Jemal M, Hawthorne D
Department of Metabolism and Pharmacokinetics, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ 08543-4500, USA.
J Pharm Biomed Anal. 1995 Dec;14(1-2):43-8. doi: 10.1016/0731-7085(95)01624-4.
BMS-186318 is a member of the recently discovered "aminodiol" class of HIV protease inhibitors. A simple but sensitive method was developed for the determination of BMS-186318 in dog plasma and then applied to monkey and rat plasma. The compound was extracted from dog plasma with methyl tert-butyl ether at basic pH. The dried extract was reconstituted in mobile phase and injected into a 150 x 2.1 mm i.d. Zorbax Rx-C18 HPLC column. A portion of the effluent was directed into the LC-ionspray MS system, where the [M+H]+ ion of the secondary amine compound was monitored. The HPLC conditions were chosen in order to achieve a short run time and large sample throughput, with both analyte and internal standard eluting within 1.5 min. The liquid-liquid extraction procedure provided very clean extracts so that sufficient signal-to-noise ratio was obtained with single-stage mass spectrometry instead of the more costly tandem mass spectrometry. The required lower limit of quantitation of 2.5 ng ml-1 was easily achieved. The method has also been validated for BMS-186318 in monkey plasma without modification. The method has been modified for rat plasma. Owing to irreproducibility observed when applying the liquid-liquid extraction method to rat plasma, a solid-phase extraction method was developed. The addition of phenylmethylsulfonyl fluoride was necessary to stabilize BMS-186318 in rat blood and plasma.
BMS-186318是最近发现的“氨基二醇”类HIV蛋白酶抑制剂的一员。开发了一种简单而灵敏的方法用于测定犬血浆中的BMS-186318，然后将其应用于猴和大鼠血浆。该化合物在碱性pH条件下用甲基叔丁基醚从犬血浆中萃取。干燥的萃取物用流动相复溶后注入内径为150×2.1 mm的Zorbax Rx-C18高效液相色谱柱。一部分流出物导入LC-离子喷雾质谱系统，监测仲胺化合物的[M+H]+离子。选择高效液相色谱条件以实现较短的运行时间和较大的样品通量，分析物和内标均在1.5分钟内洗脱。液-液萃取程序提供了非常纯净的萃取物，因此通过单级质谱法即可获得足够的信噪比，而无需使用成本更高的串联质谱法。轻松实现了所需的2.5 ng ml-1的定量下限。该方法未经修改已在猴血浆中对BMS-186318进行了验证。该方法已针对大鼠血浆进行了修改。由于将液-液萃取方法应用于大鼠血浆时观察到不可重复性，因此开发了一种固相萃取方法。添加苯甲基磺酰氟对于在大鼠血液和血浆中稳定BMS-186318是必要的。
