Yifeng Chai

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Organization: Second Military Medical University

Department: School of Pharmacy

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Chemicals
References

Time Course Analysis of Candida albicans Metabolites during Biofilm Development

Co-reporter:ZhenYu Zhu;Hui Wang;QingHua Shang;YingYing Cao;YuanYing Jiang

Journal of Proteome Research June 7, 2013 Volume 12(Issue 6) pp:2375-2385

Publication Date(Web):2017-2-22

DOI:10.1021/pr300447k

Biofilm-associated infections are difficult to treat because of their decreased susceptibility to antimicrobial therapy. Candida albicans is the most common fungal pathogen associated with colonization and biofilm formation on the surfaces of indwelling medical devices which show intrinsic resistance to many commonly used antifungal agents. In this study, a metabonomic method using gas chromatography–mass spectrometry (GC/MS) was developed to characterize metabolic profiles during the whole biofilm developmental phases compared to the planktonic mode in C. albicans. Thirty-one differentially produced metabolites between the biofilm and planktonic specimens at each time point were identified, and they were mainly involved in the tricarboxylic acid (TCA) cycle, lipid synthesis, amino acid metabolism, glycolysis, and oxidative stress. Further experiments showed that lack of trehalose, one of the metabolites differentially produced between biofilm and planktonic cells, resulted in abnormal biofilm formation and increased sensitivity to amphotericin B and miconazole. This study provides a systemic view of the metabolic pattern during the development of C. albicans biofilms, indicating that multicomponent, phase-specific mechanisms are operative in the process of biofilm formation.Keywords: biofilm; Candida albicans; drug resistance; metabolite; trehalose;

Development of APTES-Decorated HepG2 Cancer Stem Cell Membrane Chromatography for Screening Active Components from Salvia miltiorrhiza

Co-reporter:Yongfang Yuan;Yue Liu;Dan Jia;Lei Lv;Zhanying Hong;Zhenyu Zhu;Xiaofei Chen;Dongyao Wang;Guoqing Zhang;Xuan Ding;Yan Cao;Zhirong Gong;Liang Zhao

Analytical Chemistry December 20, 2016 Volume 88(Issue 24) pp:12081-12089

Publication Date(Web):November 16, 2016

DOI:10.1021/acs.analchem.6b02709

Cell membrane chromatography (CMC) is an ideal method for screening potential active components acting on target cell membranes from a complex system, such as herbal medicines. But due to the decay and falling-off of membranes, the CMC column suffers from short life span and low reproducibility. This has greatly limited the application of this model, especially when the cell materials are hard to obtain. To solve this problem, a novel type of (3-aminopropyl)triethoxysilane (APTES)-decorated silica gel was employed. The silica gel was decorated with aldehydes with the help of APTES, which react with the amino groups on cell membranes to form a covalent bond. In this way, cell membranes were immobilized on the surface of silica gel, so it is not easy for membranes to fall off. According to our investigation, the column life of the APTES-decorated group was prolonged to more than 12 days, while the control group showed a sharp decline in column efficiency in the first 3 days. To verify this model, a novel APTES-decorated HepG2 cancer stem cell membrane chromatography (CSCMC) was established and applied in a comprehensive two-dimensional chromatographic system to screen potential active components in Salvia miltiorrhiza. As a result, tanshinone IIA, cryptotanshinone, and dihydrotanshinone I were retained on this model and proved to be effective on HepG2 cancer stem cells by the following cell proliferation and apoptosis assay, with IC50 of 10.30 μM, 17.85 μM, and 2.53 μM, respectively. This improvement of CMC can significantly prolong its column life span and broaden the range of its application, which is very suitable for making invaluable or hard-to-obtain cell materials, such as stem cells, for specific drug screening.

Identification of Annexin A2 as a target protein for plant alkaloid matrine

Co-reporter:Dongyao Wang;Yan Cao;Leyi Zheng;Diya Lv;Langdong Chen;Xinrui Xing;Zhenyu Zhu;Xiaoyu Li

Chemical Communications 2017 vol. 53(Issue 36) pp:5020-5023

Publication Date(Web):2017/05/02

DOI:10.1039/C7CC02227A

Matrine is a plant alkaloid and a major active component in the Chinese medical herb Sophora flavescens. Matrine has shown potent anti-cancer activities but its molecular target(s) and mechanism are still unknown. Using the photo-affinity labeling approach, for the first time, Annexin A2 was identified as a direct-binding target of matrine in cancer cells.

A metabolomics approach for predicting the response to intravenous iron therapy in peritoneal dialysis patients with anemia

Co-reporter:Qiong Wu;Xue-li Lai;Hong-xia Zhao;Zhen-yu Zhu;Zhan-ying Hong;Zhi-yong Guo;Yi-feng Chai

RSC Advances (2011-Present) 2017 vol. 7(Issue 4) pp:1915-1922

Publication Date(Web):2017/01/04

DOI:10.1039/C6RA24152B

Anemia is an almost universal complication of chronic kidney disease (CKD), and nearly all patients with end-stage renal disease (ESRD) and approximately 70% of those with earlier stages of CKD receive treatment for anemia. Due to its significance in the treatment of anemia, there is increased reliance on iron in the renal anemia population. In clinical practice, not every patient benefits from intravenous (IV) iron therapy. In order to identify patients who will respond to IV iron therapy and who will not respond to it, our goals were to identify the potential serum biomarkers that could predict the response to IV iron therapy in renal anemia patients. The metabolic profiles of serum from 41 renal anemia patients with complete, partial or non-response to IV iron therapy were studied using a combination of liquid chromatography coupled with mass spectrometry (LC-MS) and multivariate analysis methods to identify the potential biomarkers that could predict the response to IV iron therapy in renal anemia patients. Oleamide and ascorbate 2-sulfate (AS) were identified and verified as the potential biomarkers. A prediction model constructed with oleamide and AS correctly identified approximately 83.3% of patients who were non-responsive to IV iron therapy and 87.5% of patients who had a complete response to IV iron therapy. The model has excellent discriminant performance, with an AUC of 0.901. These results show promise for larger studies that could advance more personalized treatment protocols for renal anemia patients.

Rapid identification of dual p53-MDM2/MDMX interaction inhibitors through virtual screening and hit-based substructure search

Co-reporter:Si Chen;Xiang Li;Weirong Yuan;Yan Zou;Zhongwu Guo;Wuyuan Lu

RSC Advances (2011-Present) 2017 vol. 7(Issue 16) pp:9989-9997

Publication Date(Web):2017/01/30

DOI:10.1039/C7RA00473G

Multi-target agents have garnered great interest over the past decade for their favorable therapeutic efficacy and drug resistance profiles. Recently, dual inhibition of the p53 tumor suppressor interaction with its two negative regulators MDM2 and MDMX has become an attractive anticancer approach as it can induce sustained MDM2/MDMX antagonism and robust p53 activation. However, small molecule inhibitors with dual specificity against MDM2 and MDMX are difficult to design and are still scarce. To identify novel scaffolds for dual inhibition of the p53-MDM2/MDMX interactions, we developed two five-point pharmacophore models for filtering the 2012 National Cancer Institute database, from which molecular docking was conducted to identify dual inhibitors. We found 38 virtual hits and subjected them to a fluorescence polarization-based competitive binding assay, resulting in 10 active compounds of different scaffolds. To further expand the chemical diversity of the initial hits, we performed a hit-based substructure search and identified NSC148171 from pharmacophore 1 as the most potent dual-specificity inhibitor with Ki values for MDM2 and MDMX at 0.62 and 4.6 μM. All hits were subjected to inhibition assay of cancer cellular vitality and showed anti-proliferative activity roughly correlated with their Ki values. This work not only yields several novel scaffolds for further structural and functional optimization of dual-specificity inhibitors of the p53-MDM2/MDMX interactions, but also showcases the power of our computational methods for small molecule anticancer drug discovery.

