Mechanism of Didang Decoction in prevention of anti-atherosclerosis and hyperlipidemia by HPLC-Q-TOF-MS/MS and network pharmacology based on theory of "nutrients return to heart and fat accumulates in channels" / 中国中药杂志

Atherosclerosis(AS) is caused by impaired lipid metabolism, which deposits lipids in the intima, causes vascular fibrosis and calcification, and then leads to stiffening of the vascular wall. Hyperlipidemia(HLP) is one of the key risk factors for AS. Based on the theory of "nutrients return to the heart and fat accumulates in the channels", it is believed that the excess fat returning to the heart in the vessels is the key pathogenic factor of AS. The accumulation of fat in the vessels over time and the blood stasis are the pathological mechanisms leading to the development of HLP and AS, and "turbid phlegm and fat" and "blood stasis" are the pathological products of the progression of HLP into AS. Didang Decoction(DDD) is a potent prescription effective in activating blood circulation, removing blood stasis, resolving turbidity, lowering lipids, and dredging blood vessels, with the functions of dispelling stasis to promote regeneration, which has certain effects in the treatment of atherosclerotic diseases. This study employed high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(HPLC-Q-TOF-MS/MS) to screen the main blood components of DDD, explored the targets and mechanisms of DDD against AS and HLP with network pharmacology, and verified the network pharmacological results by in vitro experiments. A total of 231 blood components of DDD were obtained, including 157 compounds with a composite score >60. There were 903 predicted targets obtained from SwissTargetPrediction and 279 disease targets from GeneCards, OMIM, and DisGeNET, and 79 potential target genes of DDD against AS and HLP were obtained by intersection. Gene Ontology(GO) analysis suggested that DDD presumably exerted regulation through biological processes such as cholesterol metabolism and inflammatory response, and Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis suggested that signaling pathways included lipid and atherosclerosis, insulin resistance, chemo-carcinogenesis-receptor activation, and AGE-RAGE signaling pathways in diabetic complications. In vitro experiments showed that DDD could reduce free fatty acid-induced lipid accumulation and cholesterol ester content in L02 cells and improve cellular activity, which might be related to the up-regulation of the expression of PPARα, LPL, PPARG, VEGFA, CETP, CYP1A1, and CYP3A4, and the down-regulation of the expression of TNF-α and IL-6. DDD may play a role in preventing and treating AS and HLP by improving lipid metabolism and inflammatory response, and inhibiting apoptosis with multi-component, multi-target, and multi-pathway characteristics.

Identification of Q-markers for Cistanches Herba based on HPLC-Q-TOF-MS/MS and network pharmacology / 中国中药杂志

This study aims to establish a method for analyzing the chemical constituents in Cistanches Herba by high performance liquid chromatography(HPLC) and quadrupole-time-of-flight tandem mass spectrometry(HPLC-Q-TOF-MS/MS), and to reveal the pharmacological mechanism based on network pharmacology for mining the quality markers(Q-markers) of Cistanches Herba. The chemical constituents of Cistanche deserticola and C. tubulosa were analyzed via HPLC-Q-TOF-MS/MS. The potential targets and pathways of Cistanches Herba were predicted via SwissTargetPrediction and DAVID. The compound-target-pathway-pharmacological action-efficacy network was constructed via Cytoscape. A total of 47 chemical constituents were identified, involving 95 targets and 56 signaling pathways. We preliminarily elucidated the pharmacological mechanisms of echinacoside, acteoside, isoacteoside, cistanoside F, 2'-acetylacteoside, cistanoside A, campneoside Ⅱ, salidroside, tubuloside B, 6-deoxycatalpol, 8-epi-loganic acid, ajugol, bartsioside, geniposidic acid, and pinoresinol 4-O-β-D-glucopyranoside, and predicted them to be the Q-markers of Cistanches Herba. This study identified the chemical constituents of Cistanches Herba, explained the pharmacological mechanism of the traditional efficacy of Cistanches Herba based on network pharmacology, and introduced the core concept of Q-markers to improve the quality evaluation of Cistanches Herba.

