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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