Dzerk A M, Carlson A, Loewen G R, Shirley M A, Lee J W
Harris Laboratories, Lincoln, NE 68502, USA.
J Pharm Biomed Anal. 1998 Feb;16(6):1013-9. doi: 10.1016/s0731-7085(97)00114-3.
A HPLC method was developed and validated for the quantitation of 9-cis-retinoic acid (ALRT1057) and its major metabolite, 4-oxo-9-cis-retinoic acid (LG100182) in human plasma. Samples were buffered and extracted with methyl-tert-butyl-ether. The analytes and an I.S. were separated on a C18 HPLC column using a shallow gradient of 70-89% organic solvent. The analytes were quantitated by UV detection at 348 nm. Selectivity against endogenous compounds and potential metabolites (retinol, all trans-, 13-cis-, and 4-hydroxy-9-cis-retinoic acid) was demonstrated. The run time was 29 min. The standard curve was linear from 2.5 to 450 ng ml-1. Interassay precision for both analytes in quality control samples was less than 5.0% RSD. Accuracy was within 11.0% RE for both compounds. Analyte stability during sample storage, extraction processing, and chromatography was established. Method ruggedness was tested by two analysts and on two HPLC systems. This method has been applied to the quantitation of clinical samples.
建立并验证了一种高效液相色谱法（HPLC），用于定量测定人血浆中9-顺式维甲酸（ALRT1057）及其主要代谢物4-氧代-9-顺式维甲酸（LG100182）。样品经缓冲后用甲基叔丁基醚萃取。分析物和内标物在C18高效液相色谱柱上分离，采用70 - 89%有机溶剂的浅梯度洗脱。通过在348 nm处的紫外检测对分析物进行定量。证明了该方法对内源性化合物和潜在代谢物（视黄醇、全反式、13-顺式和4-羟基-9-顺式维甲酸）具有选择性。运行时间为29分钟。标准曲线在2.5至450 ng ml-1范围内呈线性。质控样品中两种分析物的批间精密度RSD均小于5.0%。两种化合物的准确度均在11.0% RE以内。确定了样品储存、萃取过程和色谱分析过程中分析物的稳定性。由两名分析人员在两个高效液相色谱系统上测试了方法的耐用性。该方法已应用于临床样品的定量分析。
