinal materials were ranked by quality with principal component analysis, and the results indicated that the top 15 all came from northern Xinjiang. The quality difference of A. venetum and P. pictum had a correlation with the place of origin. This study provides a reference for the analysis and evaluation of A. venetum and P. pictum from different habitats and the selection of introduction and cultivation areas.

Determining the effective compounds of Salvia miltiorrhiza through chromatographic fingerprinting coupled with chemometrics

Herbs are rich in the active ingredients of drugs for preventing or treating various disorders. However, conventional bioactivity-guided separation is time and labor-intensive and neglects the additive effect of multiple components. These problems hinder the development of new medicines from natural products. This study established a chemometric analysis method that integrates processes based on the spectrum-effect relationship for the rapid identification of the primary active components of a plant. The high-performance liquid chromatography (HPLC) fingerprints of 171 Salvia miltiorrhiza extracts (SMEs) with varied constituent profiles were analyzed. Chemometric analysis was performed to establish an HPLC fingerprint–bioactivity relationship to explore the components of SMEs that contribute to the antioxidant activity and cytotoxicity effect, respectively. The results indicated that the developed strategy can be used to identify components largely contributing to particular bioactivities and re-evaluate the efficacy of previously neglected components. The present study identified not only the primary active components of S. miltiorrhiza but also the optimal ratios of constituents, validating the method for use in the future investigation and development of herbal medicines. Keywords: Chemometric, herb, antioxidant, cytotoxicity, fingerprint, Salvia miltiorrhiza.

Spectrum-Effect Relationship between HPLC Fingerprints and Antioxidant Activity of Yangyin Tongnao Prescription.

Yangyin Tongnao (YYTN) prescription is used as a traditional Chinese herbal formula, and it has antioxidant activity that mainly contributes in the treatment of cardiovascular and cerebrovascular diseases. However, the compounds related to its antioxidant activity are still unknown. In the present study, the fingerprints of YYTN extracts under different extraction conditions were obtained by high performance liquid chromatography (HPLC) to identify the common peaks to all the samples processed. A 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assay and ferric reducing antioxidant power (FRAP) assay were carried out to evaluate the antioxidant activity of the extracts. Spectrum-effect relationship between HPLC fingerprints and antioxidant activity of YYTN was assessed by Pearson product-moment correlation coefficient (PPMCC) and multiple linear regression analysis (MLRA). The results showed that peaks 5, 6, 13, 15, and 24 of the fingerprints were closely connected to antioxidant activity. Five peaks were identified: vanillic acid (P5), puerarin (P7), ferulic acid (P13), daidzein (P21), and formononetin (P23). Our study successfully established the spectrum-effect relationship between HPLC fingerprints and antioxidant activity of YYTN, which provided a general method for establishing quality standards with a combination of chromatography and antioxidant activity.

Screen of anti-migraine active compounds from Duijinsan by spectrum-effect relationship analysis and molecular docking

Ethnopharmacological relevanceDuijinsan (DJS) is a famous Chinese medicine prescription composed of Radix scutellariae (RS) and Rhei Radix (RRR), which has been mainly used for treating migraine. Aim of the studyThis study aimed to uncover the anti-migraine active compounds from DJS and preliminary predicted the pharmacological mechanism by evaluating the spectrum-effect relationship between high-performance liquid chromatography (HPLC) fingerprints and anti-migraine effects of Duijinsan (DJS) extract combined with molecular docking. Materials and methodsHPLC and LC-MS were applied for chemical analyses of DJS extracts in different proportions. Inhibition of DJS extracts on trigeminal nerve cell releasing calcitonin gene related peptide (CGRP) experiment was performed. The active compounds were screened by spectrum-effect relationship analysis and confirmed by molecular docking and the activities of major predicted compounds were validated in vitro. ResultsTwenty-six common peaks were assigned and identified from the fingerprints of different proportions DJS extracts. In vitro experimental results showed that DJS extracts inhibited inflammation and release of CGRP from trigeminal nerve cells. Five predicted active compounds, Chrysin 6-C-arabinoside 8-C-glucoside, Chrysin 6-C-glucoside 8-C-arabinoside, baicalin, Chrysin-7-O-Beta-D-glucoronide and Oroxylin A 7-O-glucuronide were sorted out according to spectrum-effect relationship analysis and molecular docking comprehensively. In vitro validation experiments showed that all the predicted compounds inhibited the CGRP releasing and the activation of TRPV1 channel. Baicalin, chrysin-7-O-β-D-glucuronide and Oroxylin A-7-glucoronide significantly inhibited the activation of TRPV1 channel. ConclusionChrysin 6-C-arabinoside 8-C-glucoside, Chrysin 6-C-glucoside 8-C-arabinoside, baicalin, Chrysin-7-O-Beta-D-glucoronide and Oroxylin A 7-O-glucuronide which can inhibit the CGRP releasing and the activation of TRPV1 channel were screened as the anti-migraine active compounds by spectrum-effect relationship analysis and molecular docking.

