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

Results:

Optimization of compound based mass-spec parameters:

The compound based parameters were determined by pumping the mobile phase at 0.5 mL/min through the HPLC pump without column. The analyte mixture solution was pumped into the MS source by the 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 1. Table 10: Optimization of MS conditions 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 6 442.2 176.2 75 6 6 45 6 5-CH3-THF 460.2 313.2 75 6 6 28 6 460.2 194.2 75 6 6 46 6 5-CHO-THF 474.2 327.2 75 6 6 25 6 474.2 299.2 75 6 6 40 6 THF 446.2 299.2 75 6 6 30 6 446.2 166.2 75 6 6 50 6 5,10-CH=CH- THF 456.2 412.2 75 6 6 42 6 456.2 282.2 75 6 6 70 6 10-CH3-FA 456.2 309.2 75 6 6 20 6 456.2 176.2 75 6 6 50 6 10-CHO-FA 470.2 295.2 75 6 6 35 6 470.2 176.2 75 6 6 50 6 13 C 5 -FA 447.2 295.2 75 6 6 20 6 13 C 5 -5-CH3- THF 465.2 313.2 75 6 6 28 6 DP: Declustering potential; EP Entrance potential; CEP collision cell entrance potential; CE Collision cell; CXP collision cell exit potential.

These conditions were used for analytes detection through the study. More intense transition 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 flowing section. Curtain gas (nitrogen): 25 psi; Collision Assisted Dissociation: Medium Ion Spray voltage: 5500 V; Temperature: 700 °C;

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