Effect of processing on the alkaloids in Aconitum tubers by HPLC-TOF/MS

Co-reporter:Min Liu, Yan Cao, Diya Lv, Wen Zhang, ... Yifeng Chai

Journal of Pharmaceutical Analysis 2017 Volume 7, Issue 3(Volume 7, Issue 3) pp:

Publication Date(Web):1 June 2017

DOI:10.1016/j.jpha.2017.01.001

According to the Chinese Pharmacopoeia 2015, only processed Aconitum tubers can be clinically applied, and the effect of processing is unclear. This research aimed to explore the effect of processing on cardiac efficacy of alkaloids in Aconitum tubers. First, the chemical ingredients in unprocessed and processed Aconitum tubers were identified and compared by using high performance liquid chromatography time-of-flight mass spectrometry (HPLC-TOF/MS) and multivariate pattern recognition methods. Then the representative alkaloids in Aconitum tubers, aconitine, benzoylaconine, and aconine, which belong to diester-diterpenoid alkaloids, monoester-diterpenoid alkaloids, and amine-diterpenoid alkaloids, respectively, were selected for further validation of attenuated mechanism. Subsequent pharmacological experiments with aconitine, benzoylaconine, and aconine in SD rats were used for validate the effect of processing on cardiac functions. After processing the Aconitum tubers, it was found that the contents of diester-diterpenoid alkaloids were reduced, and those of monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids were increased, suggesting that diester-diterpenoid alkaloids were transformed into monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids. Through further decocting the aconitine in boiling water, it was confirmed that the three alkaloids could be progressively transformed. Pharmacological experiments with aconitine, benzoylaconine, and aconine in SD rats showed that aconitine at a dose of 0.01 mg/kg and aconine at a dose of 10 mg/kg enhanced the cardiac function, while benzoylaconine at a dose of 2 mg/kg weakened the cardiac function. The effect of processing is attributed to the transformation of the most toxic diester-diterpenoid alkaloids into less toxic monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids.

On-line comprehensive two-dimensional HepG2 cell membrane chromatographic analysis system for charactering anti-hepatoma components from rat serum after oral administration of Radix scutellariae: A strategy for rapid screening active compounds in vivo

Co-reporter:Dan Jia, Xiaofei Chen, Yan Cao, Xunxun Wu, Xuan Ding, Hai Zhang, Chuan Zhang, Yifeng Chai, Zhenyu Zhu

Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 118() pp:27-33

Publication Date(Web):25 January 2016

DOI:10.1016/j.jpba.2015.10.013

•A comprehensive 2D CMC system was developed and applied to characterize active components from drug-containing serum for the first time.•A matrix interference deduction method effectively eliminated the interference of endogenous substances in serum.•Wogonin and oroxylin A screened both in vitro and in vivo, could be selected as anti-hepatoma leading compounds.•The methods had advantages in simplicity, sensitivity, accurate recognition and identification.Cell membrane chromatography (CMC) is a bioaffinity chromatography technique for characterizing interactions between drugs and membrane receptors and has been widely used to screen active components from complex samples such as herbal medicines (HMs). However, it has never been applied in vivo due to its relatively high limit of detection (LOD) and the matrix interferences. In this study, a novel on-line comprehensive two-dimensional HepG2/CMC/enrich columns/high performance liquid chromatography/time-of-flight mass spectrometry system was developed to rapidly screen potential anti-hepatoma components from drug-containing serum of rats after oral administration of Radix scutellariae. A matrix interference deduction method with a home-written program in MATLAB was developed, which could successfully eliminate the interference of endogenous substances in serum. Baicalein, wogonin, chrysin, oroxylin A, neobaicalein and rivularin from Radix scutellariae extraction were significantly retained in the HepG2/CMC column. Three potential active components, wogonin, oroxylin A and neobaicalein were firstly screened from the drug-containing serum as well. The cell counting kit-8 assay demonstrated that wogonin, oroxylin A and chrysin showed high inhibitory activities in a dose-dependent manner on HepG2 cells at the concentration of 12.5–200 μM (p < 0.05) and the IC50 values were 69.83, 16.66 and 51.6 μM, respectively. Wogonin and oroxylin A, which were screened both from Radix scutellariae extraction and the drug-containing serum, could be selected as lead compounds to obtain good anti-hepatoma effects. The proposed comprehensive 2D CMC system and matrix interference elimination strategy have significant advantages for in vivo screening of active components from complex biological samples and could be applied to other biochromatography models.

Comparative Normal/Failing Rat Myocardium Cell Membrane Chromatographic Analysis System for Screening Specific Components That Counteract Doxorubicin-Induced Heart Failure from Acontium carmichaeli

Co-reporter:Xiaofei Chen, Yan Cao, Hai Zhang, Zhenyu Zhu, Min Liu, Haibin Liu, Xuan Ding, Zhanying Hong, Wuhong Li, Diya Lv, Lirong Wang, Xianyi Zhuo, Junping Zhang, Xiang-Qun Xie, and Yifeng Chai

Analytical Chemistry 2014 Volume 86(Issue 10) pp:4748

Publication Date(Web):April 14, 2014

DOI:10.1021/ac500287e

Cell membrane chromatography (CMC) derived from pathological tissues is ideal for screening specific components acting on specific diseases from complex medicines owing to the maximum simulation of in vivo drug-receptor interactions. However, there are no pathological tissue-derived CMC models that have ever been developed, as well as no visualized affinity comparison of potential active components between normal and pathological CMC columns. In this study, a novel comparative normal/failing rat myocardium CMC analysis system based on online column selection and comprehensive two-dimensional (2D) chromatography/monolithic column/time-of-flight mass spectrometry was developed for parallel comparison of the chromatographic behaviors on both normal and pathological CMC columns, as well as rapid screening of the specific therapeutic agents that counteract doxorubicin (DOX)-induced heart failure from Acontium carmichaeli (Fuzi). In total, 16 potential active alkaloid components with similar structures in Fuzi were retained on both normal and failing myocardium CMC models. Most of them had obvious decreases of affinities on failing myocardium CMC compared with normal CMC model except for four components, talatizamine (TALA), 14-acetyl-TALA, hetisine, and 14-benzoylneoline. One compound TALA with the highest affinity was isolated for further in vitro pharmacodynamic validation and target identification to validate the screen results. Voltage-dependent K+ channel was confirmed as a binding target of TALA and 14-acetyl-TALA with high affinities. The online high throughput comparative CMC analysis method is suitable for screening specific active components from herbal medicines by increasing the specificity of screened results and can also be applied to other biological chromatography models.

Quality improvements of cell membrane chromatographic column

Co-reporter:Xuan Ding, Xiaofei Chen, Yan Cao, Dan Jia, Dongyao Wang, Zhenyu Zhu, Juping Zhang, Zhanying Hong, Yifeng Chai

Journal of Chromatography A 2014 Volume 1359() pp:330-335

Publication Date(Web):12 September 2014

DOI:10.1016/j.chroma.2014.07.071

•The methodology of CMC model was fully investigated to improve the technique with the option to obtain more reliable results.•This is the first time that Paraformaldehyde (PFA) was used in CMC to enhance the efficiency of columns.•Two novel different CMC models (HSC-T6/CMC and CMMC-7721/CMC) were established.•This article offers perspectives to be applied for the screening and identification of active compounds in medicines and drugs.Cell Membrane Chromatography (CMC) is a biological affinity chromatographic method using a silica stationary phase covered with specific cell membrane. However, its short life span and poor quality control was highlighted in a lot of research articles. In this study, special attention has been paid to the disruption, cell load and packing procedure in order to improve the quality of the CMC columns. Hereto, two newly established CMC models, HSC-T6/CMC and SMMC-7721/CMC have been developed and used in this research project. The optimization of the abovementioned parameters resulted in a better reproducibility of the retention time of the compound GFT (RSD < 10%) and improved significantly the quality of the CMC columns. 3.5 × 107 cells were the optimal cell load for the preparation of the CMC columns, the disruption condition was optimized to 5 cycles (400 W and 20 s interval per cycle) by an ultrasonic processor reducing the total time of cell disruption to 1.5 min and the packing flow rate was optimized by applying a linear gradient program. Additionally, 4% paraformaldehyde (PFA) was employed to improve the column quality and prolong the column life span. The results showed that the retention time was longer with PFA treated columns than the ones obtained with the control groups

A strategy for rapid analysis of xenobiotic metabolome of Sini decoction in vivo using ultra-performance liquid chromatography-electrospray ionization quadrupole-time-of-flight mass spectrometry combined with pattern recognition approach

Co-reporter:Guangguo Tan, Min Liu, Xin Dong, Si Wu, Li Fan, Youbei Qiao, Yifeng Chai, Hong Wu

Journal of Pharmaceutical and Biomedical Analysis 2014 Volume 96() pp:187-196

Publication Date(Web):5 August 2014

DOI:10.1016/j.jpba.2014.03.028

•A reliable strategy was developed for xenobiotic metabolome identification of TCM.•The S- and VIP-plot can clearly display leading contributing markers.•A total of 102 SND-related xenobiotic metabolites were characterized.•The metabolite profiles of major components from SND in rat urine were described.Xenobiotic metabolome identificatioqn of Chinese herbal prescription in biological systems is a very challenging task. In the present work, a reliable strategy based on the combination of ultra-performance liquid chromatography-electrospray ionization quadrupole-time-of-flight mass spectrometry (UHPLC-ESI-Q-TOFMS) and pattern recognition approach such as principal component analysis (PCA) and partial least squared discriminant analysis (PLS-DA) was proposed to rapidly discover and analyze the xenobiotic metabolome from Sini decoction (SND). Using the S- and VIP-plots of PLS-DA, 96 and 112 interest ions from positive and negative ion datasets were extracted as SND metabolome in rat urine following oral administration of SND. Among them, 53 absorbed prototype components of SND and 49 metabolites were identified, which provided essential data for further studying the relationship between the chemical components and pharmacological activity of SND. Our results indicated that hydrolysis and demethylation were the major metabolic pathways of diterpenoid alkaloids, while glucuronidation, sulfation, hydrolysis, reduction, demethylation, and hydroxylation were the main metabolic pathways of flavonoids, and hydrolysis was the metabolic pathway of gingerol-related compounds. No saponin-related metabolites were detected.

Investigation of the therapeutic effectiveness of active components in Sini decoction by a comprehensive GC/LC-MS based metabolomics and network pharmacology approaches

Co-reporter:Si Chen, Si Wu, Wuhong Li, Xiaofei Chen, Xin Dong, Guangguo Tan, Hai Zhang, Zhanying Hong, Zhenyu Zhu and Yifeng Chai

Molecular BioSystems 2014 vol. 10(Issue 12) pp:3310-3321

Publication Date(Web):24 Sep 2014

DOI:10.1039/C4MB00048J

As a classical formula, Sini decoction (SND) has been fully proved to be clinically effective in treating doxorubicin (DOX)-induced cardiomyopathy. Current chemomics and pharmacology proved that the total alkaloids (TA), total gingerols (TG), total flavones and total saponins (TFS) are the major active ingredients of Aconitum carmichaelii, Zingiber officinale and Glycyrrhiza uralensis in SND respectively. Our animal experiments in this study demonstrated that the above active ingredients (TAGFS) were more effective than formulas formed by any one or two of the three individual components and nearly the same as SND. However, very little is known about the action mechanisms of TAGFS. Thus, this study aimed to use for the first time the combination of GC/LC-MS based metabolomics and network pharmacology for solving this problem. By metabolomics, it was found that TAGFS worked by regulating six primary pathways. Then, network pharmacology was applied to search for specific targets. 17 potential cardiovascular related targets were found through molecular docking, 11 of which were identified by references, which demonstrated the therapeutic effectiveness of TAGFS using network pharmacology. Among these targets, four targets, including phosphoinositide 3-kinase gamma, insulin receptor, ornithine aminotransferase and glucokinase, were involved in the TAGFS regulated pathways. Moreover, phosphoinositide 3-kinase gamma, insulin receptor and glucokinase were proved to be targets of active components in SND. In addition, our data indicated TA as the principal ingredient in the SND formula, whereas TG and TFS served as adjuvant ingredients. We therefore suggest that dissecting the mode of action of clinically effective formulae with the combination use of metabolomics and network pharmacology may be a good strategy.

Effect of Amphotericin B on the Metabolic Profiles of Candida albicans

Co-reporter:YingYing Cao, ZhenYu Zhu, XiaoFei Chen, XiangWen Yao, LiuYa Zhao, Hui Wang, Lan Yan, HaiTang Wu, YiFeng Chai, and YuanYing Jiang

Journal of Proteome Research 2013 Volume 12(Issue 6) pp:2921-2932

Publication Date(Web):2017-2-22

DOI:10.1021/pr4002178

Amphotericin B (AmB) is a polyene antifungal drug widely used for systemic fungal infections. In this study, a metabonomic method using gas chromatography–mass spectrometry (GC/MS) was developed to characterize the metabolic profiles of Candida albicans cells exposed to AmB. Thirty-one differentially produced metabolites between AmB-treated and the control groups were identified, among which 10 metabolites were upregulated and 21 metabolites were downregulated. These differentially produced metabolites were mainly involved in polyamines synthesis, tricarboxylic acid (TCA) cycle, oxidative stress, glutathione metabolism, lipid synthesis and glycolysis. Further experiments showed that the polyamines including putrescine, spermidine, and spermine played an important role in the sensitivity of C. albicans cells upon AmB treatment, and combined use of AmB and inhibitors of polyamine biosynthesis pathway might be a potential antifungal strategy. This study provided a systemic view of the metabolic pattern in C. albicans upon exposure to AmB, which shed new light on the mechanisms of action of antifungal agents.

Serum metabonomics coupled with Ingenuity Pathway Analysis characterizes metabolic perturbations in response to hypothyroidism induced by propylthiouracil in rats

Co-reporter:Si Wu, Yue Gao, Xin Dong, Guangguo Tan, Wuhong Li, Ziyang Lou, Yifeng Chai

Journal of Pharmaceutical and Biomedical Analysis 2013 Volume 72() pp:109-114

Publication Date(Web):18 January 2013

DOI:10.1016/j.jpba.2012.09.030

A serum metabonomic profiling method based on ultra-performance liquid chromatography/time-of-flight mass spectrometry (UHPLC/TOF-MS) was applied to investigate the metabolic changes in hypothyroid rats induced by propylthiouracil (PTU). With Significance Analysis of Microarray (SAM) for classification and selection of biomarkers, 13 potential biomarkers in rat serum were screened out. Furthermore, Ingenuity Pathway Analysis (IPA) was introduced to deeply analyze unique pathways of hypothyroidism that were primarily involved in sphingolipid metabolism, fatty acid transportation, phospholipid metabolism and phenylalanine metabolism. Our results demonstrated that the metabonomic approach integrating with IPA was a promising tool for providing a novel methodological clue to systemically dissect the underlying molecular mechanism of hypothyroidism.

Plasma pharmacochemistry combined with microdialysis to screen potential bioactive components and their metabolites in Anemarrhena asphodeloides saponin extract using ultrahigh-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry

Co-reporter:Zhirui Liu;Min Liu;Yunpeng Qi;Zhenyu Zhu;Chao Yuan;Yang Lin

Journal of Separation Science 2013 Volume 36( Issue 9-10) pp:1659-1666

Publication Date(Web):

DOI:10.1002/jssc.201201116

Although various techniques have been employed to analyze drug metabolites, the metabolism of multicomponent herbal medicine has seldom been fully addressed. In contrast to chemical drugs, a number of compounds in herbal medicine could get into circulation and then be metabolized. Metabolism study on active constituents in herbal medicine is a good way for us to explain and predict a variety of events related to the efficacy and toxicity of herbal medicine. The present work aims to elucidate the multicomponent metabolic characteristics of a herbal medicine by the combination of plasma pharmacochemistry and microdialysis sampling. Anemarrhena asphodeloides, a well-known traditional Chinese medicine, was chosen as a model. After oral administration of A. asphodeloides saponin extract to rats, microdialysis samples were collected continuously in the jugular vein and analyzed by ultrahigh-performance LC/quadrupole-TOF MS. The identification of compounds in biosamples was achieved by accurate mass measurement and detailed fragmentation pathway analysis. The results showed that unbound constituents in blood circulation of the rat included seven parent saponins and six metabolites, which might be the potential active components in vivo. Among which, three metabolites have not been previously reported and were identified in this study. It is the first report on systemic metabolism of total saponins of A. asphodeloides in mammalian plasma.