Quality monitoring and evaluation of Powdered Poppy Capsule Extractive by multiple wavelengths matching average fusion chromatographic fingerprint combined with electrochemical fingerprint

As quality determined the effectiveness and safety of the drug, it is essential to investigate and control the quality of Powdered Poppy Capsule Extractive (PPCE). In this paper, the qualitative and quantitative evaluation results of PPCE were obtained by using electrochemical fingerprint and high performance liquid chromatographic fingerprint. In the multi-wavelength matching average fusion chromatographic fingerprint, three parameters of macro qualitative similarity (Sc), macro quantitative similarity (Pc) and the variation coefficient of fingerprint homogeneity (α) were calculated by the limited quantified ratio fingerprint method (LQRFM). The Sc values of the samples were all greater than 0.986, the Pc values between 94.1 and 104.7, and the α values all less than 0.024 calculated by LQRFM. The calibration curve method was used to determine the contents of two markers in the samples. The loading plot of contents for two markers illustrated that the samples were divided into three categories. Firstly, the contents of morphine and codeine phosphate were below 9.3% and 4.20% in Group I (S2, S3, S6, S8, S27 and S30). Secondly, the codeine phosphate contents of Group II (S7, S9, S10, S16, S20, S24, S28 and S29) were less than 4.2%, but the morphine contents were over 9.3%.Finally, Samples with codeine phosphate content higher than 4.2% and morphine higher than 9.3% were classified as the Group III. Combined with cluster analysis of the maximum amplitude in the electrochemical fingerprint and the content determination results of PPCE, the results showed a high similarity. This study demonstrated that the electrochemical fingerprint was suitable for the identification of traditional Chinese medicine, and LQRFM was reliable and accurate for the quality control of complex traditional Chinese medicine or its related products.

Chemometric analysis and quantitative determination of Fici Microcarpae Folium from different geographical regions by HPLC-DAD

Fici Microcarpae Folium, the leaves of Ficus microcarpa, is commonly used for the treatment of influenza, chronic bronchitis, and tonsillitis in Traditional Chinese Medicine (TCM). Ficus microcarpa is mainly distributed in Guangdong and Guangxi provinces. The composition of herbal medicines can be easily affected by geographical regions, the quality control of FMF is essential. In this study, the high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD) was developed to obtain a chromatographic fingerprint of Fici Microcarpae Folium from different geographical regions. Meanwhile, the contents of eight compounds of Fici Microcarpae Folium were determined. Principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares discrimination analysis (OPLS-DA) were employed to evaluate the HPLC fingerprint. The results showed that the main compositions of Fici Microcarpae Folium from different geographical origins had no difference. The content of the main compositions was different, from which the peak 8, 15, 12, 11, 16, 3, and 21 were the main components that contribute to the difference. Isovitexin, vitexin, and isoorientin were the main components reflecting the quality of Fici Microcarpae Folium. This research could contribute to the quality control of Fici Microcarpae Folium.

High performance liquid chromatography fingerprint and headspace gas chromatography-mass spectrometry combined with chemometrics for the species authentication of Curcumae Rhizoma

Curcumae Rhizoma (Ezhu), a multi-origin Chinese medicine, originates from the dry rhizomes of C. kwangsiensis, C. phaeocaulis and C. wenyujin. The three species have great variation in chemical components and therapeutic effects. To improve safety and effectiveness in clinical use, a strategy integrating chromatographic analysis and chemometrics for the species authentication of Ezhu was proposed. Firstly, systematic analysis of chemical compositions in Ezhu was achieved using high performance liquid chromatography (HPLC) fingerprint and headspace gas chromatography-mass spectrometry (HS-GC–MS). HPLC fingerprints showed that seventeen peaks in common for C. kwangsiensis and eleven peaks in common for C. wenyujin both presented a good similarity (> 0.9, only several samples < 0.8). Eleven common peaks in C. phaeocaulis and the similarity values of most samples were higher than 0.700. Additionally, there were ten common peaks in all Ezhu samples and they had relatively poor similarity with the correlation coefficients ranging from 0.364 to 0.881. For HS-GC–MS, thirty-six volatile components were identified in the three species of Ezhu, mainly monoterpenes and sesquiterpenes. Subsequently, chemometrics including unsupervised principal component analysis (PCA), supervised linear discriminant analysis (LDA), K-nearest neighbors (KNN), back propagation neural network (BP-NN) and orthogonal partial least squares-discrimination analysis (OPLS-DA) was applied to extract useful information from chromatographic profiles. Based on HPLC fingerprint data, PCA could hardly differentiate Ezhu with the three species, and LDA, KNN and BP-NN models provided more than 85 % correct identification. With HS-GC–MS data, PCA could only distinguish C. wenyujin from the other two species, and LDA, KNN, BP-NN and OPLS-DA models achieved excellent classification with 100 % accuracy. Finally, five volatile components (eucalyptol, humulene, β-elemene, (+)-2-bornanone and linalool) with variable importance for the projection (VIP) values higher than 1 in the OPLS-DA model were selected as potential chemical markers for the species authentication of Ezhu. And the constructed OPLS-DA model using these markers obtained 100 % accuracy. Consequently, a rapid, precise and feasible strategy was established for the discrimination and quality control of Ezhu with different species.

