AOAC SPIFAN ERP Meeting Book-March 16, 2016

VitK-01 w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Silliker Chem, Res. Center Crete, IL – Validation of a LC-MS/MS Method for Vitamin K Analysis

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 msec

DP

CEP

CE

Ion (Q1) Ion (Q3)

451.361 451.36 445.31 445.31

187.1

100 100 100 100 100 100 100

60.00 60.00 40.00 40.00 60.00 60.00 60.00

23.60 23.60 23.41 23.41 30.14 30.14 23.60

35.00

K1

128

110.00

187.1

35.00 60.00 52.00 90.00 35.00

K2-4

81

649.5 649.5 458.5

187.1

K2-7

81

191.1

Vit K1 Int. Std.,

d7(5,6,7,8-d4, 2-methyl-d4)

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

Silliker Laboratories, Chemistry Research Center, Date: 1/30/15

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