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

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• 作者：Lena Jafri^a ; Aysha Habib Khan^b ; ^{aysha.habib@aku.edu"" rel=""nofollow} ; Anwar A. Siddiqui^c ; Shamim Mushtaq^d ; Romaina Iqbal^e ; Farooq Ghani^a ; Imran Siddiqui^a
• 关键词：Vitamin D ; High performance liquid chromatography ; Radio immunoassay ; Methodology
• 刊名：Clinical Biochemistry
• 出版年：2011
• 出版时间：July 2011
• 年：2011
• 卷：44
• 期：10-11
• 页码：864-868
• 全文大小：512 K

文摘

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.