Sensitive Laser induced fluorescence detection of Polyamine-Fluoresceinisothiocyanate-derivatives after capillary zone electrophoretic separation

Mattusch, Jürgen; Huhn, Gernot; Wennrich, Rainer

doi:10.1007/bf00323628

Cited by 17 publications

(8 citation statements)

References 30 publications

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“…Because the reaction between FITC and the amino groups of PAs or CAs is generally faster than between FITC and amino acids (time of amino acid derivatization . 12 h), specificity can be achieved by a shorter reaction time [34,41]. Since the reaction ratio of FITC to PAs and CAs has been investigated [34] for optimal detection sensitivity, a ratio of 45:1 (FITC:PAs or CAs) was adopted in our study.…”

Section: Optimization Of Derivatizationmentioning

confidence: 99%

Simultaneous determination of polyamines and catecholamines in PC‐12 tumor cell extracts by capillary electrophoresis with laser‐induced fluorescence detection

Flanigan

2004

Electrophoresis

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A method to detect polyamines and catecholamines in PC-12 tumor cell extracts by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) is described for the first time. Both derivatization conditions and buffer concentrations and pH were optimized. Under optimized conditions the polyamines (putresine, spermine, spermidine) and catecholamines (dopamine, norepinephrine, epinephrine, serotonin) were derivatized with fluorescein isothiocyanate and separated at 25 kV in a fused-silica capillary (50 microm ID x 40 cm) with 0.1 M borate, pH 9.0, in less than 18 min. The influence of running buffer conditions, such as buffer pH and concentrations, were also investigated. Linearity of the analytes ranged from 0.05 to 1.0 micromol/L, and the detection limit (S/N = 3 ) ranged from 0.03 to 2.50 nmol/L. The concentrations of polyamines and catecholamines in PC-12 tumor cell extracts were determined with this method.

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Section: Optimization Of Derivatizationmentioning

confidence: 99%

Simultaneous determination of polyamines and catecholamines in PC‐12 tumor cell extracts by capillary electrophoresis with laser‐induced fluorescence detection

Flanigan

2004

Electrophoresis

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“…The reagents FITC and CFSE, which are fluorescein analogues that can label compounds containing primary or secondary amino groups [16], were chosen to test their suitability for the derivatization of Hy compounds. [17] or polyamines [18] at concentrations of 10 26 M. An explanation can be found in the high reactivity of Hys towards oxygen which prevents successful derivatization reactions of lower concentrations of Hys during the relatively long reaction time (6 h). The concentration of Hys required for the derivatization could be lowered to 10 27 M when CFSE was used as the fluorescent reagent.…”

Section: Mekc-lif Determination Of Hy Compoundsmentioning

confidence: 97%

Determination of hydrazine, monomethylhydrazine, 1,1‐dimethylhydrazine, and 1,2‐dimethylhydrazine by nonaqueous capillary electrophoresis with amperometric detection

et al. 2005

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The present study is concerned with the application of nonaqueous capillary electrophoresis (NACE) with electrochemical detection (ED) to the separation and quantitative determination of hydrazine (Hy) and its methyl derivatives. The best performance of NACE-ED was found when using 4 mM sodium acetate/10 mM acetic acid/methanol: acetonitrile = 1:2 as the running buffer, with a bare platinum working electrode set at +1.0 V in an end-column amperometric detection cell. The choice and ratio of suitable solvents for the separation and injection media played an essential role for the performance characteristics of the method. The limits of detection for Hy, methylhydrazine, symmetrical dimethylhydrazine, and unsymmetrical dimethylhydrazine were 5, 2, 12, and 1 ng/mL, respectively. This is between one and two orders of magnitude lower than that achieved by previously reported CE-ED methods in aqueous buffer systems in conjunction with various types of chemically modified electrodes. The practical utility of the new NACE-ED methodology is demonstrated in terms of the determination of traces of Hys in spiked environmental samples containing a wide range of explosives and related compounds.

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“…One of the most widely used fluorescent derivatization agents for primary and secondary amines in the biochemical fields is fluorescein isothiocyanate isomer I (FITC; Shintani and Polansky, 1997;Yu and Dovichi, 1989), which was first used for CE in 1988 (Cheng and Dovichi, 1988). Different CE-LIF determinations have been found in the literature using FITC as precolumn derivatization reagent to determine polyamines (Mattusch et al, 1995), biogenic amines (Nouadje et al, 1995a(Nouadje et al, ,b, 1996(Nouadje et al, , 1997Paez et al, 1996;Rodríguez et al, 1996) or some aliphatic amines in water samples (Brumley and Kelliher, 1997 We report here the separation and determination of the biogenic amine serotonin by CE-LIF prior to fluorescent derivatization reaction with FITC which, to the best of our knowledge, is the first report of derivatization of the amine to carry out fluorescent detection in capillary electrophoresis. Previous works dealing with CE demonstrate that serotonin can be analysed by this technique using other detection procedures such as electrochemical detection (Chen et al, 2001) or indirect chemiluminescence (Tsai and Whang, 1999).…”

Section: Introductionmentioning

confidence: 98%

Subminute and sensitive determination of the neurotransmitter serotonin in urine by capillary electrophoresis with laser‐induced ﬂuorescence detection

Arráez-Román

Segura‐Carretero

Blanco

et al. 2003

Biomedical Chromatography

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In this work, a sub-minute and sensitive capillary electrophoresis with laser-induced fluorescence (CE-LIF) method was developed for the analysis and quantitation of the neurotransmitter 5-hydroxytryptamine (5-HT) or serotonin in urine. The method involves precolumn derivatization with fluorescein isothiocyanate isomer I (FITC) using an excitation light from an argon ion laser of 488 nm and a 520 nm band pass emission filter. Different variables that affect derivatization (pH, FITC concentration, reaction time and temperature) and separation (buffer concentration, pH, applied voltage and injection time) were studied. The linear dynamic range obtained was between 0 and 188 nM with a detection limit of 16 nm with a RSD between 2 and 9%. The applicability of the proposed method was demonstrated by analysis of 5-HT in human urine, establishing a concentration of 57 nM in control urine. The method was validated by standard-addition methodology.

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Sensitive Laser induced fluorescence detection of Polyamine-Fluoresceinisothiocyanate-derivatives after capillary zone electrophoretic separation

Cited by 17 publications

References 30 publications

Simultaneous determination of polyamines and catecholamines in PC‐12 tumor cell extracts by capillary electrophoresis with laser‐induced fluorescence detection

Simultaneous determination of polyamines and catecholamines in PC‐12 tumor cell extracts by capillary electrophoresis with laser‐induced fluorescence detection

Determination of hydrazine, monomethylhydrazine, 1,1‐dimethylhydrazine, and 1,2‐dimethylhydrazine by nonaqueous capillary electrophoresis with amperometric detection

Subminute and sensitive determination of the neurotransmitter serotonin in urine by capillary electrophoresis with laser‐induced ﬂuorescence detection

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