尿滤纸片法气相色谱2质谱分析技术

・遗传代谢内分泌疾病研究・

基金项目:国家自然科学基金委员会国家杰出青年科学基金(30125019);卫生部临床学科重点建设项目(97070240)作者单位:430030武汉,华中科技大学同济医学院附属同济医院儿科,同济医学院遗传代谢性疾病诊断中心

尿滤纸片法气相色谱2质谱分析技术在

遗传性代谢病高危筛查诊断中的应用

罗小平　王慕逖　魏虹　梁雁　王宏伟　林汉华　董永绥　刘皖君　方俊敏　宁琴 【摘要】　目的　建立一种适合我国国情的有机酸尿症及其他遗传性代谢病的高危筛查诊断方法。方法　应用滤纸片收集尿标本,建立尿有机酸及氨基酸代谢产物的萃取、衍化和气相色谱2质谱分析方法;成立遗传代谢性疾病高危筛查诊断协作网络并开展工作。结果　352例样本中共检出阳性病例34例(9166%),病种包括甲基丙二酸血症6例,α2酮戊二酸尿症5例,酪氨酸血症Ⅰ型4例,二羧酸尿症4例,多种羧化酶缺乏症3例,苯丙酮尿症3例,高乳酸血症3例,丙酸血症2例,鸟氨酸氨甲酰基转移酶缺乏症1例,乙基丙二酸2己二酸尿症1例,戊二酸尿症Ⅱ型1例和32甲基巴豆酰羧化酶缺乏症1例。常见的临床症状和体征包括智能发育落后、惊厥、肌张力异常和黄疸。常规实验室检查多见代谢性酸中毒、低血糖和高血氨等异常。结论　尿有机酸气相色谱2质谱分析为遗传性代谢病高危筛查诊断的重要技术。应用滤纸片收集尿标本后在遗传代谢病诊断中心进行检测为实用而有效的方法。建立协作网络是成功开展遗传性代谢病高危筛查诊断工作的关键。

【关键词】　代谢缺陷,先天性;　有机酸类;　碎片质谱法;　尿分析

Application of gas chrom atography 2m ass spectrometry analysis on urine filter p aper in the high 2risk screening and diagnosis of inherited metabolic diseases LUO Xiao 2ping ,WANG Mu 2ti ,WEI Hong ,LIANG Yan ,WANG Hong 2wei ,LIN Han 2hua ,DONG Yong 2sui ,LIU Wan 2jun ,F ANG Jun 2min ,NING Qin 1Department o f Pediatrics ,Tongji Hospital ,Center for the Diagnosis o f G enetic Metabolic Diseases ,Tongji Medical College ,Huazhong Univer sity o f Science and Technology ,Wuhan 430030,China

【Abstract 】　Objective T o establish a specific procedure for the high 2risk screening and diagnosis of organic acidurias and other inherited metabolic diseases in China 1Methods A nation 2wide netw ork for the high 2risk screening and diagnosis of genetic metabolic diseases was formed to facilitate the collaboration 1Urine samples were collected using filter paper from patients with clinical sym ptoms suspicious of inherited metabolic diseases 1The samples were eluted with distilled water and internal standards were added 1Samples were treated with hydroxylamine hydrochloride to form oximes to im prove the recoveries of 22ketoacids 1Urinary organic acids were extracted with ethyl acetate and diethyl ether under acidic condition 1A fter dehydration ,the combined organic phase was evaporated to dryness with nitrogen 1The residues were added with BSTFA +1%T MCS and heat incubated to form the trimethylsilyl derivatives ,and then were analyzed on an Agilent 5890/5973N gas chromatography 2mass spectrometer (G C 2MS ),with a 7683series auto 2sam pler 1The peaks were identified by reference to a mass spectral library 1R esults T otally 352sam ples were collected from the netw ork collaborating hospitals since 20011Thirty 2four (9166%)cases of various inherited metabolic diseases were diagnosed with an age range of 2days to 14years 1The disease profile was consisted of methylmalonic acidemias (6),α2keto 2glutaric aciduria (5),tyrosinemia type Ⅰ(4),dicarboxylic aciduria (4),multiple carboxylase deficiency (3),phenylketonuria (3),lactic acidemia (3),propionic acidemia (2),ornithine transcarbam oylase deficiency (1),ethylmalonic 2adipic aciduria (1),glutaric aciduria type Ⅱ(1)and 32methylcrotyl C oA carboxylase deficiency (1)1The m ost comm on clinical sym ptoms and signs included mental and developmental retardation ,convulsion ,musculotonic abnormality and jaundice 1R outine laboratory tests often revealed metabolic acidosis ,hypoglycemia and hyperamm onemia ,etc 1Conclusion Urine organic acids analysis by G C 2MS remains to be the m ost important technique for the high 2risk screening and diagnosis of inherited metabolic diseases 1Use of urine filter paper for sample collection and analysis in advanced genetic metabolic centers is a practical approach to extend the diagnostic capacity and improve the management of such diseases in China 1C ollaborative netw ork played a critical role in the