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

Folate Compound

Concentration (ppb)

Figure 2 : Calibration Curve of different folate compounds (Dionex 3000 Ultmate HPLC and 5500 API MS detector). The color scheme for the calibration curve of different folate compounds is as the following . Folate (295) ______, 5-CH3-THF (313)_______, 5-CHO-THF (327)_______, THF (299) _____, 10-CHO-FA (295)_________, 10-CH3-FA (309) ______. CH=THF (282) _______. Conc Calibration Values: The calibration values based on the above calibration curve for different folate compounds are listed in the following Table. Table 15: Evaluation of Response Linearity against concentration of six target folate compounds the values of linear regression Folate Compound Transition Slope Intercept r2 weighing Calibration Line Color

Folic Acid 5-CH3-THF 5-CHO-THF CH=CH-THF 10-CH3-FA 10-CHO-FA THF

295 313 327 299 282 309 295

0.02807 +0.00269 0.9996 0.02453 +0.01463 0.9931 0.02746 +0.01105 0.9922

1/x 1/x 1/x 1/x 1/x 1/x 1/x

Navy Blue

Purple

Red

0.01553 -0.01530

0.9992

Orange Sky Blue

0.1050 +0.00211 0.9965

0.03783 -0.01387

0.9979

Light Green Dark Green

0.04435 +0.00650 0.9996

Effect of Methanol concentration in the injection solvent The initial calibration work and the injection solvent for LC-MS analysis of the analytes used in the AOAC proposed method contains methanol at 20% in a mixture with 80% of 1% acetic acid solution along with 1% ascorbic acid (20:80:1 v/v/w). In the subsequent work we found a way to make the method a little bit less time consuming and labor intensive and most important to improve analyte

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