Comparison of high performance liquid chromatography, radio immunoassay and electrochemiluminescence immunoassay for quantification of serum 25 hydroxy vitamin D

Lena Jafri, Aysha Habib Khan, Anwar A. Siddiqui, Shamim Mushtaq, Romaina Iqbal, Farooq Ghani, Imran Siddiqui

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Abstract

Objective: To compare the performance of radioimmunoassay (RIA) with high-performance liquid chromatography (HPLC) and electrochemiluminescence (ECLIA) for the quantification of vitamin D (25OHD). Methods: HPLC method for the determination of 25OHD in human biological samples was developed and compared in terms of accuracy and precision with a commercially available RIA assay. Performance of RIA assay with ECLIA technology for 25OHD analysis was further compared. Results: Median 25OHD levels with HPLC vs. RIA were 50.1. nmol/L (IQ = 17.7-199.4. nmol/L) and 51.1. nmol/L (IQ = 12.5-187.2. nmol/L) respectively, whereas median 25OHD concentration with RIA vs. ECLIA was 32.4. nmol/L (9.98-199.7. nmol/L) and 29.9. nmol/L (4.9-214.6. nmol/L), respectively. Comparison data for HPLC vs. RIA showed RIA = -1.13. +. 1.01 (HPLC) (RMSE = 11.2. nmol/L) and for RIA vs. ECLIA revealed, ECLIA = 3.21. +. 0.9 (RIA) (RMSE9.6. nmol/L). Conclusion: Acceptable correlation was observed among HPLC and RIA and also with RIA and ECLIA in quantification of 25OHD.

Original languageEnglish
Pages (from-to)864-868
Number of pages5
JournalClinical Biochemistry
Volume44
Issue number10-11
DOIs
Publication statusPublished - Jul 2011

Keywords

  • High performance liquid chromatography
  • Methodology
  • Radio immunoassay
  • Vitamin D

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