高效液相色谱法同时测定人血阿莫西林钠/克拉维酸钾血药浓度

摘要

Objective To establish a chromatography-based method for simultaneous analysis of the concentrations of amoxicillin and clavulanate potassium in human blood. Methods With paracetamol as the internal control, human plasma samples, after treatment with methanol for protein sedimentation and centrifugation, were loaded for analysis with high-performance liquid chromatography (HPLC). HPLC analysis was carried out using a C18 column (5 urn, 4.6 mm × 150 mm) with the mobile phase of acetonitrile-PBS (0.05 mol/L) of 10:90 (pH 2.3), UV detection wavelength of 220 ran, flow rate of 1.0 ml/min, and column temperature of 25 t. Results The retention time of acetaminophen for potassium clavulanate, amoxicillin sodium and the internal control was 5.3, 7.2, and 8.5 min, respectively, and no interference by the endogenous impurities in the plasma samples was found. Amoxicillin sodium showed a good linearity within the concentration range of 0.52-4.16 μg/ml (r2=0.9996), and potassium clavulanate had a good linearity within the range of 0.266-2.14 μg/ml (r2=0.9998). The minimum detectable concentrations of amoxicillin sodium and potassium clavulanate were 0.065 μg/ml and 0.066 μg/ml, respectively. The relative recoveries of amoxicillin sodium were 95.9%-96.5% (n=5), and those of clavulanate potassium were 92.5%-98.8% (n=5); the intra- and inter-day RSD of amoxicillin sodium was 1.84%-6.4% and 2.1%-7.8%, as compared to that of potassium clavulanate of 3.57%-8.6% and 1.8%-9.1%, respectively. Conclusion This method is simple, accurate, sensitive, specific and reproducible for analyzing the concentrations of amoxicillin and clavulanate potassium simultaneously in human plasma.%目的 建立阿莫西林钠/克拉维酸钾人血药浓度的色谱分析方法.方法 采用对乙酰氨基酚为内标,人血浆样品用甲醇沉淀蛋白后高速离心进样;色谱柱:C18色谱柱(依利特Hypersil BDS,5μm,4.6 mm×150 mm);流动相:乙腈-0.05mmol/L磷酸盐缓冲液=10∶90(V/V,磷酸调pH至2.3);紫外检测波长:220nm;流速:1.0ml/min;柱温:25℃.结果 克拉维酸钾、阿莫西林钠及内标物对乙酰氨基酚保留时间分别为5.3、7.2、8.5 min,人血浆中内源性杂质对目标物的测定没有干扰.阿莫西林钠在0.52～4.16μg/ml范围线性良好(r2=0.9996),克拉维酸钾在0.266 ～2.14μg/ml范围线性良好(r2=0.9998);阿莫西林钠和克拉维酸钾的最低检测浓度分别为0.065和0.066μg/ml.;阿莫西林钠的相对回收率分别为95.9%～96.5%(n=5),克拉维酸钾的相对回收率分别为92.5%～98.8%(n=5),阿莫西林钠日内RSD为1.84%～6.4%,日间RSD为2.1%～7.8%;克拉维酸钾日内RSD为3.57%～8.6%,日间RSD为1.8%～9.1%.结论 本方法简便、准确、灵敏、特异性强、重现性好,可作为同时测定人血浆中阿莫西林钠/束拉维酸钾浓度方法.