色谱2012，Vol.30Issue(6)：613-617,5.DOI:10.3724/SP.J.1123.2012.03026
柱前衍生高效液相色谱-荧光检测法测定血浆中同型半胱氨酸
Determination of homocysteine in plasma by precolumn derivatization-high performance liquid chromatography with fluorescence detection
摘要
Abstract
A precolumn derivatization-high performance liquid chromatographic method for the determination of homocysteine (Hey) in plasma was established. Tris (2-carboxyethyl) phos-phine hydrochloride (TCEP) and N-( 1-pyrenyl) maleimide (NPM) were used as the reduced reagent and derivatization reagent, respectively. The separation was carried out on an Agilent Hypersil C-18 column (250 mm x4. 0 mm, 5μm) in gradient elution mode. The mobile phase consisted of A (15 mmoI/L sodium acetate solution), B (acetonitrile) and C (300 mL water containing 1 mL acetic acid and 1 mL phosphoric acid). The eluate was monitored by the fluorescence detector at an excitation wavelength of 330 nm and an emission wavelength of 380 nm. The mean recovery of Hey was (102.08 ±4.94)%. The linear range was from 0.500 μmol/L to 100μmol/L, with a detection limit of 0. 016μmol/L. The intra-day and inter-day relative standard deviations (RSDs) for Hey were less than 5%. Seven plasma samples of patients with hypertension and seven plasma samples of healthy controls were tested, and the results demonstrated that the Hey in the plasma from the hypertension group was significantly different from that of the control group (p < 0. 05). The developed method is simple, fast, accurate, and suitable for clinical measurement.关键词
高效液相色谱/荧光检测/同型半胱氨酸/血浆Key words
high performance liquid chromatography ( HPLC )/ fluorescence detection/homocysteine/ plasma分类
化学化工引用本文复制引用
唐秀芳,甄乾娜,樊子勉,冯成亚,丁敏..柱前衍生高效液相色谱-荧光检测法测定血浆中同型半胱氨酸[J].色谱,2012,30(6):613-617,5.基金项目
重庆市科委自然科学基金项目(No.CSTC2011JJA0225). （No.CSTC2011JJA0225）
