摘要
目的：建立快速测定细胞色素Ｐ４５０ＣＹＰ１Ａ２酶活性的高压液相色谱方法．方法：取３００μＬ血浆样品，用β羟乙基茶碱作内标，经５ｍＬ氯仿／异丙醇（９∶１）萃取处理后，用００５％的乙酸、乙腈和甲醇作为基本流动相，采用梯度洗脱程序在ＯＤＳ柱上分离待测组分，紫外检测波长２８２ｎｍ．结果：无内源性物质干扰测定．次黄嘌呤、内标和咖啡因快速基线分离，三者的保留时间均小于１３分钟．次黄嘌呤和咖啡因的检测下限均为０１μｍｏｌ·Ｌ－１，线性范围分别为１－１００μｍｏｌ·Ｌ－１和１－２００μｍｏｌ·Ｌ－１，相关系数分别为０９９９９和０９９８７，变异系数分别小于６％和１０％．两者的平均相对回收率为９６％－１０８％．结论：本方法快速、灵敏，可用于人群ＣＹＰ１Ａ２酶活性研究．
AIM: To develop a rapid HPLC method for the determination of cytochrome P 450 CYP1A2 activity. METHODS: A 300 μL plasma was prepared by extraction with 5 mL chloroform/isopropanol (9∶1) , and β hydroxyletheophylline was added as internal standard (IS). Samples were separated on an ODS column by a gradient elution system, of which mobile phase consisted of 0 05 % acetic acid, acetonitrile, and methanol. The compounds of interest were monitored at 282 nm by UV detector. RESULTS: No potential interfering peaks were found. Paraxanthine (17X), IS and caffeine (137X) were rapidly eluted with baseline resolution, and their retention time was less than 13 min. The detection limits of both 17X and 137X were 0 1 μmol·L -1 . Linear relations ranged over 1-100 μmol·L -1 and 1-200 μmol·L -1 with correlation coefficient of 0 9999 and 0 9987, respectively, for 17X and 137X. The coefficients of variation were within 6 % for 17X, and 10 % for 137X. The average recoveries for both compounds were ranged from 96 % to 108 % . CONCLUSION: This method is sensitive and rapid, and can be used for population studies of CYP1A2.
出处
《中国药理学报》
CSCD
1998年第1期44-46,共3页
Acta Pharmacologica Sinica
关键词
咖啡因
细胞色素
P-450
CYP1A2
caffeine
xanthines
cytochrome P 450 CYP1A2
high pressure liquid chromatography
pharmacokinetics
