Poorthuis B J, Jille-Vlcková T, Onkenhout W
Department of Pediatrics, University Hospital, Leiden, The Netherlands.
Clin Chim Acta. 1993 Jul 16;216(1-2):53-61. doi: 10.1016/0009-8981(93)90138-t.
A high-performance liquid chromatographic method is presented for the determination of urinary acylcarnitines. After solid phase extraction on silica columns the acylcarnitines are converted to 4'-bromophenacyl esters with 4'-bromophenacylbromide in the presence of N,N-diisopropylethylamine. Complete derivatization was achieved at 37 degrees C within 30 min. The 4'-bromophenacyl esters were separated by high-performance liquid chromatography on a Hypersil BDS C8 reversed-phase column with a binary gradient containing varying proportions of acetonitrile, water and 0.1 M triethylamine phosphate buffer. Essentially baseline separation was obtained with a standard mixture containing 4'-bromophenacyl esters of carnitine and synthetic acylcarnitines of increasing chain length ranging from acetyl- to palmitoylcarnitine. The method was used to obtain urinary acylcarnitine profiles from patients with propionic, methylmalonic and isovaleric acidemia and with medium-chain and multiple acyl-CoA dehydrogenase deficiency. Quantification of the acylcarnitines was achieved using undecanoylcarnitine as internal standard.
本文介绍了一种用于测定尿中酰基肉碱的高效液相色谱法。在硅胶柱上进行固相萃取后，酰基肉碱在N，N-二异丙基乙胺存在下与4'-溴苯甲酰溴反应转化为4'-溴苯甲酰酯。在37℃下30分钟内可实现完全衍生化。4'-溴苯甲酰酯通过高效液相色谱在Hypersil BDS C8反相柱上进行分离，采用含有不同比例乙腈、水和0.1M磷酸三乙胺缓冲液的二元梯度洗脱。使用含有肉碱的4'-溴苯甲酰酯和链长从乙酰肉碱到棕榈酰肉碱递增的合成酰基肉碱的标准混合物，基本上实现了基线分离。该方法用于从患有丙酸血症、甲基丙二酸血症和异戊酸血症以及中链和多种酰基辅酶A脱氢酶缺乏症的患者中获取尿酰基肉碱谱。使用十一烷酰肉碱作为内标对酰基肉碱进行定量。
