同时测定血清中苯巴比妥、苯妥英及卡马西平的反相高效液相色谱法

Simultaneous determination of phenobarbital(PB), phenytoin(PHT) and carbamazepine(CBZ) in serum by reversed-phase high-performance liquid chromatography(RP-HPLC)

为了探讨高效液相色谱法同时测定苯巴比妥（ＰＢ）、苯妥英（ＰＨＴ）、卡马西平（ＣＢＺ）血药浓度的最佳实验条件，将标本在ｐＨ７．６条件下用二氯甲烷提取后经高纯度氮气在５０℃水浴中吹干，再用甲醇重溶后进行色谱分析。色谱条件：ＹＷＧ－Ｃ１８柱（２５０ｍｍ×４．６ｍｍ，１０μｍ），流动相甲醇：水为６０∶４０，流速１ｍｌ／ｍｉｎ，柱温４０℃，波长２５４ｎｍ。结果ＰＢ、ＰＨＴ、ＣＢＺ的最低检出浓度各为０．６７μｇ／ｍｌ、０．９４μｇ／ｍｌ和０．３１μｇ／ｍｌ。在三个不同水平下，ＰＢ、ＰＨＴ、ＣＢＺ的平均回收率各为９８．１４％、９６．７２％和９９．３６％，日内和日间变异均低于６．２５％，内源性物质及１０种常用药物未见干扰，在分析范围内具有良好的线性关系。该方法操作简便、快捷、准确，可满足临床和科研的需要。

o discuss the optimum experimental conditions of a simultaneous RP-HPLC determination of the concentration of PB, PHT and CBZ, these three drugs were extracted from serum at pH 7.6 with dichloromethane,blown dry at 50℃ under pure nitrogen gas, and reconstituted with methanol and injected in a YWG-C18(250mm×4.6mm, 10μm), column for analysis at wave length of 254 mm. The mobile phase used was methanol 60: water 40(V/V), with a flow rate of 1ml/min. The column temperature was 40℃. Results showed that the minimal detectable concentration for PB, PHT and CBZ were 0.67μg/ml, 0.94μg/ml and 0.31μg/ml, respectively. The average recovery rate at three different levels for PB, PHT and CBZ were 98.14%, 96.72% and 99.365, respectively. The within-day and day-to-day RSD were below 6.25%. Endogenous compounds and ten routine drugs were not found to interfere with this chromatographic procedure. The analytical range was linear from 5-80μg/ml for PB, from 5-50μg/ml for PHT and from 2-32μg/ml for CBZ. It is suggested that this manipulation is quick, simple, and accurate, and therefore suitable for clinics and scientific research.