Analytical Method Validation and Rapid Determination of Polycyclic Aromatic Hydrocarbons (PAHs) in Cocoa Butter Using HPLC-FLD

Abstract

A simple, fast and ecological analytical method using a semi-automatic fat extractor and HPLC-FLD (fluorescence detection) for determination of polycyclic aromatic hydrocarbon markers i.e. benzo(a)anthracene (BaA), chrysene (Chr), benzo(a)pyrene (BaP) and benzo(b)fluoranthene (BbF) in cocoa butter has been validated. Validation’s procedure performed out in concordance with French standard NF V03-110 (2010) was based on existing polycyclic aromatic hydrocarbon (PAH) determination methods in various smoked foodstuffs and edible vegetable oils. Determination of correlation coefficients for specific PAHs ranged from 0.9992 to 0.9998. Respective values of limits of detection were 0.010, 0.011, 0.033 and 0.029 μg kg−1 and those of quantification were 0.035, 0.038, 0.111 and 0.098 μg kg−1 for BaA, Chr, BbF and BaP. Both values of repeatability and intermediary precision tests coefficients of variation were less than 5%. Recovery scores of four PAH markers matched EU standard 836/2011 recommendations. Sum of four PAH markers (BaA, Chr, BbF, BaP) contents varied from 5.42 ± 0.58 to 11.37 ± 0.01 μg kg−1 whereas those of BaP was comprised between 0.26 ± 0.00 and 1.75 ± 0.13 μg kg−1 in 20 cocoa butter samples extracted from raw cocoa bean stored at Ivorian cocoa farmer levels.