Measurement of Vitamin D in Foods and Nutritional Supplements by Liquid Chromatography/Tandem Mass Spectrometry

Abstract

Vitamin D is a fat-soluble vitamin with great nutritional interest. An HPLC/MS/MS method was developed to measure vitamin D with atmospheric pressure chemical ionization. Under the experimental parameters used, the LOQ was 0.018 IU/g or 0.45 ng/g, which greatly enhances the capability of measurement of vitamin D at low levels in foods and supplements. This method was validated with spike recovery of 100 15 and the RSD of less than 10 for most sample matrixes, including infant formula, cheese, cereal and cerealbased foods, multivitamin supplements, and pet foods. The results for vitamin D were compared with those obtained by other methods.

Mechanism of error caused by isotope-labeled internal standard: accurate method for simultaneous measurement of vitamin D and pre-vitamin D by liquid chromatography/tandem mass spectrometry

“…It is important to note that the elution sequence of the peaks might be altered when using different columns. When using a C30 column, pre-D eluted after vitamin D. [9] However, under the LC conditions described here, with a Hypersil column, pre-D eluted before vitamin D, with retention times of 3.2 and 3.4 min, respectively. Good separation between pre-D and vitamin D was also achieved for vitamin D 2 and all of the isotope-labeled vitamin D compounds studied in this work.…”

“…High-performance liquid chromatography (HPLC) has been widely used for the measurement of vitamin D. [2,3] However, with high sensitivity and high selectivity, liquid chromatography/mass spectrometry (LC/MS) has become more popular for vitamin D measurement recently. [4][5][6][7][8][9][10][11][12][13][14][15] Heudi et al [8] reported an LC/MS method for measurement of vitamin D 3 and other fat-soluble vitamins in infant formula with vitamin D 2 as internal standard. Dimartino [5,6] used LC/MS to measure vitamin D 3 in cheese and other foods, also with D 2 as the internal standard for measurement of D 3 .…”

“…Kamao et al [12] synthesized and used an isotope-labeled vitamin D 3 to measure fat-soluble vitamins in human breast milk. Recently, several authors have used deuterium-labeled internal standards to measure vitamin D in a variety of foods and supplements by LC/MS/MS after a variety of sample preparation methods including overnight saponification at room temperature (25°C), [5,6,[9][10][11] hot saponification, [7,15] or direct extraction of vitamin D in infant formula followed by derivatisation. [4] Compared with overnight saponification at room temperature, hot saponification offers the advantage of quick sample preparation and has been frequently used for analysis of foods and supplements; i.e.…”

Survey of vitamin D and 25-hydroxyvitamin D in traditional native Alaskan meats, fish, and oils

“…The analyses were conducted according to methodology reported by Huang et al (2009). Briefly, a 2-10 g subsample of the homogenized food, with internal standard added ([ 2 H 3 )cholecalciferol for vitamin D 3 , [ 2 H 3 ]25-hydroxycholecalciferol for 25(OH)D 3 , and [ 2 H 3 ]ergocalciferol for vitamin D 2 ) (> 95%; ISO Sciences, Ambler, PA, USA), was subjected to overnight alkaline saponification in alcoholic KOH, under nitrogen and with 2% pyrogallic acid added.…”

“…A key part of this work was validating methods for vitamin D and 25(OH)D. Until recently, methods for vitamin D were generally not standardized and often gave disparate results between laboratories using similar methods Phillips et al, 2008). More recently, methodology for measurement of vitamin D and 25(OH)D by LC-MS has been validated (Byrdwell, 2009;Huang et al, 2009). The USDA Nutrient Data Laboratory (Beltsville, MD) has demonstrated that it is possible to obtain consistent results for vitamin D 3 and 25(OH)D 3 among laboratories .…”