Quantitation of Polycyclic Aromatic Hydrocarbons (PAH4) in Cocoa and Chocolate Samples by an HPLC-FD Method

Abstract

As a consequence of the PAH4 (sum of four different polycyclic aromatic hydrocarbons, named benzo[a]anthracene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene) maximum levels permitted in cocoa beans and derived products as of 2013, an high-performance liquid chromatography with fluorescence detection method (HPLC-FD) was developed and adapted to the complex cocoa butter matrix to enable a simultaneous determination of PAH4. The resulting analysis method was subsequently successfully validated. This method meets the requirements of Regulation (EU) No. 836/2011 regarding analysis methods criteria for determining PAH4 and is hence most suitable for monitoring the observance of the maximum levels applicable under Regulation (EU) No. 835/2011. Within the scope of this work, a total of 218 samples of raw cocoa, cocoa masses, and cocoa butter from several sample years (1999-2012), of various origins and treatments, as well as cocoa and chocolate products were analyzed for the occurrence of PAH4. In summary, it is noted that the current PAH contamination level of cocoa products can be deemed very slight overall.

Review of PAH contamination in food products and their health hazards

“…European No. 835/2011 amended the contamination regulation pertaining to the cocoa beans and its derivative products to a maximum concentration of 5.0 μg/kg fat for BAP and 35 μg/kg fat for PAH4 (30 μg/kg fat for PAH4 as of April 2015) (Raters and Matissek, 2014). These authors quantified the PAH contents (BAA, CHR, BBF, and BAP) in the samples of cocoa and chocolates as 12.44 μg/kg (fat) for PAH4 and 0.96 μg/kg (fat) for BAP.…”

“…Thus, relatively low contamination levels of PAH were found in raw coca and semi-finished coca products. A decrease in the content of PAH in cocoa butter samples was observed from a comparative study made over an extended period (1999-2012) from 17 μg/kg (fat) (1999) to 8.8 μg/kg (fat) (2012) in terms of PAH4 (Raters and Matissek, 2014). This decrease was due to the strict code of practices for the drying and smoking processes of cocoa (Codex Alimentarius Commision, 2009).…”

Polycyclic Aromatic Hydrocarbons in Foods: Biological Effects, Legislation, Occurrence, Analytical Methods, and Strategies to Reduce Their Formation

“…In food products consisting mostly of fats and proteins, saponification, liquid-liquid extraction, and purification steps are necessary to isolate and quantify the PAHs [89,90,92]. For example, a saponification step is necessary when analyzing the PAHs content of meat products.…”

Microbial communication during bioremediation of polyaromatic hydrocarbons

“…The peaks 1-4 are prominent with UV detector while the remaining 12 PAHs have been observed using a fluorescence detector. Following that, unknown polyaromatic hydrocarbons from contaminated soil are extracted and compared to standard combinations to determine their identity [73,99]. The principle of interactions is used in donor-acceptor complex chromatography, in which the stationary phase (DACC column) functions as an electron acceptor, retaining the PAHs (electron donors) [104].…”