HPLC Method for the Quantification of Procyanidins in Cocoa and Chocolate Samples and Correlation to Total Antioxidant Capacity

Monomeric and oligomeric procyanidins present in cocoa liquors and chocolates were separated and quantified in four different laboratories using a normal-phase high-performance liquid chromatography (HPLC) method with fluorescence detection. Procyanidin standards through decamers were obtained by ex...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 1999-10, Vol.47 (10), p.4184-4188
Main Authors: Adamson, Gary E, Lazarus, Sheryl A, Mitchell, Alyson E, Prior, Ronald L, Cao, Guohua, Jacobs, Pieter H, Kremers, Bart G, Hammerstone, John F, Rucker, Robert B, Ritter, Karl A, Schmitz, Harold H
Format: Article
Language:English
Subjects:
absorbance
analytical methods
Antioxidants - analysis
assays
Biflavonoids
Biological and medical sciences
Cacao
Catechin - analysis
chocolate
Chromatography, High Pressure Liquid - methods
cocoa (beverage)
cocoa beans
cocoa liquors
Confectionery products and chocolate industries, honey
detection
Fluorescence
Food industries
Fundamental and applied biological sciences. Psychology
General pharmacology
high performance liquid chromatography
Humans
inhibition
Mass Spectrometry
Medical sciences
oxidation
oxygen radical absorbance capacity assay
Pharmacognosy. Homeopathy. Health food
Pharmacology. Drug treatments
Plant Extracts - chemistry
polyphenols
Proanthocyanidins
spectrofluorimetry
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Summary:Monomeric and oligomeric procyanidins present in cocoa liquors and chocolates were separated and quantified in four different laboratories using a normal-phase high-performance liquid chromatography (HPLC) method with fluorescence detection. Procyanidin standards through decamers were obtained by extraction from cocoa beans, enrichment by Sephadex LH-20 gel permeation chromatography, and final purification by preparative normal-phase HPLC. The purity of each oligomeric fraction was assessed using HPLC coupled to mass spectrometry. A composite standard was then prepared, and calibration curves were generated for each oligomeric class using a quadratic fit of area sum versus concentration. Results obtained by each of the laboratories were in close agreement, which suggests this method is reliable and reproducible for quantification of procyanidins. Furthermore, the procyanidin content of the samples was correlated to the antioxidant capacity measured using the ORAC assay as an indicator for potential biological activity. Keywords: HPLC; procyanidins; cocoa; chocolate; quantification; antioxidant
ISSN:0021-8561
1520-5118
DOI:10.1021/jf990317m