高效液相色谱法测定人血浆中双氯西林和阿莫西林的含量

HPLC determination of dicloxacillin and amoxicillin in human plasma

目的:建立人血浆中双氯西林和阿莫西林的测定方法。方法:高效液相色谱法,血浆样品以甲醇除蛋白,色谱柱为岛津Shim-pack VP-ODS C_(18)(150mm×4.6mm,5μm),双氯西林的测定以0.05mol·L^(-1)磷酸盐缓冲液(pH 4.5)-乙腈(68:32),加0.3%三乙胺为流动相,检测波长为225nm,柱温为25℃;阿莫西林的测定以0.033mol·L^(-1)磷酸盐缓冲液(pH7.2)-甲醇(88:12)为流动相,检测波长为229nm,柱温为20℃,流速均为1.0mL·min^(-1),进样量均为20μL。结果:在不同的色谱条件下,血浆样品中双氯西林和阿莫西林无干扰,双氯西林的线性范围为0.2～102.4μg·mL^(-1),r=0.9999,最低检测浓度为0.2μg·mL^(-1);阿莫西林的线性范围为0.25～128.0μg·mL^(-1),r=0.9998,最低检测浓度为0.25μg·mL^(-1)。双氯西林和阿莫西林的平均回收率分别为104.2%,101.6%。结论:这种以相同的样品预处理方法,不同的色谱分离条件分析双氯西林和阿莫西林的方法简便、准确、可靠,结果稳定,可用于阿双西林胶囊或片剂的药代动力学研究和生物利用度研究。

Objective： To establish HPLC methods for determination of dicloxacillin and amoxicillin in human plasma. Methods：Plasma samples were pretreated by direct deproteinization with methanol. The chromatographic separation dicloxacillin and amoxicillin from human plasma were achieved using C18 （150 mm ×4. 6 mm,5 μm）column and a mobile phase, consisting of acetonitrile - phosphate solution - triethylamine solution（ 65： 32： 0. 3 ） and methanol - phosphate solution（88： 12） , respectively. They were detected at 225 nm and 229 nm, respectively, and the column temperature was 25 ℃ and 20 ℃ , respectively, and flow rate was 1.0 mL · min^- 1 with the sampling volume 20 μL. Results：In the different chromatographic conditions dicloxacillin and amoxicillin were separated completely without interference. The average recovery of dicloxacillin and amoxicillin was 104.2% and 101.6% ,respectively, and the minimum detection concentration of dicloxacillin and amoxicillin was 0. 2 μg · mL^-1 and 0. 25 μg · mL^-1, respectively. The calibration curves were linear over the concentration range of 0. 2 - 102.4 μg · mL^-1 for dicloxacillin and 0. 25 - 128.0 μg · mL^- 1 for amoxicillin with correlation coefficients greater than 0. 999. Conclusion： Both high -performance liquid chromatographic methods by the same sample pretreatment are simple, accurate, stable and suitable for studying the pharmacokinetics and relative bioavailability of dicloxacillin and amoxicillin capsules or tablets.