Quantification and clinical performance of serum parathyroid hormone 1-84 via immunocapture coupled to LC-MS/MS in chronic renal failure

Abstract

Accurate measurement of serum parathyroid hormone (PTH) is crucial for diagnosing and managing endocrine and osteological diseases. Conventional immunoassay methods struggle with cross reactivity issues between full-length PTH and truncated fragments or post-translationally modified forms. Both the standardization of PTH assays and the peptide's stability are concerning. This study addresses these issues by establishing an immunocapture coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to precisely quantify PTH1-84. PTH1-84 was isolated from one mL serum samples by immunocapture on a polystyrene bead and eluted from matrix, then quantitated by LC-MS/MS. The results from 268 serum samples were compared to an intact PTH immunoassay. The assay's linear range was 5.0-1000.0 pg/mL. The intra-assay coefficients of variation (CVs) ranged from 3.2 % to 6.8 %, and the inter-assay CVs ranged from 4.6 % to 9.5 %. The extraction efficiencies were 98.0 %-100.5 %, with no significant matrix effects observed after internal standard correction. The correlation coefficient between LC-MS/MS and immunoassay was 0.989, but the bias between the methods was substantial. Nevertheless, the immunocapture purification coupled LC-MS/MS method offers a promising approach for accurate PTH measurement.

Keywords: Immunoassay; Immunocapture purification; LC–MS/MS; Parathyroid hormone.