Qualitative and quantitative analysis of aristolochic acid components in nine Chinese medicinal materials containing toxic aristolochic substances / 药学学报

The objective of this work was to explore the content and composition of aristolochic acid compounds in Chinese medicinal materials containing toxic aristolochic chemicals, so as to ensure the safety of these medicinal materials and their related products. Nine Chinese medicinal materials were selected for study, including the tuber of Aristolochia cinnabarina, the herbs of Asarum forbesii, the stems of Aristolochia manshuriensis., the fruits of Aristolochia debilis, the roots of Aristolochia debilis, the stems and leaf of Aristolochia debilis, the herbs of Aristolochia mollissima, the roots of Aristolochia fangchi, and the roots of Asarum heterotropoides var. mandshuricum. The aristolochic acid components in the nine Chinese medicinal materials were analyzed by high performance liquid chromatography-quadrupole time of flight mass spectrometry (HPLC-Q-TOF-MS) combined with high performance liquid chromatography diode-array detection. The separation was performed on an Agilent ZORBAX SB-Aq column (250 mm×4.6 mm, 5 μm) with gradient elution using a mobile phase consisting of acetonitrile and 0.2% acetic acid. ESI positive ion mode MS was used to investigate the ionization pathways of aristolochic acid Ⅰ, Ⅱ, Ⅲa, Ⅳa, Ⅶa, and aristololactam Ⅰ, Ⅱ using seven reference standards, and the structures of the components with UV spectrasimilar to those of the seven reference standards in the selected medicinal materials were qualitatively analyzed by following the investigated ionization pathways. The identified aristolochic acid components were quantified using an external standard method by HPLC-UV with detection at 254 nm. Twenty-two aristolochic acid components including 11 aristolochic acids and 11 aristololactams were identified from the nine selected medicinal materials; 15 aristolochic acids were found in the tuber of Aristolochia cinnabarina and the roots of Aristolochia debilis, followed by 14 aristolochic acids in the fruits of Aristolochia debilis and the stems of Aristolochia manshuriensis. The greatest content of aristolochia components was found in the tuber of Aristolochia cinnabarina and the stems of Aristolochia manshuriensis, ranging from 8.91 mg·g-1 to 13.40 mg·g-1, and the least amount was in the herbs of Asarum forbesii, at less than 0.10 mg·g-1 and containing only two aristolochia components. This study systematically explored the quantity and composition of aristolochic acid components in selected Chinese medicinal materials believed to contain toxic aristolochic compounds, providing a basis for follow-up studies on the toxicity of these substances that can lead to safety standards for their use.

Qualitative and quantitative analysis of alkaloids in Eurycoma longifolia by HPLC-Q-TOF-MS combined with UPLC-QQQ-MS/MS / 中国中药杂志

A sensitive and efficient method was established and validated for qualitative and quantitative analysis of total alkaloids from the extract of Eurycoma longifolia by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS) combined with ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS). The HPLC-Q-TOF-MS conditions are as follows: Welch Ultimate XB-C_(18) column(4.6 mm×250 mm, 5 μm) with acetonitrile(containing 0.1% formic acid)-0.1% formic acid in water as mobile phase for gradient elution. The UPLC-QQQ-MS/MS conditions are as below: Agilent Eclipse Plus C_(18) column(2.1 mm×50 mm, 1.8 μm) with acetonitrile(containing 0.1% formic acid) and 0.1% formic acid in water as mobile phase for gradient elution. MS data were collected by electrospray ionization in positive ion mode. According to the comparison with reference standards and the accurate masses of molecules, a total of 17 alkaloids in E. longifolia extract were identified by HPLC-Q-TOF-MS. The UPLC-QQQ-MS/MS quantitative analysis result of 3 alkaloids showed that the linear ranges of them were good(r≥0.999 7) and the overall recoveries ranged from 108.8%-110.2%, with RSDs of 2.9%-5.3%. The method is accurate, reliable, and efficient, which can comprehensively reflect the constituents and content of alkaloids in E. longifolia. The result can serve as a reference for further elucidating its therapeutic material basis and quality control.

