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Silliker
Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analysis
13
6.50
0.500
100.0
6.55
0.500
0.0
9.00
0.500
0.0
A = 1%Acetic Acid in Water
B = 100% Methanol
Column Temperature: 40
°
C
Autosampler Temperature: 6
°
C
Injection Solvent: Std., samples and blanks injection solvent is 90% MeOH + 10% Formic acid +
1% ascorbic acid neutralized to pH 7 with ammonium hydroxide (29%) containing 50 uL/mL.
Injection Volume = 2 uL
Table 6:
LC Conditions using Shimadzu LC in front of API 3200 MS detector
:
Time (min)
%B
Flow
(mL/min)
0.10
0.1
0.2
1.00
0.1
0.2
8.00
100
8.01
0.2
8.02
0.3
10.00
100
10.10
0.1
11.00
0.3
11.01
0.2
12.00
0.1
15.00
(Stop)
A = 1%Acetic Acid in Water
B = 100% Methanol
Column Temperature: 40
°
C
Autosampler Temperature: 6
°
C
Injection Solvent: Std., samples and blanks injection solvent is 90% MeOH + 10% Formic acid +
1% ascorbic acid neutralized to pH 7 with ammonium hydroxide (29%) containing 50 uL/mL.
Injection Volume = 2 uL
MS Conditions for API 5500 (Qtrap):
MS conditions API 5500 (Qtrap) for detection of different transitions of the analytes in the scope
Table 7: MS conditions API 5500 (Qtrap) for detection of different transitions of the analytes in the
scope
Comp
ound
Q1
Q3
DP
EP
CEP
CE
CXP
FA
442.2
295.2
75
6
6
20
6
2011.06 (Fol-22) w/SLV
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