Shaik Abdul Naveed, Altomare Deborah A, Lesko Lawrence J, Trame Mirjam N
Center for Pharmacometrics and Systems Phamacology, Department of Pharmaceutics, College of Pharmacy, University of Florida, Orlando, FL, USA.
Burnett School of Biomedical Sciences, University of Central Florida, Orlando, FL, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Mar 1;1046:243-249. doi: 10.1016/j.jchromb.2016.11.027. Epub 2016 Nov 23.
Till date, no analytical method published to detect Cisplatin has been validated according to the U.S. Food and Drug Administration (FDA) guidance using liquid chromatography mass spectrometry (LC-MS/MS). We report, a validated LC-MS/MS method for quantitative determination of cisplatin in rat plasma and urine according to FDA guidlines. Cisplatin is a platinum containing compound used for the treatment of different types of cancers. Quantitative determination of cisplatin has been carried out using atomic absorption spectroscopy, high pressure liquid chromatography with phosphorescence, ultra-violet detection, or with inductively coupled plasma mass spectrometry. Few LC-MS/MS methods have been reported for the analysis of cisplatin either for direct quantification or indirect by derivatizing with organic compounds but none of the reported methods have validated the method. The developed and validated assay presented here is a highly sensitive LC-MS/MS method developed and validated for the quantitative determination of cisplatin following derivatization with diethyldithiocarbamate (DDTC) in order to detect platinum (Pt) of cisplatin, suitable for pharmacokinetic studies in rats and to further use it to study human toxicology. Chromatographic separation was achieved using a Poroshell 120 EC-C column (3×50mm, 2.7μm) with a binary gradient mobile phase. Quantification was performed on a triple quadruple with electrospray ionization and detection was performed using multiple reaction monitoring. The method has a limit of detection of 1ng/mL, and the quantifiable range was 3-3000ng/mL in rat plasma and urine. The method was accurate and precise with an accuracy and precision for intra-day and inter-day of ±20% for lower limit of quantitation and of ±15% for low, mid and high quality control samples. This method was successfully applied to study the pharmacokinetic profile of cisplatin in rat plasma and urine given a range of doses from 0.5 to 3.5mg/kg.
迄今为止，尚未有已发表的用于检测顺铂的分析方法依据美国食品药品监督管理局（FDA）的指导原则通过液相色谱-质谱联用（LC-MS/MS）进行验证。我们报告了一种依据FDA指导原则对大鼠血浆和尿液中的顺铂进行定量测定的经过验证的LC-MS/MS方法。顺铂是一种含铂化合物，用于治疗不同类型的癌症。已使用原子吸收光谱法、带磷光检测的高压液相色谱法、紫外检测法或电感耦合等离子体质谱法对顺铂进行了定量测定。很少有LC-MS/MS方法被报道用于顺铂的分析，无论是直接定量还是通过与有机化合物衍生化进行间接分析，但所报道的方法均未对该方法进行验证。此处所展示的已开发并验证的分析方法是一种高度灵敏的LC-MS/MS方法，该方法通过用二乙基二硫代氨基甲酸盐（DDTC）衍生化以检测顺铂中的铂（Pt）来开发并验证用于顺铂的定量测定，适用于大鼠的药代动力学研究，并进一步用于研究人体毒理学。使用Poroshell 120 EC-C柱（3×50mm，2.7μm）和二元梯度流动相实现色谱分离。在配备电喷雾电离的三重四极杆上进行定量，使用多反应监测进行检测。该方法的检测限为1ng/mL，在大鼠血浆和尿液中的可定量范围为3 - 3000ng/mL。该方法准确且精密，定量下限的日内和日间准确度和精密度为±20%，低、中、高质量控制样品的准确度和精密度为±15%。该方法成功应用于研究给予0.5至3.5mg/kg一系列剂量的顺铂在大鼠血浆和尿液中的药代动力学特征。
