An LCMS Method for the Detection of Cocoa Butter Substitutes, Replacers, and Equivalents in Commercial Chocolate-like Products

There is increasing demand for genuine cocoa butter (CB) in chocolate products in developed nations, however, this demand has created a shortage of CB and raised its costs. To overcome this, chocolate manufactures sometimes add vegetable-derived fats to some chocolate products to reduce costs while still maintaining desirable physical characteristics. It is of current interest to have a reliable method to detect, identify, and quantify the triacylglycerol (TAG) components of cocoa butter substitutes, replacers, and equivalents (CBEs) in chocolate products. Traditionally GC was used for this task, but due to the low volatility of triacylglycerides and their susceptibility to thermal decomposition, retention time is the only identifying factor for the TAGs and typical GC analyses of this type can take 40 minutes. LCMS is able to not only provide faster throughput, but also has the additional advantage of allowing characterization of the TAG, including qualitative regiospecific analysis. We have developed a single, UHPLC column-based LCMS method to analyze the TAG components in commercial chocolate and chocolate-like products. This analysis has a runtime of 17minutes, making it suitable for relatively high throughput. Additionally, the method was very repeatable, with an interday variability of <7% for the absolute area counts of the three major TAGs in CB (POP,POS,SOS).