| Objective A liquid chromatography-tandem mass spectrometry(LC-MS/MS)method was established for the detection of parathyroid hormone(PTH)in human serum,and the methodological comparison of different methodological detection systems for PTH were conducted.Methods The serum was immunologically enriched with polystyrene beads coated with anti-PTH antibody and then the serum was discarded.The enriched PTH was hydrolyzed by trypsin and tested by LC-MS/MS.In the positive ion mode,PTH was detected by electrospray ionization(ESI)and multiple reaction monitoring(MRM).We have evaluated the interference of PTH 1-44.Serum with different PTH concentration levels were detected for 5 consecutive days to investigate the precision of the method.The limits of detection(LOD)and quantitation(LOQ),matrix effects,recovery rate,linear range and stability of serum were evaluated.The concentration of PTH in serum of 50 patients was determined to evaluate the consistency of LC-MS/MS and electrochemiluminescence immunoassay(ECLIA).Results Serum PTH can be detected by the LC-MS/MS method established in this study.The retention time of the compounds was about 4.0 min.The method was with high specificity and no significant interference.In the range of 30-4000pg/ml,the linearity showed well,and the correlation coefficient was greater than 0.99.The reproducibility ranged from 5.23%to 6.59%and laboratory imprecision ranged from 7.55%to 13.53%.The LOD and LOQ were 25 pg/ml and 30 pg/ml,respectively.The recoveries of PTH were 94.19%～98.36%.The regression equation of comparison between ECLIA and LC-MS/MS was y=1.06 x-24.57,and average percentage bias was-14.72%.Conclusion This study successfully established a method for detection of serum PTH by LC-MS/MS,which was specific,no significant interference and can be used for clinical evaluation of human serum PTH concentration.The correlation between ECLIA and LC-MS/MS was fine. |
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