Sotnichenko A I, Serdiuk O A, Kilesso T Iu, Sukhanov V A, Saprin A N
Antibiot Med Biotekhnol. 1985 Aug;30(8):584-8.
A highly sensitive method for determination of the activity of epoxide hydrolase (EH) (EC 3.3.2.3) in the mammalian lymphocytes was developed. 9,10-Epoxy-9,10-dihydrophenanthrene was used as substrate. For elimination of the enzyme latency the lymphocytes were solubilized with lubrol PX, a nonionic detergent. 9,10-Hydroxy-9,10-dihydrophenanthrene was estimated quantitatively by the method of the internal standard (1-naphthol) after reversed phase HPLC. The time of chromatographic matographic separation was 2.5 minutes. By the pH optimum EH in the lymphocytes differed from microsomal EH. However, the structure of their active sites was similar. Among the species studied (mice, rats, guinea-pigs, man), the lowest activity of EH was detected in the lymphocytes of mice and the highest activity in the lymphocytes of guinea-pigs. The activity of EH in the lymphocytes of man was 10-20 times higher than the sensitivity limit of the method (5 pkmol/mg/min). By its sensitivity and reproducibility and the level of the activity determined the method is much more superior to the methods described in the literature.
建立了一种用于测定哺乳动物淋巴细胞中环氧化物水解酶（EH）（EC 3.3.2.3）活性的高灵敏度方法。使用9,10-环氧-9,10-二氢菲作为底物。为消除酶的潜伏性，用非离子去污剂卢勃罗尔PX溶解淋巴细胞。反相高效液相色谱后，采用内标法（1-萘酚）对9,10-羟基-9,10-二氢菲进行定量测定。色谱分离时间为2.5分钟。淋巴细胞中的EH最适pH与微粒体EH不同。然而，它们活性位点的结构相似。在所研究的物种（小鼠、大鼠、豚鼠、人）中，小鼠淋巴细胞中EH的活性最低，豚鼠淋巴细胞中EH的活性最高。人淋巴细胞中EH的活性比该方法的灵敏度极限（5皮摩尔/毫克/分钟）高10 - 20倍。该方法的灵敏度、重现性以及所测定的活性水平均远优于文献中描述的方法。
