Sensitive LC-MS/MS Method for the Simultaneous Determination of Bendamustine and its Active Metabolite, -Hydroxybendamustine in Small Volume Mice and Dog Plasma and its Application to a Pharmacokinetic Study in Mice and Dogs
Sensitive LC-MS/MS Method for the Simultaneous Determination of Bendamustine and its Active Metabolite, -Hydroxybendamustine in Small Volume Mice and Dog Plasma and its Application to a Pharmacokinetic Study in Mice and Dogs
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DOI：
10.1055/s-0043-108124
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发表时间：
2017-09-01
期刊：
影响因子：
2.2
通讯作者：
Srinivas, Nuggehally R.
中科院分区：
文献类型：
--
作者：
Chandrashekar, Devaraj V.;Suresh, Ponnayyan S.;Srinivas, Nuggehally R.
根据监管指南，采用非那西丁作为内标物（IS），开发并验证了一种高灵敏度、特异性和快速的LC-ESI-MS/MS方法，用于同时定量小体积（20 μ L）小鼠和犬血浆中的苯达莫司汀（BM）和β-羟基苯达莫司汀（HBM）。使用液-液萃取法从小鼠和犬血浆中萃取分析物和IS。色谱条件：Atlantis dC（18）柱，等度移动的流动相（0.2%甲酸：乙腈，25：75），流速0.40 mL/min，总色谱运行时间3.0 min，BM、HBM和IS的洗脱时间分别为1.2、1.2和2.0 min。小鼠和犬血浆中分析物的线性响应函数均为0.11- 518 ng/mL。小鼠血浆中BM和HBM的日内和日间准确度和精密度分别为3.46-12.9和3.63-8.23%; 1.15-9.00和7.86-9.49%，犬血浆中BM和HBM的日内和日间准确度和精密度分别为2.15-6.49和1.73-13.1%; 4.35-13.9和4.33-10.5%。这种新方法已应用于小鼠和犬的药代动力学研究。
A highly sensitive, specific and rapid LC-ESI-MS/MS method has been developed and validated for the simultaneous quantification of bendamustine (BM) and -hydroxybendamustine (HBM) in small volume (20 mu L) mice and dog plasma using phenacetin as an internal standard (IS) as per regulatory guidelines. Both the analytes and IS were extracted from mice and dog plasma using a liquid-liquid extraction method. Chromatography was achieved on Atlantis dC(18) column using an isocratic mobile phase (0.2% formic acid:acetonitrile, 25:75) at a flow rate of 0.40mL/min. The total chromatographic run time was 3.0min and the elution of BM, HBM and IS occurred at similar to 1.2, 1.2 and 2.0min, respectively. A linear response function was established 0.11-518ng/mL for both the analytes in mice and dog plasma. The intra- and inter-day accuracy and precisions were in the range of 3.46-12.9 and 3.63-8.23%; 1.15-9.00 and 7.86-9.49% for BM and HBM, respectively in mice plasma and 2.15-6.49 and 1.73-13.1%; 4.35-13.9 and 4.33-10.5% for BM and HBM, respectively in dog plasma. This novel method has been applied to a pharmacokinetic study in mice and dogs.