Review Team (February 2016)

BVit-03 (February 2016) FOR ERP USE ONLY DO NOT DISTRIBUTE

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

Table-4 QQQ Parameters: Compund Name Precursor Ion

Dwell Time

Fragmentor Voltage

Cell Acc. Voltage

Product ion

CE

Thiamine

265.2

122.2, 81.3 243.2, 198.1, 172.1 80.1, 78.1, 53.1 202.1, 90.0, 71.9 152.1, 134.1, 77.3 227.3, 105.2, 97.1 295, 176, 120 80.3, 53.3

40

92

10, 30

7

Riboflavin

377.1

40

100

20, 35, 40

7

Niacinamide Nicotinic acid Pantothenic acid

123.2

40 40

85 50

20, 30

7 7

124.2

20, 20, 32

220.1

40

100

10, 10, 15

7

Pyridoxine

170.2

40

55

10, 20, 35

7

Biotin

245.2

45

110

10, 30, 30

7

Folic acid

442.2

45

110

10, 40, 40

7

d) Calculations:

- A b

) * V s

* DF

(A s

Result (mg/100g) =

* 10000

W s

= Sample concentration in μg/L, from calibration curve = Blank concentration in μg/L, from calibration curve

A s A b V s W s

= Sample Makeup Volume = Sample weight in grams

= Sample dilution factor (Other than V s and W s )

DF S

G. References: a) AOAC official methods of analysis (AOAC 2004.07 , AOAC 2012.16 ) b) BS EN 14122:2014, Food Stuffs – Determination of Vitamin B1 by HPLC c) BS EN 14164:2014, Food Stuffs – Determination of Vitamin B6 by HPLC d) BS EN 15652:2009, Food Stuffs – Determination of Niacin by HPLC e) Agilent application note for “Determination of multi water soluble vitamins by LC - MS/MS”.

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