Kamimori H, Hamashima Y, Konishi M
Shionogi Research Laboratories, Shionogi & Co., Ltd., Osaka, Japan.
Anal Biochem. 1994 May 1;218(2):417-24. doi: 10.1006/abio.1994.1201.
A high-performance liquid chromatographic method was developed for the determination of carnitine and 11 acylcarnitines (acetyl-, propionyl-, butyryl-, isovaleryl-, hexanoyl-, octanoyl-, decanoyl-, lauroyl-, myristoyl-, palmitoyl-, and stearoylcarnitine) in human urine. Urine samples were treated with a disposable cation-exchange column to extract carnitine and acyl-carnitines using cyclopentanoylcarnitine as an internal standard. The extracted carnitine and acylcarnitines were reacted with 3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone as a fluorescence derivatization reagent and were separated and detected by high-performance liquid chromatography with gradient elution. Calibration curves gave good linearities at the individual assay range. Analytical recoveries were 70-76% for palmitoylcarnitine and stearoylcarnitine, and were more than 96% for carnitine and the 9 other acylcarnitines. The lower limits of quantitation were 103 nmol/ml for carnitine, 19 nmol/ml for acetylcarnitine, and 0.5-10.5 nmol/ml for other acetylcarnitines. Precision and accuracy including within- and between-run were < or = 19% and -17 approximately +13% at the lowest concentrations of the individual assay ranges. The coefficient of correlation between this method and the radioenzymatic method was found to be 0.988 for the measured concentration of carnitine. We concluded that the present method can be used to measure carnitine and acylcarnitines in human urine.
建立了一种高效液相色谱法，用于测定人尿中的肉碱和11种酰基肉碱（乙酰基、丙酰基、丁酰基、异戊酰基、己酰基、辛酰基、癸酰基、月桂酰基、肉豆蔻酰基、棕榈酰基和硬脂酰基肉碱）。尿样用一次性阳离子交换柱处理，以环戊酰基肉碱为内标提取肉碱和酰基肉碱。提取的肉碱和酰基肉碱与3-溴甲基-6,7-二甲氧基-1-甲基-2(1H)-喹喔啉酮作为荧光衍生试剂反应，通过梯度洗脱的高效液相色谱进行分离和检测。校准曲线在各个测定范围内具有良好的线性。棕榈酰基肉碱和硬脂酰基肉碱的分析回收率为70-76%，肉碱和其他9种酰基肉碱的回收率超过96%。肉碱的定量下限为103 nmol/ml，乙酰基肉碱为19 nmol/ml，其他酰基肉碱为0.5-10.5 nmol/ml。在各个测定范围的最低浓度下，包括批内和批间的精密度和准确度分别≤19%和-17%至+13%。该方法与放射酶法测定肉碱浓度的相关系数为0.988。我们得出结论，本方法可用于测定人尿中的肉碱和酰基肉碱。
