Chwatko G, Bald E
Department of Environmental Chemistry, University of Lódz, Poland.
J Chromatogr A. 2002 Mar 8;949(1-2):141-51. doi: 10.1016/s0021-9673(01)01343-7.
This assay measures reduced, free oxidized, protein-bound, and total homocysteine in human plasma. Oxidized species of homocysteine are converted to reduced form by sodium borohydride, and, after precolumn derivatization with 2-chloro-1-methylquinolinium tetrafluoroborate, homocysteine 2-S-quinolinium derivative is separated from those of other plasma thiol derivatives, and quantitated by ion-paired reversed-phase high-performance liquid chromatography with ultraviolet detection. The reduced homocysteine sulfhydryl groups are trapped with minimal oxidation by derivatizing blood samples at the time of collection. With the use of this precise and sensitive HPLC method utilizing popular ultraviolet detection, homocysteine in plasma can be detected and quantitated at the level of 0.1 and 0.2 for reduced fraction, and 0.3 and 0.5 nmol/ml for total homocysteine, respectively. The method is applied for determination of different fractions of homocysteine in plasma of apparently healthy men and women.
该检测方法可测定人血浆中还原型、游离氧化型、蛋白结合型和总同型半胱氨酸。同型半胱氨酸的氧化型物种通过硼氢化钠转化为还原型，在用四氟硼酸2-氯-1-甲基喹啉鎓进行柱前衍生化后，同型半胱氨酸2-S-喹啉鎓衍生物与其他血浆硫醇衍生物分离，并通过离子对反相高效液相色谱法结合紫外检测进行定量。在采集血样时对其进行衍生化，可使还原型同型半胱氨酸巯基以最小程度的氧化被捕获。通过使用这种利用常见紫外检测的精确且灵敏的高效液相色谱法，血浆中的同型半胱氨酸可以分别在还原部分为0.1和0.2，总同型半胱氨酸为0.3和0.5 nmol/ml的水平下进行检测和定量。该方法用于测定看似健康的男性和女性血浆中不同组分的同型半胱氨酸。
