Brunner L J, Yau J C, Lautersztain J, Luke D R
Department of Pharmaceutics, University of Houston.
Clin Chem. 1990 Feb;36(2):329-31.
We describe a modification of the commercially available cyclosporine (CAS) 3H-RIA kit, allowing detection of drug in lipoprotein fractions obtained by ultracentrifugation. Sodium bromide solutions containing the lipoprotein fractions were dried and reconstituted with drug-free plasma. Methanolic extractions were centrifuged, and 250 microL of the supernate was dried and reconstituted with 50 microL of methanol. Buffer, radioactive tracer, and monoclonal antibody were then added and incubated for 2 h at 4 degrees C. Samples were subjected to charcoal de-activation and then centrifuged, and the radioactivity in the supernate was counted. The limit of detection of CSA was 2.5 micrograms/L; analytical recovery was between 97.5% and 101.3%. Within- and between-day CVs were less than 9%. We conclude that the present method may be beneficial for therapeutic drug monitoring of CSA in lipoprotein fractions.
我们描述了一种对市售环孢素（CAS）3H-RIA试剂盒的改进方法，该方法可检测通过超速离心获得的脂蛋白组分中的药物。将含有脂蛋白组分的溴化钠溶液干燥，并用无药物血浆重新溶解。甲醇提取物经离心后，取250微升上清液干燥，并用50微升甲醇重新溶解。然后加入缓冲液、放射性示踪剂和单克隆抗体，在4℃下孵育2小时。样品经活性炭失活处理后再离心，测定上清液中的放射性。环孢素A的检测限为2.5微克/升；分析回收率在97.5%至101.3%之间。批内和批间变异系数均小于9%。我们得出结论，本方法可能有助于脂蛋白组分中环孢素A的治疗药物监测。
