关键词: 双波长差示一阶导数分光光度法, 一氧化碳, 定量分析, 药物代谢, 肝微粒体, 三氟苯胺, 细胞色素P-450

Abstract: A dual wavelength differential first derivative spectrophotometric method has been developed to standardize the concentration of a saturated aqueous solution of carbon monoxide (CO) as the standard and to identify and to determine CO formed during the microsomal metabolism of xenobiotics in vitro. The method can significantly eliminate the background interference in the assay media and increase the quantitative accuracy and the sensitivity. There is a good linear relationship between CO concentration in the range of 2~10 μmol·L^-１ CO and the distance D between the first derivative peak at 415 nm amd valley at 426 nm with r = 0.9999 (n = 5), the regression equation being C (mmol·L^-１) = 17.6 D-0.4, the detection limit lower than 0.1 μmol·L^-１ CO. The average recoveries of CO from the assay system and the sample were 102.1%, RSD = 2.9% (n = 7) and 79.7%, RSD = 6.8% (n = 12),respectively. The RSD of within day was 4.4% (n = 18), and the RSD of day to day was 6.1% (n = 16). By this method, four trihaloanilines and one trihalobenzene were tested, the results showed that only 2,4,5 trifluoroaniline could be converted to CO by the incubation with rat hepatic microsomes, NADPH and oxygen, the ability of phenobarbital or dexamethasone to induce rat hepatic microsomes to catalyze CO formation was 3 or 8 times higher than that of the control.

Key words: Dual wavelength differential first derivative spectrophotometry, Carbon monoxide, Trifluoroaniline, Drug metabolism, Hepatic microsomes, Cytochrome P-450