To establish the reversed phase high-performance liquid chromatography(RP-HPLC)method for determination of plasma concentration of acetaminophen in Bactrian camel,the chromatography was carried on a C18 column(150 mm× 4.6 mm,5 μm).The mobile phase was composed of methanol-water(20:80),the flow rate was 0.5 mL/min,and the detection wavelength was set at 248 nm for acetaminophen.The retention time of acetaminophen was 6.5 min,and was not disturbed by other miscellaneous peaks.The calibration curve was linearity in the range of 0.08 to 10.24 μg/mL(R2=0.9997),and the quantitation limit of acetaminophen was 0.10 μg/mL.Both the inter-day and intra-day RSD were under 7.0％,the absolute recovery of the method was 80.39％to 93.56％,the relative recovery of the method was 94.66％to 104.73％.Therefore,the established method was simple,reproducible and accurate,which could be used for determination of plasma acetaminophen concentration and in vivo activities of CYP1A enzyme in Bactrian camel.%试验旨在建立测定双峰驼血浆中对乙酰氨基酚浓度的反相高效液相色谱法(reversed phase high-performance liquid chromatography,RP-HPLC).选用Supelco Discovery C18色谱柱(150 mm×4.6 mm,5 μm),以甲醇:水=20:80为流动相,流速为0.5 mL/min,紫外检测波长为248 nm,采用甲醇直接沉淀蛋白方法处理血浆样品.结果表明,对乙酰氨基酚的保留时间为6.5 min,分离度良好,无血浆杂峰干扰,且在0.08～10.24 μg/mL浓度范围内线性关系良好(R2=0.9997,n=8),检测限为0.10 μg/mL,日内和日间精密度RSD均小于7.0％,绝对回收率为80.39％～93.56％,相对回收率为94.66％～104.73％.因此,本试验所建立的RP-HPLC法操作简便、准确,重复性好,专属性强,适用于检测双峰驼血浆中对乙酰氨基酚浓度,为后续的借助特异性探针药物——对乙酰氨基酚研究双峰驼CYP1A酶的体内活性提供了可靠而准确的定量方法.
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