摘要
目的建立高效液相荧光检测法(HPLC-FLD)来同时测定人体血浆、血小板、脑脊液中5-羟色胺、5-羟吲哚乙酸的含量。方法样本经预处理后用高效液相色谱仪进行测定。色谱柱为KromasilC18(250mm×4.6mm,7μm),流动相:甲醇和0.1mol/L磷酸二氢钾(15:85),流速1.5ml/min,Ex=254.nm,EM=338nm,柱温为35℃。结果5-羟色胺和5-羟吲哚乙酸得到良好分离,线性范围5-羟色胺为1.1328～1160ng/ml,5-羟吲哚乙酸为0.9765～2000ng/ml,日内RSD分别为0.743%和1.470%,日间RSD分别为1.310%和2.043%;5-羟色胺和5-羟吲哚乙酸的平均回收率:血浆中分别为98.32%、98.50%,血小板中分别为91.41%、88.05%,脑脊液中分别为92.84%、93.51%。结论该法具有取样量少、简单、快速、准确、灵敏等特点,可作为临床检测的常规方法。
Objective To establish a method for simultaneous determination of the concentration of serotonin (5-HT) and 5-hydroxy-indoleacetic acid (5-HIAA) in human plasma, platelet, CSF by HPLC-FLD. Methods Use HPLC system, the Kromasil C18 column(250 mm× 4.6 mm, 7 μm)was used as analytical column with a mobile phase consisting of methanol-KH2PO4(15:85), the flow rate was of 1.5 ml/min, with fluorescence detection set at EX = 254 nm, EM = 338 nm, the column temperature was 35 ℃ . Results The chromatography was good and was not interfered by the components of the sample. The linear range of 5-HT was from 1.1328 to 1160 ng/ml, The linear range of 5-HIAA was from 0. 9765 to 2000 ng/ml. The RSD of inter-day and intra-day of 5-HT was 0. 743% and 1.310%. The RSD of inter-day and intra-day of 5-HIAA was 1.470 % and 2.043 % . The average recovery rate of 5-HT and 5-HIAA, in human plasma was 98. 32 %, 98. 50 % ; in human platelet was 91.41%, 88. 05% ; in human CSF was 92. 84%, 93. 51% . Conclusion The method is simple, rapid, accurate with good sensitivity; it is stuitable for the clinical monitoring.
出处
《苏州大学学报（医学版）》
CAS
北大核心
2005年第1期84-87,共4页
Suzhou University Journal of Medical Science
