A Sensitive Liquid Chromatography-Tandem Mass Spectrometry Method for the Determination of Nimbolide in Mouse Serum: Application to a Preclinical Pharmacokinetics Study
Lingzhi Wang,
Do-Dang Khoa Phan,
Nicholas Syn,
Xiaoqiang Xiang,
Hongyan Song,
Win Lwin Thuya,
Shili Yang,
Andrea Li-Ann Wong,
Alan Prem Kumar,
Wei Peng Yong,
Gautam Sethi,
Paul Chi-Lui Ho,
Boon Cher Goh
Affiliations
Lingzhi Wang
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
Do-Dang Khoa Phan
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
Nicholas Syn
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
Xiaoqiang Xiang
Department of Clinical Pharmacy, School of Pharmacy, Fudan University, Shanghai 201203, China
Hongyan Song
Institute of Materials Research and Engineering (IMRE), ASTAR, Singapore 138634, Singapore
Win Lwin Thuya
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
Shili Yang
Department of Pharmacy, National University of Singapore, Singapore 117543, Singapore
Andrea Li-Ann Wong
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
Alan Prem Kumar
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
Wei Peng Yong
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
Gautam Sethi
Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117600, Singapore
Paul Chi-Lui Ho
Department of Pharmacy, National University of Singapore, Singapore 117543, Singapore
Boon Cher Goh
Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599, Singapore
A sensitive and robust liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the determination of nimbolide in mouse serum. Exemestane was used as the internal standard (IS). Here, we employed acetonitrile-based protein precipitation (PPT) for serum sample preparation, and performed chromatographic separation using an ODS Hypersil C18 column (100 mm × 2.1 mm, 5 µm) with gradient elution (0.1% formic acid in water vs 100% acetonitrile). The run time was 6 min. Instrumental analysis was performed by electrospray ionization tandem mass spectrometry (ESI-MS/MS) in the multiple-reaction monitoring (MRM) under positive mode. A good linear calibration was achieved in the 5–1000 ng/mL range. The intra- and inter-day precisions for nimbolide were ≤12.6% and ≤13.9% respectively. Intra-day accuracy ranged from 96.9–109.3%, while inter-day accuracy ranged from 94.3–110.2%. The matrix effect of nimbolide, detected but consistent at low and high concentrations, do not affect linearity of standard curve. In conclusion, we have developed and validated a sensitive analytical method for determination of a novel natural compound nimbolide in mouse serum, and it has been successfully applied to our preclinical study in investigating the pharmacokinetic properties of nimbolide, which could greatly facilitate the preclinical development of the promising lead compound for anticancer therapy.
