Selective Determination of Tryptophan-containing Peptides through Precolumn Derivatization and Liquid Chromatography Using Intramolecular Fluorescence Resonance Energy Transfer Detection

Yoshitake, Makoto; Sejima, Naoko; Yoshida, Hideyuki; Todoroki, Kenichiro; Nohta, Hitoshi; Yamaguchi, Masatoshi

doi:10.2116/analsci.23.949

Cited by 8 publications

(6 citation statements)

References 25 publications

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“…o -Phthalaldehyde (OPA) is a fluorogenic reagent, which reacts with primary amino group of compounds in the presence of 2-mercaptoethanol at room temperature, and then provides FL products for amino acids, amines, peptides and proteins36373839. Herein, we examined on the specificity and sensitivity of the OPA reaction for the determination of collagens, since a large number of α-amino group from one molecule of collagens would be produced by the enzymatic digestion with collagenase.…”

Section: Resultsmentioning

confidence: 99%

Amplified and selective assay of collagens by enzymatic and fluorescent reactions

Yasmin

Kabashima

Rahman

et al. 2014

Sci Rep

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Sensitive and selective assay of collagen is of substantial importance to the diagnostic study of health- and aging-related failures. In this paper, we describe a highly specific and sensitive method for the assay of whole collagens in biological samples using a novel fluorogenic reagent, 3,4-dihydroxyphenylacetic acid (3,4-DHPAA). The 3,4-DHPAA reagent can selectively detect N-terminal Gly-containing peptides (NGPs) in the presence of sodium borate and NaIO4. Under conditions optimized, this assay format for collagen, termed 3,4-DHPAA assay method showed a good linear relationship between the amplified FL signals and the collagen concentrations from 0.18 to 12 μg/ml. Therefore the sensitive determination of intracellular collagens in cheek tissue and HeLa cells was individually possible without any separation protocol. The dual recognitions of the collagens in the samples could be performed by the enzymatic digestion and the FL reaction. The proposed assay method enables the determination facile, specific, sensitive and quantitative for biogenic collagens.

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Section: Resultsmentioning

confidence: 99%

Amplified and selective assay of collagens by enzymatic and fluorescent reactions

Yasmin

Kabashima

Rahman

et al. 2014

Sci Rep

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“…OPA in combination with a thiol compound, such as 2‐ME, is widely utilized as a fluorescent derivatizing agent for amino compounds. Several pharmaceutical compounds have been determined through this approach . OST was found to react with OPA in the presence of 2‐ME to form a highly fluorescent derivative with maximum fluorescence at 450 nm after excitation at 336 nm (Fig.…”

Section: Resultsmentioning

confidence: 99%

Spectrofluorimetric determination of oseltamivir phosphate through derivatization with o‐phthalaldehyde. Application to pharmaceutical preparations with a preliminary study on spiked plasma samples

et al. 2012

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A simple and sensitive spectrofluorimetric method has been developed and validated for the determination of oseltamivir phosphate (OST) in pharmaceutical preparations. The method is based on the reaction between oseltamivir phosphate and o-phthalaldehyde in presence of 2-mercapto-ethanol in borate buffer, pH 10.8, to give a highly fluorescent product measured at 450 nm after excitation at 336 nm. The different experimental parameters affecting the development and stability of the reaction product were studied and optimized. The fluorescence intensity-concentration plot is rectilinear over the range 0.05-1.0 µg/mL, with a lower detection limit of 5 ng/mL and limit of quantitation of 16 ng/mL. The developed method was successfully applied to the analysis of the drug in its commercial capsules and suspension, mean recoveries of OST were 99.97 ± 1.67% and 100.17 ± 1.18%, respectively (n = 3). Statistical comparison of the results obtained by the proposed and comparison method revealed no significant difference in the performance of the two methods regarding accuracy and precision. The proposed method was further extended to in vitro determination of the studied drug in spiked human plasma as a preliminary investigation; the mean recovery (n = 3) was 98.68 ± 5.8%. A reaction pathway was postulated.

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“…After the pretreatments, the calibration curve was not significantly altered by co-existing proteins (Fig. 4) that may exert intra-molecular FRET effect if the N-terminal amino acid residue happens to be tryptophan (Yoshitake, Sejima, et al, 2007;. the flow-injection system, and the difference in peak heights at 280 and 300 nm excitation wavelengths (as in Fig.…”

Section: Effect Of Sample Pretreatmentmentioning

confidence: 99%

“…Yoshitake, Sejima, et al (2007), labeled the bioamines in urine by a pre-column derivatization reaction with OPA, and practically all primary amino groups will become isoindoles and fluoresce under UV illumination (Roth, 1971). In the case of tryptophan, an intra-molecular FRET phenomenon between the inherent indole moiety and the newly formed isoindole group on derivatized tryptophan molecule occurred efficiently, which made the fluorescent detection sensitive and selective for tryptophan.…”

Section: Introductionmentioning

confidence: 99%

Judgment of pure fermented soy sauce by fluorescence resonance energy transfer of OPA-tryptophan adduct

et al. 2015

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Selective Determination of Tryptophan-containing Peptides through Precolumn Derivatization and Liquid Chromatography Using Intramolecular Fluorescence Resonance Energy Transfer Detection

Cited by 8 publications

References 25 publications

Amplified and selective assay of collagens by enzymatic and fluorescent reactions

Amplified and selective assay of collagens by enzymatic and fluorescent reactions

Spectrofluorimetric determination of oseltamivir phosphate through derivatization with o‐phthalaldehyde. Application to pharmaceutical preparations with a preliminary study on spiked plasma samples

Judgment of pure fermented soy sauce by fluorescence resonance energy transfer of OPA-tryptophan adduct

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