Qualitative analysis and quality control of Traditional Chinese Medicine preparation Tanreqing injection by LC-TOF/MS and HPLC-DAD-ELSD

Co-reporter:Liang Sun, Hua Wei, Feng Zhang, Shouhong Gao, Qinghua Zeng, Wenquan Lu, Wansheng Chen and Yifeng Chai

Analytical Methods 2013 vol. 5(Issue 22) pp:6431-6440

Publication Date(Web):03 Sep 2013

DOI:10.1039/C3AY40681D

Tanreqing injection, a classical Traditional Chinese Medicine formulation, consists of Radix Scutellariae baicalensis, Fel selenarcti, Cornu naemorhedi, Flos lonicerae and Forsythiae fructus, having a marked curative effect on the diseases of the upper respiratory tract. In order to clarify its chemical profile, a method based on liquid chromatography coupled with time-of-flight mass spectrometry was proposed, and 53 compounds were identified on a formula database of 515 known compounds with the aid of accurate mass measurement for molecular ions and characteristic fragment ions. Using the chemical fingerprints for the purpose of quality control, a high performance liquid chromatography coupled with photodiode array detection and evaporative light scattering detection (HPLC-DAD-ELSD) was developed in this study. The chromatographic fingerprints were evaluated by the dual similarities (S and S′) of an ultra-violet chromatogram, in combination with SE of an ELSD chromatogram, reflecting the complexity of Tanreqing injection with the consideration of both large and small peaks. In addition, the established HPLC-DAD-ELSD method was successfully applied to the quantitative analysis of the five ingredients (chlorogenic acid, caffeic acid, baicalin, ursodeoxycholic acid and chenodeoxycholic acid) and fingerprint analysis in 11 batches of Tanreqing injections. The results obtained in this research will provide the basis for quality control of Tanreqing injections.

Comparative pharmacokinetics of timosaponin B-II and timosaponin A-III after oral administration of Zhimu–Baihe herb-pair, Zhimu extract, free timosaponin B-II and free timosaponin A-III to rats

Co-reporter:Zhirui Liu, Xin Dong, Xuan Ding, Xiaofei Chen, Lei Lv, Yueyue Li, Yifeng Chai

Journal of Chromatography B 2013 Volume 926() pp:28-35

Publication Date(Web):1 May 2013

DOI:10.1016/j.jchromb.2013.03.003

•It firstly developed an LC–MS method for simultaneous measurement of timosaponin B-II and timosaponin A-III.•The method is simple, sensitive and specific, with a short analysis time (8 min).•There were obvious differences in the pharmacokinetics between timosaponin B-II and timosaponin A-III.•This study revealed the practical value of the compatibility of herb-pair remedy in clinic.The aim of this study was to compare the pharmacokinetics of timosaponin B-II and timosaponin A-III in rat plasma after oral administration of Zhimu–Baihe herb-pair, Zhimu extract, free timosaponin B-II and free timosaponin A-III. After addition of internal standard (IS) ginsenoside Rh1, plasma samples were pretreated by protein precipitation. Chromatographic separation was carried out on a SHISEIDO CAPCELL PAK C18 column (100 mm × 3 mm i.d., 3.0 μm) with the mobile phase consisting of acetonitrile and 0.05% (v/v) formic acid by linear gradient elution. The detection was performed on an Agilent G1946D single quadrupole mass spectrometer with negative electrospray ionization (ESI) interface in select-ion-monitoring (SIM) mode. The following ions: m/z 919 for timosaponin B-II, m/z 739 for timosaponin A-III and m/z 683 for the IS were used for quantitative determination. Good linearity was achieved over the concentration ranged from 4.0–793.3 ng/mL to 3.9–781.3 ng/mL for the two saponins. The precision of the in vivo study was evaluated by intra- and inter-day assays and the percentages of relative standard deviation were all within 15%. The plasma concentrations of timosaponin B-II and timosaponin A-III in rats at designated time periods were successfully determined using this fully validated method, and statistically significant differences (p < 0.5) in pharmacokinetic parameters including AUC0−t, AUC0−∞ and MRT (mean residence time) were obtained. Compared to these pharmacokinetic parameters after oral administration of Zhimu extract and monomer solution, higher peak concentration (Cmax is higher), slower elimination (MRT is longer) and larger AUC values could be observed after giving Zhimu–Baihe herb-pair in our study. Therefore, this result not only elucidated the steady and long-lasting pharmacological properties but also revealed the practical value of the compatibility of herb-pair remedy.

Metabonomic studies on potential plasma biomarkers in rats exposed to ionizing radiation and the protective effects of Hong Shan Capsule

Co-reporter:Yue Liu;Ze-Bin Lin;Guang-Guo Tan;Zhi-Yong Chu;Zi-Yang Lou

Metabolomics 2013 Volume 9( Issue 5) pp:1082-1095

Publication Date(Web):2013 October

DOI:10.1007/s11306-013-0529-6

An ultra performance liquid chromatography coupled to mass spectrometry-based metabonomic approach, combined with pattern recognition methods including PCA, PLS-DA, RF and heatmap, has been developed to characterize the global serum metabolic profile associated with ionizing radiation (IR). As the VIP-value threshold cutoff of the metabolites was set to 2, metabolites above this threshold were filtered out as potential target biomarkers. Nineteen distinct potential biomarkers in rat plasma were identified. To further elucidate the pathophysiology of IR, related metabolic pathways have been studied. It was found that IR was closely related to disturbed fatty acid metabolism, taurine and hypotaurine metabolism, sphingolipid metabolism, purine metabolism, pyrimidine metabolism, phospholipid catabolism, tryptophan metabolism, phenylalanine metabolism, and bile acid metabolism. With the presented metabonomic method, we systematically analyzed the protective effects of Traditional Chinese Medicine Hong Shan Capsule (HSC). The results demonstrated that HSC administration could provide satisfactory effects on IR through partially regulating the perturbed metabolic pathways.

Computer-aided molecular modeling study of enantioseparation of iodiconazole and structurally related triadimenol analogues by capillary electrophoresis: Chiral recognition mechanism and mathematical model for predicting chiral separation

Co-reporter:Wuhong Li, Guangguo Tan, Liang Zhao, Xiaofei Chen, Xinrong Zhang, Zhenyu Zhu, Yifeng Chai

Analytica Chimica Acta 2012 Volume 718() pp:138-147

Publication Date(Web):9 March 2012

DOI:10.1016/j.aca.2012.01.007

Chiral separation of iodiconazole, a new antifungal drug, and 12 new structurally related triadimenol analogues had been developed by capillary electrophoresis (CE) using hydroxypropyl-γ-cyclodextrin (HP-γ-CD) as the chiral selector. The effect of structural features of analytes on Δt and Rs was studied under the optimum separation conditions. Using molecular docking technique and binding energy calculations, the inclusion process between HP-γ-CD and enantiomers was investigated and chiral recognition mechanisms were discussed. The results suggest that hydrogen bonding between fluorine at position 4 of the phenyl group beside the chiral carbon and the hydroxyl group on the HP-γ-CD rim and face to face π–π interactions between two phenyl rings highly contributed to the enantiorecognition process between HP-γ-CD and iodiconazole. The N-methyl group beside chiral carbon also played an important role in enantiomeric separation. Additionally, the big difference in binding energy (ΔΔE) highly contributed to good separation in the presence of HP-γ-CD chiral selector, which may be a helpful initial guide for chiral selector selection and predicting the result of enantioseparation. Furthermore, the new mathematical equation established based on the results of molecular mechanics calculations exhibited good capability in predicting chiral separation of these triadimenol analogues using HP-γ-CD mediated CE.Graphical abstractHighlights► Molecular modeling studies chiral recognition mechanism of iodiconazole with HP-γ-CD. ► The good separation obtained is due to the big binding energy difference. ► A new mathematical equation is established to predict chiral separation. ► Molecular modeling serves as a useful method for studying chiral separation.

