Chittur S V, Tracy T S
Department of Basic Pharmaceutical Sciences, School of Pharmacy, West Virginia University, Morgantown 26506, USA.
J Chromatogr B Biomed Sci Appl. 1997 Jun 6;693(2):479-83. doi: 10.1016/s0378-4347(97)00024-8.
A high-performance liquid chromatographic assay was developed for the quantitation of chlorzoxazone and its major metabolite 6-hydroxychlorzoxazone. These compounds along with phenacetin, the internal standard, were extracted from incubation mixtures using ether extraction. The extracts were analyzed on a Brownlee Spheri-5 C8 column with a mobile-phase of acetonitrile-0.5% phosphoric acid (30:70, v/v). The assay utilized UV detection at 287 nm which provided sensitivity and specificity to simultaneously quantify chlorzoxazone and 6-hydroxychlorzoxazone from liver microsomal samples at amounts of 10 ng and greater. The mean correlation coefficient of the standard curves for 6-hydroxychlorzoxazone and chlorzoxazone was 0.998 and 0.993, respectively, over the range of 25-400 ng, and the regression curves were found to be linear at least through 1600 ng. All components eluted within 7 min, resulting in a total analysis time of 8 min. The inter-day and intra-day coefficients of variation were <7 and <3%, respectively. This method provides a rapid, sensitive and cost-effective assay for 6-hydroxychlorzoxazone and chlorzoxazone in liver microsomal incubations.
开发了一种高效液相色谱法用于定量氯唑沙宗及其主要代谢物6-羟基氯唑沙宗。这些化合物与内标非那西丁一起，使用乙醚萃取法从孵育混合物中提取。提取物在Brownlee Spheri-5 C8柱上进行分析，流动相为乙腈-0.5%磷酸（30:70，v/v）。该测定法利用287 nm处的紫外检测，可从肝微粒体样品中同时定量10 ng及以上的氯唑沙宗和6-羟基氯唑沙宗，具有灵敏度和特异性。在25 - 400 ng范围内，6-羟基氯唑沙宗和氯唑沙宗标准曲线的平均相关系数分别为0.998和0.993，并且发现回归曲线至少在1600 ng范围内呈线性。所有成分在7分钟内洗脱，总分析时间为8分钟。日间和日内变异系数分别<7%和<3%。该方法为肝微粒体孵育中的6-羟基氯唑沙宗和氯唑沙宗提供了一种快速、灵敏且经济高效的测定方法。
