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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 7 of 10

d

CE = collision energy

e

CXP = collision cell exit potential

Mass Spectrometry: Compound Parameters (Vitamin D

3

instrument method only)

Vitamin D

3

Type

a

Precursor

ion

(

m/z

)

Product

ions

(

m/z

)

DP

c

(V)

EP

d

(V)

CE

e

(V)

CXP

e

(V)

Dwell

time

(ms)

Analyte quantifier ion

560.2

298.0

151

10

21

18

120

Analyte qualifier ion

560.2

279.8

37

18

80

Internal standard quantifier ion

566.2

298.0

21

18

120

Internal standard qualifier ion

566.2

279.8

37

18

80

a

Analyte = vitamin D

3

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

3

-PTAD adduct

b

DP = declustering potential

c

EP = entrance potential

d

CE = collision energy

e

CXP = collision cell exit potential

Calculation and Reporting

Concentration of stable isotope labelled vitamin D

2

in stock standard

SILD

2

SS

:

10000

x

1%

1cm

max)

abs(

λ

2

D2conc

2

E

SS SILD

SS SILD

where:

SILD

2

SS

D2conc

= concentration of SIL d6-vitamin D

2

in stock standard (

units

: μg mL

-1

)

SILD

2

SS

abs(

λ

max)

= UV absorbance of stock standard at 265 nm (

units

: none)

= extinction coefficient for vitamin D

2

in ethanol [461] (

units

: dL g

-1

cm

-1

)

10000 = concentration conversion factor (

units

: g dL

-1

to μg mL

-1

)

Concentration of stable isotope labelled vitamin D

3

in stock standard

SILD

3

SS

:

10000

x

1%

1cm

max)

abs(

λ

3

D2conc

3

E

SS SILD

SS SILD

where:

SILD

3

SS

D3conc

= concentration of SIL d6-vitamin D

3

in stock standard (

units

: μg mL

-1

)

SILD

3

SS

abs(

λ

max)

= UV absorbance of stock standard at 265 nm (

units

: none)

= extinction coefficient for vitamin D

3

in ethanol [485] (

units

: dL g

-1

cm

-1

)

10000 = concentration conversion factor (

units

: g dL

-1

to μg mL

-1

)

Concentration of stable isotope labelled vitamin D

2

in internal standard

SILIS

:

1000

10

1.0 x

SS SILD

SILIS

D3conc

2

D2conc

u

where:

SILIS

D2conc

= concentration of SIL d6-vitamin D

2

in internal standard (

units

: ng mL

-1

)

SILD

2

SS

D2conc

= concentration of SIL d6-vitamin D

2

in stock standard (

units

: μg mL

-1

)

1000 = concentration conversion factor (

units

: μg mL

-1

to ng mL

-1

)

Concentration of stable isotope labelled vitamin D

3

in internal standard

SILIS

:

1%

1cm

E

1%

1cm

E

VitD-18 (February 2016)

FOR ERP USE ONLY

DO NOT DISTRIBUTE