液相色谱串联质谱检测尿黏多糖在黏多糖贮积症患者诊断与随访中的应用  

Detection of urine glycosaminoglycans by liquid chromatography-tandem mass spectrometry in the diagnosis and follow-up of patients with mucopolysaccharidosis

作　　者：占霞 高晓岚[1] 季文君[1] 常思宇 刘丹 张惠文[1] ZHAN Xia;GAO Xiaolan;JI Wenjun;CHANG Siyu;LIU Dan;Zhang Huiwen(Department of Pediatric Endocrinology and Genetics,Xinhua Hospital,Shanghai Institute for Pediatric Research,Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China;Shanghai AB Sciex Analytical Instrument Trading Co.Ltd,Shanghai 20050,China)

机构地区：[1]上海交通大学医学院附属新华医院儿内分泌遗传代谢科上海市儿科医学研究所,上海200092 [2]上海爱博才思分析仪器贸易有限公司,上海200050

出　　处：《临床儿科杂志》2024年第5期399-406,共8页Journal of Clinical Pediatrics

基　　金：上海市卫生健康委员会临床专项(No.202240361);上海市“科技创新行动计划”医学创新研究专项(No.20MC1920400)。

摘　　要：目的运用液相色谱串联质谱(LC-MS/MS)技术建立尿液中硫酸软骨素(CS)、硫酸皮肤素(DS)和硫酸乙酰肝素(HS)含量的检测方法,探究该方法在黏多糖贮积症(MPS)患者诊断及治疗随访中的应用。方法收集20例MPS患儿及37例正常儿童尿液样本,测定尿液中CS、DS和HS等浓度。尿样氮气吹干,盐酸甲醇衍生化,再次氮气吹干,复溶后LC-MS/MS分析;评价方法的线性、定量限、精密度、加样回收率和基质效应。结果LC-MS/MS检测CS、DS和HS的线性均大于0.99。CS、DS和HS的定量下限依次为:0.5、1.0和1.0 mg/L。批内不精密度为1.9%~10.4%,批间不精密度为2.6%~9.8%。加标回收率为85.6%~110.4%,相对基质效应为84.9%~115.5%。CS、DS和HS在正常儿童尿液中呈正态分布,以x+1.64 SD计算正常儿童尿液中的参考区间上限,分别为16.5、1.8、1.4 mg/mmol。通过分析MPS患儿尿液黏多糖特点,LC-MS/MS法能有效检出MPS I、MPSⅡ、MPSⅢ和MPSⅥ等MPS患儿。已接受造血干细胞移植的MPSⅠ和MPSⅡ患儿尿DS、HS水平降低。结论LC-MS/MS检测尿黏多糖性能良好,有望用于MPS患者的精准诊断和治疗随访监测。Objective To establish a method to determine chondroitin sulfate(CS),dermatan sulfate(DS) and heparan sulfate(HS) in urine by liquid chromatography tandem mass spectrometry(LC-MS/MS),and to investigate the application of urine glycosaminoglycans detection in the diagnosis and follow-up of mucopolysaccharidosis(MPS) patients.Methods Twenty urine samples from MPS patients and 37 urine samples from normal children were collected,and glycosaminoglycans levels in these samples were determined.Urine samples were added and dried under nitrogen.Then the samples were derivatized with methanol hydrochloride and dried under nitrogen.After reconstituted,the glycosaminoglycans were analyzed by LC-MS/MS.The linearity,limit of quantification,precision,spiked recoveries,and matrix effect were evaluated.Results The linearity of CS,DS and HS by LC-MS/MS was>0.99.The limits of quantification for CS,DS and HS were 0.5 mg/L,1 mg/L and 1 mg/L,respectively;the intra-day imprecision ranged from 1.9% to 10.4%,and the interday imprecision ranged from 2.6% to 9.8%;and the recovery ranged from 85.6% to 110.4%,the relative matrix effect ranged from 84.9% to 115.5%.CS,DS and HS in urine samples of normal children were normally distributed,then the upper limit of reference interval were preliminarily determined by mean+ 1.64 SD,which were 16.5 mg/mmol,1.8 mg/mmol and 1.4 mg/mmol,respectively.Urine GAGs of MPS patients were analyzed,the LC-MS/MS method was effective in detecting MPSⅠ,MPSⅡ,MPSⅢand MPSⅥpatients.Urine samples of MPSⅠor MPSⅡpatients who had hematopoietic stem cell transplantation showed reduced DS and HS concentration.Conclusion LC-MS/MS has good performance in detecting urinary mucopolysaccharides and is expected to be used for accurate diagnosis and treatment follow-up monitoring of MPS patients.

关 键 词：液相色谱串联质谱 黏多糖 黏多糖贮积症 

分 类 号：R725[医药卫生—儿科]