Convenient Analysis of Vitamin D in Cheese and other food matrices by Liquid Chromatography/Mass Spectrometry

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Slide 1 : Rapid Determination of Cholecalciferol (Vitamin D3) in Different Food Matrices by LC/APCI/MS Gianluca Dimartino Kraft Foods Inc., East Hanover, NJ In Blood In Food Vitamin D As Cancer Crusher (American Journal of Public Health). Researchers found that Vitamin D can dramatically lower the risk of developing common types of cancers. In fact, they say taking the right amount of Vitamin D can cut the chances of getting breast, ovarian, even colon cancer in half…. Breathing Easier with Vitamin D (Science News). Physicians in New Zealand have now linked the vitamin to yet one more apparent advantage: improved lung function…. Health Benefits
Slide 2 : Official Procedure To Quantify Vitamin D 3 times extraction Wash until pH=7 saponification Conc. N2 Direct-phase Chromatography (multiple solvents) Reverse-phase Chromatography (multiple solvents) Conc. At 500C and sonicate HPLC RP-HPLC
Slide 3 : Kraft’s procedure to determine vitamin D Concentrated Saponification at room temperature (50g) and one time extraction Reverse-phase Single solvent * After saponification Dimartino, G. J. of AOAC INTERNATIONAL, 2007, 90, 5, 1340-1345
Slide 4 : Vitamin D3 and D2 calibration curves Calculated by looking at the area obtained by positive APCI detection D3 D2 Vitamin D2 is used as an internal standard for the analysis. It is more unstable than vitamin D3 and solutions need to be prepared fresh every couple of weeks
Slide 5 : Left: A response factor of 1.4 was chosen as an average. Concentration range includes usual fortification range for cheese (160IU/100 g to 320 IU/100 g) Vitamin D3/D2 response factor and blending experiment on Control/Fortified Cheddar Right: “0” indicates experiment carried out with control non-fortified shredded cheddar while “100” indicates experiment carried out with fortified shredded cheddar 16 IU/100g 224 IU/100g
Slide 6 : Recovery and Accuracy experiments * Experiments performed on mild cheddar
Slide 7 : Precision was determined over two months period for cheddar. Sample was split in 20 containers; each containing 50g of cheese for one test. a results obtained from an external Lab b analysis performed on cheese directly from bag Experiments on Precision and results from Silliker Lab Process Cheese results by LC/MS
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Slide 9 : Experiments on Orange Juice 40 ml extra volume of extracting hexane was required for this product Preextraction/Extraction: 200 ml of EtOH 130 ml of water Stirr 10 min Add 100 ml Hex+40 ml Hex Withdraw 70 ml hexane Federal labelling regulations require that added nutrients in fortified foods shall be 100-150% of label claims. 50 ml initial sample Added 80 IU of IS No D3 added 100-150 IU/240 ml of D3 Orange juice
Slide 10 : Application of vitamin D methodology to beverages Experiment was performed on Capri Sun Raspberry-passion fruit using the usual vitamin D procedure: No extra volume of extracting hexane was required for this product
Slide 11 : beverage; 2. orange juice; 3-5. Cereals; 6-7. Natural shredded cheese; 8-10. Process cheese Applicability of the method to various matrices Results on Y-axis are obtained from an external lab; results on x-axis are obtained at Kraft Dimartino, G. J. of AOAC INTERNATIONAL, 2007, 90, 5, 1340-1345

 


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