Zhang Xiqian, Li Ruina, Hu Wenya, Zeng Jin, Jiang Xuehua, Wang Ling
Department of Clinical Pharmacy and Pharmacy Administration, Key Laboratory of Drug Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu, Sichuan, China.
Biomed Chromatogr. 2018 Nov;32(11):e4331. doi: 10.1002/bmc.4331. Epub 2018 Sep 6.
A rapid, specific, and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated to simultaneously quantify N-acetyl-p-benzoquinoneimine (NAPQI), acetaminophen-glutathione (acetaminophen-glut) and acetaminophen-glucuronide (acetaminophen-gluc) in mouse plasma, liver and kidney homogenates. Analytes were eluted by a binary gradient mobile phase composed of water (phase A) and methanol containing 0.1% formic acid (phase B) at a flow rate of 0.3 mL/min, which was performed on a CAPCELL PAK C MG II column. It took 3.2 min to detect three analytes in a single run. Quantification was carried out in positive mode combined with multiple reaction monitoring. The validation of the LC-MS/MS method consisted of specificity, linearity, precision, accuracy, protein precipitation recovery, matrix effect, dilution integrity and stability. The plasma and tissue homogenate calibration curves were linear over concentration ranges of 0.050-5.00, 0.050-5.00 and 0.100-40.0 μg/mL, with a lower limit of quantification of 0.050, 0.050, and 0.100 μg/mL for NAPQI, acetaminophen-glut and acetaminophen-gluc, respectively. The intra- and inter-run precision values were within 12.47% for NAPQI, 12.11% for acetaminophen-glut and 11.86% for acetaminophen-gluc at their lower limit of quantitation levels. The samples were stable under all tested conditions. This method was successfully applied to study the pharmacokinetics of NAPQI, acetaminophen-glut and acetaminophen-gluc in ICR mice following oral administration of 200 mg/kg of acetaminophen suspension.
已开发并验证了一种快速、特异且灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量小鼠血浆、肝脏和肾脏匀浆中的N-乙酰对苯醌亚胺（NAPQI）、对乙酰氨基酚-谷胱甘肽（对乙酰氨基酚-谷胱）和对乙酰氨基酚-葡萄糖醛酸苷（对乙酰氨基酚-葡糖醛）。分析物通过由水（A相）和含0.1%甲酸的甲醇（B相）组成的二元梯度流动相以0.3 mL/min的流速洗脱，该过程在CAPCELL PAK C MG II柱上进行。单次运行检测三种分析物耗时3.2分钟。定量采用正离子模式结合多反应监测进行。LC-MS/MS方法的验证包括特异性、线性、精密度、准确度、蛋白沉淀回收率、基质效应、稀释完整性和稳定性。血浆和组织匀浆校准曲线在0.050 - 5.00、0.050 - 5.00和0.100 - 40.0 μg/mL的浓度范围内呈线性，NAPQI、对乙酰氨基酚-谷胱和对乙酰氨基酚-葡糖醛的定量下限分别为0.050、0.050和0.100 μg/mL。在定量下限水平，NAPQI的批内和批间精密度值在12.47%以内，对乙酰氨基酚-谷胱为12.11%，对乙酰氨基酚-葡糖醛为11.86%。样品在所有测试条件下均稳定。该方法成功应用于研究200 mg/kg对乙酰氨基酚悬浮液口服给药后ICR小鼠体内NAPQI、对乙酰氨基酚-谷胱和对乙酰氨基酚-葡糖醛的药代动力学。
