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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