Review Team (February 2016)

Bio-03 (February 2016) FOR ERP USE ONLY DO NOT DISTRIBUTE

ABBOTT NUTRITION DIVISION TITLE

PAGE NO.

Determination of Biotin in Infant, Pediatric and Adult Nutritionals by HPLC and Fluorescence Detection

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ss: Sample size in g D 1 : Unit conversion (from per g to per 100 g sample and from ng to P g of biotin) D 1 = (100g/100g)(1 P g/1000ng) = 1/10 ( P g/ng/100g) D 2 : Conversion from sample to reconstituted finish product For powder: D 2 = 25 g powder y (25 g powder + 200 g water) = 25 g y 225 g = 1/9 For liquid (2 times dilution by weight): D 2 = 2 For ready to feed (RTF, as is): D 2 = 1

3. Example: Sample Calculation a) Analysis of SRM 1849a (1.0020 g) Sample peak area = 5.8505, C i

= 40.21 ng/mL, ss = 1.0020 g

Final result: C p

= C i

x 50 y ss x D 1

x D 2 = 40.21 (ng/mL) x 50 (mL) y 1.0020 x (1/10) ( P g/ng/100g)

x (1/9) = 22.29 P g/100g reconstituted finish product b) Analysis of Infant Formula RTF (20.0333 g) Sample peak area = 1.7494, C i

= 15.78 ng/mL, ss = 20.0333 g

= C i

x 50 y ss x D 1

x D 2 = 15.78 (ng/mL) x 50 (mL) y 20.0333 x (1/10) ( P g/ng/100g)

Final result: C p

x 1 = 3.94 P g/100g RTF