AOAC SPIFAN Review Team Meeting Book-July 22, 2015

B-VIT-01 FOR ERP/WORKING GROUP USE ONLY DO NOT DISTRIBUTE

Simultaneous Determination of Thiamine HCl, Riboflavin, Niacin (Nicotinic Acid and Nicotinamide), Pantothenic acid, Vitamin B6 (Pyridoxine, Pyridoxal and Pyridoxamine) and Biotin in Infant Formula and Adult Nutritionals

CONFIDENTIAL METHOD FOR ERP ASSESSMENT ONLY

IV. CALCULATION and REPORT The LCMSMS software will automatically calculate the concentration of the sample in ng/mL for Thiamin HCl, Riboflavin, Nicotinic Acid, Nicotinamide, Pantothenic acid, Pyridoxine, Pyridoxal, Pyridoxamine and Biotin, provided the concentration of the working standards in ng/mL, sample weight / powder equivalent (g) or sample volume (mL), dilution factor and conversion to 100g or 100 mL are entered correctly. Report in 2 decimal places or as customer requirement.

100 1000 W CxV ))ml or(g 100 / mcg ( WSV .  xDFx

Conc

Where: C

= = = = = =

Concentration of water-soluble vitamin from working curve (ng/mL)

V

Sample volume (ml)

= 100 mL

DF

Dilution Factor

= 1/0.5

W

Sample weight (3.125 g for powder or 1 g for SRM1849a) or sample volume (25 mL)

1000 100

conversion factor from ng to mcg

conversion factor from g to 100 g or from mL to 100 mL

V. DISCUSSION a. Niacin (mcg/100g(or mL)) = Nicotinamide(mcg/100g(or mL))+Nicotinic acid (ng/100g (or mL)) b. Vitamin B6 (mcg/100g(or mL)) = Pyridoxine (mcg/100g(or ml)) + Pyridoxal (mcg/100g(or ml)) + Pyridoxamine (mcg/100g(or mL)) c. Vitamin B6 (as Pyridoxine) (mcg/100g(or mL)) = Pyridoxine (mcg/100g(or ml)) + Pyridoxal (mcg/100g(or ml))x(169.18/167.162) + Pyridoxamine (mcg/100g(or mL))x(169.18/168.19) Where Molecular weight of Pyridoxine = 169.18 g/mol Molecular weight of Pyridoxal = 167.162 g/mol Molecular weight of Pyridoxamine = 168.19 g/mol d. If Thiamine (C12H17N4OS) is reported, Thiamine Hydrochloride (C12H18Cl2N4OS) is converted by multiplying with 0.787. Thiamine (mcg/100g (or mL) = Thiamine HCl (mcg/100g (or mL)) x 0.787 Note : 0.787 = MW of Thiamine (265.4 g/mol)/MW of Thiamine HCl (337.3 g/ mol) e. Comments:

Comment No. 1

The gradient given can be altered as required to maximize the analytical separation and avoid interferences. This should only be done by an authorized analyst.

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