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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j) pH Meter k) Disposable transfer pipettes l) Reagent reservoirs: assorted sizes

m) Volumetric flasks: Class A, assorted sizes n) Volumetric pipettes: Class A, assorted sizes o) Weighing boats or equivalent p) Yellow lights or yellow shields with a cutoff of 385 nm q) Microcentrifuge r) Microcentrifuge tubes: 2 mL

s) Magnetic stirring plate t) Magnetic stirring bars

D. CHEMICAL, STANDARD AND SOLUTION 1. Chemicals/Standards

Unless otherwise noted, the following chemicals should be stored at room temperature. Their - expiration date is one year after the date they are opened. Upon expiration, the chemicals should be either discarded or re-evaluated. Discontinue use of any chemicals or solutions whenever indications of contamination, chemical degradation, or changes in concentration are evident . a) Biotin Standard: USP current official lot, reference standard stored as indicated on label. Store at room temperature unless otherwise stated. Regardless of expiration date, a standard solution should be discarded if it is suspected to contain chemical or micro biological contamination, chemical degradation products, or of a change in concentration is evident. b) Ethanol (Reagent Alcohol): ACS or equivalent c) Streptavidin fluorescein conjugate: 1mg/mL, GeneTex Streptavidin-FITC, CAT# GTX30950 or AnaSpec Streptavidin-FITC, CAT# AS-60659-FITC, or equivalent. Store in freezer. d) Laboratory water e) Methanol: HPLC grade or equivalent f) pH Meter buffer solutions for pH meter standardization g) Meta-phosphoric acid: ACS or equivalent

j) Sodium phosphate monobasic: ACS or equivalent k) Sodium phosphate dibasic: ACS or equivalent

2. Solutions

a) 0.2 M Sodium Phosphate Monobasic (SPM)