HPLC-UV法测定大鼠脑组织中5-羟色胺含量

Determination of 5-hydroxytryptamine(5-HT) in brain tissue of rats by HPLC-UV

目的建立大鼠脑组织中5-羟色胺(5-HT)含量的HPLC-UV测定方法。方法采用SHIMADZU VP-ODS C18反相柱(250×4.6mm,5μm),流动相:甲醇-0.01mol/L的醋酸钾缓冲液(10:90,V/V,用0.2mol/L柠檬酸调pH至4.00);流速:1.0ml/min;柱温:25℃;检测波长:275nm。结果 5-羟色胺在0.1～20μg/ml范围内,回归方程为A=46008.89C+1886.58,相关系数r=0.9999,最低检测限为40ng/ml,最低定量限为100ng/ml。加样回收率为89.62%～101.60%,日内、日间精密度均小于3%。结论该方法准确可靠,简单易行,可重复性强,适用于多样本5-HT的快速测定。

Objective To establish a HPLC-UV method for determining the concentration of 5-HT in brain tissue of rats. Methods Adopted a SHIMADZU VP-ODS C18 reversed-phase column （250×4. 6 mm,5 μm）. Methanol-0.01mol/L CH3COOK buffer solution containing 10 % citric acid （ 1：9, V/V, pH 4.00） as mobile phase, with the flow rate was 1.00 ml/min, and the detection way wavelength of 275 nm, The column temperature was 25 ℃. Results A excellent linearity of 5-HT concentration curve was obtained in a rang of 0. 1 - 20 μg/ml, and the regression equation was A = 46008. 89C ＋ 1886.58 （ r = 0. 9999）. The detection limit was 40 ng/ml, the lowest detection limits of 5-HT was 100 ng/ml. And the spotting recovery was within the range of 89.62% - 101.6%. Intra -day and inter -day precisions were less than 3 %. Condusion The meth- od is accurate, reliable, simple and feasible, which is accepted to determine the concentration of 5-HT.