Analysis of Chemical Constituents in Daishenning by HPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:High performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry （HPLC-Q-TOF-MS/MS） was used to identify the main chemical constituents of Daishenning. Method:Cosmosil 5 C₁₈-AR-Ⅱ column （4.6 mm×250 mm， 5 μm） was employed for chromatographic separation with mobile phase of acetonitrile （A）-0.5% formic acid aqueous solution （B） for gradient elution （0-10 min， 5%A； 10-20 min， 5%-20%A； 20-30 min， 20%A； 30-55 min， 20%-35%A； 55-65 min， 35%-55%A； 65-75 min， 55%-100%A； 75-80 min， 100%A； 80-85 min， 100%-5%A； 85-90 min， 5%A）， the flow rate was 1 mL·min^-1， column temperature was 40 ℃， and injection volume was 10 μL. Electrospray ionization （ESI）， positive and negative ion detection modes and mass scanning range of <italic>m</italic>/<italic>z</italic> 100-2 000 were selected for mass spectrometry. The main chemical constituents in Daishenning were identified by MassHunter B.06.00 software in combination with PubChem， MassBank， ChemicalBook and other databases， and reference information. Result:A total of 96 components were identified from Daishenning， including 32 flavonoids， 19 organic acids， 6 glycosides， 6 terpenoids， 5 phenylpropanoids， 8 phenols， 14 other components and 6 unknown components. Conclusion:The established method can simultaneously analyze different types of compounds in Daishenning， it is helpful for further research on the extraction and separation of main chemical components and quality control of this preparation. In addition， through the rapid identification of the chemical constituents in Daishenning， it is speculated that the main effective substances of Daishenning may be flavonoids and organic acids.

Research of tissue metabolomics in papillary thyroid carcinoma based on HPLC/Q-TOF-MS / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：用组织代谢组学方法，探讨甲状腺乳头状癌（papillary thyroid carcinoma, PTC）组织及癌旁组织的代谢差异，寻找PTC的潜在生物标志物，探索PTC的发病机制与治疗策略。方法：收集2018年10月至2020年2月期间湖南省人民医院乳甲外科手术切除的40例PTC患者的癌组织及癌旁组织标本。利用HPLC-MS技术平台对PTC组织及癌旁组织样本的差异性代谢物进行多维统计分析，寻找与PTC相关的异常代谢通路。结果：经PCA、PLS-DA、OPLS-DA分析得知癌组织和癌旁组织的代谢轮廓具有显著性差异。经OPLS-Loading plot分析，结合VIP>1、FC>2，且P<0.05，筛选出76个潜在差异性代谢物。其中亮氨酸、2-褪黑素、香草酸等33种代谢物在PTC组织中表达上调；3-葡萄糖苷、甘油磷脂、磷脂酰胆碱、乳糖等43代谢物在PTC癌组织中表达下调。寻找到与差异性代谢物相关的13条异常代谢通路，如半胱氨酸与蛋氨酸代谢、与甘油磷脂代谢、嘧啶代谢、半乳糖代谢以及丙氨酸、天冬氨酸和谷氨酸代谢、柠檬酸循环等，这些代谢通路可能参与PTC代谢的病理生理过程。ROC曲线下面积大于0.9的差异性代谢物有5种，分别是庚二酸、糖醇、辛二酸、乳糖和L-丝氨酸。结论：PTC组织中半乳糖代谢和氨基酸代谢发生改变，PTC组织细胞中存在沃伯格效应(Warburg effect)。庚二酸、糖醇、辛二酸、乳糖、L-丝氨酸五种差异性代谢物可以用来区分PTC患者与正常人。

Main chemical constituents of Changyanning Tablets based on HPLC-Q-TOF-MS/MS / 中草药

Objective: To study the chemical constituents of Changyanning Tablets, a high performance liquid chromatography-quadrupole time of flight mass spectrometry (HPLC-Q-TOF-MS/MS) was established to recognize and classify the ingredients accurately and rapidly. Methods: Agilent ZORBAX Eclipse XDB-C18 chromatographic column (250 mm × 4.6 mm, 5 μm) was employed and the separation was performed with the mobile phase consisting of methanol-0.05% acetic acid aqueous solution. The information of accurate mass and multistage fragment ions were obtained by the monitored simultaneously for positive and negative ions. The main chemical constituents of Changyanning Tablets were identified by high resolution mass spectrometry data, combining with Pubmed, Hmdb, Massbank network database, reference literature and comparing the reference. Results: Fifty-one chemical components were finally identified in this study, including two phenylpropanoids, eight iridoids, 12 flavonoids, four tannins, 23 organic acids, and two other classes. Conclusion: This study comprehensively studies the material basis in Changyanning Tablets, which provides a basis for improving the quality evaluation system of Changyanning Tablets and lays the foundation for elucidating the active components mechanism.

