Simultaneous RP-HPLC-DAD quantification of benzaldehyde, 4-methoxybenzaldehyde, and 3,4-dimethoxybenzaldehyde in
Sabrina Stýblová1, Sonja Berensmeier2
1Chair of Bioseparation Engineering, TUM School of Engineering and Design, Technical University of Munich, Boltzmannstraße 15, Garching 85748, Germany.
Abstract:
A reversed-phase high-performance liquid chromatography method with a diode-array detector (RP-HPLC-DAD) was developed and validated for the simultaneous determination of 3,4-dimethoxybenzaldehyde (3,4-DMBA), benzaldehyde (BA), and 4-methoxybenzaldehyde (4-MBA) in acid whey fermented with Ischnoderma benzoinum. These structurally related aromatic aldehydes are valuable aroma compounds formed during basidiomycete-mediated fermentation of dairy by-products; however, no method enabling their simultaneous quantification in this complex matrix without prior derivatization has been reported to date. Chromatographic separation was achieved on an XBridge BEH C18 column (250 × 4.6 mm, 5 μm) using gradient elution with water and methanol as mobile phases, with a total runtime of 30 min. The method was validated following the relevant ICH Q2(R2) validation characteristics and showed linearity (R2 > 0.999) over ranges of 0.1-100 mg L-1 (BA, 4-MBA) and 0.1-75 mg L-1 (3,4-DMBA). Limits of detection and quantification ranged from 0.32 to 0.66 mg L-1 and 0.96-1.98 mg L-1, respectively. Accuracy (94.9-111.5%), intra- and inter-day precision (RSD ≤ 3.16%), and matrix recovery (90.7-105.5%) confirmed the reliability of the method for quantitative analysis in real fermentation samples. The developed method provides a practical, direct analytical tool for monitoring aromatic aldehyde production at mg L-1 level in complex aqueous fermentation matrices, rather than for trace-level determination.
