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Abbott Nutrition Division
Simultaneous Determination of Total Vitamin B
6,
B
2,
B
3
and B
1
in Infant
Formula Products by LC-MS/MS Using Enzymatic Digestion
c)
Mass Spectrometer Data Acquisition Parameters
Table 3. Mass Spectrometer data collection parameters.
Compound
Start
(min)
End
(min)
SRM Transition Typ. RT
(min)
Mass Spec Conditions
Molecular
Ion
Fragment
Ion
Cone
Voltage
CE
Voltage
Dwell Time
(msec)
Pyridoxamine
0.3
3.0
169.1
151.8
0.70
20
15
40
2
H
3
-Pyridoxamine
0.3
3.0
172.1
154.6
0.70
20
15
40
Pyridoxal
0.3
3.0
168.1
149.8
1.2
15
15
40
2
C
3
-Pyridoxal
0.3
3.0
171.1
153.1
1.2
15
15
40
Nicotinic Acid
0.3
3.0
124.0
80.4
1.25
50
16
40
2
H
4
- Nicotinic Acid
0.3
3.0
128.0
84.0
1.25
44
18
40
Pyridoxine
0.3
3.0
170.2
151.7
1.5
25
13
40
13
C
4
-Pyridoxine
0.3
3.0
174.2
155.7
1.5
25
13
40
Thiamine
0.8
1.75
265.3
122.0
1.56
20
15
40
13
C
3
-Thiamine
0.8
1.75
268.3
122.0
1.56
20
15
40
Niacinamide
1.75
3.5
122.9
96.0
2.67
8
16
40
2
H
4
-Niacinamide
1.75
3.5
127.0
100.0
2.67
8
16
40
Riboflavin
1.75
3.5
377.2
242.8
3.67
30
20
40
13
C
4
,
15
N
2
-Riboflavin
1.75
3.5
383.2
248.6
3.67
30
20
40
4.
UPLC-MS System Equilibration
a)
Place freshly prepared mobile phases onto the system and purge old solvents from the solvent
lines and needle washes using the UPLC Control Panel.
b)
The sample cone must be cleaned prior to each sample set. Cleaning can only be performed by
trained personnel and requires less than 10 minutes.
c)
Start gas flow into the mass spectrometer. Once gas flow has stabilized load the mass
spectrometer Tune Page and allow the source temperatures to equilibrate.
NOTE: It is very important to complete this step before proceeding. Failure
to do so could result in the premature introduction of UPLC eluate
into the Mass Spectrometer Ionization source resulting in potential
damage to the Ionization source.
d)
After confirming equilibration of the Mass Spectrometer source, start LC flow to the column.
Monitor the column backpressure and allow it to stabilize too less than 50-psi deviation for a
minimum of one minute. Once equilibration has been established, allow the system to equilibrate
for 10 minutes with flow into the source before beginning the sample set.
29
BVit-02
FOR ERP USE ONLY
DO NOT DISTRIBUTE