Innovative microwave-assisted oximation and silylation procedures for metabolomic analysis of plasma samples using gas chromatography–mass spectrometry

Co-reporter:Zhanying Hong, Zebin Lin, Yue Liu, Guangguo Tan, Ziyang Lou, Zhenyu Zhu, Yifeng Chai, Guorong Fan, Junping Zhang, Liming Zhang

Journal of Chromatography A 2012 Volume 1254() pp:14-22

Publication Date(Web):7 September 2012

DOI:10.1016/j.chroma.2012.07.033

Analysis of plasma metabolomic samples by gas chromatography–mass spectrometry always requires comprehensive pretreatment including oximation and silylation. Although heating block (HB) is a commonly used method, it is time consuming. This study describes an extremely time-effective microwave-assisted (MA) oximation and silylation approach for metabolomic study of plasma samples. The Box–Behnken design was employed to optimize the MA conditions by means of oximation at 65 W for 100 s and then silylation through 180 s incubation with 230 W microwave irradiation. The results showed that microwave irradiation decreased the sample preparation time from approximately 180 min to 5 min without loss of information for the metabolites in plasma samples. Both the HB method and the developed MA method were applied in plasma metabolomic study of sulfur mustard intoxication. Partial least-squares discriminant analysis (PLS-DA) was used to globally understand the metabolic changes, and multi-criteria assessment was used to select the most significant and reliable variables as potential biomarkers. The data obtained by the MA method were in good correlation with the HB method. Compared with HB method, the newly developed MA oximation and silylation of plasma metabolome samples was more efficient and time-effective and may prove to be an attractive alternative for high-throughput sample preparation in plasma metabolomics.Highlights► Microwave-assisted oximation and silylation for plasma metabolomic was conducted. ► Box–Behnken design was employed to optimize the microwave-assisted conditions. ► Metabolomic approach was used to discover the sulfur mustard intoxication biomarkers. ► The optimized microwave-assisted method improved the time efficiency about 36 times.

Comprehensive two-dimensional HepG2/cell membrane chromatography/monolithic column/time-of-flight mass spectrometry system for screening anti-tumor components from herbal medicines

Co-reporter:Xiaofei Chen, Yan Cao, Diya Lv, Zhenyu Zhu, Junping Zhang, Yifeng Chai

Journal of Chromatography A 2012 Volume 1242() pp:67-74

Publication Date(Web):15 June 2012

DOI:10.1016/j.chroma.2012.04.034

Cell membrane chromatography (CMC) is a biological affinity chromatographic method using specific cell membrane as stationary phase. It has been proved to be a practical tool for investigating binding interactions between drugs and membrane receptors. In this study, a novel comprehensive two-dimensional (2D) chromatography approach was established for screening anti-tumor components from herbal medicines (HMs). HepG2/CMC model was first developed and applied as the first dimensional column. Using an automatic ten-port switching valve equipped with two sample loops, the fractions of the first-dimension were introduced in the second-dimension consists of a monolithic column and a time-of-flight mass spectrometry (TOFMS) with high resolving ability. Based on the stability, selectivity and suitability assays of the HepG2/CMC/monolithic column/TOFMS system, berberine (BBR) and tetrahydropalmatine (THP) from Cortex phellodendri amurensis, oxymatrine and matrine from Radix sophorae ﬂavescentis were screened and identified as potential active components. The competitive displacement assay suggested that the four components could act on epidermal growth factor receptor region on the HepG2 cell membrane in similar manner of gefitinib. Furthermore, their inhibiting effects on cell proliferation in vitro were also confirmed and, BBR and THP showed concentration dependently inhibitory ability on HepG2 cell proliferation (p < 0.05). The result demonstrated that the proposed comprehensive 2D HepG2/CMC/monolithic column/TOFMS system has the advantages of strong recognition and rapid analysis abilities for the total screening procedure, which will be selectable and practical in drug discovery from complex HM samples and can also be applied to other biochromatography models.Highlights► A new HepG2 cell membrane model was developed. ► Novel comprehensive two-dimensional chromatography approach was established and optimized for fast screening procedure. ► Four anti-tumor active components from herbal medicines were screened and reliably identified by TOFMS. ► EGFR was preliminary selected as the target membrane receptor via competitive displacement assays. ► BBR and THP revealed inhibiting effects on cell proliferation in vitro.

Combined Metabonomic and Quantitative Real-Time PCR Analyses Reveal Systems Metabolic Changes in Jurkat T-Cells Treated with HIV-1 Tat Protein

Co-reporter:Wenting Liao, Guangguo Tan, Zhenyu Zhu, Qiuli Chen, Ziyang Lou, Xin Dong, Wei Zhang, Wei Pan, and Yifeng Chai

Journal of Proteome Research 2012 Volume 11(Issue 11) pp:5109-5123

Publication Date(Web):2017-2-22

DOI:10.1021/pr300173c

HIV-1 Tat protein is released by infected cells and can affect bystander uninfected T cells and induce numerous biological responses which contribute to its pathogenesis. To elucidate the complex pathogenic mechanism, we conducted a comprehensive investigation on Tat protein-related extracellular and intracellular metabolic changes in Jurkat T-cells using combined gas chromatography–mass spectrometry (GC-MS), reversed-phase liquid chromatography–mass spectrometry (RPLC-MS) and a hydrophilic interaction liquid chromatography–mass spectrometry (HILIC-MS)-based metabonomics approach. Quantitative real-time PCR (qRT-PCR) analyses were further employed to measure expressions of several relevant enzymes together with perturbed metabolic pathways. Combined metabonomic and qRT-PCR analyses revealed that HIV-1 Tat caused significant and comprehensive metabolic changes, as represented by significant changes of 37 metabolites and 10 relevant enzymes in HIV-1 Tat-treated cells. Using MetaboAnalyst 2.0, it was found that 11 pathways (Impact-value >0.10) among the regulated pathways were acutely perturbed, including sphingolipid metabolism, glycine, serine and threonine metabolism, pyruvate metabolism, inositol phosphate metabolism, arginine and proline metabolism, citrate cycle, phenylalanine metabolism, tryptophan metabolism, pentose phosphate pathway, glycerophospholipid metabolism, glycolysis or gluconeogenesis. These results provide metabolic evidence of the complex pathogenic mechanism of HIV-1 Tat protein as a “viral toxin”, and would help obligate Tat protein as “an important target” for therapeutic intervention and vaccine development.

Elucidation of steroid glycosides in Anemarrhena asphodeloides extract by means of comprehensive two-dimensional reversed-phase/polyamine chromatography with mass spectrometric detection

Co-reporter:Zhirui Liu;Dongliang Zhu;Yunpeng Qi;Xiaofei Chen;Zhenyu Zhu

Journal of Separation Science 2012 Volume 35( Issue 17) pp:2210-2218

Publication Date(Web):

DOI:10.1002/jssc.201200236

In the work presented here, a novel approach to comprehensive two-dimensional liquid chromatography is evaluated. Reversed-phase liquid chromatography was employed for the first-dimension separation and polyamine chromatography was chosen for the second-dimension separation mode. The two dimensions are highly orthogonal and the separation efficacy of the developed octadecylsilica × polyamine was tested by separating an extract from Anemarrhena asphodeloides. The steroid glycosides identified by comprehensive two-dimensional liquid chromatography in this experiment were compared to those obtained for monodimensional liquid chromatography. The comprehensive two-dimensional liquid chromatography system, thanks to the complementary separation selectivity and enhanced peak capacity provided by the two columns, allowed to distribute five compounds of low amounts otherwise unachievable by monodimensional liquid chromatography. In addition, four steroid isomers with similar fragmentation characteristics in MS/MS spectra, were newly separated based on their different chemical structures.

Hydrophilic interaction and reversed-phase ultraperformance liquid chromatography TOF-MS for serum metabonomic analysis of myocardial infarction in rats and its applications

Co-reporter:Guangguo Tan, Ziyang Lou, Wenting Liao, Xin Dong, Zhenyu Zhu, Wuhong Li and Yifeng Chai

Molecular BioSystems 2012 vol. 8(Issue 2) pp:548-556

Publication Date(Web):31 Oct 2011

DOI:10.1039/C1MB05324H

An ultra performance liquid chromatography coupled to mass spectrometry-based metabonomic approach, which utilizes both reversed-performance (RP) chromatography and hydrophilic interaction chromatography (HILIC) separations, has been developed to characterize the global serum metabolic profile associated with myocardial infarction (MI). The HILIC was found necessary for a comprehensive serum metabonomic profiling, providing complementary information to RP chromatography. By combining with partial least squares discriminant analysis, 21 potential biomarkers in rat serum were identified. To further elucidate the pathophysiology of MI, related metabolic pathways have been studied. It was found that MI was closely related to disturbed sphingolipid metabolism, phospholipid catabolism, fatty acid transportation and metabolism, tryptophan metabolism, branched-chain amino acids metabolism, phenylalanine metabolism, and arginine and proline metabolism. With the presented metabonomic method, we systematically analyzed the therapeutic effects of Traditional Chinese Medicine Sini decoction (SND). The results demonstrated that SND administration could provide satisfactory effects on MI through partially regulating the perturbed metabolic pathways.

