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

iii) Pyridoxamine Stock Solution; Approximate Concentration: 1.2 mg/mL Weigh approximately 18 mg ( ± 0.50mg) of Pyridoxamine dihydrochloride 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. iv) Riboflavin Stock Standard Solution: 30 mg/L Dry an appropriate amount of the USP riboflavin reference standard at 105 ( ± 1) °C for 2 hours ( ± 10 minutes) prior to making the stock standard. All work with riboflavin standards must be conducted under yellow lighting as specified in Apparatus section. Accurately weigh 0.033000 (± 0.00010) g of the reference standard and quantitatively transfer to a 1000-mL volumetric flask. Add approx. 800 mL water and 1.2 ( ± 0.1) mL acetic acid, glacial. Mix to dissolve. Bring to volume with water. Mix well and store under yellow lights. Store refrigerated (2˚-8˚C). Expiration: 3 months. v) Thiamine Stock Standard Solution: 100 mg/L Determine the moisture of the USP thiamine HCL reference standard as directed on the container just prior to making the stock standard. Using the percent moisture of the reference standard, determine the amount of reference standard to accurately weigh out to equal 0.025000g (± 0.0010) g per 500250mL. Quantitatively transfer to a 500250-mL volumetric flask. Add approximately 300150 mL 0.15 M HCl. Mix to dissolve and bring to volume with 0.15 M HCl. Mix well and Store under yellow lights. Store refrigerated (2˚-8˚C). Expiration: 3 months. h) Intermediate Working Standard Solution (IWSS) Combine the following volumes of the stock standard solutions in a 10-mL volumetric flask containing approximately 5 mL of laboratory water. Table 4. Stock Standard solutions volumes needed to make IWSS. Vitamin Stock Standard Solution Volume (µL) Pyridoxine 250 Pyridoxal 50 Pyridoxamine 50 35