Remmel R P, Huang S H, Hoff D, Zimmerman C L
Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota, Minneapolis 55455.
J Chromatogr. 1990 Dec 14;534:109-18. doi: 10.1016/s0378-4347(00)82153-2.
Carbovir is a carbocyclic guanosine analogue with potent in vitro activity against the human immunodeficiency virus. All of the activity resides in the (-)-enantiomer. An ion-paired liquid chromatographic assay for (-)-carbovir was developed on a Spherisorb C8 column with fluorescence detection (275 nm excitation, 345 nm emission). Guanosine nucleosides are fluorescent at a pH less than 2.5, and fluorescence detection resulted in a four-fold improvement in the limit of quantitation (0.039 microgram/ml) compared to the previously developed assay with ultraviolet detection. Standard curves were processed with an internal standard at (-)-carbovir concentrations of 0.039-40 micrograms/ml in whole rat blood with a solid-phase extraction technique. Total variability was less than 16% at all concentrations and less than 10% at concentrations greater than 0.3 microgram/ml. Within-day variability was less than 7.5% at concentrations greater than 0.3 microgram/ml. Urine was analyzed directly after dilution and an diethyl ether wash to remove impurities. The total coefficients of variation were less than 10% from 0.5-20 micrograms/ml in urine. The concentrations of (-)-carbovir in rat blood were detectable for as long as 8 h after intravenous and oral doses of 20 and 60 mg/kg, respectively.
卡波韦是一种碳环鸟苷类似物，在体外对人类免疫缺陷病毒具有强大的活性。所有活性都存在于（-）-对映体中。在Spherisorb C8柱上开发了一种用于（-）-卡波韦的离子对液相色谱法，并采用荧光检测（激发波长275nm，发射波长345nm）。鸟苷核苷在pH值小于2.5时具有荧光性，与之前开发的紫外检测法相比，荧光检测使定量限（0.039微克/毫升）提高了四倍。采用固相萃取技术，在全血中（-）-卡波韦浓度为0.039 - 40微克/毫升时，用内标处理标准曲线。在所有浓度下，总变异系数均小于16%，在浓度大于0.3微克/毫升时，总变异系数小于10%。在浓度大于0.3微克/毫升时，日内变异系数小于7.5%。尿液经稀释和乙醚洗涤以去除杂质后直接进行分析。在尿液中，0.5 - 20微克/毫升的（-）-卡波韦总变异系数小于10%。分别静脉注射和口服20毫克/千克和60毫克/千克剂量后，大鼠血液中（-）-卡波韦的浓度在长达8小时内均可检测到。
