Li Ke, Sun Qingbao
Department of Medical Laboratory Science, Jinling Hospital, Clinical School of Medical College, Nanjing University, Nanjing 210002, Jiangsu, PR China.
J Chromatogr Sci. 2010 May-Jun;48(5):371-4. doi: 10.1093/chromsci/48.5.371.
A simple and reliable high-performance liquid chromatographic (HPLC) method for simultaneous determination of free and total carnitine in human serum has been developed, and its clinical significance has been investigated. After proteins in serum were precipitated, carnitine in serum was derivatized to form its ester. HPLC separation of the sample solution was performed on a SiO(2) column and detected by UV absorbance at 260 nm. A mobile phase composed of acetonitrile-citric acid-triethanolamine was found to be the most suitable for this separation. The free and total carnitine levels in serum were studied in 347 subjects. The method proved to be linear in the range of carnitine from 5 micromol/L to 400 micromol/L. The relative standard deviations of within-assay for free and total carnitine analyses were 3.36% and 1.97%, respectively; and between-assay for free and total carnitine analyses were 3.34% and 1.77%, respectively. The average recovery was 98.2% for free carnitine and 96.3% for total carnitine, respectively. The method has been applied to the simultaneous determination of free and total carnitine in serum. Statistics analysis showed that total and free carnitine levels of males were higher than that of females (p < 0.01) while acylcarnitine level had no statistically significant difference (p > 0.05).
已开发出一种简单可靠的高效液相色谱（HPLC）方法，用于同时测定人血清中的游离肉碱和总肉碱，并对其临床意义进行了研究。血清中的蛋白质沉淀后，血清中的肉碱被衍生化形成其酯。样品溶液的HPLC分离在硅胶柱上进行，并通过260nm处的紫外吸光度检测。发现由乙腈 - 柠檬酸 - 三乙醇胺组成的流动相最适合这种分离。对347名受试者的血清中游离和总肉碱水平进行了研究。该方法在5μmol/L至400μmol/L的肉碱范围内被证明是线性的。游离和总肉碱分析的批内相对标准偏差分别为3.36%和1.97%；批间游离和总肉碱分析的相对标准偏差分别为3.34%和1.77%。游离肉碱的平均回收率分别为98.2%，总肉碱的平均回收率为96.3%。该方法已应用于血清中游离和总肉碱的同时测定。统计分析表明，男性总肉碱和游离肉碱水平高于女性（p < 0.01），而酰基肉碱水平无统计学显著差异（p > 0.05）。
