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Sensitive determination of pheomelanin after 5-carboxyfluorescein succinimidyl ester precapillary derivatization and micellar electrokinetic capillary chromatography with laser-induced fluorescence detection.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Jan 01; 861(1):136-9.JC

Abstract

The suitability of micellar electrokinetic capillary chromatography (MEKC) with laser-induced fluorescence (LIF) detection for the determination of pheomelanin in biological materials has been investigated. 5-Carboxyfluorescein succinimidyl ester was chosen as the labeling reagent to precapillary derivatize the two marker aminohydroxyphenylalanine (AHP) isomers produced after reductive hydrolysis of pheomelanin with hydriodic acid (HI). Various parameters affecting derivatization and separation were systematically studied. Under optimal conditions, the analytes could be separated within 18 min, and the relative standard deviations (R.S.D.) of migration time and corrected peak areas were less than 5.5%. Compared with the conventional high-performance liquid chromatography (HPLC) method with electrochemical detection, the 100-fold improvements in sensitivity were achieved by applying LIF detection. As a preliminary application, this method has been successfully applied to the determination of pheomelanin in two human melanoma cell cultures, black hair, melanoma tissue and urine samples of human melanoma patients with the spiked recoveries in the range of 88-96%.

Authors+Show Affiliations

College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310029, China.No affiliation info availableNo affiliation info availableNo 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

18088575

Citation

Zhang, Xiao-Ling, et al. "Sensitive Determination of Pheomelanin After 5-carboxyfluorescein Succinimidyl Ester Precapillary Derivatization and Micellar Electrokinetic Capillary Chromatography With Laser-induced Fluorescence Detection." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 861, no. 1, 2008, pp. 136-9.
Zhang XL, Yang Q, Jiao BH, et al. Sensitive determination of pheomelanin after 5-carboxyfluorescein succinimidyl ester precapillary derivatization and micellar electrokinetic capillary chromatography with laser-induced fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2008;861(1):136-9.
Zhang, X. L., Yang, Q., Jiao, B. H., Dai, J., Zhang, J. X., & Ni, W. Z. (2008). Sensitive determination of pheomelanin after 5-carboxyfluorescein succinimidyl ester precapillary derivatization and micellar electrokinetic capillary chromatography with laser-induced fluorescence detection. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 861(1), 136-9.
Zhang XL, et al. Sensitive Determination of Pheomelanin After 5-carboxyfluorescein Succinimidyl Ester Precapillary Derivatization and Micellar Electrokinetic Capillary Chromatography With Laser-induced Fluorescence Detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Jan 1;861(1):136-9. PubMed PMID: 18088575.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Sensitive determination of pheomelanin after 5-carboxyfluorescein succinimidyl ester precapillary derivatization and micellar electrokinetic capillary chromatography with laser-induced fluorescence detection. AU - Zhang,Xiao-Ling, AU - Yang,Qiao, AU - Jiao,Bing-Hua, AU - Dai,Jun, AU - Zhang,Jun-Xiang, AU - Ni,Wu-Zhong, Y1 - 2007/12/04/ PY - 2007/08/07/received PY - 2007/11/20/revised PY - 2007/11/23/accepted PY - 2007/12/20/pubmed PY - 2008/4/15/medline PY - 2007/12/20/entrez SP - 136 EP - 9 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 861 IS - 1 N2 - The suitability of micellar electrokinetic capillary chromatography (MEKC) with laser-induced fluorescence (LIF) detection for the determination of pheomelanin in biological materials has been investigated. 5-Carboxyfluorescein succinimidyl ester was chosen as the labeling reagent to precapillary derivatize the two marker aminohydroxyphenylalanine (AHP) isomers produced after reductive hydrolysis of pheomelanin with hydriodic acid (HI). Various parameters affecting derivatization and separation were systematically studied. Under optimal conditions, the analytes could be separated within 18 min, and the relative standard deviations (R.S.D.) of migration time and corrected peak areas were less than 5.5%. Compared with the conventional high-performance liquid chromatography (HPLC) method with electrochemical detection, the 100-fold improvements in sensitivity were achieved by applying LIF detection. As a preliminary application, this method has been successfully applied to the determination of pheomelanin in two human melanoma cell cultures, black hair, melanoma tissue and urine samples of human melanoma patients with the spiked recoveries in the range of 88-96%. SN - 1570-0232 UR - https://www.unboundmedicine.com/medline/citation/18088575/Sensitive_determination_of_pheomelanin_after_5_carboxyfluorescein_succinimidyl_ester_precapillary_derivatization_and_micellar_electrokinetic_capillary_chromatography_with_laser_induced_fluorescence_detection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(07)00828-8 DB - PRIME DP - Unbound Medicine ER -