In vitro Cytotoxicity, Antimicrobial, Antioxidant Activities and HPLC Finger Print Analyses of the Extracts of Ceiba insignis Leaves Growing in Egypt

Polyphenols - rich plants can be used to reduce health disorder resulting from cancer and infectious diseases. Methodology: Different solvent extracts of Ceiba insignis leaves were evaluated for their in vitro cytotoxic, antimicrobial, and antioxidant activities, while their chemical profiles were investigated via high‑performance liquid-chromatography (HPLC) fingerprint analyses. Results: Showed that methanol, butanol, and dichloromethane extracts of Ceiba insignis leaves exhibited a moderate cytotoxicity against HepG2 with IC50 values of 98.54, 75.38, 40.71 µg/ml respectiviely, while petroleum ether and water exhibited weak activity with IC50 values of 118.15 and 170.03 µg/ml respectively and very weak activity was recorded with ethyl acetate extract with IC50 value of 924.05µg. Moreover, methanol, dichloromethane, petroleum ether, and ethyl acetate exhibited strong antimicrobial activities with inhibition zones (20-25 mm), (15-30 mm), (14-16 mm), and (16-28 mm) respectively, while butanol and water exhibited low to moderate activity with inhibition zones (0-18 mm), and (0-16 mm) respectively. On the other side, strong DPPH antioxidant scavenging activity was recorded within petroleum ether extract with IC50 (24.72 μg/ml), while the least antioxidant activity was recorded within ethyl acetate with IC50 (97.50 μg/ml). HPLC finger-print analyses revealed the presence of major compounds; syringic acid in dichloromethane extract, gallic acid, chlorogenic acid & syringic acid in ethyl acetate extract, and naringenin & gallic acid in methanol, butanol and water extracts, this finding provides an insight into the usage of the tested species as a source of naturally occurring antioxidant, cytotoxic and antimicrobial agents.

HPLC fingerprint and UV–Vis spectroscopy coupled with chemometrics for Curcumae radix species discrimination and three bioactive compounds prediction

Curcumae Radix with four botanical origins (Wenyujin, Huangyujin, Guiyujin, Lvyujin) has a long medical history in China. The four species have not been differentiated well in clinical application, though there were differences in pharmacological effects among them. For the aim of fast discrimination and quantitative analysis of Yujin, HPLC (high performance liquid chromatography) fingerprint and UV–Vis (ultraviolet–visible) spectroscopy technologies coupled with chemometrics were employed. The HPLC fingerprints of 26 Guiyujin, 11 Huangyujin, 11 Lvyujin, 29 Wenyujin and full 77 samples were separately established, with good similarities among same species except Wenyujin and poor consistence in full samples. UV–Vis spectra showed obvious three absorption peaks in the spectra of Huangyujin samples and only one in that of the rest samples. LDA (linear discriminant analysis) and Bagged Trees models established based on HPLC fingerprint data attained higher than 90% discrimination accuracy. Based on the spectral data, complete separation for the four species of Yujin was failed to achieve by PCA (principal component analysis), and the KNN (k-nearest neighbors) and SVM (support vector machine) models achieved no<90% discrimination accuracies. Furthermore, the SVMR (support vector machine regression) and Bagged Trees Regression models used to quickly predict the contents of germacrone, curcumenol and curdione performed well, with relatively lower RMSE (root mean square error) and higher R2 (coefficient of determination) values. The present study proposed relatively rapid methods for the determination of bioactive compounds and discrimination of Yujin.