Rapid identification of classical herbal formulae Baoyinjian's chemical substance based by HPLC-Q/TOF-MS / 中草药

Objective: To analyze the chemical constituent cluster of classical herbal formulae Baoyinjian systemically by HPLC-Q/TOF-MS. Methods: The seperation was performed on Diamonsil C18 (250 mm × 4.6 mm, 5 μm) column with gradient elution with acetonitrile-0.1% formic acid. The column temperature was 30 ℃, the flow rate was 1 mL/min, the injection volume was 10 μL, and the mass spectrometry condition was X500R QTOF mass spectrometer, electrospray ion source, positive and negative mode scanning. Results: A total of 52 chemical constituents were identified by reference confirmation, literature comparison, and high mass spectrometry data analysis. The chemical constituent cluster was composed of 17 flavonoids, six phenolics, 12 iridoid glycosides, eight alkaloids, one phenethyl alcohol glycosides, four monoterpene glycoside, two triterpenes and two other compound. Conclusion: This study can identify various chemical constituents of Baoyinjian systematically, accurately, and rapidly, which provides a basis for the determination of the quality attributes of Baoyinjian.

Qualitative Analysis of Multiple Cumarins in Angelicae Sinensis Radix Based on HPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To systematically and comprehensively analyze coumarin components in Angelicae Sihensis Radix by an efficient and stable HPLC-Q-TOF-MS/MS method,in order to offer the theoretical basis to develop coumarin,establish the quality control standard and apply in clinic. Method:The separation effect of coumarin components was extracted by adjusting the column,temperature,mobile phase,flow rate,sample concentration and other conditions,and various coumarin components in Angelicae Sihensis Radix were identified by corresponding standards,precise molecular mass,polarity,pyrolysis pattern. Result:In this study,a high-efficiency and stable coumarin separation method was established that can be used to separate complex components,and 14 coumarin components were identified in this study,including phellopterin and osthenol that were rarely reported as effective components in Angelicae Sihensis Radix. Major fragment ions of coumarin components were analyzed. The cleavage in methoxy bond or anisole bond on the parent nucleus was the primary pattern for coumarin components, which was summarized for detecting unknowing coumarins. Conclusion:Abundant coumarins were contained in Angelicae Sihensis Radix. Further qualitative and quantitative analysis of coumarins are conducive to improving the quality standards of Angelicae Sihensis Radix,and providing reference for the development of coumarins and clinical application of Angelicae Sinensis Radix.

Qualitative analysis of metabolites of ethanol extract of Cuscuta chinensis by HPLC-Q/TOF MS/MS / 中草药

Objective To qualitatively analyze the chemical constituents in rats after intragastric administration of estrogenically active ethanol extract of Cuscuta chinensis. Methods The high performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q/TOF MS/MS) was used to identify the prototypes and metabolites in rat urine. Results Twelve chemical constituents were identified in the drug-containing urine, including six prototypes and six metabolites. The prototypes are kaemoferol-3-β-D-glucuronide, 6-O-(E)-p-coumaroyl-β-D-fructofuranosyl-(2→1)-α-D-glucopyranoside, aempferol-7-rhamnoside, chlorogenic acid, and apigenin. The metabolites are p-hydroxycinnamic acid, p-hydroxyphenylpropionic acid, isorhamnetin, kaempferol-3-O-glucoside, acetyl caffeic acid, and quercetin sulfate. Conclusion The method of HPLC-Q/TOF MS/MS is simple and rapid for the analysis of prototype components and metabolites in rats urine after oral administration of ethanol extract of C. chinensis, providing the basis for further clarification of the estrogen material basis of C. chinensis.

