Schmidt-Sommerfeld E, Zhang L, Bobrowski P J, Penn D
Department of Pediatrics, Louisiana State University Medical Center, New Orleans 70112-2822, USA.
Anal Biochem. 1995 Oct 10;231(1):27-33. doi: 10.1006/abio.1995.1498.
A method for the quantitation of short- and medium-chain acylcarnitines in plasma and its clinical application are described. The method is based on enzymatic exchange of L-[3H]carnitine into the acylcarnitine pool, subsequent separation of labeled acylcarnitines by high-performance liquid chromatography, and quantitation of the radioactivity by a beta flowthrough detector. Since only acylcarnitines are detected, no sample cleanup procedure is required. Isotopic equilibrium, a prerequisite for accurate quantitation, was reached in plasma after 1 h of incubation for all acylcarnitines except isovalerylcarnitine which required a longer incubation time. No significant hydrolysis of acylcarnitines occurred during the incubation. Linearity was demonstrated after adding increasing amounts of individual acylcarnitines to plasma. The method is highly sensitive requiring no L-carnitine administration to the patient and differentiates short-chain acylcarnitine isomers. It is suitable for the detection of a number of inborn errors of organic acid and fatty acid metabolism.
本文描述了一种定量测定血浆中短链和中链酰基肉碱的方法及其临床应用。该方法基于L-[3H]肉碱与酰基肉碱池的酶促交换，随后通过高效液相色谱法分离标记的酰基肉碱，并通过β流通检测器对放射性进行定量。由于仅检测酰基肉碱，因此无需样品净化程序。除异戊酰肉碱需要更长的孵育时间外，所有酰基肉碱在孵育1小时后血浆中均达到了准确定量的前提条件——同位素平衡。孵育过程中酰基肉碱未发生明显水解。向血浆中添加越来越多的单个酰基肉碱后证明了线性关系。该方法高度灵敏，无需向患者施用L-肉碱，并且能够区分短链酰基肉碱异构体。它适用于检测多种有机酸和脂肪酸代谢的先天性缺陷。
