AOAC SPIFAN ERP Meeting Book-March 16, 2016

2011.06 (Fol-22) w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Silliker Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analysis

6.50 6.55 9.00

0.500 0.500 0.500

100.0

0.0

0.0 A = 1%Acetic Acid in Water B = 100% Methanol

Column Temperature: 40 ° C Autosampler Temperature: 6 ° C

Injection Solvent: Std., samples and blanks injection solvent is 90% MeOH + 10% Formic acid + 1% ascorbic acid neutralized to pH 7 with ammonium hydroxide (29%) containing 50 uL/mL. Injection Volume = 2 uL

Table 6: LC Conditions using Shimadzu LC in front of API 3200 MS detector :

Flow (mL/min)

Time (min)

%B

0.10 1.00 8.00 8.01 8.02

0.1 0.1 100

0.2 0.2

0.2 0.3

10.00 10.10 11.00 11.01 12.00 15.00

100 0.1

0.3 0.2

0.1

(Stop)

A = 1%Acetic Acid in Water B = 100% Methanol

Column Temperature: 40 ° C Autosampler Temperature: 6 ° C

Injection Solvent: Std., samples and blanks injection solvent is 90% MeOH + 10% Formic acid + 1% ascorbic acid neutralized to pH 7 with ammonium hydroxide (29%) containing 50 uL/mL. Injection Volume = 2 uL MS Conditions for API 5500 (Qtrap): MS conditions API 5500 (Qtrap) for detection of different transitions of the analytes in the scope Table 7: MS conditions API 5500 (Qtrap) 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

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