人血浆中全反式维甲酸、13-顺式维甲酸和9-顺式维甲酸浓度的监测

Determination of all-trans,13-cis-and 9-cis-retinoic acid in human plasma by HPLC

目的建立高效液相色谱法同时测定人血浆中全反式维甲酸、13-顺式维甲酸和9-顺式维甲酸浓度。方法色谱柱为KromasilC18柱(4.6mm×250mm,5μm);柱温为室温。流动相A:甲醇;流动相B:0.01mol/L醋酸钠缓冲液(pH=5.7),梯度洗脱,流速1ml/min;检测波长340nm。结果全反式维甲酸、13-顺式维甲酸和9-顺式维甲酸血药浓度在1～200ng/ml范围内,浓度与峰面积比有良好的线性关系,最低检测浓度为0.5ng/ml。全反式维甲酸方法回收率为97.22%～108.80%,日内RSD≤8.24%,日间RSD≤11.34%;13-顺式维甲酸方法回收率为98.62%～104.80%,日内RSD≤8.02%,日间RSD≤11.70%;9-顺式维甲酸方法回收率为97.74%～102.24%,日内RSD≤7.72%,日间RSD≤9.17%。结论本方法简单、快速、灵敏、重现性好,适用于全反式维甲酸、13-顺式维甲酸和9-顺式维甲酸临床血药浓度监测及人体药代动力学研究。

Objective To establish a HPLC method for determination of all-trans,13-cis-and 9-cis-retinoic acid in human plasma.Methods A Kromasil C18 column(4.6 mm×250 mm,5 μm)was adopted.The mobile phase of gradient elution was composed of 0.01 mol/L sodium acetate buffer(pH=5.7)and methanol.The flow rate was 1 ml/min.The UV detector was operated at 340 nm.Results The assay procedure was shown to produce linear calibration curves over the range of 1-200 ng/ml of retinoic acid in plasma.The detect limitation were 0.5 ng/ml.The recovery of all-trans,13-cis-and 9-cis-retinoic acid were 97.22%-108.80%,98.62%-104.80% and 97.74%-102.24%,respectively.The RSD within day and betten days were less than 8.24% and 11.34% for all-trans retinoic acid,less than 8.02% and 11.70% for 13-cis-retinoic acid and less than 7.72% and 9.17% for 9-cis-retinoic acid.Conclusion This method is found to be reproducible,convenient and sensitive for plasma concentration monitoring and clinical pharmacokinetics study of retinoic acid.