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Silliker
Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analys
5
Results
Optimization of compound based mass-spec (Sciex 3200/ 5500/6500) parameters:
The compound based parameters were determined by pumping the mobile phase at 0.5 mL/min
through the HPLC pump without column. The analyte mixture solution was pumped into the MS
source by the syringe pump in the Mobile phase. The compound dependent parameters were ramped
with infusion of the analytes and the optimum values were determined for the corresponding Q1/Q3
transitions providing the strongest response. Results of the optimization of the parameters Sciex 3200
are presented in Table 1. Parameters later on optimized for the Sciex 5500 and 6500 LC-MS/MS are
provided in the method.
Table 1: Optimization of MS conditions 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
442.2 176.2
75
6
6
45
6
5-CH3-THF
460.2 313.2
75
6
6
28
6
460.2 194.2
75
6
6
46
6
5-CHO-THF
474.2 327.2
75
6
6
25
6
474.2 299.2
75
6
6
40
6
THF
446.2 299.2
75
6
6
30
6
446.2 166.2
75
6
6
50
6
5,10-CH=CH-
THF
456.2 412.2
75
6
6
42
6
456.2 282.2
75
6
6
70
6
10-CH3-FA
456.2 309.2
75
6
6
20
6
456.2 176.2
75
6
6
50
6
10-CHO-FA
470.2 295.2
75
6
6
35
6
470.2 176.2
75
6
6
50
6
13
C
5
-FA
447.2 295.2
75
6
6
20
6
13
C
5
-5-CH3-THF 465.2 313.2
75
6
6
28
6
13C5-5-CHO-THF
479.2 327.2
75
6
6
25
6
DP: Declustering potential;
EP Entrance potential;
CEP collision cell entrance potential;
CE Collision cell;
CXP collision cell exit potential.
These conditions were used for analytes detection through the study. More intense transition was used
for quantification and the other for qualification.
2011.06/Fol-22 (February 2016)
FOR ERP USE ONLY
DO NOT DISTRIBUTE