Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Fluorescamine”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 91 records · Page 5Linked to original sources
PubMed1977

Thin-layer chromatography of histidine, histamine and histidyl peptides at picomole level using a unique fluorogenic reaction with fluorescamine.

A thin-layer chromatography procedure has been devised for detecting histidine, histamine, histidyl peptides and related imidazole compounds; it is based on a unique fluorogenic reaction, in which the compounds are derivatized with fluorescamine, converted into different fluorescent products by heating in strong acid medium, and separated on silica gel plates with an appropriate solvent system. As little as 4 to 60 pmole of the histidine derivative could be detected under long-wave (366 nm) ultraviolet radiation. Other classes of compounds gave no fluorescent spots.

Chemical Phenomena↗
PubMed1978

Detection and determination of N-nitrosamino acids by thin-layer chromatography using fluorescamine.

A novel procedure is described for the detection and determination of N-nitrosamino acids (NAAs) on activated silica gel thin-layer chromatographic plates. N-Nitrososarcosine, N-nitrosoproline, and N-nitroso-4-hydroxyproline could be detected as fluorophors at the 200-pmole level (20-30 ng) after being irradiated with ultraviolet light and sprayed with fluorescamine reagent. Spectrophotometric determination of the relative fluorescence of 0.4-40 nmoles of NAAs gave rise to similar calibration curves when plotted on a log-log scale. An application of this method to the detection of NAAs in uncooked bacon is described.

Amino Acids↗
PubMed1976

Detection and determination of N-nitrosamines by thin-layer chromatography using fluorescamine.

A novel procedure is described for the detection and determination of N-nitrosamines (NAs) on thin-layer chromatographic plates. Ultraviolet irradiation of NAs on activated plates (silica gel or aluminum oxide) yields primary or secondary amines, which after spraying with fluorescamine reagent give fluorescent or non-fluorescent products, respectively. The majority of the 24 NAs examined afforded NAs, although the volatile dimethyl-, diethyl- and pyrrolidine-N-nitrosamines gave limits of 500, 40, and 40 ng, respectively. Spectrophotometric determinations of the relative fluorescence of 0.1-40 nmoles of NAs gave rise to nearly linear calibration curves when plotted on a log-log scale.

Chromatography, Thin Layer↗
PubMed1975

Fluorimetric assay of dopamine, norepinephrine and their 3-O-methyl metabolites by using fluorescamine.

Fluorescamine was subjected to reaction with dopamine and norepinephrine (catecholamines) and with 3-methoxytyramine and normetanephrine (3-methyl metabolites of catecholamines) in phosphate or borate buffer. Catecholamines gave the highest fluorescent intensity at pH 8.0 in phosphate buffer but lower fluorescence in borate buffer. The fluorophores produced in phosphate or borate buffer were the same but the fluorescence intensities were suppressed in borate buffer. The dopamine and norepinephrine fluorophores were separated by high-pressure liquid chromatography on Hitachi 3011 gel with methanol-0.10 M Tris buffer of pH 8.0 (7:3). They were measurable at the 100-pmole level. The metabolites were also measurable by the same chromatography. By using methanol-0.15 M borate buffer of pH 8.0, cate-chol-O-methyltransferase activity might be assayed.

Boric Acids↗
PubMed1990

Determination of the monoamine oxidase B inhibitor Ro 19-6327 in plasma by high-performance liquid chromatography using precolumn derivatization with fluorescamine and fluorescence detection.

A specific high-performance liquid chromatographic (HPLC) method using precolumn derivatization and fluorescence detection was developed for the determination of the monoamine oxidase B inhibitor Ro 19-6327 in human plasma. After extraction of the basified plasma with tert.-butyl methyl ether-1-butanol (8:2, v/v) and back-extraction into dilute phosphoric acid, the solution was neutralized with phosphate buffer and the drug derivatized with fluorescamine. The derivative was chromatographed on a reversed-phase C8 column, using phosphate buffer-acetonitrile (68:32, v/v) as mobile phase, and fluorescence detection (excitation 370 nm, emission 485 nm). The limit of quantification was 1 ng/ml using 1 ml of plasma. The recovery was 79% in the range 5-200 ng/ml and the inter-assay precision was 3.1-7.9% in the range 2-500 ng/ml. The compound proved to be stable in human plasma. Moderate instability was found in rat plasma and, surprisingly, severe instability in dog plasma. Measures for handling unstable dog plasma samples are described. An HPLC method with UV detection was used for the analysis of dog and rat plasma samples, which is also described briefly. The fluorescence method, which was five times more sensitive than the UV method, was successfully applied to a human tolerance study.

Chromatography, High Pressure Liquid↗
PubMed1991

Determination of 3-methylhistidine in hydrolysed proteins by fluorescamine derivatization and high-performance liquid chromatography.

A method is described for the determination of the 3-methylhistidine content in myofibrilar proteins (myosin and actin) using reversed-phase high-performance liquid chromatography with ultraviolet detection. Proteins were hydrolysed and free amino acids were derivatized with fluorescamine. Elution was performed isocratically with acetonitrile-water (24:76). This method allows the detection of picomole amounts of 3-methylhistidine in myosin and actin.

