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Silliker
Chem, Res. Center Crete, IL – Validation of a LC-MS/MS Method for Vitamin K Analysis
Silliker Laboratories, Chemistry Research Center, Date: 1/30/15
8
Results:
Optimization of compound based mass-spec parameters:
The compound based MS parameters were determined by pumping the mobile phase at 0.4 mL/min
through the HPLC pump without a HPLC column. The analyte mixture solution was pumped into the
MS source by a 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 are presented in Table 2.
Table 2: Mass spec parameters determined to be optimum for the targeted vitamin K analytes
Standard
Parent
Daughter
Dwell
Time
DP
CEP
CE
Ion (Q1) Ion (Q3)
msec
K1
451.361
187.1
100
60.00
23.60
35.00
451.36
128
100
60.00
23.60
110.00
K2-4
445.31
187.1
100
40.00
23.41
35.00
445.31
81
100
40.00
23.41
60.00
K2-7
649.5
187.1
100
60.00
30.14
52.00
649.5
81
100
60.00
30.14
90.00
Vit K1 Int.
Std.,
d7(5,6,7,8-d4,
2-methyl-d4)
458.5
191.1
100
60.00
23.60
35.00
DP: Declustering potential;
EP Entrance potential;
CEP collision cell entrance potential;
CE Collision cell;
CXP collision cell exit potential. = 4.0 Volt for all thetransitions
These conditions were used for analytes detection through the study. More intense transitions was used
for quantification and the other for qualification.
The General MS parameters:
The MS parameters applicable for detection of all the analytes (all transitions) in our study are provided
in the following section.
Ion Source: positive
Curtain Gas: 20.00 psi
Ionspray Voltage: 5500 V
TEMP: 350°C
VitK-01 w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE