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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
7
Gradient: Time
%A %B
Flow Rate (mL/min)
0.10
10 90
0.4
5.00
0 100
0.4
10.00
0 100
0.4
10.10
10 90
0.4
15.00 System Controller
STOP
Temperature: 35
o
C (column compartment will not cool below 35
o
C)
Injection Volume: 10uL
Autosampler Temperature: 5°C
Stop Time: 15min
MS Conditions
Ion Source: positive
Curtain Gas: 20.00 psi
Ionspray Voltage: 5500 V
TEMP: 350°C
Ion Source Gas 1 (GS1): 35.00 psi
Ion Source Gas 2 (GS2): 30.00 psi
Inter Face Heater: on
Entrance Potential: 5.40 V
Collisionally Activated Dissociation Potential (CAD): 4.00 V
Collision Cell Exit Potential (CXP): 4.00 V
Standard
Parent Daughter
Dwell
Time
DP
CEP
CE
Ion
Ion
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
Calculations:
Draw Calibration curve for Vitamin K1 and Vitamin K2-4 & Vitamin K2-7 based on concentration of standard
solutions used and obtained respective peak areas. Calculate vitamin K1 and K2 in the sample extract based on
their respective peak areas in the standards processed like a sample. Multiply by extract dilution and
concentrations folds and final volume. Adjust concentration in the extract by the sample aliquot weight and
calculate analytes in the sample as µg/100g (w/w).
VitK-01 w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE