Hoizey Guillaume, Lamiable Denis, Frances Carole, Trenque Thierry, Kaltenbach Matthieu, Denis Jacky, Millart Hervé
Laboratoire de Pharmacologie et Toxicologie, Hôpital Maison Blanche, CHU de Reims, 45, rue Cognacq-jay, 51092 Reims, Cedex, France.
J Pharm Biomed Anal. 2002 Oct 15;30(3):661-6. doi: 10.1016/s0731-7085(02)00289-3.
A simple and accurate high-performance liquid chromatographic method with ultraviolet detection at 220 nm has been validated for the simultaneous determination of amoxicillin and clavulanic acid in human plasma. Plasma samples were pretreated by direct deproteinization with methanol. A good chromatographic separation between both compounds was achieved using a reversed phase C8 column and a mobile phase, consisting of acetonitrile-phosphate solution-tetramethyl ammonium chloride solution. The calibration curves were linear over the concentration range of 0.625-20 mg l(-1) for amoxicillin and 0.3125-10 mg l(-1) for clavulanic acid with determination coefficients > 0.998. The method is accurate (bias < 7%) and reproducible (intra- and inter-day R.S.D. < 15%), with a quantitation limit of 0.625 and 0.3125 mg l(-1) for amoxicillin and clavulanic acid, respectively. Analytical recoveries from human plasma ranged from 91 to 102% for both components. This fully validated method, which allows the simultaneous measurement of amoxicillin and clavulanic acid in biological samples, is rapid (total run time < 10 min) and requires only a 100 microl sample. This assay is suitable for biomedical applications and was successfully applied to a pilot pharmacokinetics study in healthy volunteers after a single-oral administration of amoxicillin/clavulanic acid combination (500/125 mg).
已验证一种简单准确的高效液相色谱法，该方法采用220 nm紫外检测，可同时测定人血浆中的阿莫西林和克拉维酸。血浆样品通过用甲醇直接脱蛋白进行预处理。使用反相C8柱和由乙腈 - 磷酸盐溶液 - 四甲基氯化铵溶液组成的流动相，实现了两种化合物之间良好的色谱分离。阿莫西林的校准曲线在0.625 - 20 mg l(-1)浓度范围内呈线性，克拉维酸的校准曲线在0.3125 - 10 mg l(-1)浓度范围内呈线性，测定系数> 0.998。该方法准确（偏差< 7%）且可重现（日内和日间相对标准偏差< 15%），阿莫西林和克拉维酸的定量限分别为0.625和0.3125 mg l(-1)。两种成分在人血浆中的分析回收率在91%至102%之间。这种经过充分验证的方法能够同时测量生物样品中的阿莫西林和克拉维酸，速度快（总运行时间< 10分钟），仅需100微升样品。该测定方法适用于生物医学应用，并已成功应用于健康志愿者单次口服阿莫西林/克拉维酸组合（500/125 mg）后的初步药代动力学研究。
