title: Identification, bioactivity evaluation and pharmacokinetics of multiple components in rat serum after oral administration of Xian-Ling-Gu-Bao capsule by ultra performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry.
abstract: The Xian-Ling-Gu-Bao capsule (XLGB) is a famous traditional Chinese medicine prescription (TCMP), which has proven effective in osteoporosis treatment. However, due to the lack of a dynamic XLGB profile, the in vivo pharmacokinetics of multiple bioactive components within this medicine remains unknown. In the present study, ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC/Q-TOF-MS) identified a total of eighteen prototypes (using reference standards) in rat serum after oral administration of XLGB. These prototypes were subsequently evaluated to ascertain their effects on the proliferation and alkaline phosphatase activity of UMR106 cells and the adipogenesis of 3T3-L1 cells. Furthermore, a rapid and sensitive UPLC/Q-TOF-MS method was developed and validated for simultaneous quantitative analysis of 11 prototypes in rat serum. Chromatographic separation was achieved using a Waters Acquity BEH C18 column (2.1×100mm, 1.7μm) and linear gradient elution employing a mobile phase consisting of water and acetonitrile (both containing 0.1% formic acid). All calibration curves showed excellent linearity (r2>0.99) within the sampling ranges considered. The assay was accurate, precise and reproducible, as demonstrated by the obtained intra- and inter-day precisions (less than 12.3%) and accuracies (between -12.7% and 11.0%), and the matrix effects, extraction recoveries and stabilities were all satisfactory. Moreover, pharmacokinetic parameters were calculated from the plasma concentration-time data. Compared to single-compound dosing, significantly enhanced responses were obtained when several analytes were administered simultaneously, indicating possible drug-drug interactions among the complex ingredients of TCMP. This work provides an experimental baseline regarding the clinical applications and medicinal effectiveness of XLGB in the treatment of osteoporosis.
authors.affiliation: College of Pharmacy, Jinan University, Guangzhou, 510632, PR China. Electronic address: tyaozh@jnu.edu.cn.
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Yao
authors.affiliation: College of Pharmacy, Jinan University, Guangzhou, 510632, PR China.
authors.forename: Zi-Fei
authors.initials: ZF
authors.lastname: Qin
authors.affiliation: College of Pharmacy, Jinan University, Guangzhou, 510632, PR China.
authors.forename: Liang-Liang
authors.initials: LL
authors.lastname: He
authors.affiliation: Musculoskeletal Research Laboratory, Department of Orthopaedics and Traumatology, Chinese University of Hong Kong, Shatin, N.T. Hong Kong SAR, PR China; Translational Medicine R&D Centre, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518057, PR China.
authors.forename: Xin-Luan
authors.initials: XL
authors.lastname: Wang
authors.affiliation: College of Pharmacy, Jinan University, Guangzhou, 510632, PR China. Electronic address: daiyi1004@163.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Dai
authors.affiliation: Musculoskeletal Research Laboratory, Department of Orthopaedics and Traumatology, Chinese University of Hong Kong, Shatin, N.T. Hong Kong SAR, PR China; Translational Medicine R&D Centre, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518057, PR China.
authors.forename: Ling
authors.initials: L
authors.lastname: Qin
authors.affiliation: Laboratory of Metabolism, Centre for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Gonzalez
authors.affiliation: College of Pharmacy, Jinan University, Guangzhou, 510632, PR China.
authors.forename: Wen-Cai
authors.initials: WC
authors.lastname: Ye
authors.affiliation: College of Pharmacy, Jinan University, Guangzhou, 510632, PR China.
authors.forename: Xin-Sheng
authors.initials: XS
authors.lastname: Yao
mesh_terms: D000284:Administration, Oral; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000032:analysis* / Q000493:pharmacokinetics*; D016014:Linear Models; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004365:Drugs, Chinese Herbal; C572489:xian ling gu bao
keywords: Bioactivity evaluation; Main prototypes; Pharmacokinetics; Rat serum; UPLC/Q-TOF-MS; Xian-Ling-Gu-Bao capsule