串联质谱分析干血滤纸片酰基肉碱方法的建立

Establishing a tandem mass spectrometry method for the analysis of acylcarnitines from dry blood spots

目的 建立串联质谱方法进行干血滤纸片酰基肉碱 (acylcarnitine)分析 ,以便应用于临床相关疾病诊治和新生儿遗传代谢病筛查。方法 取含不同浓度的 8种酰基肉碱的干血滤纸片 (直径 3mm) ,包括乙酰肉碱 (C2 )、丙酰肉碱 (C3)、丁酰肉碱 (C4 )、异戊酰肉碱 (C5 )、己酰肉碱 (C6 )、辛酰肉碱 (C8)、肉豆蔻酰肉碱 (C14 )和棕榈酰肉碱 (C16 )。用含已知量相应酰基肉碱内标的甲醇萃取、盐酸正丁醇衍生后 ,用串联质谱仪检测酰基肉碱及其浓度。分析该方法的精确性、样品回收率及实测值与加入浓度之间的相关性。结果 酰基肉碱批内变异系数为 6 3%～ 9 9% ,批间变异为 10 3%～13 3%。低浓度酰基肉碱回收率为 98%～ 10 6 % ,中等浓度酰基肉碱回收率为 97%～ 10 9% ,高浓度酰基肉碱回收率为 94 %～ 10 8%。实测浓度与加入浓度之间的相关系数为 0 9986～ 0 9999。结论 串联质谱技术进行血滤纸片酰基肉碱分析 ,有较高的回收率 ,能够达到较高的精确性和准确性 ,为临床进行酰基肉碱分析提供了一项新技术。

Objective To establish a method for the analysis of acylcarnitine in dry blood spots by use of tandem mass spectrometry and apply to newborn screening and diagnosis for high risk children with inborn error of metabolism Method The diameter of 3 mm dry blood spots with different concentration of C2、C3、C4、C6、C8、C14 and C16 were punched and extracted into methanol solution with stable isotope labeled internal standards, then derivatized with butanolic HCI After preparation, the samples were analyzed by tandem mass spectrometry The method precision (coefficient of variation , CV ), recovery, and linearity were analyzed Results The intraassay CVs were 6 3 %-9 9 %,The interassay CVs were 10 3 %-13 3 % The recoveries of low, middle and high level of acylcarnitines were 98 %-106 %, 97 %-109 %and 94 %-108 %, respectively。The linearity of different acylcarnitines were 0 998 6-0 999 9 respectively Conclusion The precision, recovery and linearity of the method are good enough for acylcarnitines analysis Tandem mass spectrometry method for acylcarnitines analysis may be applied on newborn screening and diagnosis for high risk children with inborn error of metabolism