Automated assay of methylmalonic acid in serum and urine by derivatization with 1-pyrenyldiazomethane, liquid chromatography, and fluorescence detection.
Automated assay of methylmalonic acid in serum and urine by derivatization with 1-pyrenyldiazomethane, liquid chromatography, and fluorescence detection.
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通过 1-芘基重氮甲烷衍生化、液相色谱和荧光检测自动测定血清和尿液中的甲基丙二酸。
DOI：
10.1093/clinchem/39.3.392
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发表时间：
1993
影响因子：
9.3
通讯作者：
P. Ueland
中科院分区：
文献类型：
--
作者：
J. Schneede;P. Ueland
测定血清中甲基丙二酸（MMA）已被确立为诊断钴胺素缺乏症的准确试验。我们在这里描述的发展和性能的MMA在血液和尿液中的液相色谱测定。该测定基于我们最近的发现，即一些短链二羧酸的羧酸部分之一与荧光试剂1-芘基重氮甲烷在水性介质中反应，而另一个保持未衍生化（Anal Chem 1992; 63：315-9）。1-芘基甲基单酯的游离羧酸基团的pH依赖性电离允许保留在用于固相萃取的阴离子交换柱上。使用与十八烷基二甲基甲硅烷基色谱柱串联的氰丙基色谱柱进行分析。固相萃取和进样由Gilson ASPEC样品处理器自动进行。在0.1-1000 μ mol/L范围内，测定响应值与血清中MMA浓度呈线性关系。日内和日间CV为2.8- 10.9%，进样5 fmol（血清中约20 nmol/L）的检测限足够低，可测定健康受试者血清（平均值0.187 mumol/L，SD 0.084，范围0.044-0.431，n = 44）和尿液中的MMA。
Determination of methylmalonic acid (MMA) in serum has been established as an accurate test for the diagnosis of cobalamin deficiency. We describe here the development and performance of a liquid-chromatographic assay of MMA in blood and urine. The assay is based on our recent finding that one of the carboxylic acid moieties of some short-chain dicarboxylic acids reacts with the fluorogenic reagent 1-pyrenyldiazomethane in an aqueous medium, whereas the other remains underivatized (Anal Chem 1992; 63:315-9). The pH-dependent ionization of the free carboxylic acid group of 1-pyrenylmethyl monoesters permits retention on anion-exchange columns, which are used for solid-phase extraction. The analysis is done with a cyanopropyl column coupled in series with an octadecyldimethylsilyl column. Solid-phase extraction and sample injection are carried out automatically by a Gilson ASPEC sample processor. The assay response varies linearly with MMA concentration in the range 0.1-1000 mumol/L in serum. The within-day and between-day CVs are 2.8-10.9%, and the detection limit of 5 fmol injected (approximately 20 nmol/L in serum) is sufficiently low to determine MMA in serum (mean 0.187 mumol/L, SD 0.084, range 0.044-0.431, n = 44) and urine from healthy subjects.
钴胺素缺乏症的临床谱和诊断。
影响因子：
20.3
作者：
Stabler,SP;Allen,RH;Savage,DG;Lindenbaum,J
通讯作者：
Lindenbaum,J
影响因子：
158.5
作者：
LINDENBAUM, J;HEALTON, EB;ALLEN, RH
通讯作者：
ALLEN, RH