title: Determination of deltonin in rat plasma by using HPLC-MS/MS and the application of this method in pharmacokinetic studies.
abstract: Deltonin is a naturally occurring spirostanol glycoside from Dioscorea zingiberensis C.H. Wright, which is used in traditional Chinese medicine. It exerts strong cytotoxic effect on C26 cells, inhibits C26 derived-tumor growth, and prolongs the survival of tumor-bearing mice after its oral administration, indicating its potential for use as an anti-tumor drug. To investigate the pharmacokinetic profiles of deltonin, a rapid, sensitive, and simplified high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay was developed and validated for the determination of deltonin in rat plasma. After acetonitrile-mediated plasma protein precipitation, chromatographic separation of deltonin was achieved using a reversed phase Hypersil Gold column (150mm×2.1mm, 5μm), with gradient elution using 0.1% formic acid and acetonitrile. Thereafter, deltonin was quantified using MS/MS with electrospray ionization (ESI) in positive multiple reaction monitoring (MRM) mode. The flow rate of the mobile phase was 200μL/min, and the retention time was 9.03min for deltonin and 6.31min for the internal standard (IS: 20(S)-ginsenoside Rb1). The linear range of the calibration curve was 2-5000ng/mL (r(2)>0.99), and the limit of detection (LOD) was 0.46ng/mL. The intra- and inter-day accuracies ranged from -2.8% to 11.1% and precisions (RSD) were within 13.1%. Deltonin was found to be stable under short-term temperature conditions, post-preparative temperature conditions, and after 3 freeze-thaw cycles conditions. The validated method was successfully applied to a pharmacokinetic study in rats after oral administration of deltonin (50 and 100mg/kg). The pharmacokinetics is characterized by high apparent clearance (CL/F) and apparent volume of distribution (Vd/F).
authors.affiliation: Laboratory of Ethnopharmacology, Regenerative Medicine Research Center, West China Hospital/West China Medical School, and Institute for Nanobiomedical Technology and Membrane Biology, Sichuan University, Chengdu 610041, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Du
authors.forename: Bo
authors.initials: B
authors.lastname: Gao
authors.forename: Guang
authors.initials: G
authors.lastname: Xin
authors.forename: Aimin
authors.initials: A
authors.lastname: Sun
authors.forename: Baozhan
authors.initials: B
authors.lastname: Huang
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Xing
authors.forename: Qianming
authors.initials: Q
authors.lastname: Chen
authors.forename: Yang
authors.initials: Y
authors.lastname: He
authors.forename: Wen
authors.initials: W
authors.lastname: Huang
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D016018:Least-Squares Analysis; D057230:Limit of Detection; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D013150:Spirostans / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D013150:Spirostans; C016347:deltonin