Chandrasekhar Devaraj V, Suresh Ponnayyan Sulochana, Dittakavi Sreekanth, Hiremath Rakesh A, Bhamidipati Ravi Kanth, Richter Wolfgang, Srinivas Nuggehally R, Mullangi Ramesh
Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Yeshwanthpur, Bangalore, India.
TUBE Pharmaceuticals GmbH, Wien, Austria.
Biomed Chromatogr. 2018 Feb;32(2). doi: 10.1002/bmc.4065. Epub 2017 Sep 25.
A simple, specific, sensitive and rapid LC-ESI-MS/MS method has been developed and validated for the quantification of 4-methylpyrazole in dog plasma using N-methylnicotinamide-d as an internal standard (IS) as per regulatory guidelines. Sample preparation was accomplished through a simple protein precipitation. Chromatographic separation of 4-methylpyrazole and the IS was performed on a monolithic (Chromolith RP ) column using an isocratic mobile phase comprising 0.2% formic acid in water and acetonitrile (20:80, v/v) at a flow rate of 1.0 mL/min. Elution of 4-methylpyrazole and the IS occurred at ~1.60 and 1.56 min, respectively. The total chromatographic run time was 3.2 min. A linear response function was established in the concentration range of 4.96-4955 ng/mL. The intra- and inter-day accuracy and precision were in the ranges 1.81-12.9 and 3.80-11.1%, respectively. This novel method has been applied to a pharmacokinetic study in dogs.
已根据监管指南开发并验证了一种简单、特异、灵敏且快速的液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）方法，用于定量犬血浆中的4-甲基吡唑，以内标N-甲基烟酰胺-d（IS）进行测定。通过简单的蛋白沉淀完成样品制备。4-甲基吡唑和内标的色谱分离在整体式（Chromolith RP）柱上进行，使用由0.2%甲酸水溶液和乙腈（20:80，v/v）组成的等度流动相，流速为1.0 mL/min。4-甲基吡唑和内标的洗脱时间分别约为1.60分钟和1.56分钟。总色谱运行时间为3.2分钟。在4.96 - 4955 ng/mL的浓度范围内建立了线性响应函数。日内和日间准确度及精密度分别在1.81 - 12.9%和3.80 - 11.1%的范围内。该新方法已应用于犬的药代动力学研究。
