高效液相色谱法测定大鼠血浆中全反式维甲酸  

HPLC determination of all-trans retinoic acid in rat plasma

作　　者：卢燕雯[1] 忻箐[1] 朱秋毓[1] 李曼[1] 顾勇[1] 林善锬[1] 

机构地区：[1]复旦大学附属华山医院肾病科,上海200040

出　　处：《药物分析杂志》2008年第9期1446-1449,共4页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立大鼠血浆游离维甲酸浓度的测定方法。方法:用正丁醇-乙腈-正庚烷-磷酸盐混合溶液提取大鼠血浆中的全反式维甲酸,用 HPLC 定量检测。色谱柱:Hypersil C_(18)(100 mm×2.1 mm,3μm);流动相:60 mmol·L^(-1)醋酸铵(pH 4.5)溶液,内含78%甲醇,用前经0.22 μm滤膜(Millipore,USA)过滤;流速:0.20 mL·min^(-1);检测波长:350 nm;柱温:40℃。结果:全反式维甲酸标准曲线的线性范围为0.1～100 ng·mL^(-1),相关系数大于0.998;平均回收率大于94.9%;日内和日间的RSD 分别小于6.8%和11.3%;全反式维甲酸平均保留时间为(7.09±0.04)min,总分析时间少于8.2 min。结论:本法所需样本量低,预处理步骤简便,适用于临床大批量样本的检测及药代动力学研究等工作之需。Objective： To develop a quantitative analysis method for the determination of free all - trams retinoic acid （ATRA） in rat plasma. Method： ATRA in rat plasma was extracted with a mixed solvent consisted of n - butanol- acetonitril -heptane plus saturated phosphate buffer and assayed by reversed -phase HPLC. Hypersil C8 column （100 mm× 2. 1 mm,3 μm） was adopted;The mobile phase was 60 mmol· L^-1 ammonium acetate solution（ pH 4.5 ,containing 78% methanol） at a flow rate of 0.20 mL · min ^-1 ;The detection wavelength was 350 nm and the column temperature was 40℃. Results： The linearity of the method was in the range of 0. 1 to 100 ng · mL^- 1 of ATRA with a con＇elation coefficient of more than 0. 998. Retention time for ATRA was （7.09± 0.04 ） min. A single run for the measurement of ATRA was only 8.2 min. RSD of the method was less than 6. 8 % for within - day and 11.3% for between -day, respectively. The average recoveries of ATRA in plasma was over 94.9% at low, middle and high concentrations. Conclusion：Presented method is reliable, fast and simple with applicability to both diagnosis and research in clinic for the determination of ATRA from biological specimen.

关 键 词：高效液相色谱 全反式维甲酸 血药浓度 窄径柱 

分 类 号：R917[医药卫生—药物分析学]