高效液相色谱法测定人血浆中头孢吡肟浓度及其药动学

HPLC Method for the Determination of Cefepime in Human Plasma and its Pharmacokinetics

目的建立人血浆中头盐酸孢吡肟浓度的HPLC-UV测定方法，研究静脉注射盐酸头孢吡肟在健康人体中药动学行为。方法血浆经高氯酸沉淀蛋白后进行HPLC-UV分析，色谱柱为岛津Shim-packODS，5μm，250mm×4．6mm I．D，流动相为0．025mol／L的NaH2PO4：乙腈（89：11），磷酸调pH=3．0，流速1．0mL／min，紫外检测波长270nm．测定健康志愿者30min内静滴盐酸头孢吡肟500mg及停药后12h的血药浓度．时间过程。结果头孢吡肟在血浆中的线性范围为0．2～50．0μg／mL，LLOQ为0．2μg／mL，批内和批间的精密度（RSD）均小于10％，准确度（Relative error，R．E）为-3．48％～0．06％．血浆中回收率大于85％．健康志愿者单次静滴500mg盐酸头孢吡肟30min后，实测得tmax和Cmax分别为（0．51±0．02）h和（24．96±3．50）μg／mL，估算的t1／2β和MRT分别为（2．03±0．18）h和（2．65±0．29）h，V1和CL分别为（0．168±0．068）L／kg和（0．207±0．026）L／（kg·h），AUC0-∞为（50．88±6．10）μg·h／mL。结论该方法经考察符合生物样品的测定要求，可应用于人血浆中头孢吡肟血药浓度的测定和药代动力学研究，盐酸头孢吡肟在中国人和美国人中的动力学行为相近。

OBJECTIVE To develop an HPLC method with ultraviolet detection for determining cefepime in human plasma and study pharmacokinetic behavior of cefepime following intravenous administration. METHODS 12 healthy subjects received 500 mg of cefepime hydrochloride via a 30min intravenous infusion, blood samples were collected at designed times, following protein precipitation with HC104, the analytes were separated on a Shim-pack ODS C18, 5μm, 250 mm × 4.6 ram I.D. column with the mobile phase consisting of 0. 025 mol/L phosphate buffer （ pH = 3.0） -acetonitrile （ 89 ： 11 ） at a flow rate of 1.0 mL/min and the eluent was detected at 270 nm. Concentrations of cefepime in plasma were determined and pharmacokinetic parameters were estimated. RESULTS Linear quantitative response curve was generated over a concentration range of 0.2 - 50.0μg/mL for plasma. Intra- and inter-batch precision and accuracy were acceptable for all quality control samples with relative error ranging from-3.48% to 0.06%, including lowest limit of quantification of 0.2μg/mL. The mean recovery of cefepime from plasma was 〉 85%. Pharamaeokinetic parameters after iv infusion dose were evaluated as follows ： Cmax and tmax were （ 24.96 ± 3.50）μg/mL and （0.51± 0.02 ） h, respectively; AUC0-∞ （50.88 ±6.10） μg · h/mL, MRT（2.65 ±0.29）h and V1（0. 168 ±0.068）L/kg, t1/2β（2.03 ±0. 18）h and CL （0.207 ±0. 026） L/（h · kg）. CONCLUSION This specific, sensitive and precise method is suitable for monitoring of cefepime in human and its pharmacokinetic investigation. The pharmacokinetic behavior of cefepime in Chinese is similar to that of in American.