Chan Vivien, Charles Bruce G, Tett Susan E
School of Pharmacy, The University of Queensland, Steele Building, Brisbane, Qld 4072, Australia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Apr 25;803(2):331-5. doi: 10.1016/j.jchromb.2004.01.016.
A simple method for the measurement of the active leflunomide metabolite A77 1726 in human plasma by HPLC is presented. The sample workup was simple, using acetonitrile for protein precipitation. Chromatographic separation of A77 1726 and the internal standard, alpha-phenylcinnamic acid, was achieved using a C(18) column with UV detection at 305 nm. The assay displayed reproducible linearity for A77 1726 with determination coefficients (r2) > 0.997 over the concentration range 0.5-60.0 microg/ml. The reproducibility (%CV) for intra- and inter-day assays of spiked controls was <5%. The limit of quantification was 0.8 microg/ml. The average absolute recovery was approximately 100%. This assay is suitable for the determination of A77 1726 in plasma of patients taking leflunomide, and is simpler to use than other HPLC methods reported previously.
介绍了一种用高效液相色谱法测定人血浆中活性来氟米特代谢物A77 1726的简单方法。样品处理简单，用乙腈进行蛋白质沉淀。使用C(18)柱在305 nm处进行紫外检测，实现了A77 1726与内标α-苯基肉桂酸的色谱分离。该测定法在0.5 - 60.0微克/毫升的浓度范围内，A77 1726显示出可重复的线性，测定系数（r2）> 0.997。加标对照品日内和日间测定的重现性（%CV）<5%。定量限为0.8微克/毫升。平均绝对回收率约为100%。该测定法适用于测定服用来氟米特患者血浆中的A77 1726，且比先前报道的其他高效液相色谱法更易于使用。
