Watanabe T, Hammock B D
Department of Entomology and Cancer Research Center, University of California, Davis, CA 95616, USA.
Anal Biochem. 2001 Dec 15;299(2):227-34. doi: 10.1006/abio.2001.5423.
A rapid and reliable electrospray tandem mass spectrometric method for soluble epoxide hydrolase (sEH) inhibitors in rat hepatic microsomes is described. Four synthesized sEH inhibitors were extracted from rat hepatic microsomes with ethyl acetate and were determined by HPLC using positive ion electrospray tandem mass spectrometry within 7 min. The relationship between signal intensity and concentration of sEH inhibitors was linear over the concentration range of 2.0 to 500 ng/mL per 5-microL injection with the use of a noncoeluting internal standard with a similar chemical structure. The intraassay precision was less than 12.4% relative standard deviation and accuracy ranged from -7.0 to 11.3% deviation from the theoretical values with five duplicate assays. The recovery of sEH inhibitors from rat hepatic microsomes, fortified at levels of 50, 100, and 250 ng/mL, averaged 74.2-107.7% with a RSD of 2.1-7.6%. This method was successfully applied to the quantification of residual sEH inhibitors in rat hepatic microsomes without interference.
本文描述了一种用于测定大鼠肝微粒体中可溶性环氧化物水解酶（sEH）抑制剂的快速可靠的电喷雾串联质谱方法。从大鼠肝微粒体中用乙酸乙酯提取4种合成的sEH抑制剂，并通过高效液相色谱法，采用正离子电喷雾串联质谱在7分钟内进行测定。使用具有相似化学结构的非共洗脱内标物时，每5微升进样中，sEH抑制剂的信号强度与浓度在2.0至500纳克/毫升的浓度范围内呈线性关系。在五次重复测定中，批内精密度的相对标准偏差小于12.4%，准确度与理论值的偏差范围为-7.0%至11.3%。在添加水平为50、100和250纳克/毫升的情况下，大鼠肝微粒体中sEH抑制剂的回收率平均为74.2%-107.7%，相对标准偏差为2.1%-7.6%。该方法成功应用于大鼠肝微粒体中残留sEH抑制剂的定量分析，无干扰。
