RP-HPLC法测定大鼠体内对乙酰氨基酚的血药浓度

Determination of paracetamol in rats plasma with RP-HPLC

目的建立测定大鼠血浆中对乙酰氨基酚血药浓度的RP-HPLC法。方法以茶碱为内标,甲醇沉淀蛋白提取血浆中对乙酰氨基酚,色谱柱为C18柱(250mm×4.6mm,5μm),流动相为甲醇-水(20∶80),流速1.0mL·min-1,测定波长为248nm。结果血浆中杂质不干扰样品的测定,对乙酰氨基酚血药浓度在2.0～700.0mg·L-1范国内线性关系良好(r=0.9997),最低定量限为2.0mg·L-1,高、中、低浓度的日内和日间变异均小于10.0%,提取回收率在95.31%～98.61%,相对回收率在99.87%～104.67%。结论该方法能准确简便地测定对乙酰氨基酚血药浓度,符合生物样品分析要求,能满足药代动力学研究要求。

Objective To establish a RP-HPLC method for the determination of plasma concentration of paracetamol in rats. Methods Theophylline was used as an internal standard, methanol was used for the protein sedimentation. The chromatography was carried on a Cls column （250 mm×4.6 mm,5 μm）, the mobile phase was composed of methanol-water（20 ： 80） ,the flow rate was 1.0 mL · min^-1 ,and the detection wavelength was set at 248 nm for paracetamol. Results Under the condition of this test, paraceta tool in plasma was well separated, and no interference from plasma matrix was observed. The linear ranges for paracetamol were 2.0-700.0 mg· L^-1 （ r =0. 999 7）, the lowest limit of quantitation was 2.0 mg· L^-1 ,and the ingredients showed good relationships between the peak area and the concentration. The absolute recovery of the method was between 95.31% and 98.61%,and the relative recovery was between 99.87% and 104.67%. Inter-day, intra-day RSD were both under 10%. Conclusion The method is proved to be simple, sensitive, and reproducible. It could be used for the determination of rats plasma paracetamol and pharmacokinetics researches.