Animals↗
PubMed2002

Spectrofluorimetric determination of vigabatrin and gabapentin in urine and dosage forms through derivatization with fluorescamine.

A stability-indicating, sensitive, simple and selective spectrofluorimetric method was developed for the determination of vigabatrin (VG) and gabapentin (GB). The method is based on the reaction between the two drugs and fluorescamine in borate buffer of pH 8.2 to give highly fluorescent derivatives that are measured at 472 nm using an excitation wavelength of 390 nm for both drugs. The optimum conditions were ascertained and the method was applied for the determination of VG and GB over the concentration range of 0.20-4.00 and 0.1-1.0 microg/ml, respectively with detection limits of 0.05 microg/ml (2.9 x 10(-7) M) and 0.06 microg/ml (2.3 x 10(-7) M) for VG and GB, respectively. The suggested method was applied, without any interference from the excipients, to the determination of the two drugs in their pharmaceutical formulations. Furthermore, the method was extended to the in-vitro determination of both drugs in spiked human urine. Interference from endogenous amino acids could be eliminated through selective complexation with copper acetate, the % recovery (n=4) is 98.0 +/- 7.05. Co-administered drugs such as lamotrigine, phenobarbitone, valproic acid, clopazam, carbamazepine, clonazepam and cimitidine did not interfere with the assay. The method is also stability-indicating; as the degradation product of vigabatrin: 5-vinylpyrrolidin-2-one, produced no interference with its analysis.

Acetates↗
PubMed1999

Fluorimetric determination of procaine in pharmaceutical preparations based on its reaction with fluorescamine.

A simple spectrofluorimetric analysis of a local anaesthetic named procaine using a specific labelling reagent for primary amino groups, has been developed. Because procaine shows very weak native fluorescence, the technique of spectrofluorimetry has been very much limited for its determination. A detail study of the variables affecting the derivatisation reaction (pH, fluorescamine (FC) concentration, temperature, reaction time), have minuciously been studied. The minimum detectable quantity is estimated as 7.7 ng ml(-1), with a relative standard deviation of 2.16% (ten determinations) for a procaine concentration of 100 ng ml(-1). The present method can be employed for the analysis of procaine by direct fluorescence measurements, without the interference from other compounds. The applicability of the present methodology have been demonstrated analysing three pharmaceutical preparations containing the analyte with satisfactory results.

Anesthetics, Local↗
PubMed1996

Combined liquid chromatography/mass spectrometry of the radical adducts of a fluorescamine-derivatized nitroxide.

Dynamic liquid secondary ion mass spectrometry (dyn-LSIMS) was employed to acquire continuous, on-line mass spectral data from the effluent of a reversed-phase high-performance liquid chromatograph (HPLC) used to separate a broad suite of carbon-centered radicals trapped as the O-alkylhydroxylamine adducts of an amino nitroxide that was subsequently derivatized with fluorescamine. Data obtained by the use of these combined techniques (LC/MS) can be employed to elucidate radical adduct structures; elemental compositions of the adducts can be confirmed by acquiring mass spectra at high resolution. At low resolution, introduction into the source of < 1 pmol of adduct yielded usable spectra. The first application of this technique to the identification of photochemically generated radicals in natural water samples is presented.

Chromatography, Liquid↗
PubMed1979

A fluorescamine-based sensitive method for the assay of proteinases, capable of detecting the initial cleavage steps of a protein.

The properties of the reaction of fluorescamine with proteins are the basis for the development of a sensitive, general and simple method for the assay of proteolytic activities. More importantly, the assay measures the initial step(s) of proteolytic attack, making it specially suitable for the examination of the controlling factors that regulate proteolytic degradation and/or the detection of 'specific' proteinases. The method allows the simple determination of the general parameters of enzyme action, V and Km, using proteins, i.e. the physiological substrates of the proteinases. The more appropriate proteins to be used as substrates are the N-amino-terminally blocked ones. Many proteins fulfill this requirement. If the particular protein whose degradation has to be studied lacks this modification, three different approaches can be used to study its degradation: (a) the accumulation of N-amino termini in excess over that of the intact substrate; (b) a gel filtration/continuous method and (c) the chemical blockage of its amino groups. The particular advantages of each of these approaches are discussed.

Endopeptidases↗
PubMed1989

Determination of cephalosporin-C amidohydrolase activity with fluorescamine.

A spectrophotometric procedure for the assay of cephalosporin-C amidohydrolase activity, based on the determination of the 7-aminocephalosporanic acid (7-ACA) produced in the hydrolysis of cephalosporin-C by the enzyme, is described. This procedure can be used to detect 7-ACA over a range of 10 to 200 micrograms mL-1. The same method can be used as a fluorometric procedures with a 100-fold greater sensitivity. At pH 4.5 7-ACA produces a strong fluorophor with fluorescamine, detectable spectrophotometrically at 378 nm and fluorometrically at an excitation of 378 nm and emission of 495 nm. At this pH the fluorophors formed with cephalosporin-C, proteins and aminoadipic acid present minimal absorbance values. The conditions for maximal detection of 7-ACA in the presence of proteins, cephalosporin-C and aminoadipic acid have been determined.

