Simultaneous Determination of Vanillin and o-Vanillin by High Performance Liquid Chromatography

A high performance liquid chromatographic method was developed for the simultaneous separation and determination of vanillin and o-vanillin.Influences of mobile phase composition and column temperature on the separation were investigated.Under the condition of 5% acetic acid-acetonitrile(60∶40,by volume) as mobile phase at a flow rate of 1.0 mL/min and 25 ℃,the vanillin and o-vanillin were separated in less than 5 min.The calibration curves of vanillin and o-vanillin showed good linearities in the range of 10-240 mg/L with both correlation coefficients of 0.999 7.The limits of detection(S/N=3) for vanillin and o-vanillin were 0.06 mg/L and 0.02 mg/L,respectively.The average recoveries at three spiked levels were in the range of 99%-102% with relative standard deviations of 0.2%-0.6%.The method was successfully used for the determination of vanillin and o-vanillin in real samples with satisfactory results.