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Analysis of Vitamin D

2

and Vitamin D

3

by LC-MS/MS in Milk

Powders, Infant Formulas, and Adult Nutritionals

Page 6 of 10

MS Detector:

ESI

+

mode (see detector details below)

Chromatography: Mobile Phase Gradient

Time

(min)

Flow rate

(mL min

-1

)

Mobile phase composition

%A

%B

0 START

0.6

25

75

3.3 PUMP

0.6

0

100

3.7 PUMP

1.0

0

100

4.8 PUMP

1.0

0

100

4.9 PUMP

0.6

25

75

5.5 STOP

0.6

25

75

Chromatography: Switching Valve

Time

(min)

Setting

Description

0

B

Mobile phase flow to waste

1.5

A

Mobile phase flow to ion source

3.7

B

Mobile phase flow to waste

Mass Spectrometry: Source/Gas Parameters

Source/Gas

Parameter

Value

Source/Gas

Parameter

Value

Source

ESI

+

Collision gas

N

2

Curtain gas

30

Temperature

300 °C

Nebulizer gas GS1

40

CID gas

Medium

Heater gas GS2

40

Ion spray voltage

5500 V

Mass Spectrometry: Compound Parameters (Vitamin D

2

instrument method only)

Vitamin D

2

Type

a

Precursor

ion

(

m/z

)

Product

ions

(

m/z

)

DP

b

(V)

EP

c

(V)

CE

d

(V)

CXP

e

(V)

Dwell

time

(ms)

Analyte quantifier ion

572

298.0

81

10

23

22

120

Analyte qualifier ion

572

279.8

39

16

80

Internal standard quantifier ion

578

298.0

23

22

120

Internal standard qualifier ion

578

279.8

39

16

80

a

Analyte = vitamin D

2

-PTAD adduct, Internal standard ion = d6-vitamin D

2

-PTAD adduct

b

DP = declustering potential

c

EP = entrance potential

VitD-18 (February 2016)

FOR ERP USE ONLY

DO NOT DISTRIBUTE