Identification and screening of cardiac glycosides in Streptocaulon griffithii using an integrated data mining strategy based on high resolution mass spectrometry / 中国天然药物

The present study was designed to develop a practical strategy to tackle the problem of lacking standard compounds and limited references for identifying structure-related compounds in Streptocaulon griffithii Hook. f., especially those in trace concentrations, with a focus on antitumor activity. The cardiac glycosides (CGs)-enriched part was determined using in vitro bioactive assays in three cancer cell lines and then isolated using macroporous resins. The MS and MS/MS data were acquired using a high performance liquid chromatography coupled with hybrid quadrupole-time of flight (HPLC-Q-TOF-MS) system. To acquire data of trace compound in the extract, a multiple segment program was applied to modify the HPLC-Q-TOF-MS method. A mass defect filter (MDF) approach was employed to make a primary MS data filtration. Utilizing a MATLAB program, the redundant peaks obtained by imprecise MDF template calculated with limited references were excluded by fragment ion classification, which was based on the ion occurrence number in the MDF-filtered total ion chromatograms (TIC). Additionally, the complete cleavage pathways of CG aglycones were proposed to assist the structural identification of 29 common fragment ions (CFIs, ion occurrence number ≥ 5) and diagnostic fragment ions (DFIs, ion occurrence number < 5). As a result, 30 CGs were filtered out from the MDF results, among which 23 were identified. This newly developed strategy may provide a rapid and effective tool for identifying structure-related compounds in herbal medicines.

Identification and screening of cardiac glycosides in Streptocaulon griffithii using an integrated data mining strategy based on high resolution mass spectrometry / 中国天然药物

The present study was designed to develop a practical strategy to tackle the problem of lacking standard compounds and limited references for identifying structure-related compounds in Streptocaulon griffithii Hook. f., especially those in trace concentrations, with a focus on antitumor activity. The cardiac glycosides (CGs)-enriched part was determined using in vitro bioactive assays in three cancer cell lines and then isolated using macroporous resins. The MS and MS/MS data were acquired using a high performance liquid chromatography coupled with hybrid quadrupole-time of flight (HPLC-Q-TOF-MS) system. To acquire data of trace compound in the extract, a multiple segment program was applied to modify the HPLC-Q-TOF-MS method. A mass defect filter (MDF) approach was employed to make a primary MS data filtration. Utilizing a MATLAB program, the redundant peaks obtained by imprecise MDF template calculated with limited references were excluded by fragment ion classification, which was based on the ion occurrence number in the MDF-filtered total ion chromatograms (TIC). Additionally, the complete cleavage pathways of CG aglycones were proposed to assist the structural identification of 29 common fragment ions (CFIs, ion occurrence number ≥ 5) and diagnostic fragment ions (DFIs, ion occurrence number < 5). As a result, 30 CGs were filtered out from the MDF results, among which 23 were identified. This newly developed strategy may provide a rapid and effective tool for identifying structure-related compounds in herbal medicines.

Analysis of Related Substances in Minoxidil and Minoxidil Gel by HPLC-Q-TOF-MS / 中国药师

Objective:To analyze the related substances in the domestic products of minoxidil and minoxidil gel for the purpose of standard improvement. Methods:An HPLC-Q-TOF-MS method was developed. The test was performed on a Thermo Scientific Accu-core C18 column (100 mm × 2. 1 mm, 2. 6 μm) with a mobile phase of acetonitrile-water (7:93) (containing 0. 1% formic acid) at a flow rate of 0. 2 ml·min-1 , the detection wavelength was 230 nm. The full scan and two stage scanning was employed for detection with ESI source positive ion mode. The mass scan range was m/z 50-500. Results:14 related substances were separated and detected under the established HPLC-Q-TOF-MS condition. The structures and resource of 8 related substances were analyzed preliminarily. Conclusion:The HPLC-Q-TOF-MS technology has been proved to be effective in separation and identification of the related substances in minoxidil and minoxidil gel. The results were useful for quality control.

