高效液相色谱荧光法检测血小板中5-羟色胺

Determination of platelet serotonin by high performance liquid chromatography with fluorometric detector

目的 :建立一种简便、快速、准确的血小板 5 羟色胺 (5 HT)测定方法。方法 :标本去蛋白后 ,应用反相离子对高效液相色谱荧光法检测 5 HT ,N 甲基 5 HT为内标 ,色谱柱为Nova pakC18柱 (15 0mm× 4 .6mm ,颗粒 4 μm)。结果 :本法批内CV是 2 .2 9% ,批间CV是 3.88% ,回收率是 97.5 % ,线形范围 2 0～ 2 0 0 0 0nmol/L ,检测限是 1nmol/L ,5 羟吲哚乙酸、色氨酸、苯丙氨酸、酪氨酸对本法无干扰。结论 :高效液相色谱荧光法测定血小板 5 HT简便快速、灵敏特异 ,适宜于临床和科研工作。

Objective To establish a simple, rapid and precise method to determine platelet serotonin. Methods Serotonin (5-hydroxytryptamine;5-HT)was determined in platelet pellet after simple deproteinization, following ion-pair high performance liquid chromatography (HPLC) on reverse-phase column: Nova-pak C 18 column (150 mm×4.6 mm, 4 μm), serotonin and internal standard (N-methylserotonin) were detected by fluorometry. Results The intra-day CV was 2.29% and inter-day CV was 3.88% . The mean recovery of serotonin in platelet was 97.5%, the linear range was 20～20 000 nmol/L,and the detection limit was 1 nmol/L. 5-hydroxyindole acetic acid(5-HIAA), Trp, Phe and Tyr had no interference with the chromatographic condition used in this trial.Conclusion The metod described here is simple, convenient, rapid, sensitive, and accurate for clinical and scientific research.