HPLC测定8－CL－CAMP血药浓度的方法研究

Determination of 8-CL-CAMP in Rabbit Serum and Human Serum

本文介绍用反相ＨＰＬＣ法测定兔血清及人血清中８－ＣＬ－ＣＡＭＰ的浓度。以化合物ＣＡＭＰ［Ⅱ］为内标，血样用甲醇作蛋白沉淀剂，使用ＫｅｓｈｅｎｇＯＤＳＣ_（１８）（４．６ｍｍ＝×２５０ｍｍ，１０μｍ）色谱柱，流动相为０．０２ｍｏｌ／Ｌ磷酸二氢钾－甲醇（９２∶８），检测波长为２５９ｎｍ，药［Ⅰ］和内标［Ⅱ］峰的分离度好。保留时间分别为３６ｍｉｎ和２２ｍｉｎ，血药浓度的线性范围在１００～１０００ｎｇ／ｍｌ，最小检测浓度为５０ｎｇ／ｍｌ。此方法简便、灵敏度高。

A modified RP-HPLC method has been developed in the determination of 8-CL-CAMP in rabbit serum and in human serum. Methanol was used as the protein precipitant in the preparation of serum samples that contained CAMP as internal standard. Kesheng ODS C18 column was used. The mobile phase included 92% 0. 02mol/L KH2PO4 and 8% methanol (V/V) .Detection was at 259nm. The linear relationship between peak heights and the concentrations (100￣1000ng/ml) was obtained. The recovery was 96. 83%±2. 69% (rabbit serum) adn 95. 13%±1.13% (human serum). The minimal detectable concentration limit of 50ng/ml of 8-CL-CAMPwas achieved. The method is simple, sensitive and accurate.