Analysis on pharmacodynamic constituents in Gualou Guizhi Decoction by serum pharmacochemistry / 中草药

Objective: To study the serum pharmacochemistry of Gualou Guizhi Decoction (GLGZD) as well as the material basis through analyzing the constituents absorbed in blood. Methods: A high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (HPLC-Q-TOF-MS) method was developed for the analysis on absorbed components in serum after ig administration of GLGZD to rats. The separation was achieved on a C18 column by gradient elution with acetonitrile-aqueous solution (containing 0.1% formic acid) as mobile phase. The ion acquisition was performed by full scan MS and MS/MS in negative ion mode.The serum after ig administration of GLGZD to rats, blank serum of rats, and GLGZD extracts sample were analyzed by HPLC-Q-TOF-MS. Results: Forty-two prototype compounds including monoterpene glycosides, flavonoids, phenolic acids, gingerols, triterpenoid saponins, galloylglucoses, and others were confirmed and identified in serum by comparing the retention time and mass spectrometry fragmentation characteristics with reference standards or literatures. Conclusion: The analysis of absorbed components of GLGZD in serum after administration by serum pharmacochemistry method would provide an experimental basis for revealing the real neuroprotective constituents of GLGZD in treating dysfunction after stroke.

Analysis of chemical constituents in Liujing Toutong Tablets by HPLC-Q-TOF/MS / 中草药

Objective To separate and identify the chemical constituents in Liujing Toutong Tablets (LTT) by HPLC-Q-TOF/MS. Methods The main components were separated by using the gradient elution method with RP-HPLC, Diamonsil C18 (250 mm × 4.6 μm, 5 μm). The positive and negative electro spray ionization (ESI) source was adopted for determine the chromatographic effluent, the main chromatographic peaks are assigned and distinguished by Q-TOF. These components were further analyzed by accurately relative molecular mass of compounds, and Combined with Positive and negative ions MS information and related literature data. Results According to the MS principle, the corresponding reference substances and literature datas, Ninety-five compounds of LTT were identified, mainly including isoflavones, coumarins, iridoids and phthalides. Furthermore, all of the constituents were surveyed and classified according to their medicinal materials derivation. Conclusion In this study, the main components in LTT were elucidated completely by HPLC-Q-TOF/MS. The results could provide the evidence for the research on active constituents and quality control of LTT.

Identification of absorbed components and their metabolites in rat plasma after oral administration of Liujing Toutong Tablets by HPLC-Q/TOF-MS / 中草药

Objective To study the serum pharmacochemistry of Liujing Toutong Tablets (LTT). This study could be helpful to elucidate the bioactive constituents of LTT. Methods HPLC-Q/TOF-MS was used to analyze the ingredients in rats blood plasma with LTT (ig). Results By comparing the information on the total ion chromatogram, extraction chromatogram and the mass spectrogram of LTT, rat serum with drug and blank serum sample was used to confirm the constituents of LTT in vivo. As a result, a total of 46 compounds were detected in rat plasma, including 24 absorbed prototype constituents, such as puerarin, and 22 of the metabolites. Conclusion The compounds are absorbed into the blood might function as real bioactive constituents and this study can provide a scientific fundament for bioactive substances of LTT.

Analysis on chemical constituents from Glycyrrhizae Radix et Rhizoma by HPLC-Q-TOF-MS / 中草药

Objective: To analyze the main chemical constituents of Glycyrrhizae Radix et Rhizoma by HPLC-Q-TOF-MS in the positive and negative ions mode. Methods: The chromatographic fingerprint was obtained with Diamonsil II C18 column (250 mm×4.6 mm, 5 μm) and gradient elution with 0.05% H2O-formic acid (A)-acetonitrile (B), and the flow rate was 1 mL/min. The column temperature was maintained at 35℃. The detection wavelength was of 200-600 nm. Positive and negative ions MS information of Bruker Daltonics 1200 HPLC-Q-TOF was coupled with elemental analysis and compared with the literature data to analyze the compounds information. Results: Combined with the accurate relative molecular mass of compounds provided by HPLC-Q-TOF-MS, 40 compounds were identificated in the ethanolic extract from Glycyrrhizae Radix et Rhizoma, which include 28 flavonoids, 11 triterpenoids saponins, and one coumarin. Conclusion: The method provides the technical support for the quality control of Glycyrrhizae Radix et Rhizoma, and contributes the reference data to elucidating the potential basis of Glycyrrhizae Radix et Rhizoma

