串联质谱法快速检测干血滤纸片酰基肉碱含量  

Rapid Measurement of Blood Spot Acyl Carnitines by Tandem Mass Spectrometry Method

作　　者：蔡燕娜 彭敏芝 赵小媛 吴冬燕 盛慧英 范莉萍 刘丽 

机构地区：[1]广州市妇女儿童医疗中心内分泌代谢科,广州510623

出　　处：《现代检验医学杂志》2011年第4期16-20,共5页Journal of Modern Laboratory Medicine

基　　金：基金项目：国家十一五科技支撑计划（2006BA105A07）;广州市科技局支撑计划（2010J-E231-1）.

摘　　要：目的建立一种串联质谱法对干血滤纸片中的酰基内碱进行快速分析，以用于脂肪酸代谢障碍性疾病和有机酸血症诊断及治疗效果的监测。方法取8种不同浓度的酰基肉碱的干血滤纸片（直径5mm），用甲醇（含已知浓度的相应酰基肉碱同位素内标）对其进行萃取，不进行街生化，萃取物过滤后直接进入ESI^＋-MS／MS上机分析测定，采用多反应监测模式（MRM），以标准品制作标准曲线，同住素内标法定量建立分析方法，进行回收试验、精密度试验、干扰试验及准确度试验。结果各标准曲线相关系数r2〉0．994。低浓度各种酰基内碱回收率为85％～103．5％，准确度在91．8％～110．0％；中浓度各种酰基肉碱回收率为85％～96％，准确度在87．6％～110．1％；高浓度酰基内碱回收率为94％～100．1％，准确度在95．7％～110．1％。实测值与加入浓度之间的相关系数（r）为：0．997－1．000。各种酰基内碱批内变异系数（CV）1％～10％和批间变异系数（CV）5％～13％。内标液保存1～21天的检洲均值均在CDC提供的参考范围内，而批内CV％均〈10％。263例O～6岁健康儿童血滤纸片中酰基肉碱浓度因受年龄因素影响，按不同年龄组划分各种内碱参考范围。结论非衍生化串联质谱法分析干血滤纸片中的酰基内碱含量，样本前处理简单，能够达到较高的精确度和准确度，无常规串联质谱中的衍生化、氮吹和复溶等步骤，大大节省了的标本处理时间，是一项简便、快捷、准确的检测方法，可以用于脂肪酸代谢障碍性疾病和有机酸血症诊断及治疗效果监测。Objective To establish a rapid method for the analysis of acylcarnitine in dry blood spots by use of tandem mass spectrometry and apply to diagnosis for high risk children with inborn error of metabolism. Methods The diameter of Smm dry blood spots with different concentration of were punched and extracted into methanol solution with stable isotope labeled internal standards and underivatized. After preparation,the samples were analysed by tandem mass spectrometry, multi-reactions monitor. The concentration of acylcarnitines were detected with the isotope internal method. The stand curve was employed with a series of calibrators. The method precision （coefficient of variation,CV）,recovery,accuracy,linearity and Interference experiment were analysed with quality control samples. Results The correlation of these method coefficient were more than 0. 994. The recoveries of low, middle and high level of acylearnitines were 85 % - 103. 5 %, 85 % -96% and 94%-100. 1% respectively. The accuracies of method were 91.8%-110.0% ,87.6%-110. 1% and 95. 7%- 110. 1% respectively. The linearity of different acylcarnitines were 0. 997-1. 000 respectively. The intrassay CVs were 1% -10% ,the interassay CVs were 5%-13%. Internal standard solution save 1-21 d of the detection means were provided in the CDC reference range ,and both intra CV % -10 %. 263 cases of healthy children aged from 0 to 6 years old were detected acyl carnitines concentration of blood spots. According to different age groups were divided into different age groups to make acyl carnitines reference range. Conclusion The accuracy,recovery,precision and linearity of the method were good enough for cylcarnitines analysis. Tandem mass spectrometry method for acylcarnitines analysis may be applied ondiagnosis for high risk children with inborn error of metabolism.

关 键 词：串联质谱 酰基肉碱 非衍生化 干血滤纸片 

分 类 号：Q503[生物学—生物化学]