title: Development and validation of liquid chromatography-tandem mass spectrometry method for quantification of a potential anticancer triterpene saponin from seeds of Nigella glandulifera in rat plasma: application to a pharmacokinetic study.
abstract: Nigella glandulifera Freyn et Sit is a folk medicinal plant, whose seeds show significant anticancer activities attributed to triterpene saponins and volatile oil. In this study, an in vitro cytotoxicity assay demonstrated that Nigella A, the major component of triterpene saponins extracted from N. glandulifera, exhibited growth inhibition in the human lung carcinoma A-549 cell line. Due to this potential activity, a reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify Nigella A in rat plasma for a pharmacokinetic study was developed. Nigella A and pravastatin, as internal standard (IS), were extracted from rat plasma using acetonitrile to precipitate protein. Separation was performed on an Agilent Zorbax SB-Aq (3.0 × 150 mm, 3.5 μm) column using a gradient elution method with acetonitrile-0.1% formic acid in water at a flow rate of 0.35 mL/min. Detection was performed using an electrospray ionization in a negative ion multiple reaction monitoring mode. The deprotonated precursor to product ion transitions monitored for Nigella A and IS was at m/z 1352.7→882.6 and m/z 423.1→321.0, respectively. The linear range was 0.240-120 μg/mL with a square regression coefficient (r=0.9996). The intra-day and inter-day precision was less than 6.93%. The simple extraction procedure provided recovery ranged from 92.32 to 95.44% for both analyte and IS. The method was proved to be reliable, precise, and accurate, and was successfully applied to a pharmacokinetic study of Nigella A in rats after i.v. administration via the tail vein at doses of 10, 20, and 30 mg/kg.
authors.affiliation: Research Center for Clinical Pharmacy, First Affiliated Hospital, Zhejiang University, Hangzhou 310003, PR China.
authors.forename: Xingjiang
authors.initials: X
authors.lastname: Hu
authors.affiliation: Institute of Clinical Pharmaceutical Research, Hunan Provincial Tumor Hospital/Affiliated Tumor Hospital of Xiangya Medical School of Central South University, Changsha 410013, PR China.
authors.forename: Xiaobao
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, PR China.
authors.forename: Minghua
authors.initials: M
authors.lastname: Gong
authors.affiliation: Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, PR China.
authors.forename: Ming
authors.initials: M
authors.lastname: Luan
authors.affiliation: Research Center for Clinical Pharmacy, First Affiliated Hospital, Zhejiang University, Hangzhou 310003, PR China.
authors.forename: Yunliang
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Research Center for Clinical Pharmacy, First Affiliated Hospital, Zhejiang University, Hangzhou 310003, PR China.
authors.forename: Guolan
authors.initials: G
authors.lastname: Wu
authors.affiliation: Research Center for Clinical Pharmacy, First Affiliated Hospital, Zhejiang University, Hangzhou 310003, PR China. Electronic address: stjz@zju.edu.cn.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shentu
authors.affiliation: Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, PR China. Electronic address: zhanglin@zju.edu.cn.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D005260:Female; D006801:Humans; D016014:Linear Models; D008297:Male; D031844:Nigella / Q000737:chemistry*; D010936:Plant Extracts / Q000008:administration & dosage / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012503:Saponins / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D012639:Seeds / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014315:Triterpenes / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D010936:Plant Extracts; D012503:Saponins; D014315:Triterpenes
keywords: LC–MS/MS; Nigella A; Pharmacokinetics; Rat plasma