AOAC SPIFAN Review Team Meeting Book-July 22, 2015

BVIT-02 FOR ERP/WORKING GROUP USE ONLY DO NOT DISTRIBUTE

Abbott Nutrition Division Simultaneous Determination of Total Vitamin B 6, B 2, B 3 in Infant Formula Products by LC-MS/MS Using Enzymatic Digestion and B 1

Weigh 14.0 ± 0.1 mg into a tarred weighing vessel. Quantitatively transfer to a 25-mL volumetric flask with laboratory water and dilute to volume with laboratory water. Mix well and transfer to a 50-mL amber bottle and Store refrigerated (2˚-8˚C). Expiration: Until exhausted or evidence of contamination.

f) Stable-Isotope Labeled Compounds – Internal Standard Stock Mixture (ISSM) i) Combine the stable-isotope labeled internal standard stock solutions in a 5:1:1:1:1:1 volume ratio of Lab Water: 2 H 3 -Pyridoxal: 13 C 4 -Pyridoxine: 2 H 3 -Pyridoxamine: 13 C 3 -Thiamine Chloride: 13 C 4 , 15 N 2 -Riboflavin Stock Solution For example, a sample set containing 18 samples and two sets of 6 working standards, a minimum ISSM volume of approx. 1.5 mL is required. A 2.0 mL (allowing for excess) volume of ISSM is prepared by mixing 1250 m L Lab Water with 250 m L of 2 H 3 -Pyridoxal, 250 m L of 13 C 4 -Pyridoxine, and 250 m L of 2 H 3 -Pyridoxamine Labeled Internal Standard Stock Solutions. Mix solution well prior to use. Expiration: Prepare fresh daily. NOTE: Prepare a sufficient volume of this mixture on each day of analysis to allow for all sample preparations and standard solutions to be inoculated with 50 m L from one ISSM solution.

g) Native Compounds – Stock Standard Solutions i) Pyridoxine Stock Solution; Approximate Concentration: 1.2 mg/mL

Weigh approximately 15 mg ( ± 0.50mg) of Pyridoxine Hydrochloride into a weighing funnel or other appropriate weighing vessel. Quantitatively transfer to a 100-mL volumetric flask with laboratory water. After mixing thoroughly, QS to volume with laboratory water. Store refrigerated (2˚-8˚C). Expiration: 3 months.

ii) Pyridoxal Stock Solution; Approximate Concentration: 1.2 mg/mL Weigh approximately 15 mg ( ± 0.50mg) of Pyridoxal Hydrochloride into a weighing funnel or other appropriate weighing vessel. Quantitatively transfer to a 100-mL volumetric flask with laboratory water. Mix thoroughly, then QS to volume with laboratory water. Store refrigerated (2˚-8˚C). Expiration: 3 months.

34