Study on substance basis of Yinhuang Qingfei Capsule in vitro by HPLC-Q-TOF-MS/MS / 中草药

Objective: To identify and analyze the chemical constituents in Yinhuang Qingfei Capsule by HPLC-Q-TOF-MS/MS method. Methods: The HPLC method was used with the conditions that the column was Inertsil ODS-2 C18 (250 mm × 4.6 mm, 5 μm). Columu and electrospray ion (ESI) source was employed for the qualitative analysis under positive ion mode. These components were further analyzed by MS spectra, and by comparing with the corresponding reference substances and literature data. Results: According to the MS principle and literature data, 54 compounds were identified from the sample of Yinhuang Qingfei Capsule. Conclusion: An efficient HPLC-Q-TOF-MS/MS approach has been established for studying the chemical constituents in Yinhuang Qingfei Capsule, which paves a way for the quality control and further substance basis studies of the preparation.

Analysis of main active components in Desmodii Styraciflii Herba by HPLC-Q- TOF-MS and HPLC-DAD / 中草药

Objective: This study is aimed to establish a high performance liquid chromatography coupled with quadrupole-time-of- flight mass spectrometric (HPLC-Q-TOF-MS) method for analyzing the main active components in Desmodii Styraciflii Herba, and to establish an HPLC-DAD method for simultaneously determining its seven flavonoids compounds (schaftoside, isoschaftoside, vicenin-2, isoorientin, isovitexin, luteolin, and apigenin). Methods: The analysis was performed on a Phenomenex Kinetex C18 column with a gradient elution of methanol-0.2% aqueous formic acid at the flow rate of 1.0 mL/min. The column temperature was 40 oC. The Q-TOF-MS discriminant analysis was performed under negative electrospray ion mode and the split ratio was 1∶1. Quantitative analysis of seven compounds in Desmodii Styraciflii Herba was carried by HPLC-DAD. The determination wavelength was at 272 nm. Results: Forty-one main active constituents were separated and identified by HPLC-Q-TOF-MS, including 37 flavonoids compounds and four phenolic acids. A HPLC-DAD method was successfully developed for determining seven flavonoids compounds. The standard curves for all seven compounds showed good linearity with correlation coefficients higher than 0.999 0 within the test ranges. The average recoveries (n = 6) were between 96.7%-102.4%, and RSD ≤ 4.94%. Conclusion: Analyzing the main active components provides a significant guidance for the substance basis research of Desmodii Styraciflii Herba. The HPLC-DAD method for simultaneously determining seven flavonoids compounds is accurate, reproducible, and provides the basis for the quality control of Desmodii Styraciflii Herba.

Qualitative and quantitative analysis of components of Radix Polygalae in herbal pair of Radix Polygalae and Rhizoma Acori Tatarinowii / 中国药科大学学报

@#The purpose of this research was to explore the differences of the components of Radix Polygalae in herbal pair of Radix Polygalae and Rhizoma Acori Tatarinowii. An HPLC-Q-TOF-MS/MS method and an HPLC-UV method were established for the identification and determination of the components of Radix Polygalae, respectively. HPLC separation was carried out on a C18 column(250 mm×4. 6 mm, 5 μm)with linear gradient elution using a mobile phase consisting of acetonitrile and 0. 1% formic acid aqueous solution. Mass spectrometry with ESI source was performed in the positive ion mode to scan MS data including total ion chromatograms and ion peaks of Radix Polygalae. Eight components including 3, 4, 5-trimethoxycinnamic acid, p-methoxycinnamic acid, tenuifolin, sibiricose A5, polygalaxanthone III, tenuifoliside B, 3, 6′-disinapoly sucrose, and tenuifoliside A were identified according to the reference substance retention time, MS data and literatures. There was no significant variation found in the contents of eight chemical constituents of Radix Polygalae. The qualification and quantitation study of the components in herbal pair of Radix Polygalae and Rhizoma Acori Tatarinowii provide the methodological basis for compatibility mechanism exploration in vivo.