Amidohydrolases↗
PubMedDate not supplied

Release of amino acids, fluorescamine-reactive substances and substance P from the epidermis of the living animal.

The present study demonstrates a procedure for extraction and determination of stratum corneum amines in the living animal. A nonleaky well, containing 10 mM Tris-HCl buffer, pH 7.0, was constructed on the shaved backs of anesthetized animals. It was found that Ser, Ala, Gly and Pro are mainly released from the stratum corneum of 4-month-old guinea pigs, and in 2-month-old rats, Gly, Ser and Arg show the highest degree of release. Much lower amino acid concentrations were observed in 20-month-old rats. This was also reflected by the high levels of fluorescamine-reactive substances released from young rat skin as compared to the old animals. The release of the neuropeptide substance P into the aqueous medium was increased 3.2 times upon heat stimulus as compared to control skin. Amines and other compounds released from the skin may serve as markers for skin aging or for certain skin disorders, leading to a new approach for their treatments.

Aging↗
PubMed1976

Fluorometry of plasma amino nitrogen, with use of fluorescamine.

We describe a fluorometric procedure, with use of fluorescamine, for determining plasma amino nitrogen in a 10-mul sample of deproteinized plasma. Values for apparently healthy human subjects agree well with those obtained by spectrophotometry. The simplicity and speed of the assay recommends it as a screening procedure for detecting aminoacidopathies.

Adult↗
PubMed1986

Use of a rapid and highly sensitive fluorescamine-based procedure for the assay of plasma lipoproteins.

A rapid and sensitive method for determining protein concentrations using fluorescamine has been characterized for use in the analysis of intact lipoproteins. It was shown that there is no interference with the assay due to the presence of lipid-associated turbidity or primary amine content. The assay was shown to be sensitive to as little as 0.3 microgram of lipoprotein and to yield similar results when compared to the Lowry method.

Colorimetry↗
PubMedDate not supplied

[Determination of primary amines and protein by using fluorescamine in microorganism suspensions].

The paper describes a fluorimetric technique for determining the overall amount of primary amines and protein using fluorescamine directly in suspensions of yeast and bacterial microorganisms. The optimal conditions are selected for the reaction. The technique is highly sensitive and has an advantage over biochemical methods when one has to make many assays of amine containing compounds in biological material. The technique can find wide application in industrial practice.

Amines↗
PubMed1986

Determination of a prodrug of tranexamic acid in whole blood by reversed-phase liquid chromatography after pre-column derivatization with fluorescamine.

A sensitive high-performance liquid chromatographic (HPLC) method for a prodrug of tranexamic acid (KABI 2161) in whole blood is described. Since KABI 2161 is rapidly hydrolysed in whole blood the samples are collected directly into the extraction tubes and extracted immediately. After pre-column derivatization with fluorescamine the derivatives are analysed by reversed-phase liquid chromatography on a C(8)-Nucleosil column using an eluent mixture of phosphate buffer and acetonitrile (pH 3). The eluent is monitored by a fluorescence detector. Determinations as low as 10 ng ml(-1) of KABI 2161 in whole blood can be made when 0.5 ml blood is analysed. The precision of the method is 4.1% (RSD) at the 300 ng ml(-1) level and 6.9% (RSD) at the 50 ng ml(-1) level.

Journal Article↗
PubMed2004

Determination of 4-amino-m-cresol and 5-amino-o-cresol by high performance liquid chromatography and fluorescence derivatization using fluorescamine.

For the determination of two oxidation hair dyes, 4-amino-m-cresol (4-AC) and 5-amino-o-cresol (5-AC), a sensitive isocratic high performance liquid chromatography (HPLC) method using the reversed phase mode was developed. The hair dyes were pre-column derivatized with fluorescamine prior to injection. Sensitivity could be improved 10-fold for 4-AC and 50-fold for 5-AC by fluorescence detection compared to UV detection. The limit of detection was 1 ng/injection for 4-AC and 100 pg/injection for 5-AC, respectively. For the determination of both compounds in aqueous biological matrices in order to simulate conditions for penetration studies with pig skin, a solid phase extraction procedure using C18 cartridges and acetonitrile (ACN) for elution could be developed. Average recovery was 83.4% with a coefficient of variation (CV) of 2.64% for intra-day assay and 3.20% for inter-day assay for 5-AC and 2.89% and 3.41% for 4-AC, respectively.

Animals↗
PubMed1982

Simultaneous analyses of phenethylamine, phenylethanolamine, tyramine and octopamine in rat brain using fluorescamine.

A fluorometric method for the simultaneous analyses of phenethylamine, phenylethanolamine, tyramine and octopamine has been developed. The method involves ion-exchange chromatography, derivatization with fluorescamine, solvent extraction and then separation by thin-layer chromatography. The fluorescent spots are then quantitated by scanning. The detection limits of this method are about 10 pmoles for phenethylamine, phenylethanolamine and tyramine, and 20 pmoles for octopamine. The method was used for simultaneous analyses of putative neurotransmitter amines in whole rat brain.

Animals↗