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Sensitive micellar electrokinetic chromatography-laser-induced fluorescence method to analyze chiral amino acids in orange juices.
J Agric Food Chem. 2002 Sep 11; 50(19):5288-93.JA

Abstract

In this work a new method to detect the existence of chiral amino acids in orange juice is presented. The method employs beta-cyclodextrins and micellar electrokinetic chromatography with laser-induced fluorescence (MEKC-LIF) to separate and detect L- and D-amino acids (L-aa and D-aa) previously derivatized with fluorescein isothiocianate (FITC). A systematic optimization of the chiral-MEKC conditions is done bringing about in less than 20 min a good separation of the main amino acids found in orange juice (i.e., Pro, Asp, Ser, Asn, Glu, Ala, Arg, and the nonchiral GABA, i.e., gamma-aminobutyric acid). Using this procedure, the analysis time reproducibility for the 15 standard compounds (L-aa, D-aa, and GABA) has been determined to be better than 0.2% (n = 5) for the same day and better than 0.7% (n = 15) for three different days. Corrected peak area reproducibility is somewhat lower, providing values better than 3.3% (n = 5) for the same day and 6.9% (n = 15) for three different days. The limit of detection using this procedure was determined to be 0.86 attomoles for L-Arg. The optimized FITC derivatization method allows the easy and straightforward detection of amino acids in orange concentrates and juices (i.e., only centrifugation of diluted samples for 5 min is needed prior to their derivatization). D-Ala, D-Asp, D-Arg, and D-Glu were determined in orange juices and orange concentrates from different geographical origins using this new method. Moreover, the effect of different temperature treatments (50, 92, and 150 degrees C) on the content of D-aa in orange juice was evaluated.

Authors+Show Affiliations

Simó C
Institute of Industrial Fermentations (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain.
Barbas C
No affiliation info available
Cifuentes A
No affiliation info available

MeSH

Amino AcidsBeveragesChromatography, Micellar Electrokinetic CapillaryCitrusFluorescein-5-isothiocyanateFluorescenceFluorescent DyesLasersReproducibility of ResultsStereoisomerism

Pub Type(s)

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

Language

eng

PubMed ID

12207463

Citation

Simó, Carolina, et al. "Sensitive Micellar Electrokinetic Chromatography-laser-induced Fluorescence Method to Analyze Chiral Amino Acids in Orange Juices." Journal of Agricultural and Food Chemistry, vol. 50, no. 19, 2002, pp. 5288-93.
Simó C, Barbas C, Cifuentes A. Sensitive micellar electrokinetic chromatography-laser-induced fluorescence method to analyze chiral amino acids in orange juices. J Agric Food Chem. 2002;50(19):5288-93.
Simó, C., Barbas, C., & Cifuentes, A. (2002). Sensitive micellar electrokinetic chromatography-laser-induced fluorescence method to analyze chiral amino acids in orange juices. Journal of Agricultural and Food Chemistry, 50(19), 5288-93.
Simó C, Barbas C, Cifuentes A. Sensitive Micellar Electrokinetic Chromatography-laser-induced Fluorescence Method to Analyze Chiral Amino Acids in Orange Juices. J Agric Food Chem. 2002 Sep 11;50(19):5288-93. PubMed PMID: 12207463.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Sensitive micellar electrokinetic chromatography-laser-induced fluorescence method to analyze chiral amino acids in orange juices. AU - Simó,Carolina, AU - Barbas,Coral, AU - Cifuentes,Alejandro, PY - 2002/9/5/pubmed PY - 2002/11/26/medline PY - 2002/9/5/entrez SP - 5288 EP - 93 JF - Journal of agricultural and food chemistry JO - J Agric Food Chem VL - 50 IS - 19 N2 - In this work a new method to detect the existence of chiral amino acids in orange juice is presented. The method employs beta-cyclodextrins and micellar electrokinetic chromatography with laser-induced fluorescence (MEKC-LIF) to separate and detect L- and D-amino acids (L-aa and D-aa) previously derivatized with fluorescein isothiocianate (FITC). A systematic optimization of the chiral-MEKC conditions is done bringing about in less than 20 min a good separation of the main amino acids found in orange juice (i.e., Pro, Asp, Ser, Asn, Glu, Ala, Arg, and the nonchiral GABA, i.e., gamma-aminobutyric acid). Using this procedure, the analysis time reproducibility for the 15 standard compounds (L-aa, D-aa, and GABA) has been determined to be better than 0.2% (n = 5) for the same day and better than 0.7% (n = 15) for three different days. Corrected peak area reproducibility is somewhat lower, providing values better than 3.3% (n = 5) for the same day and 6.9% (n = 15) for three different days. The limit of detection using this procedure was determined to be 0.86 attomoles for L-Arg. The optimized FITC derivatization method allows the easy and straightforward detection of amino acids in orange concentrates and juices (i.e., only centrifugation of diluted samples for 5 min is needed prior to their derivatization). D-Ala, D-Asp, D-Arg, and D-Glu were determined in orange juices and orange concentrates from different geographical origins using this new method. Moreover, the effect of different temperature treatments (50, 92, and 150 degrees C) on the content of D-aa in orange juice was evaluated. SN - 0021-8561 UR - https://www.unboundmedicine.com/medline/citation/12207463/Sensitive_micellar_electrokinetic_chromatography_laser_induced_fluorescence_method_to_analyze_chiral_amino_acids_in_orange_juices_ DB - PRIME DP - Unbound Medicine ER -