采用反相高效液相色谱法测定尿液中的肌酐、紫外吸收氨基酸和有机酸。
Determination of creatinine and ultraviolet-absorbing amino acids and organic acids in urine by reversed-phase high-performance liquid chromatography.
作者信息
Yokoyama Y, Tsuchiya M, Sato H, Kakinuma H
机构信息
Laboratory of Analytical Chemistry, Faculty of Engineering, Yokohama National University, Japan.
出版信息
J Chromatogr. 1992 Nov 27;583(1):1-10. doi: 10.1016/0378-4347(92)80338-q.
A simple and reliable method for the determination of urinary creatinine, amino acids and organic acids was developed. A urine sample was preliminarily separated into an organic acid fraction (including neutral species) and an amino acid fraction by cation-exchange chromatography. Both fractions were analysed by reversed-phase high-performance liquid chromatography, with a phosphoric acid-methanol gradient elution system and ultraviolet detection at 210 nm. Relationships between concentrations and peak heights were linear from 2 to 500 microM for the analytes. Overall recoveries were ca. 100%. The concentrations of creatinine for 37 urine samples, from 20 healthy newborns and from 17 patients with several inherited metabolic disorders, were 2.35 +/- 2.29 mM (ranging between 0.27 and 10.15 mM). The method was applied to the determination of several diagnostically useful metabolites in urine. The concentrations of phenylalanine and phenylacetic acid for five urine samples from patients with phenylketonuria were 347 +/- 177 and 282 +/- 224 microM/mM creatinine, respectively. The concentrations of tyrosine and 4-hydroxyphenyllactic acid in the urine of a patient with tyrosinemia were 112 and 1871 microM/mM creatinine, respectively.
开发了一种简单可靠的测定尿肌酐、氨基酸和有机酸的方法。通过阳离子交换色谱将尿样初步分离为有机酸部分（包括中性物质）和氨基酸部分。两部分均采用反相高效液相色谱分析，使用磷酸 - 甲醇梯度洗脱系统，并在210 nm处进行紫外检测。分析物浓度与峰高之间的关系在2至500 microM范围内呈线性。总体回收率约为100%。来自20名健康新生儿和17名患有几种遗传性代谢疾病的患者的37份尿样中肌酐浓度为2.35±2.29 mM（范围在0.27至10.15 mM之间）。该方法应用于尿中几种具有诊断价值的代谢物的测定。苯丙酮尿症患者的5份尿样中苯丙氨酸和苯乙酸浓度分别为347±177和282±224 microM/mM肌酐。酪氨酸血症患者尿中酪氨酸和4 - 羟基苯乳酸浓度分别为112和1871 microM/mM肌酐。
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