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

Gradient: Time

%A %B

Flow Rate (mL/min)

0.10 5.00

10 90 0 100 0 100 10 90

0.4 0.4 0.4

10.00 10.10

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 msec

DP

CEP

CE

Ion

Ion

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)

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).

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

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