HPLC测定人血浆中亚胺培南的浓度

Determination of Imipenem in Human Plasma by HPLC

目的 建立测定人血浆中亚胺培南浓度的HPLC方法。方法 以0.5 mol·L^-1尿素为稳定剂，以5-溴脲嘧啶为内标，血浆样品经乙腈沉淀蛋白，二氯甲烷2次提取去杂质，取水相进样。色谱柱为Atlantis C18（4.6 mm×150 mm，5 μm）；流动相为甲醇-10 mmol·L^-1磷酸二氢钾溶液（pH=6.0）（4∶96）；流速为1.0 mL·min^-1；柱温：25 ℃；检测波长：300 nm。结果 亚胺培南在0.5-100 mg·L^-1内线性关系良好，r=0.999 7；定量下限为0.5 mg·L^-1；平均绝对回收率 82.72%，方法回收率为87.60%-96.36%；日内、日间RSD均〈10%。结论 本方法简单、快捷、灵敏、准确，适用于亚胺培南临床血药浓度的监测。

OBJECTIVE To establish an HPLC method for the detection of imipenem in human plasma. METHODS Samples were spiked with 0.5 mol·L^-1 urea as stabilizer solution, 5-bromine urea pyrimidine as internal standard, proteins were precipitated with acetonitrile followed by extraction with dichloromethane and the upper aquous phase was injected. Separation was achieved on an Atlantis C18 column （4.6 mm×150 mm, 5 μm）. The mobile phase composed of methanol and 10 mmol·L^-1 monopotassium phosphate （pH=6.0）（4∶96） with a flow rate of 1.0 mL·min^-1. The column temperature was 25 ℃. The UV detection wavelength was 300 nm. RESULTS Calibration curves of imipenem showed good linear regression in the range of 0.5-100 mg·L^-1（r=0.999 7）. The lower limit of quantification of imipenem was 0.5 mg·L^-1. The mean absolute recovery was 82.72%, and the method recovery was 87.60%-96.36%. Intra- and inter-day variations were 〈10%. CONCLUSION The established method is simple, sensitive and accurate for determining imipenem in human plasma.