非衍生化串联质谱法检测酰基肉碱方法的应用

摘要

目的 建立非衍生化串联质谱检测酰基肉碱的方法,以便用于有机酸和脂肪酸代谢病的检测和筛查.方法 使用含13种稳定同位素标记的酰基肉碱内标准品的溶剂萃取滤纸干血片中的游离肉碱和酰基肉碱,串联质谱仪直接进样,检测游离肉碱(CO)、乙酰基肉碱(C2)、丙酰基肉碱(C3)、丁酰基肉碱(C4)、异戊酰基肉碱(CS)、戊二酰基肉碱(C5DC)、己酰基肉碱(C6)、辛酰基肉碱(C8)、葵酰基肉碱(C10)、十二烷酰基肉碱(C12)、十四烷酰基肉碱(C14)、十六烷酰基肉碱(C16)和十八烷酰基肉碱(C18)的浓度.分析该方法的精确性、准确性及稀释回收率.结果 低浓度酰基肉碱的批内变异系数(CV)为6.61%～10.97%之间,批间CV为13.31%～18.98%;高浓度酰基肉碱的批内CV为5.74%～12.13%,批间CV为12.70%～18.88%.除C5DC外,其余酰基肉碱的偏倚为-15%～20%;酰基肉碱不同稀释浓度的回收率在90%～115%之间,实测值与预测值的相关系数(r)为0.97～0.99.结论 非衍生化串联质谱检测酰基肉碱有较高的精确性和准确度,有临床应用价值.此技术可以作为新生儿遗传代谢病筛查的常规技术.%Objective To establish the non-derivatized tandem mass spectrometry( MS/MS ) method to detect acylcarnitines for the detection and screening of the organic acid and fatty acid metabolism disorders. Methods The measurement of free carnitine and acylcarnitines, including free carnitine ( CO ), acetylcarnitine ( C2 ), propionylcarnitine( C3 ), butyrylcarnitine( C4 ), is oval eryl carnitine ( C5 ), glutaryl carnitine ( C5DC ), hexanoyl carnitine ( C6 ), octanoylcarnitine ( C8 ), decanoylcarnitine ( CIO ), dodecanoylcarnitine( C12 ), tetradecanoylcarnitine( C14 ), hexadecanoylcarnitine ( C16 ) and octadecanoylcarnitine ( C18 ), involved extraction of dried blood spots with a solution containing 13 stable-isotope labeled internal standards and analysis by MS/MS system. The precision, accuracy and recovery of the method were analyzed. Results The within-run coefficient of variation ( CV) of the acylcarnitines with low concentration was 6. 61%-10. 97%, and the between-run CV was 13. 31%-18. 98% . The CV of the acylcarnitines with high concentration were 5.74%-12. 13% (within-run) and 12. 70%-18. 88% (between-run), respectively. The bias, except for C5DC, varied from - 15% to 20%. The recoveries of the assay from different concentration levels varied from 90% to 115% . The correlation coefficien< r ) between detection value and prediction value varied from 0. 97 to 0. 99. Conclusions The high precision and accuracy non-derivatized MS/MS method has the value of clinical application. The method should be applied as a regular technique in the newborn screening for inherited metabolic disorders.