人血浆中全反式维甲酸、13-顺式维甲酸和9-顺式维甲酸浓度监测.docVIP

人血浆中全反式维甲酸、13-顺式维甲酸和9-顺式维甲酸浓度监测 　　【摘要】 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（4．6 mm×250 mm，5 μm）；柱温为室温。流动相A：甲醇；流动相B：0．01 mol/L醋酸钠缓冲液（pH=5．7），梯度洗脱，流速1 ml/min；检测波长340 nm。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1～200 ng/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0．5 ng/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。 　　【关键词】全反式维甲酸；13-顺式维甲酸；9-顺式维甲酸；血药浓度；高效液相色谱法 　　 　　Determination of all-trans,13-cis-and 9-cis-retinoic acid in human plasma by HPLC 　　 　　TANG Lei,HU Jun-yong,CUI Ying-peng,et al.Department of Pharmacy,the First Affiliated Hospital of Sun Yat-Sen University,Guangzhou 510080,China 　　 　　【Abstract】 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. 　　【Key words】 　　All-trans retinoic acid;13-cis-retinoic acid;9-cis-retinoic acid;Plasma concentration;HPLC 　　 　　维甲酸是维生素A 的天然氧化代谢产物，有多种立体异构体，包括全反式维甲酸(all-trans retinoic acid,ATRA)、13