High‑Performance Liquid Chromatography‑Fingerprint Analyses, In vitro Cytotoxicity, Antimicrobial and Antioxidant Activities of the Extracts of Ceiba speciosa Growing in Egypt

ackground: Cancer diseases and microbial resistance are serious health diseases related to oxidative stress and infectious diseases. The risk can be reduced by using plants rich in polyphenols. Methodology: Different solvent extracts from leaves of Ceiba speciosa (C.s) were evaluated for their biological and chemical activities. Also, the chemical profiles were investigated via high‑performance liquid-chromatography (HPLC) fingerprint analyses. Results: the leaves from Ceiba speciosa collected from the Zoo garden in Egypt, showed a moderate cytotoxicity against HepG2 in extracts; petroleum ether, ethyl acetate, dichloromethane, while weak-cytotoxicity in butanol, methanol extracts and non-cytotoxicity in water extract. Moreover, high antimicrobial activities were showed within dichloromethane, petroleum ether extracts, while a moderate antimicrobial activities were showed in methanol, ethyl acetate extracts, and low antimicrobial activities were showed in both of butanol and water extracts. On the other side, a strong antioxidant activities were recorded within extracts; dichloromethane and methanol while the least antioxidant activity was recorded within water extract. High performance liquid-chromatography (HPLC) fingerprint analyses is done for all fractions, this finding provides an insight into the usage of the tested species as a source of naturally occurring cytotoxic, antimicrobial and antioxidant agents

Antitumor chemical constituents of Toddalia asiatica (Linn) Lam root bark and its rational alternative medicinal parts by multivariate statistical analysis

AbstractToddalia asiatica(Linn) Lam (T. asiatica) as a traditional Miao medicine was investigated to find rational alternative medicinal parts forT. asiaticaroot bark and its antitumor chemical constituents by quantitative pharmacognostic microscopy, high performance liquid chromatography (HPLC) fingerprint and multivariate statistical analysis. A bivariate correlation analysis method based on microscopic characteristics and content of chemical constituents was established for the first time, there were some regular discoveries between powder microscopic characteristics and common chromatographic peaks ofT. asiaticathrough quantitative pharmacognostic microscopy, cork cells, calcium oxalate square crystal, brown clump, starch granule and phloem fiber, as powder microscopic characteristics may be placed where the main chemical constitutes were enriched. Scores plot of principal component analysis (PCA) and dendrogram of hierarchical clustering analysis (HCA) showed that 18T. asiaticasamples were distinguished correctly, clustered clearly into two main groups as follows: S01∼S03 (root bark) and S07∼S09 (stem bark) in cluster 1, S04∼S06 and S10∼S18 in cluster 2. Nineteen common peaks were obtained in HPLC fingerprint ofT. asiatica, loadings plot of PCA indicated seven compounds played important roles in different part of samples (P10 &gt; P08 &gt; P07 &gt; P14 &gt; P16 &gt; P17 &gt; P19), peaks 04, 06, 07, 08, 10 were identified as hesperidin, 4-methoxycinnamic acid, toddalolactone, isopimpinlline and pimpinellin. MTT assay was used to determine the inhibitory activity of different medicinal parts ofT. asiaticaon human breast cancer MCF-7 cells, all parts ofT. asiaticahad different inhibitory effects on MCF-7 cell lines, root and stem barks ofT. asiaticashowed the best inhibitory activity. The relationship between chemical constituents and the inhibitions on MCF-7 cell had been established, significant antitumor constituents ofT. asiaticawere identified by correlation analysis, the order of the antitumor effect of the main compounds was P07 (toddalolactone) &gt; P16 &gt; P06 (4-methoxycinnamic acid), P11 &gt; P18 &gt; P10 (pimpinellin) &gt; P08 (isopimpinellin) &gt; P01 &gt; P19 &gt; P14 &gt; P04 (hesperidin) &gt; P17, which were antitumor chemical constituents ofT. asiaticaroot bark.T. asiaticastem bark was the most rational alternative medicinal part forT. asiaticaroot bark.

Study on the Spectrum-Effect Relationship of the Traditional Effect of Saponins in Glycyrrhiza uralensis Fisch.

Licorice is a traditional Chinese medicine that has been used for a long time in China and still in great use today. The effect of licorice on tonifying spleen and invigorating qi has been proved for thousands of years, but the material basis of its effect is not clear. In this paper, we established the fingerprints of 21 batches of licorice collected from different origins in China with High-Performance Liquid Chromatography (HPLC) to identify the common peaks. Its effect of tonifying spleen and invigorating qi was confirmed through a series of praxiology experiments. The spectrum-effect relationship between HPLC fingerprints and its effect of tonifying spleen and invigorating qi of licorice was examined by gray relational analysis and partial least squares regression analysis. Results showed that the effect of licorice on tonifying spleen and invigorating qi resulted from various compounds and peaks. X2–X6 is presumed to be the main pharmacological substance base. This research successfully identified the spectrum-effect relationship between HPLC fingerprints and the effect of licorice on tonifying spleen and invigorating qi. The research method based on the spectrum-effect relationship helps provide new research ideas and strategies for the study of the basis of the medicinal materials and quality control of traditional Chinese medicine.