title: HPLC-DAD-MS(n) analysis and HPLC quantitation of chemical constituents in Xian-ling-gu-bao capsules.
abstract: In this study, a systematic method was established for the global quality control of Xian-ling-gu-bao capsules (XLGB), a popular six-herb Traditional Chinese Patent Medicine (TCPM) for the treatment of osteoporosis. Both qualitative and quantitative analyses were conducted. In qualitative analysis part, a fast and sensitive method based on high-performance liquid chromatography coupled with diode-array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-MS(n)) was established for rapid separation and sensitive identification. Samples were separated on a Waters Symmetry C(18) column (250mm×4.6mm, 5μm) by gradient elution using acetonitrile (A) and water-formic acid (B; 0.03%, v/v) as mobile phase at a flow rate of 1.0ml/min. Based on the mass spectra, UV spectra and retention time, 47 compounds were identified or tentatively characterized, including 27 flavonols (all from Epimedii Herba, the major component herb), 4 coumarins, 3 flavonones, 1 chalcone, 3 isoflavones, 1 coumestrol, 3 triterpenoid saponins, 1 iridoid, 3 steroidal saponins, and 1 phenolic acid. Among them, 18 compounds were confirmed by comparing with reference standards. In quantitative analysis part, 10 major compounds in 18 batches of XLGB were simultaneously determined by HPLC/UV detected at 270nm. The method was validated with respect to intra- and inter-day precision, repeatability and stability, with RSD less than 1.0%, 1.5%, 2.9% and 1.8%, respectively. All the 10 analytes showed good linearity in wide linear ranges (r(2)=0.9999), and their average recoveries varied between 97.8% and 104.9%.
authors.affiliation: The State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors.forename: Xiang-Yu
authors.initials: XY
authors.lastname: Guan
authors.forename: Hui-Fang
authors.initials: HF
authors.lastname: Li
authors.forename: Wen-Zhi
authors.initials: WZ
authors.lastname: Yang
authors.forename: Chang-Hu
authors.initials: CH
authors.lastname: Lin
authors.forename: Chao
authors.initials: C
authors.lastname: Sun
authors.forename: Bao-Rong
authors.initials: BR
authors.lastname: Wang
authors.forename: De-An
authors.initials: DA
authors.lastname: Guo
authors.forename: Min
authors.initials: M
authors.lastname: Ye
mesh_terms: D002138:Calibration; D002214:Capsules; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002845:Chromatography / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D047309:Flavones / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*; D008516:Medicine, Chinese Traditional; D028321:Plant Preparations / Q000032:analysis; D011786:Quality Control; D012044:Regression Analysis; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D002214:Capsules; D047309:Flavones; D028321:Plant Preparations