Qualitative and quantitative analysis of Fructus Corni using ultrasound assisted microwave extraction and high performance liquid chromatography coupled with diode array UV detection and time-of-flight mass spectrometry

Co-reporter:Zhirui Liu, Zhenyu Zhu, Hai Zhang, Guangguo Tan, Xiaofei Chen, Yifeng Chai

Journal of Pharmaceutical and Biomedical Analysis 2011 55(3) pp: 557-562

Publication Date(Web):

DOI:10.1016/j.jpba.2011.02.007

An Optimized Ion-Pair HPLC Method for Simultaneous Analysis of Nucleoside Triphosphate Levels in Hepatoma Cell Line

Co-reporter:Jing Jia;Hai Zhang;Liang Zhao;Zhen-yu Zhu;Guo-qing Zhang

Chromatographia 2011 Volume 73( Issue 7-8) pp:755-759

Publication Date(Web):2011 April

DOI:10.1007/s10337-010-1881-6

An improved ion-pair HPLC method was developed for the simultaneous determination of ribonucleoside triphosphates and their corresponding deoxyribonucleoside triphosphates in HepG2 cell extracts. HPLC conditions, flow rate and column temperature were optimized and good linearity (r2 > 0.9993) was obtained over the investigated concentration ranges. Reproducibility was evaluated by intra- and inter-day assays and RSD values were below 5.39%. Recoveries ranged from 98.2 ± 3.49% to 103.1 ± 1.75%, respectively. Finally, the method was successfully applied to the analysis of eight compounds present in HepG2 cell extracts.

Comparative pharmacokinetics of baicalin and wogonoside by liquid chromatography–mass spectrometry after oral administration of Xiaochaihu Tang and Radix scutellariae extract to rats

Co-reporter:Zhenyu Zhu, Liang Zhao, Xiaofan Liu, Jun Chen, Hai Zhang, Guoqing Zhang, Yifeng Chai

Journal of Chromatography B 2010 Volume 878(Issue 24) pp:2184-2190

Publication Date(Web):15 August 2010

DOI:10.1016/j.jchromb.2010.06.021

The aim of this study was to compare the pharmacokinetics of baicalin and wogonoside in rats following oral administration of Xiaochaihu Tang (Minor Radix Bupleuri Decoction) and Radix scutellariae extract. Thus, a specific LC–MS method was developed and validated for the determination of these flavonoids in the plasma of rats after oral administration Xiaochaihu Tang and Radix scutellariae extract. Chromatographic separation was performed on a Zorbax SB C18 column (150 mm × 4.6 mm, i.d.: 5 μm) with 0.1% formic acid in water and acetonitrile by linear gradient elution. Baicalin, wogonoside and carbamazepine (internal standard, I.S.) were detected in select-ion-monitoring (SIM) mode with a positive electrospray ionization (ESI) interface. The following ions: m/z 447 for baicalin, m/z 461 for wogonoside and m/z 237 for the I.S. were used for quantitative determination. The calibration curves were linear over the concentration ranges from 0.1231 to 6.156 μg mL−1 for baicalin and 0.08832 to 4.416 μg mL−1 for wogonoside. The lower limit of detection (LLOD) based on a signal-to-noise ratio of 2 was 0.06155 μg mL−1 for baicalin and 0.04416 μg mL−1 for wogonoside. Intra-day and inter-day precisions (RSD%) were within 10% and accuracy (RE%) ranged from −6.4 to 4.4%. The extraction recovery at three QC concentrations ranged from 74.7 to 86.0% for baicalin and from 71.3 to 83.7% for wogonoside. The plasma concentrations of baicalin and wogonoside in rats at designated time periods after oral administration were successfully determined using the validated method, pharmacokinetic parameters were estimated by a non-compartment model. Following oral administration of Xiaochaihu Tang and Radix scutellariae extract, the t1/2 of baicalin was 3.60 ± 0.90 and 5.64 ± 1.67, the Cmax1 was 1.64 ± 0.99 and 5.66 ± 2.02, the tmax1 was 0.13 ± 0.05 and 0.20 ± 0.07, the Cmax2 was 2.43 ± 0.46 and 3.18 ± 1.66, and the tmax2 were 6.40 ± 1.67 and 5.66 ± 2.02, respectively. Following oral administration of Xiaochaihu Tang and Radix scutellariae extract, the t1/2 of wogonoside was 4.97 ± 1.68 and 7.71 ± 1.55, the Cmax1 was 1.39 ± 0.83 and 1.45 ± 0.37, the tmax1 was 0.21 ± 0.20 and 0.17 ± 0.01, the Cmax2 was 1.90 ± 0.55 and 1.42 ± 0.70, and the tmax2 was 5.60 ± 1.67 and 5.20 ± 1.79, respectively. A significant difference (p < 0.05) was observed for t1/2, and the elimination of baicalin and wogonoside in Xiaochaihu Tang was increased.

Determination of six lignans in Schisandra chinensis (Turcz.) Baill. Fruits and related Chinese multiherb remedies by HPLC

Co-reporter:Hai Zhang, Guoqing Zhang, Zhenyu Zhu, Liang Zhao, Yang Fei, Jing Jing, Yifeng Chai

Food Chemistry 2009 Volume 115(Issue 2) pp:735-739

Publication Date(Web):15 July 2009

DOI:10.1016/j.foodchem.2008.12.010

A simple, rapid and specific HPLC method was established for simultaneous determination of six major lignans in Schisandra chinensis and related Chinese multiherb remedies (CMRs) containing this herb. The six lignans were successfully separated on a C18 column by gradient elution using acetonitrile and water as the mobile phase, and detection wavelength was set at 225 nm. The method was validated through the following performance criteria: linearity, precision, repeatability, stability, accuracy, limit of detection (LOD) and quantification (LOQ). This assay was successfully used for determination of six lignans in 10 raw herbs collected from different regions in China and five Chinese multiherb remedies. Significant variations were demonstrated in the contents of six compounds in these samples. This HPLC method established is suitable for routine quantitative analysis of S. chinensis and multiherb remedies containing this herb.

New mathematic model for predicting chiral separation using molecular docking: Mechanism of chiral recognition of triadimenol analogues

Co-reporter:Guoqing Zhang;Qingyan Sun;Ying Hou;Zhanying Hong;Jun Zhang;Liang Zhao;Hai Zhang

Journal of Separation Science 2009 Volume 32( Issue 14) pp:2401-2407

Publication Date(Web):

DOI:10.1002/jssc.200900012

Abstract

The purpose of this paper was to study the enantioseparation mechanism of triadimenol compounds by carboxymethylated (CM)-β-CD mediated CE. All the enantiomers were separated under the same experimental conditions to study the chiral recognition mechanism using a 30 mM sodium dihydrogen phosphate buffer at pH 2.2 adjusted by phosphoric acid. The inclusion courses between CM-β-CD and enantiomers were investigated by the means of molecular docking technique. It was found that there were at least three points (one hydrophobic bond and two hydrogen bonds) involved in the interaction of each enantiomer with the chiral selectors. A new mathematic model has been built up based on the results of molecular mechanics calculations, which could analyze the relationship between the resolution of enantioseparation and the interaction energy in the docking area. Comparing the results of the separation by CE, the established mathematic model demonstrated good capability to predict chiral separation of triadimenol enantiomers using CM-β-CD mediated CE.

