[关键词]
[摘要]
目的 建立高效液相色谱法测定大鼠血清中视黄醇和视黄酸浓度的方法。方法 色谱柱为Phenomenex Luna C18柱(150 mm×4.6 mm,5 μm);流动相为甲醇-10 mmol/L乙酸铵缓冲盐(90:10,乙酸调整水相pH值至4);体积流量为1.0 mL/min;检测波长为340 nm;柱温为40℃;进样量为20 μL。结果 大鼠血清样品中视黄醇和视黄酸均分离良好,视黄醇和视黄酸质量浓度在31.25~1 000 ng/mL(r=0.999 9)与峰面积均呈良好的线性关系。平均回收率大于90%,RSD值均小于10%。结论 本法操作简便、准确、灵敏、专属性强,为建立无干扰的测定大鼠血清中视黄醇和视黄酸的方法及其在大鼠体内药动学研究提供参考。
[Key word]
[Abstract]
Objective To establish an efficient and accurate method for detecting the content of retinol and retinoic acid in rat serum by HPLC. Methods The chromatographic separation was carried out on a Phenomenex C18 column (150 mm × 4.6 mm, 5 μm) at a flow rate of 1.0 mL/min. Methanol-10 mmol/L ammonium acetate buffer solution (90:10, pH 4 adjusted with acetic acid) was used as mobile phase. It was monitored at a wavelength of 340 nm, the column temperature was 40℃, and the injection volume was 20 μL. Results The retinol and retinoic acid in rat serum also had a good linearity at 31.25-1 000 ng/mL (r=0.999 9). The average recoveries were above 90% with RSD values less than 10%. Conclusion This method is accurate, simple, and sensitive,which is suitable for the establishment of non-interference determination of retinol and retinoic acid in rat serum.
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[基金项目]
中华医学会医学教育研究课题资助项目(2012-SY-44)