Analysis of Vitamin D2 and Vitamin D2 in Infant and Adult Nutritional Formulas by Liquid Chromatography-Tandem Mass Spectrometry: A Multilaboratory Testing Study *

Brendon D. Gill, Harvey E. Indyk

A multilaboratory testing study was conducted on AOAC First Action Method 2016.05: "Analysis of Vitamin D2 and Vitamin D3 in Fortified Milk Powders, Infant Formulas, and Adult/Pediatric Nutritional Formulas by Liquid ChromatographyTandem Mass Spectrometry". Nine laboratories participated in the analysis of duplicate samples of 20 nutritional products. The samples were saponified at high temperature with lipid-soluble components extracted into isooctane; an aliquot was washed and vitamin D derivatized with 4-phenyl-1,2,4-triazoline-3,5-dione to form a highmolecular mass, easily ionizable adduct, extracted into acetonitrile and analyzed by reversed-phase LC-tandem MS. Stable isotopelabeled internal standards were used for quantitation to correct for losses in extraction and variation in derivatization and ionization efficiencies. Acceptable precision as RSD was demonstrated; repeatability ranged from 1.9 to 5.8% RSDr and reproducibility values ranged from 6.4 to 12.7% RSDR, with samples meeting the precision limits specified in the vitamin D Standard Method Performance Requirements and the guidelines recommended for the Horwitz ratio. Method accuracy was assessed using NIST 1849a Standard Reference Material, with a p-value of 0.32, indicating an absence of bias against the certified value. As expected, placebo samples not fortified with vitamin D returned negligible results.

* This method was awarded the 2017 Method of the Year Award by AOAC International at the AOAC Annual Meeting & Conference, Atlanta, GA, USA, 25th September 2017.

MLT Collaborators
Andrew Todd and Mark Reynolds, Fonterra, New Zealand
Carolyn Burdette and Melissa Phillips, NIST, United States of America
Danny Samson and Maurice Seegers, Mead-Johnson, The Netherlands
Emeline Tissot, Frédéric Martin, and Esther Campos-Giménez, Nestlé, Switzerland
Greg Hostetler and Steve Tennyson, Perrigo Nutritionals, United States of America
Isabelle Malaviole and Marlène Daminato, Aquanal, France
Shane Wei, Fonterra, New Zealand
Tiffany Gallegos-Peretz and Sneh Bhandari, Merieux Nutrisciences, United States of America
Vicky Manti, Danone Nutricia, The Netherlands
Yang Zhou, Eurofins, United States of America

Gill et al. (2018) J. AOAC Int., 101, 256–263.Journal Link
Gill et al. (2018) J. AOAC Int., 101, 256–263.Article PDF
Reference Style

Journal: Journal of AOAC International
Publisher: AOAC International

Gill, B.D., & Indyk, H.E. (2018) J. AOAC Int. 101, 256‑263
doi: 10.5740/jaoacint.17-0149

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