Ultrasensitive detection of indoleamines by combination of nanoparticle-based extraction with capillary electrophoresis/laser-induced native fluorescence |
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Authors: | Mu-De Li Wei-Lung Tseng Tian-Lu Cheng |
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Affiliation: | 1. Department of Chemistry, National Sun Yat-sen University, 70, Lien-hai Road, Kaohsiung, Taiwan;2. Kaohsiung Medical University Joint Research Center, National Sun Yat-Sen University, Kaohsiung, Taiwan;3. Faculty of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan |
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Abstract: | For the first time, citrate-capped gold nanoparticles (citrate-AuNPs) have been used for the selective extraction of indoleamines – 5-hydroxytryptophan (5-HTP), tryptophan (Trp), tryptamine (TA), 5-hydroxytryptamine (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) – prior to their analysis by capillary electrophoresis/laser-induced native fluorescence (CE/LINF). The extinction spectra obtained for the citrate-AuNPs in the presence of indoleamines revealed that 5-HTP, 5-HT, and 5-HIAA were extracted mainly because of van der Waals interactions between the indole ring and the citrate-AuNPs (hydrophobic surface), while 5-HT and TA were extracted by electrostatic attractions between the amine group of the indoleamines and the citrate ligands adsorbed on the AuNP surface. The extracted indoleamines could be liberated from the AuNP surface by the addition of high concentrations of 2-mercaptoethanol (2-ME), which binds strongly to the AuNPs. The sensitivity of this method to indoleamines could be significantly enhanced by increasing the AuNP concentration, incubation time, and sample volume. Under optimal extraction and separation conditions, the combination of NP-based extraction and CE-LINF provided 48-, 4077-, 985-, 920-, and 4030-fold improvements in the limits of detection (signal-to-noise ratio of 3) for 5-HTP, Trp, TA, 5-HT, and 5-HIAA as compared to the analysis of five indoleamines by CE-LINF. In addition, this proposed method was successfully used for the determination of TA and 5-HT in urine. |
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Keywords: | Capillary electrophoresis Gold nanoparticles Extraction Indoleamines Native fluorescence |
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