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Determination of antioxidants by a novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) assay with post-column detection.
Anal Chim Acta. 2010 Jul 26; 674(1):79-88.AC

Abstract

A novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) method was developed for the selective determination of polyphenols (flavonoids, simple phenolic and hydroxycinnamic acids) in complex plant matrices. The method combines chromatographic separation, constituent analysis, and post-column identification of antioxidants in plant extracts. The separation of polyphenols was performed on a C18 column using gradient elution with two different mobile phase solutions, i.e., MeOH and 0.2% o-phosphoric acid. The HPLC-separated antioxidant polyphenols in the extracts react with copper(II)-neocuproine (Cu(II)-Nc) reagent in a post-column reaction coil to form a derivative. The reagent is reduced by antioxidants to the copper(I)-neocuproine (Cu(I)-Nc) chelate having maximum absorption at 450 nm. The negative peaks of antioxidant constituents were monitored by measuring the increase in absorbance due to Cu(I)-Nc. The detection limits of polyphenols at 450 nm (in the range of 0.17-3.46 microM) after post-column derivatization were comparable to those at 280 nm UV detection without derivatization. The developed method was successfully applied to the identification of antioxidant compounds in crude extracts of Camellia sinensis, Origanum marjorana and Mentha. The method is rapid, inexpensive, versatile, non-laborious, uses stable reagents, and enables the on-line qualitative and quantitative estimation of antioxidant constituents of complex plant samples.

Authors+Show Affiliations

Department of Chemistry, Faculty of Engineering, Istanbul University, Avcilar 34320, Istanbul, Turkey.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20638503

Citation

Celik, Saliha Esin, et al. "Determination of Antioxidants By a Novel On-line HPLC-cupric Reducing Antioxidant Capacity (CUPRAC) Assay With Post-column Detection." Analytica Chimica Acta, vol. 674, no. 1, 2010, pp. 79-88.
Celik SE, Ozyürek M, Güçlü K, et al. Determination of antioxidants by a novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) assay with post-column detection. Anal Chim Acta. 2010;674(1):79-88.
Celik, S. E., Ozyürek, M., Güçlü, K., & Apak, R. (2010). Determination of antioxidants by a novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) assay with post-column detection. Analytica Chimica Acta, 674(1), 79-88. https://doi.org/10.1016/j.aca.2010.06.013
Celik SE, et al. Determination of Antioxidants By a Novel On-line HPLC-cupric Reducing Antioxidant Capacity (CUPRAC) Assay With Post-column Detection. Anal Chim Acta. 2010 Jul 26;674(1):79-88. PubMed PMID: 20638503.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determination of antioxidants by a novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) assay with post-column detection. AU - Celik,Saliha Esin, AU - Ozyürek,Mustafa, AU - Güçlü,Kubilay, AU - Apak,Reşat, Y1 - 2010/06/16/ PY - 2010/03/26/received PY - 2010/05/31/revised PY - 2010/06/09/accepted PY - 2010/7/20/entrez PY - 2010/7/20/pubmed PY - 2010/12/14/medline SP - 79 EP - 88 JF - Analytica chimica acta JO - Anal. Chim. Acta VL - 674 IS - 1 N2 - A novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) method was developed for the selective determination of polyphenols (flavonoids, simple phenolic and hydroxycinnamic acids) in complex plant matrices. The method combines chromatographic separation, constituent analysis, and post-column identification of antioxidants in plant extracts. The separation of polyphenols was performed on a C18 column using gradient elution with two different mobile phase solutions, i.e., MeOH and 0.2% o-phosphoric acid. The HPLC-separated antioxidant polyphenols in the extracts react with copper(II)-neocuproine (Cu(II)-Nc) reagent in a post-column reaction coil to form a derivative. The reagent is reduced by antioxidants to the copper(I)-neocuproine (Cu(I)-Nc) chelate having maximum absorption at 450 nm. The negative peaks of antioxidant constituents were monitored by measuring the increase in absorbance due to Cu(I)-Nc. The detection limits of polyphenols at 450 nm (in the range of 0.17-3.46 microM) after post-column derivatization were comparable to those at 280 nm UV detection without derivatization. The developed method was successfully applied to the identification of antioxidant compounds in crude extracts of Camellia sinensis, Origanum marjorana and Mentha. The method is rapid, inexpensive, versatile, non-laborious, uses stable reagents, and enables the on-line qualitative and quantitative estimation of antioxidant constituents of complex plant samples. SN - 1873-4324 UR - https://www.unboundmedicine.com/medline/citation/20638503/Determination_of_antioxidants_by_a_novel_on_line_HPLC_cupric_reducing_antioxidant_capacity__CUPRAC__assay_with_post_column_detection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2670(10)00751-8 DB - PRIME DP - Unbound Medicine ER -