Kaartinen V, Mononen I
Department of Clinical Chemistry, Kuopio University Central Hospital, Finland.
J Chromatogr. 1989 May 30;490(2):293-9. doi: 10.1016/s0378-4347(00)82787-5.
A sensitive method for quantitative analysis of aspartylglucosamine as its dabsyl chloride derivative by high-performance liquid chromatography is described. Precolumn-derivatized aspartylglucosamine and internal standard (carboxymethylcysteine) are separated on a reversed-phase column with a mobile phase consisting of phosphate buffer and acetonitrile and monitored by UV-VIS detection at 436 nm. Aspartylglucosamine acts in the assay like a polar amino acid, and it can be separated from interfering substances in urine with a retention time of ca. 13 min. Its detection limit is ca. 0.3 microM in water and 0.5-1.0 microM in urine and other biological samples, which permits its quantitation in normal urine, for example. The within-day coefficient of variation is less than 4.7% and the day-to-day coefficient of variation is less than 8.3%. The present method is applicable to the direct analysis of aspartylglucosamine in body fluids and tissues without any prepurification and, in combination with automated liquid chromatography, allows rapid assay of a large number of samples in the detection of aspartylglycosaminuria. The sensitivity of the assay also allows direct quantitation of aspartylglucosamine in normal urine and leukocytes of aspartylglycosaminuria patients, and may thus be used in metabolic studies of the compound.
本文描述了一种通过高效液相色谱法对天冬氨酰葡糖胺的丹磺酰氯衍生物进行定量分析的灵敏方法。柱前衍生化的天冬氨酰葡糖胺和内标（羧甲基半胱氨酸）在反相柱上分离，流动相由磷酸盐缓冲液和乙腈组成，并通过436nm的紫外-可见检测进行监测。天冬氨酰葡糖胺在测定中表现得像一种极性氨基酸，它可以与尿液中的干扰物质分离，保留时间约为13分钟。其检测限在水中约为0.3μM，在尿液和其他生物样品中为0.5 - 1.0μM，例如这使得它能够在正常尿液中进行定量。日内变异系数小于4.7%，日间变异系数小于8.3%。本方法适用于体液和组织中天冬氨酰葡糖胺的直接分析，无需任何预纯化，并且与自动液相色谱相结合，能够在检测天冬氨酰葡糖胺尿症时快速测定大量样品。该测定方法的灵敏度还允许直接定量正常尿液和天冬氨酰葡糖胺尿症患者白细胞中的天冬氨酰葡糖胺，因此可用于该化合物的代谢研究。
