摘要
目的:建立高效液相色谱-荧光检测测定大鼠血小板内5-羟色胺含量的方法。方法:色谱柱为Nucleosil-C18(4mm×250mm,10μm),流动相:磷酸二氢钾(10mmol/L,内含EDTA-Na20.3mmol,pH=3.5):甲醇:乙腈=85:10:5,流速:0.5ml/min,荧光激发波长:254nm,发射波长:360nm,增益:32。血小板沉淀用300μl纯水悬浮,加入15μl内标溶液(30μg/ml,α-甲基-5-HT水溶液),100μl蛋白沉淀液,混匀,离心,取上清液20μl,进样。结果:5-羟色胺在0.25～4.00μg/ml浓度范围内,线性关系良好。5-羟色胺测定的批内和批间相对标准差(RSD)分别小于4.29%和4.44%,回收率为84.3%～88.7%。正常雄性SD大鼠血小板内5-羟色胺的平均含量分别为(1.65±0.14)μg/109plts。结论:本法简便、准确、快捷,可用于大鼠血小板内5-羟色胺的测定。
Objective： To establish a method for the contend determination of serotonin in blood platelets by high performance liquid chromatography with fluorescence detector （HPLC-FD）. Methods： Preparating platelets samples commonly with 2% EDTA-Na2 anticoagulated blood, we determinated them by HPLC. The nucleosil-Cls column （4 mm×250 mm, 10 μm） was used. The mobile phases consisted of mobilphase of 0.01mol/L-potassium dihydrogen phosphate, acetonitrile and methanol （85：5：10）, the flow rate was 0.5 ml/min and the detection wavelength was at 254 nm, the excitation wavelength was at 360 nm for emission, quantitating in internal standard method. Results： The results showed that the 5-Hydroxytryptamine peak was chromatographically resolved from the internal satandard; a good linearity range from 0.25-4.00 μg/ml; the RSD of within- batch and between-batch of the method was 4.29% and 4.44%; the average recovery was 84.3%-88.7%. The average contents of 5-Hydroxytryptamine in platelets of normal male Sprague-Dawley rats was （1.65±0.14） μg/109 pits. Conclusion： The method is simple, accurate, rapid, and can be used for the determination of 5-Hydroxytryptamine in rates platelets.
出处
《中国医药导报》
CAS
2010年第13期51-53,共3页
China Medical Herald
基金
广东省自然科学基金(项目编号:04010029)
国家自然科学基金(项目编号:30701100)
