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A collaborative study of a method for analysis of vanillin and ethyl vanillin in fortified and imitation vanilla flavors was performed. The method, which is an extension of AOAC Official Method 990.25, Vanillin, Vanillic Acid, p-Hydroxybenzaldehyde, and p-Hydroxybenzoic Acid in Vanilla Extract, Liquid Chromatographic Method, involves reversed-phase high-pressure liquid chromatography (LC) of the sample on a Cβ column with a water-methanol-acetic acid (89 + 10 + 1) mobile phase and UV detection at 254 nm. The method yields good recoveries of vanillin at 484 and 723 mg/100 mL and of ethyl vanillin from 37 to 400 mg/100 mL. For vanillin, repeatability (r) values were 8 mg/100 mL at a level of 155 mg/100 mL, 12 mg/100 mLat a level of 484 mg/100 mL, and 31 mg/100 mL at a level of 723 mg/100 mL. Reproducibility (R) values were 20 mg/100 mL, 55 mg/100 mL, and 137 mg/100 mL over the same range. For ethyl vanillin, r values were 2.4 mg/100 mL at a level of 37 mg/100 mL, 3.2 mg/100 mL at a level of 74 mg/100 mL, and 8.6 mg/100 mL at a level of 180 mg/100 mL. R values were 6.4 mg/100 mL, 5.4 mg/100 mL, and 22.0 mg/100 mL over the same range. AOAC Official Method 990.25 has been modified to include determination of ethyl vanillin in vanilla extract and artificial vanilla flavor.

journal of aoac international

Liquid Chromatographic Method for Determination of Vanillin and Ethyl Vanillin in Imitation Vanilla Extract (Modification of AOAC Official Method 990.25): Collaborative Study