EVALUATION OF A NOVEL LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY BASED METHOD FOR VITAMIN D ANALYSIS IN BLOOD

  • Sehrish Naz Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Muhammad Aamir Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Zujaja Hina Haroon Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Sobia Irum Kirmani Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Nisar Ahmed Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Sadiq Ali Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
Keywords: 25 Hydroxy vitamin d2, 25 Hydroxy vitamin d3, Liquid chromatography mass spectrometry (LCMS), Validation

Abstract

Objective: To evaluate a novel liquid chromatography tandem mass spectrometry based method for serum 25 hydroxy vitamin D (D2 and D3 metabolites) analysis.

Study Design: Cross sectional validation study.

Place and Duration of Study: Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Pakistan, from Mar 2019 to Mar 2020.

Methodology: Samples were extracted and 25 OH vitamin-D was separated by means of chromatography and finally quantified via mass spectrometer. A quadrupole- tandem mass spectrometer with Electron spray Ionization coupled to high performance liquid chromatography was adopted for detection and quantitation of 25-hydroxyvitamin D2 and D3 in serum.

Results: Limit of detection (LOD) was at the level of 2.49 ng/ml and limit of quantitation (LOQ) was estimated to be 3.9 ng/ml for both the metabolites. The correlation coefficient was 0.99. For D3 observed recovery was 98% and 97.5% respectively while for D2 the recovery was calculated to be 97% and 98%. Percentage relative standard deviation (RSD) was 0.8% and 1.3% respectively. This method has an advantage of less matrix effects, minimal cross-reactivity with 24, 25 hydroxy vit D and 25, 26 di-hydroxy vitamin D metabolite than the routinely used antibody-based assays.

Conclusion: This LC-MS/MS methodology is sensitive, specific and can quantitate Vitamin D2 and D3 quite efficiently. This method may be employed for vitamin D analysis in clinical laboratories.

Downloads

Download data is not yet available.
Published
2021-08-27
How to Cite
Naz, S., Aamir, M., Haroon, Z., Kirmani, S., Ahmed, N., & Ali, S. (2021). EVALUATION OF A NOVEL LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY BASED METHOD FOR VITAMIN D ANALYSIS IN BLOOD. PAFMJ, 71(4), 1355-59. https://doi.org/10.51253/pafmj.v71i4.6090
Section
Original Articles

Most read articles by the same author(s)