Nation R L, Peng G W, Chiou W L
J Chromatogr. 1978 Jul 1;146(1):121-31. doi: 10.1016/s0378-4347(00)81296-7.
A rapid high-pressure liquid chromatographic (HPLC) assay is described for the quantitative analysis of tolbutamide and its major metabolite, carboxy tolbutamide in plasma. An aliquot (25--100 microliter) of plasma was prepared for chromatography by deproteinization as follows. One volume of plasma and 2.5 volumes of acetonitrile were vortex mixed for a few seconds and then centrifuged for approx. 1 min. A 50-microliter sample of the clear supernatant was injected into the chromatograph. A mu Bondapak C18 reversed-phase column was used with a mobile phase acetonitrile--0.05% phosphoric acid (45:55) at a flow-rate of 1.5 ml/min. The column effluent was monitored by a variable-wavelength UV detector set at 200 nm. Tolbutamide and its metabolite had retention times of 5.75 and 3.25 min, respectively. The procedure yuelds reproducible results with sensitivity adequate for routine clinical monitoring of plasma levels or for single-dose pharmacokinetic studies. A number of commonly used drugs do not interfere with the method. A single plasma sample can be analyzed in approx. 9 or 10 min.
本文描述了一种快速高压液相色谱（HPLC）分析法，用于定量分析血浆中的甲苯磺丁脲及其主要代谢产物羧基甲苯磺丁脲。取一份血浆样本（25 - 100微升），按如下方法进行脱蛋白处理以制备用于色谱分析的样品。将一份血浆与2.5份乙腈涡旋混合数秒，然后离心约1分钟。取50微升清亮上清液注入色谱仪。使用μ Bondapak C18反相柱，流动相为乙腈 - 0.05%磷酸（45:55），流速为1.5毫升/分钟。用可变波长紫外检测器在200纳米处监测柱流出物。甲苯磺丁脲及其代谢产物的保留时间分别为5.75分钟和3.25分钟。该方法结果可重现，灵敏度足以用于血浆水平的常规临床监测或单剂量药代动力学研究。许多常用药物不干扰该方法。单个血浆样本大约可在9或10分钟内分析完毕。
