Minkler P E, Hoppel C L
Medical Research Service, Department of Veterans Affairs Medical Center, Cleveland, OH 44106.
J Chromatogr. 1993 Apr 2;613(2):203-21. doi: 10.1016/0378-4347(93)80135-q.
This paper describes the development of a high-performance liquid chromatographic method for the quantitation of free carnitine, total carnitine, acetylcarnitine, propionylcarnitine, isovalerylcarnitine, hexanoylcarnitine and octanoylcarnitine in human urine. Carnitine and acylcarnitines were isolated from 10 or 25 microliters of urine using 0.5-ml columns of silica gel, derivatized with 4'-bromophenacyl trifluoromethanesulfonate and separated by high-performance liquid chromatography. Using 4-(N,N-dimethyl-N-ethylammonio)-3-hydroxybutanoate ("e-carnitine") as the internal standard, standard curves (10-300 nmol/ml) were generated. Carnitine and acylcarnitines were quantified (when they were present) in normal human urine and the urine of patients diagnosed with one of three different disorders of organic acid metabolism: methylmalonic aciduria, isovaleric aciduria, isovaleric acidemia, and medium-chain acyl-CoA dehydrogenase deficiency.
本文描述了一种高效液相色谱法的开发，用于定量测定人尿液中的游离肉碱、总肉碱、乙酰肉碱、丙酰肉碱、异戊酰肉碱、己酰肉碱和辛酰肉碱。使用0.5毫升硅胶柱从10或25微升尿液中分离肉碱和酰基肉碱，用4'-溴苯甲酰三氟甲磺酸酯进行衍生化，然后通过高效液相色谱法进行分离。以4-(N,N-二甲基-N-乙基铵基)-3-羟基丁酸酯（“e-肉碱”）作为内标，生成标准曲线（10-300纳摩尔/毫升）。对正常人尿液以及被诊断患有三种不同有机酸代谢紊乱之一（甲基丙二酸尿症、异戊酸尿症、异戊酸血症和中链酰基辅酶A脱氢酶缺乏症）的患者尿液中的肉碱和酰基肉碱进行了定量（如果存在）。
