LC–MS/MS method development for quantification of busulfan in human plasma and its application in pharmacokinetic study

• Published in: Journal of pharmaceutical and biomedical analysis 2016-02, Vol.120, p.168-174
• Main Authors: Nadella, Taraka Ramarao, Suryadevara, Vidyadhara, Lankapalli, Sasidhar Reddyvallam, Mandava, Venkata Basaveswara Rao, Bandarupalli, Deepti
• Format: Article
• Language: English
• Subjects: Administration, Oral; Busulfan; Busulfan - blood; Busulfan - pharmacokinetics; Busulfan d8; Chromatography, Liquid - methods; Chromatography, Liquid - trends; Human Plasma; Humans; LC–MS/MS; Male; Mass Spectrometry - methods; Mass Spectrometry - trends; Tandem Mass Spectrometry - methods; Tandem Mass Spectrometry - trends; Validation
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2015.12.024

[Display omitted] •A simple, cost effective rugged and a high throughput method was developed for estimation of busulfan in human plasma.•The method consists of a simple sample pretreatment by protein precipitation to give consistent and reproducible recoveries of busulfan.•In the present work a new LC–MS/MS method for the quantification of busulfan in human plasma was developed for clinical study sample analysis.•The proposed method is suitable for comparing the pharmacokinetic profiles of formulations. A simple, rapid, specific and precise liquid chromatography—tandem mass spectrophotometric (LC–MS/MS) method was developed and validated for quantification of busulfan, in human plasma. busulfan d8 was used as internal standard, added to plasma sample prior to extraction using acetonitrile as a precipitating agent. Chromatographic separation was achieved on phenomenex kinetex C18 column (50mm×2.1mm, 2.6μm) with acteonitrile: 10mM ammonium formate buffer (80:20v/v) as an isocratic mobile phase with a flow rate of 0.5mLmin−1. Quantitation was performed by transition of 264.1→151.1 (m/z) for busulfan and 272.1→159.1 (m/z) for busulfan d8. The lower limit of quantitation was 0.2ngmL−1 with a 100μL plasma sample. The concentrations of nine working standards showed linearity between 0.2 and 100ngmL−1 (r2≥0.9986). Chromatographic separation was achieved within 2.0min. The average extraction recoveries of 3quality control concentrations were 92.52% for busulfan and 90.75% for busulfan d8. The coefficient of variation was ≤15% for intra- and inter-batch assays. The developed method was successfully applied for the determination of Busulfan pharmacokinetics after oral administration.