Background: Vanillin is a key constituent of natural vanilla. Usage of natural vanilla is
affected due to its high price and limited supply, which leads to the use of artificial vanilla flavoring
substances. Coumarin is a commonly encountered adulterant in beverage, food, and cosmetics as a
flavoring and fragrance enhancer. However, coumarin has been banned for use as a food additive due
to its toxic effects. To comply with the quality of vanillin in food and food products needs to be ensured.
Methods: A rapid, simple and selective analytical method has been developed and validated using
ultra-high performance liquid chromatography-tandem mass spectrometry for quantitative analysis of
vanillin and coumarin. We also optimized fragmentation pattern of these metabolites while increasing
collision energy to elucidate its structural information. The suitability and robustness of the
method was checked by Zorbax Eclipse XDB C8 column (4.6 × 150 mm, 5 μm) using mobile phase
comprising of methanol (A) and water with 0.1% formic acid (B) (90:10) with a flow rate
200 μL/min. The separation was achieved within 4.2 min with total run time of 5.0 min. The analysis
was done by multiple reaction monitoring using 153/93 and 147/91 pair transition in positive electrospray
ionization for vanillin and coumarin respectively.
Results: The lower limit of quantification of vanillin and coumarin was 0.39 ng/mL and 3.9 ng/mL
respectively. The intra and inter-day precisions for vanillin and coumarin were lower than 8.87 and
8.62 whereas, accuracy was within ± 2.13 and ± 1.53 respectively. The vanillin and coumarin was
found to be stable under the examined conditions. This method was successfully applied for quantification
of vanillin and coumarin in mangrove species and commercial food products.
Conclusion: The described method and fragmentation pattern could be useful to direct confirmation
and quality monitoring of a commercial food products assimilated with vanillin.