LC-MS/MS检测血清25-羟基维生素D替代校准品基质的选择与评价

目的探讨如何选择和评价液相色谱-串联质谱(LC-MS/MS)检测血清25-羟基维生素D25(OH)D]替代校准品基质。方法分别用4%牛血清白蛋白(BSA)溶液和30%乙醇水溶液2种替代基质,配制25(OH)D的标准溶液并绘制标准曲线。通过比较内标在不同基质中的响应值差异及基质效应混合实验、准确度验证,对替代基质进行全面的评价。结果内标在4%BSA和血清样本中的响应值差异无统计学意义(P>0.05),在30%乙醇水溶液和血清样本中的响应值差异有统计学意义(P<0.05)。4%BSA溶液通过了基质效应混合实验验证,30%乙醇水溶液未通过;使用4%BSA基质的标准曲线对室间质量评价样本的定量结果满足准确度要求;30%乙醇水基质未满足要求,有2个样本的定量结果偏移>25%。结论4%BSA可作为LCMS/MS检测人血清25(OH)D的替代校准品基质。

Evaluation of surrogate matrices for serum 25-hydroxyvitamin D calibrators by LC-MS/MS

Objective To explore the selection and evaluation of a suitable surrogate matrix for preparing 25-hydroxyvitamin D25(OH)D]calibrators by liquid chromatography tandem-mass spectrometry(LC-MS/MS).Methods 4%bovine serum albumin(BSA)buffer and 30%ethanol-water were used to prepare 25(OH)D calibrators.They were thoroughly assessed by comparing the peak areas of internal standards in different matrices,testing their matrix effects and accuracy validation.Results There was no significant difference in response between 4%BSA matrix and serum(P>0.05),while there was statistical difference between 30%ethanol water and serum(P<0.05).The validation of matrix effect showed that 4%BSA-based calibrator was acceptable,while 30%ethanol-water-based calibrator was not.The external quality assessment(EQA)samples quantified by 4%BSAbased calibrator were well consistent with the target values,but the EQA samples quantified by 30%ethanol-waterbased calibrator did not meet accuracy requirements.The bias between the two samples was over 25%.Conclusion 4%BSA can be used as substitute calibrator matrix for the measurement of serum 25(OH)D by LC-MS/MS.