高效液相色谱法快速测定生物样品中的视黄酸  

Rapid quantification of retinoic acid in biological samples by high-performance liquid chromatography

作　　者：曲斐[1] 方志娥[1] 尚京川[1] 

机构地区：[1]重庆医科大学药学院,重庆400016

出　　处：《分析试验室》2009年第7期5-8,共4页Chinese Journal of Analysis Laboratory

摘　　要：建立了快速测定生物样品中全反式（all—trans．），9-顺式（9-cis-），13-顺式（13-cis-）-视黄酸（RA）的高效液相色谱分析方法。经2次液．液萃取提取生物样品中的视黄酸后，直接应用高效液相色谱法同时测定了3种视黄酸同分异构体的含量。采用Waters C18反相柱（3．9×150mm），V（乙腈）：V（0．1％冰醋酸溶液）：86-14为流动相，流速1．0mL/min，柱温为25℃，检测波长为350nm。3种视黄酸同分异构体的线性范围均为5—500ng／mL，r^2均大于0．999；检出限均为1．g／mL；提取回收率为92．7％～101．8％，方法回收率为102．4％-104．4％：日内精密度小于8．3％，日间精密度小于11％。本方法适用于不同生物样品中视黄酸的定量研究。To establish a rapid and simultaneous determination of atRA, 9-cis-RA and 13-cis-RA in biological samples by high-performance liquid chromatography （HPLC）. AtRA and its isomers from the biological samples through liquidliquid extraction can be quantified for HPLC analysis. The column was Waters btBondapak ClS column （3.9 ×150 mm）. The mobile phase was acetonitrile/water containing 1% glacial acetic acid with the volume ratio of 86：14 （V/V）. The flow rate was 1.0 mL/min, and the colmnn temperature was maintained at 25℃ with a column oven. Absorbance of the eluent was monitored at 350 nm. The assay quantified over a linear range of 5 to 500 ng/mL （ r^2 〉 0.999） with a minimum detectable limit of 1 ng/mL at a signal/noise ratio of 3. The recoveries of extraction were 92.66% - 101.79%, and the recoveries of the method were 102.4% - 104.4%. The intm-assay RSD （same-day） and the inter-assay RSD （different day） were lower than 8.3% and 11%. The HPLC method to quantif atRA and its isomers in this assay is rapid, direct and sensitive. At the same time, isocratic elution and the mobile phase system are simple. The developed analytical method is practicable for analyses in every laboratory equipped with basic HPLC equipments. It has been successfully applied to diverse limited biological samples and it is suitable for application in clinical detection of retinoic acids.

关 键 词：全反式视黄酸 9-顺式-视黄酸 13-顺式-视黄酸 高效液相色谱 

分 类 号：O657.7[理学—分析化学]