Development of a rapid and sensitive LC–MS/MS assay for the determination of combretastatin A4 phosphate, combretastatin A4 and combretastatin A4 glucuronide in beagle dog plasma and its application to a pharmacokinetic study

Co-reporter:Xiaojing Wang, Zhihang Chen, Jinjing Che, Qingfang Meng, Chengqi Shan, Yunan Hou, Xiaolei Liu, Yifeng Chai, Yuanguo Cheng

Journal of Chromatography B 2009 Volume 877(Issue 30) pp:3813-3821

Publication Date(Web):15 November 2009

DOI:10.1016/j.jchromb.2009.09.027

This study details the development and validation of a simple and sensitive liquid chromatography/tandem mass spectrometry (LC–MS/MS) method for the quantification of combretastatin A-4 3-O-phosphate (CA4P), combretastatin A4 (CA4) and its main metabolite, combretastatin A4 glucuronide (CA4G), in beagle dog plasma. Sample pretreatment includes simple protein precipitation by adding methanol to the plasma sample containing an internal standard (colchicine). LC separation was successfully accomplished on a Waters RP8 Symmetryshield™ column (3.0 mm×150 mm, i.d., 5 μm) with a gradient mobile phase of methanol (0.1% formic acid, v/v) and water (20 mM ammonium acetate) at a flow rate 0.8 mL min−1. The three analytes were detected in the positive/negative ion alternate mode, negative ion mode for CA4G and positive ion mode for CA4P and CA4. Multiple reaction monitoring (MRM) was used for determination of three analytes. Calibration curves were linear in the concentration range of 5–5000 ng mL−1 for CA4P (r ≥ 0.999), 5–3000 ng mL−1 for CA4 (r ≥ 0.999) and 5–5000 ng mL−1 for CA4G (r ≥ 0.999). All the validation data, such as accuracy, precision, and inter-day repeatability, were within the required limits. The method was reliable and has been successfully applied to a pharmacokinetic study of CA4P in beagle dogs via intravenous drop infusion at dose rates of 1, 3 and 9 mg kg−1. After daily intravenous drop infusions at 1 mg kg−1 for 7 consecutive days, the accumulation ratio was approximately 1.0, indicating no accumulation from multiple doses in beagles.

Rapid LC–TOFMS Separation and Identification of Diarylheptanoids and Gingerol-Related Compounds in Dried Ginger

Co-reporter:Xiang Li;Ziyang Lou;Hai Zhang;Liang Zhao;Hao Wu;Guoqing Zhang

Chromatographia 2009 Volume 69( Issue 5-6) pp:531-536

Publication Date(Web):2009 March

DOI:10.1365/s10337-008-0934-6

A tandem liquid chromatographic–time-of-flight mass spectrometric (LC–TOFMS) method has been developed for rapid separation and identification of diarylheptanoids and gingerol-related compounds in aqueous extracts of dried ginger. Total-ion-current chromatograms and mass spectra were acquired. A formula database of known compounds was established, against which components of dried ginger could be rapidly identified by matching their exact masses with theoretical masses of compounds calculated from their empirical formulae. Identification of 20 compounds was accomplished with error of 4 ppm, and further confirmation of elemental composition was obtained from the abundance of the isotope peaks. LC–TOFMS has been shown to be a useful tool for rapid identification of compounds in aqueous extracts of dried ginger.

LC Separation and Determination of Five Diester-Diterpenoid Alkaloids in the Unprocessed and Processed Aconite Roots

Co-reporter:Min Liu;Hai Zhang;Liang Zhao;Baiyun Zhao;Lingling Dong;Zhenyu Zhu

Chromatographia 2008 Volume 67( Issue 11-12) pp:

Publication Date(Web):2008 June

DOI:10.1365/s10337-008-0624-4

A reliable liquid chromatographic method with photodiode array detector (DAD) was developed and validated for simultaneous separation and determination of five diester-diterpenoid alkaloids in the aconite roots. The separation was successfully performed on a Zorbax Extend-C18 column with a mobile phase gradient prepared from methanol and ammonia solution at a flow rate of 1.0 mL min−1. Good linearity (r > 0.999) was observed over the concentration ranges investigated, and intra-day and inter-day precision were high. The mean recoveries of five components ranged from 90.45 to 102.63% and relative standard deviations were always <5%. The validated method was successfully used for simultaneous determination of the five diester-diterpenoid alkaloids of unprocessed and processed aconite roots. The quantitative method provided a scientific basis for safety assurance and clinical application of aconite roots.

Liquid chromatography coupled with time-of-flight and ion trap mass spectrometry for qualitative analysis of herbal medicines

Co-reporter:Xiao-Fei Chen, Hai-Tang Wu, Guang-Guo Tan, Zhen-Yu Zhu, Yi-Feng Chai

Journal of Pharmaceutical Analysis (November 2011) Volume 1(Issue 4) pp:235-245

Publication Date(Web):1 November 2011

DOI:10.1016/j.jpha.2011.09.008

With the expansion of herbal medicine (HM) market, the issue on how to apply up-to-date analytical tools on qualitative analysis of HMs to assure their quality, safety and efficacy has been arousing great attention. Due to its inherent characteristics of accurate mass measurements and multiple stages analysis, the integrated strategy of liquid chromatography (LC) coupled with time-of-ﬂight mass spectrometry (TOF-MS) and ion trap mass spectrometry (IT-MS) is well-suited to be performed as qualitative analysis tool in this field. The purpose of this review is to provide an overview on the potential of this integrated strategy, including the review of general features of LC-IT-MS and LC-TOF-MS, the advantages of their combination, the common procedures for structure elucidation, the potential of LC-hybrid-IT-TOF/MS and also the summary and discussion of the applications of the integrated strategy for HM qualitative analysis (2006–2011). The advantages and future developments of LC coupled with IT and TOF-MS are highlighted.

Using cell membrane chromatography and HPLC-TOF/MS method for in vivo study of active components from roots of Aconitum carmichaeli

Co-reporter:Yan Cao, Xiao-Fei Chen, Di-Ya Lü, Xin Dong, ... Yi-Feng Chai

Journal of Pharmaceutical Analysis (May 2011) Volume 1(Issue 2) pp:125-134

Publication Date(Web):1 May 2011

DOI:10.1016/S2095-1779(11)70022-3

An offline two-dimensional system combining a rat cardiac muscle cell membrane chromatography time-of-flight mass spectrometry (CMC-TOF/MS) with a high Performance liquid chromatography time-of-flight mass spectrometry (HPLC-TOF/MS) was established for investigating the parent components and metabolites in rat urine samples after administration of the roots of Aconitum carmichaeli. On the basis ofthe analysis of the first dimension, retention components of the urine sample were collected into 30 fractions (one fraction per minute). Then offline analysis of the second dimension was carried out. 34 compounds including 24 parent alkaloids and 10 potential metabolites were identified from the dosed rat urine, and then binding affinities of different compounds on cell membranes were compared and influences of some functional groups on activity were estimated with the semi-quantification and curve fitting method. As a result, binding affinities decreased along with the process of deacylation, debenzoylation and demethylation, which may be related to the alleviation of toxicity in the procedure of herb processing or metabolism. Moreover, some minor components in rat urine (Songorine, 14-benzoylneoline, Deoxyaconitine, etc.) exerted relatively strong affinity on cell membranes are worth exploring. The results delivered by the System suggest that the CMC can be applied to in vivo study.

UPLC-Q-TOF/MS based metabolomic profiling of serum and urine of hyperlipidemic rats induced by high fat diet

Co-reporter:Qiong Wu, Hai Zhang, Xin Dong, Xiao-Fei Chen, ... Yi-Feng Chai

Journal of Pharmaceutical Analysis (December 2014) Volume 4(Issue 6) pp:360-367

Publication Date(Web):1 December 2014

DOI:10.1016/j.jpha.2014.04.002

Hyperlipidemia is considered to be a high lipid level in blood, can induce metabolic disorders and dysfunctions of the body, and results in some severe complications. Therefore, hunting for some metabolite markers and clarifying the metabolic pathways in vivo will be an important strategy in the treatment and prevention of hyperlipidemia. In this study, a rat model of hyperlipidemia was constructed according to histopathological data and biochemical parameters, and the metabolites of serum and urine were analyzed by UPLC-Q-TOF/MS. Combining pattern recognition and statistical analysis, 19 candidate biomarkers were screened and identified. These changed metabolites indicated that during the development and progression of hyperlipidemia, energy metabolism, lipid metabolism, amino acid metabolism and nucleotide metabolism were mainly disturbed, which are reported to be closely related to diabetes, cardiovascular diseases, etc. This study demonstrated that a UPLC-Q-TOF/MS based metabolomic approach is useful to profile the alternation of endogenous metabolites of hyperlipidemia.

