Determination of urinary sulfatides and other lipids by combination of reversed-phase and thin-layer chromatographies
Determination of urinary sulfatides and other lipids by combination of reversed-phase and thin-layer chromatographies
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DOI：
10.1006/abio.1999.4002
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发表时间：
1999-05-01
影响因子：
2.9
通讯作者：
Ledvinová, J
中科院分区：
文献类型：
--
作者：
Berná, L;Asfaw, B;Ledvinová, J
已经开发出一种快速、简便的方法用于测定异染性脑白质营养不良（MLD，芳基硫酸酯酶A缺乏症）患者尿液中的硫脂。该过程包括两个步骤：通过反相色谱法提取尿液中的总脂质，并对其进行高效薄层色谱（HPTLC）分离。对基于不同基质的两种吸附剂进行了比较，发现甲基丙烯酸羟乙酯C - 18型吸附剂效果更佳。20毫升尿液样本足以进行分析。该技术适用于同时进行定性鉴定和半定量光密度测定，适合常规工作。硫脂的量是相对于鞘磷脂来表示的，鞘磷脂与硫脂共同纯化，并且比常用参数（肌酐、尿量等）更能反映尿液中膜脂质的水平。对照组个体的硫脂/鞘磷脂范围为0.15 - 0.68纳摩尔/纳摩尔，MLD患者为3.5 - 27.2纳摩尔/纳摩尔。硫脂的排泄对于真正的MLD具有诊断意义（由于在肾脏中积累），因此该分析对于评估疑似MLD病例（包括临床和酶学非典型病例）非常重要。该方法对于其他尿液中鞘脂排泄量高的脂质沉积症（例如法布里病）作为补充分析也很有用。（C）1999年学术出版社
A fast and simple method for determination of sulfatides in the urine of patients with metachromatic leukodystrophy (MLD, arylsulfatase A deficiency) has been developed. The procedure consists of two steps: extraction of total urinary lipids by reversed-phase chromatography and their HPTLC separation. Two types of sorbents based on different matrixes were compared, of which the hydroxyethyl methacrylate C-18 type sorbent was found to be superior. Twenty-milliliter aliquots of urine are sufficient for the analysis. The technique is appropriate for simultaneous qualitative identification and semiquantitative densitometric determination and is suitable for routine work. The amount of sulfatides is expressed in relation to sphingomyelin, which copurifies with sulfatides and better reflects the level of membrane lipids in urine than commonly used parameters (creatinine, urine volume, etc.). The ranges were found to be 0.15-0.68 nmol sulfatide/nmol sphingomyelin for control individuals and 3.5-27.2 nmol sulfatide/nmol sphingomyelin for MLD patients. The excretion of sulfatides is pathonognomic for true MLD (due to the accumulation in kidney) and therefore Ms analysis is important for evaluation of suspected MLD cases including clinically and enzymatically atypical cases. The method is also useful as a complementary analysis for other lipidoses with high excretion of sphingolipids in urine (e.g., Fabry disease). (C) 1999 Academic Press.