HPLC method for the quantification of procyanidins in cocoa and chocolate samples and correlation to total antioxidant capacity
Abstract
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.
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- Authors
- Adamson, G. E., Lazarus, S. A., Mitchell, A. E., Prior, R. L., Cao, G., Jacobs, P. H., Kremers, B. G., Hammerstone, J. F., Rucker, R. B., Ritter, K. A., & Schmitz, H. H.
- Affiliations
- Tufts University (US)
- Venue
- Journal of Agricultural and Food Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1999-09-14
- Volume
- 47
- Issue
- 10
- Pages
- 4184-4188
- ISSN
- 0021-8561 (ISSN-L)
- 1520-5118
- OpenAlex
- W2007545619
- Cited by
- 404
- References
- 17
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 8 · 2023: 2 · 2022: 5 · 2021: 13 · 2020: 11 · 2019: 11 · 2018: 11 · 2017: 15 · 2016: 19 · 2015: 19 · 2014: 21 · 2013: 15 · 2012: 33
- FWCI
- 42.18
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Agricultural and Food Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Agricultural and Biological Sciences › Food Science › Food Chemistry and Fat Analysis
- Health Sciences › Medicine › Biochemistry › Phytochemicals and Antioxidant Activities
- Health Sciences › Medicine › Pharmacology › Coffee research and impacts
- Keywords
- procyanidins, Theobroma cacao, chocolate, oligomeric procyanidins, antioxidant capacity, ORAC, fluorescence detection, normal-phase HPLC, Sephadex LH-20, calibration standards, interlaboratory agreement, cocoa liquor
- MeSH terms
- Antioxidants, Cacao, Catechin, Chromatography, High Pressure Liquid, Fluorescence, Humans, Plant Extracts, Mass Spectrometry, Proanthocyanidins, Biflavonoids
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