Comparative analysis of essential oils found in Rhizomes Curcumae and Radix Curcumae by gas chromatography–mass spectrometry

Co-reporter:Di-Ya Lü, Yan Cao, Ling Li, Zhen-Yu Zhu, ... Zi-Yang Lou

Journal of Pharmaceutical Analysis (August 2011) Volume 1(Issue 3) pp:203-207

Publication Date(Web):1 August 2011

DOI:10.1016/j.jpha.2011.05.001

A comparison of the volatile compounds in Rhizomes Curcumae (Ezhu) and Radix Curcumae (Yujin) was undertaken using gas chromatography–mass spectrometry (GC–MS). Ultrasonic extraction and GC–MS methods were developed for the simultaneous determination of five sesquiterpenes, namely, α-pinene, β-elemene, curcumol, germacrone and curdione, in Ezhu and Yunjin. Good linearity (r>0.999) and high inter-day precision were observed over the investigated concentration ranges. The validated method was successfully used for the simultaneous determination of five sesquiterpenes in Ezhu and Yujin. The quantitative method can be effectively used to evaluate and monitor the quality of Chinese curcuma in clinical use.

Development and validation of liquid chromatography–tandem mass spectrometry method for simultaneous determination of six steroidal saponins in rat plasma and its application to a pharmacokinetics study

Co-reporter:Zhirui Liu, Wenxing Qin, Zhenyu Zhu, Yao Liu, Fengjun Sun, Yifeng Chai, Peiyuan Xia

Steroids (April 2015) Volume 96() pp:21-29

Publication Date(Web):1 April 2015

DOI:10.1016/j.steroids.2015.01.006

•Development and validation of an UHPLC–MS/MS method for selected steroidal saponins.•The method is simple, sensitive and specific, with a short analysis time (10 min).•Application to rat plasma demonstrates the method is fit-for-purpose.A specific and reliable liquid chromatography–electrospray ionization-tandem mass spectrometry method was developed for the simultaneous determination of timosaponin H1 (TH1), timosaponin E1 (TE1), timosaponin E (TE), timosaponin B-II (TB-II), timosaponin B-III (TB-III) and anemarrhenasaponin I (AS-I) in rat plasma. After addition of internal standard (IS) ginsenoside Rh1, plasma samples were pretreated by protein precipitation with acetonitrile. Chromatographic separation was performed on a reverse phase ACQUITY™ BEH C18 column (100 mm × 2.1 mm i.d., 1.7 μm) using a gradient mobile phase system of acetonitrile–water containing 0.05% formic acid and 5 mM ammonium formate. The triple quadruple mass spectrometer was set in negative electrospray ionization mode and multiple reaction monitoring (MRM) was used for six steroidal saponins quantification. The precursors to produce ion transitions monitored for TH1, TE1, TE, TB-II, TB-III, AS-I and IS were m/z 1211.5 > 1079.6, 935.5 > 773.4, 935.4 > 773.5, 919.6 > 757.4, 901.5 > 739.3, 757.4 > 595.3 and 637.3 > 475.3, respectively. The method validation was conducted over the curve range of 0.5–400 ng/mL for the six saponins. The intra- and inter-day precisions (RSD%) were less than 9.4% and the average extraction recoveries ranged from 82.5% to 97.8% for each analyte. Six steroidal saponins were proved to be stable during sample storage, preparation and analytical procedures. The validated method was successfully applied for the first time to determine the concentrations of six main steroidal saponins in incurred rat plasma samples, after intragastric administration of the extract of Anemarrhena asphodeloides Bge. for a rat pharmacokinetic study.Download full-size image

Absorption and metabolism of three monoester-diterpenoid alkaloids in Aconitum carmichaeli after oral administration to rats by HPLC–MS

Co-reporter:Hai Zhang, Qiong Wu, Wuhong Li, Sen Sun, Wen Zhang, Zhenyu Zhu, Guoqing Zhang, Yifeng Chai

Journal of Ethnopharmacology (3 July 2014) Volume 154(Issue 3) pp:645-652

Publication Date(Web):3 July 2014

DOI:10.1016/j.jep.2014.04.039

Ethnopharmacological relevance: Aconitum carmichaelii (AC) is a well-known herbal medicine for its excellent pharmacological effects and toxicity. The monoester-diterpenoid alkaloids (MDAs), including benzoylmesaconine (BMC), benzoylaconine (BAC) and benzoylhyaconine (BHC), are the main active components in AC. It was found that the diester alkaloids could be transformed into monoester-diterpenoid alkaloids after being decocted. In Chinese pharmacopoeia, the MDAs are also used as phytochemical markers for the quality control of AC. Benzoylmesaconine, benzoylaconine and benzoylhyaconine are representatives of monoester-diterpenoid alkaloids. It was reported that the absolute bioavailability of MDAs was very low but there was toxicity often occurred in AC. Because most of DDAs are transformed into MDAs after decoction, we speculate that some other components may promote the bioavailability of MDAs but result in toxicity by enhancing their absorption. To demonstrate the dynamic changes of MDAs in vivo and reveal the causes of low bioavailability and toxicity, this study will explore the mechanisms of absorption and metabolism of 3 MDAs.Materials and methods: A sensitive, accurate and specific LC-MS method was developed to determine the three MDAs in rat plasma. The pharmacokinetic parameters were estimated after orally administered 3 MDAs to the Male Sprague-Dawley rats, and the metabolism stability was calculated after incubating with rat liver microsomes, finally, the absorption characteristics of the 3 MDAs were investigated using Caco-2 transwell model.Results: It was found that the pharmacokinetic parameters of 3 MDAs were similar, Cmax and Tmax were very small, and t1/2 was large, which indicated 3 MDAs can be absorbed rapidly and is difficult to be metabolized or excreted. However, the low Cmax indicated that the bioavailability of 3 MDAs will be very low and their absorption may be inhibited by some transport proteins. By incubating three MDAs in rat liver microsomes, it was proved that they almost can’t be metabolized in vivo. The Caco-2 transwell experiments reveal that the P-gp inhibits the absorption of MDAs.Conclusions: LC-MS combined with a direct precipitation method for the simultaneous quantification of 3 MDAs in rat plasma has been developed and validated and successfully used in pharmacokinetic study of 3 MDAs. It was proved that the three components almost can’t be metabolized in vivo, and P-gp inhibits the absorption of MDAs.Download high-res image (161KB)Download full-size image

HIV-1 Tat induces biochemical changes in the serum of mice

Co-reporter:Wenting Liao, Guangguo Tan, Zhenyu Zhu, Qiuli Chen, Ziyang Lou, Xin Dong, Wei Zhang, Wei Pan, Yifeng Chai

Virology (20 January 2012) Volume 422(Issue 2) pp:288-296

Publication Date(Web):20 January 2012

DOI:10.1016/j.virol.2011.11.005

The HIV-1 Tat protein is released by infected cells and has numerous biological activities which might contribute either to the impairment of the immune response or to viral dissemination and pathogenesis. To date, the effects of Tat protein on metabolites remain unclear. In this study, a metabolomic study on serum of HIV-1 Tat-induced ICR mice was performed to research the pathologic mechanism of Tat protein by using gas chromatography coupled to mass spectrometry (GC/MS). Clear separations among the HIV-1 Tat-induced mice and the inaTat-induced or control mice were observed by principal component analysis and partial least-squares discriminant analysis based on the GC/MS spectral data. Additionally, 16 significantly changed metabolites in HIV-1 Tat-induced mice were identified that are involved in multiple perturbed metabolic pathways, which contributed to the elucidation of the complex pathogenic mechanism of Tat protein and may shed new lights on future improvement of HIV-1 therapy.Highlights► We create a mouse model to study Tat-induced biochemical changes in serum. ► We first report a metabolomic study on the bioactivity of Tat protein. ► We find 16 changed metabolites which involve in multiple metabolic pathways. ► Our findings help in elucidating the pathogenic mechanism of HIV infection.

Identification of Annexin A2 as a target protein for plant alkaloid matrine

Co-reporter:Dongyao Wang, Yan Cao, Leyi Zheng, Diya Lv, Langdong Chen, Xinrui Xing, Zhenyu Zhu, Xiaoyu Li and Yifeng Chai

Chemical Communications 2017 - vol. 53(Issue 36) pp:NaN5023-5023

Publication Date(Web):2017/04/11

DOI:10.1039/C7CC02227A

Qualitative analysis and quality control of Traditional Chinese Medicine preparation Tanreqing injection by LC-TOF/MS and HPLC-DAD-ELSD

Co-reporter:

Analytical Methods (2009-Present) 2013 - vol. 5(Issue 22) pp:

Publication Date(Web):

DOI:10.1039/C3AY40681D