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Title: Vitamin D levels in fish and shellfish determined by liquid chromatography with ultraviolet detection and mass spectrometry

item Byrdwell, W Craig
item HORST, RONALD - Heartland Assays, Inc
item PHILLIPS, KATHERINE - Virginia Bioinformatics Institute
item Holden, Joanne
item PATTERSON, KRISTINE - US Department Of Agriculture (USDA)
item Harnly, James - Jim
item EXLER, JACOB - US Department Of Agriculture (USDA)

Submitted to: Journal of Food Composition and Analysis
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/4/2013
Publication Date: 3/21/2013
Citation: Byrdwell, W.C., Horst, R.L., Phillips, K.M., Holden, J.M., Patterson, K.Y., Harnly, J.M., Exler, J. 2013. Vitamin D levels in fish and shellfish determined by liquid chromatography with ultraviolet detection and mass spectrometry. Journal of Food Composition and Analysis. 10:1016/j.jfca.2013.01.005.

Interpretive Summary: There are very few natural sources of vitamin D, among which are salmon, tuna, and other fatty fishes. Unfortunately, there are relatively few reports in the literature on the amounts of vitamin D3 in fish and shellfish, and many of these are older reports that did not employ highly selective and sensitive instrumentation. This article reports vitamin D levels (g/100g) determined in a range of fish species, including catfish, cod, flounder, haddock, halibut, herring, ocean perch, pollock, rockfish, salmon, sardines, swordfish, trout, and tuna, as well as shellfish species, including clams, crabs, mussels, oysters, scallops, and shrimp. The data were obtained using both ultraviolet detection and mass spectrometry. A sample of a certified reference material (NIST 1849 Infant/Adult Nutritional Formula) was analyzed to validate the method and confirm that it gave good results (within expected uncertainty limits) for a sample with a known amount of vitamin D3. To provide further method validation, salmon control composite (CC) samples that had been previously analyzed were included as 'blinded' control samples, and were analyzed without knowing which samples were commercial samples and which were control samples. The data presented here are included in the National Nutrient Database.

Technical Abstract: Vitamin D3 (cholecalciferol) levels were determined in finfish and shellfish using UV detection at 265nm (combined with auxiliary full scan UV detection) and selected ion monitoring (SIM) mass spectrometry (MS), using vitamin D2 (ergocalciferol) as an internal standard. Analysis of standard reference material (SRM) NIST 1849 (Infant/Adult Nutritional Formula) was included to validate the method. Three-point calibration curves were employed, allowing values to be determined over a range of species, from those having little or no detectable vitamin D3 (e.g., pollock, shrimp) to those with high levels (e.g., salmon with up to 33.23 ug/100g). The limit of detection (LOD) and limit of quantitation (LOQ) calculated from the uncertainty and intercept of the calibration curves were 1.22 ug/100g and 5.3 ug/100g, respectively, based on all analyses (n=27 sequences). Use of response factors (RF) allowed quantitation at lower levels of vitamin D3, with an LOQ of < 0.25 ug/100g. The values obtained using the validated methodology agreed well with literature and tabulated database results for most species. However, much lower average vitamin D3 concentrations were found for oysters (0.05 ug/100g, raw) and clams (0.18 ug/100g, cooked) compared to other reports for these products.