小鼠脑组织中组胺含量的HPLC-UV检测

目的:建立一种测定小鼠脑组织匀浆中组胺含量的柱前衍生高效液相色谱-紫外检测法。方法:雄性昆明种小鼠8只,在乙醚麻醉下断头处死后,迅速取出脑组织,分离出皮层、海马和下丘脑,称重并制成匀浆。匀浆中组胺用高氯酸提取,经丹磺酰氯衍生后,用高效液相色谱-紫外(HPLC-UV)检测法检测含量。结果:组胺在0.1～10μg/ml范围内线性关系良好,相关系数γ2=0.9993,最低检出限为0.1μg/ml,保留时间约6.1 min。小鼠下丘脑中组胺含量最高,约为0.21μg/g。结论:本实验所建立的HPLC-UV检测法快速简便,敏感实用,可用于脑组织中组胺含量的测定。

Determination of histamine in the mouse brain by HPLC-UV

Objective:To develop a pre-column derivatization followed by high performance liquid chromatography with ultraviolet detection(HPLC-UV) method for determining the histamine in the mouse brain.Methods:Eight male mice of Kunming strain were anesthetized with diethyl ether and killed by decapitation.The mouse brains were quickly removed out and dissected on ice into cortex,hippocampus and hypothalamus,and homogenized.The histamine in the homogenates was extracted with perchloric acid aqueous solution,derivatized by dansyl chloride and determined by HPLC-UV for its level.Results:Histamine reagent showed a good linear relationship within the range of 0.1-10 μg/ml(γ2=0.9993).The minimal detection limit was 0.1 μg/ml and the retention,6.1 min.The mean content of histamine in the hypothalamus measured approximately 0.21 μg/g,higher than that in the cortex or hippocampus.Conclusion:The pre-column derivatization followed HPLC-UV determination appears fast,sensitive and practical for analyzing histamine in the mouse brain.