Measurement of epsilon-N-trimethyllysine in human blood plasma and urine.
Measurement of epsilon-N-trimethyllysine in human blood plasma and urine.
复制标题
人血浆和尿液中ε-N-三甲基赖氨酸的测量。
DOI：
10.1016/0003-2697(87)90018-2
复制
发表时间：
1987
影响因子：
2.9
通讯作者：
Rebouche,CJ
中科院分区：
文献类型：
--
作者：
Lehman,LJ;Olson,AL;Rebouche,CJ
本文描述了一种测定人血浆和尿液中ϵ-N-trimethyllysine的方法。将内标δ- n -三甲基鸟氨酸加入血浆和尿液标本中，并对混合物进行脱蛋白和/或水解。ϵ-N-trimethyllysine和δ- n-三甲基鸟氨酸的初步纯化采用了顺序阳离子交换阴离子交换色谱法。用邻苯二醛和巯基乙醇对柱洗脱液中的氨基酸进行衍生化，在离子配对试剂存在下，采用等容反相高效液相色谱分离。采用柱后荧光法进行定量。检出限为5 pmol ϵ-N-trimethyllysine注入色谱仪。该方法适用于1 ml血浆或0.2-0.4 ml尿液中ϵ-N-trimethyllysine的测定。应用该方法测定了4例全身性肉碱缺乏症患者和6例正常人血浆和尿液中的ϵ-N-trimethyllysine。与正常人相比，全身性肉碱缺乏症患者血浆ϵ-N-trimethyllysine浓度显著降低，但两组之间尿ϵ-N-trimethyllysine排泄量无显著差异。
A method for measurement of ϵ-N-trimethyllysine in human blood plasma and urine is described. An internal standard, δ-N-trimethylornithine, was added to plasma and urine specimens and the mixtures were deproteinized and/or hydrolyzed. Preliminary purification of ϵ-N-trimethyllysine and δ-N-trimethylornithine was achieved by sequential cation-exchange-anion-exchange chromatography. Amino acids in the column eluates were derivatized with o-phthalaldehyde and mercaptoethanol, and were separated by isocratic reversed-phase high-performance liquid chromatography in the presence of an ion-pairing reagent. Quantitation was achieved by post-column fluorometry. The limit of detection was 5 pmol of ϵ-N-trimethyllysine injected into the chromatograph. The procedure was suitable for determination of ϵ-N-trimethyllysine in 1 ml of plasma or 0.2–0.4 ml of urine. The method was applied to measurements of ϵ-N-trimethyllysine in plasma and urine of four systemic carnitine deficiency patients and six normal subjects. Plasma ϵ-N-trimethyllysine concentration was significantly lower in systemic carnitine deficiency patients compared to normal individuals, but no significant difference in urinary ϵ-N-trimethyllysine excretion was observed between the two groups.