High Pressure Liquid Chromatographic Determination of Theobromine and Caffeine in Cocoa and Chocolate Products

Abstract

A method was developed for determining theobromine and caffeine in cocoa and chocolate products by high pressure liquid chromatography. After a simple hot water extraction, both theobromine and caffeine were separated by using a reverse phase Cl s column and a mobile phase of methanol-water-acetic acid (20+ 79+1). Theobromine and caffeine were quantitated at 280 nm; average recoveries were 98.7 and 95.0%; and coefficients of variation were 2.31 and 3.91%, respectively.

Fermentation of cocoa beans: influence of microbial activities and polyphenol concentrations on the flavour of chocolate

“…Concentrations of theobromine and caffeine were determined using a modified version of the HPLC method described by Kreiser and Martin 23. A mobile phase of water/acetonitrile (80:20, vol/vol) at a flow rate of 1 mL min −1 and UV detection at 254 nm were applied.…”

The influence of carbon source on the growth and development of asexual embryos of Theobroma cacao

“…Alkaloids were extracted and analyzed using HPLC as described by Kreiser and Martin (1978), Theobromine and caffeine were identified using known standards, and quantified with the inclusion of theophylline as an internal standard.…”

Effect of Caffeine on Aflatoxin Production on Cocoa Beans

“…Aflatoxins were assayed according to the technique described by Hurst and Toomey (1978). Caffeine was analyzed by the method of Kreiser and Martin (1978).…”

“…They hypothesized that the varying levels of aflatoxin produced on three cocoa bean varieties were the result of varying levels of caffeine. Kreiser and Martin (1978) reported that cocoa beans from different geographic origins have varying levels of caffeine. The objective of this study was to examine the aflatoxin production by A. parasiticus on cocoa beans having a wide range of caffeine content.…”