AOAC SPIFAN Review Team Meeting Book-July 22, 2015

Abbott Nutrition Division

Simultaneous Determination of Total Vitamin B

and B

in Infant

Formula Products by LC-MS/MS Using Enzymatic Digestion

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 (

°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.

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.

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

Pyridoxamine

BVIT-02

FOR ERP/WORKING GROUP USE ONLY

DO NOT DISTRIBUTE