ABSTRACT
Objective:To develop a national secondary reference material of Urea and Creatinine in frozen human serum as a standard for metrological traceability.Methods:According to JJF1343-2012 "General and Statistical Principles for Characterization of Reference Materials" and JJF 1006-1994 " Technical Norm of Primary Reference Material ", the homogeneity, stability, and commutability were evaluated;Using the JCTLM recommended methods, the value of the reference materials was assigned through collaboration with 6 accredited reference laboratories from Guangdong Provincial Hospital of Chinese Medicine, Beijing Aerospace General Hospital, Shenzhen Mindray Bio-Medical Electronics, Maccura Biotechnology, Beijing Leadman Biochemistry, and Zhejiang MedicalSystem Biotechnology. Uncertainty components including inhomogeneity, stability and value assignment were evaluated.Results:The results of one-way analysis of variance of homogeneity for the reference materials showed P>0.05, and the stability evaluation was less than the critical value of the t-test. The measured values were in the 95% confidence interval in the four conventional detection systems for commutability, and the certified values and expanded uncertainties were urea:(14.7±0.3) mmol/L ( k=2),Cr:(313.9±14.5) μmol/L ( k=2). Conclusion:The prepared secondary reference materials of urea and creatinine had promising homogeneity, stability, and commutable, the values of urea and creatinine concentration in reference materials were accurate and reliable.
ABSTRACT
Objective To develop an isotope dilution liquid chromatography tandem mass spectrometry (ID-LC-MS/MS) method for determining the concentration of 5-methyltertrahydrofolate (5MT) in human serum.Methods After qualitative analysis of 5MT reference,its purity was identified by two methods with different principle,i.e.,mass balance and quantitative nuclear magnetic resonance.The definite value and uncertainty of 5MT concentrations in 6 serum samples containing same level of 5MT were assessed by ultra performance liquid chromatography-triple quadrupole tandem mass spectrometer equipped with electrospray ionization in monitoring mode of multiple reactions,in which 13C5-5MT was used as the internal standard.The precision,accuracy,limit of detection (LOD) and limit of quantification (LOQ) of the method were also evaluated.Results After qualitative analysis for confirming the principal component in the sample,the purity of 5MT reference was measured as 76.65％ by mass balance and quantitative nuclear magnetic resonance method.The definite value of 5MT in the serum sample was 8.22 ng/mL,the extended uncertainty was 0.54 ng/mL,LOD was 0.05 ng/mL,LOQ was 0.50 ng/mL and the repeatability and inter-batch precision CV were within 5.0％.The accuracy of measured results was verified by using NIST SRM 1955 since all the values of 3 levels were within the range of certified value with the extended uncertainty.Conclusion A accurately quantitative determination for the concentration of 5 MT in human serum with high specificity and sensitivity was established by using ID-LC-MS/MS method.