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Determination of B-Vitamins in Infant Formula and Adult/Pediatric Formulas

ESI LC-MSMS

2

c) Standards Preparation:

1) Preparation of Stock Standards:

Table-1

Standard Name

Standard

Weight

(mg)

Standard

Makeup volume

(mL)

Diluent

Thiamine Hydrochloride

10.0

10.0

0.1M Hydrochloric acid

Riboflavin

10.0

100.0

0.1 M Acetic acid

Niacinamide

10.0

10.0

Milli-Q water

Nicotinic acid

10.0

10.0

Milli-Q water

Calcium Pantothenate

10.0

10.0

Milli-Q water

Pyridoxine hydrochloride

10.0

10.0

Milli-Q water

Biotin

10.0

50.0

Milli-Q water

Folic acid

10.0

10.0

0.02 M Potassium Hydrogen

Carbonate

Note 1:

Individual stock standards can be used for 30 days when stored in dark under refrigerated

conditions.

Note 2:

Apply appropriate salt and purity corrections to the prepared stock standards using the

formula.

Conc. after salt correction (mg/L) =

Molecular weight of compound x Wt of Reference Materials x % Purity X 10

-----------------------------------------------------------------------------------------------------------------

Formula weight of CRM X Makeup volume (ml)

2) Preparation of Intermediate standards:

Prepare 10 mg/L of mix standard by transferring required volume of each stock solution into

a 100 mL low actinic volumetric flask and makeup with laboratory water.

3) Preparation of working standards:

Prepare 1.0 mg/L and 0.1 mg/L working standards from 10 mg/L standard using laboratory

water as diluent.

4) Preparation of calibration standards:

Table-2:

Working

Standard

Conc. (mg/L)

Volume of

Working Std

(mL)

Volume of

Diluent (mL)

Final Volume

(mL)

Final Conc.(μg/L)

10.0

0.500

9.500

10.000

500.00

10.0

0.200

9.800

10.000

200.00

10.0

0.100

9.900

10.000

100.00

1.0

0.500

9.500

10.000

50.00

1.0

0.200

9.800

10.000

20.00

1.0

0.100

9.900

10.000

10.00

0.10

0.500

9.500

10.000

5.00

0.10

0.200

9.800

10.000

2.00

Diluent:

Laboratory water

.

BVit-03 (February 2016)

FOR ERP USE ONLY

DO NOT DISTRIBUTE