Sphingosine mediates the immediate negative inotropic effects of tumor necrosis factor-alpha in the adult mammalian cardiac myocyte.
AuthorsOral H,Dorn GW 2nd,Mann DL
JournalThe Journal of biological chemistry
PubMed ID9030540
Rapid and sensitive method for the determination of arterial-venous differences and leg efflux of 3-methylhistidine using ion-pair high-performance liquid chromatography and post-column fluorescence derivatization.
AuthorsAndersson E, Håkanson E, Larsson J, Mårtensson J
JournalJ Chromatogr
PubMed ID3571381
A fluorometric assay for glutathione.
AuthorsCohn VH, Lyle J
JournalAnal Biochem
PubMed ID5944947
Evaluation of methods for measuring cellular glutathione content using flow cytometry.
AuthorsHedley DW, Chow S
JournalCytometry
PubMed ID8026225
'The currently available flow cytometric stains for cellular glutathione were evaluated, examining the labelling of both human and rodent cell lines under various conditions of concentration, time, and temperature. Procedures were used that depleted glutathione (GSH) while having a minimal effect on other cellular sulphydryls in order to estimate linearity ... More
Derivatization in capillary electrophoresis.
AuthorsBardelmeijer HA, Lingeman H, de Ruiter C, Underberg WJ
JournalJ Chromatogr A
PubMed ID9646486
'In recent years capillary electrophoresis (CE) has been developed into a versatile separation technique, next to gas and liquid chromatography (LC), well suited for the determination of a wide variety of e.g., pharmaceutical, biomedical and environmental samples. The main advantages of CE over chromatographic separation techniques are its simplicity and ... More
Determination of arginine, asymmetric dimethylarginine, and symmetric dimethylarginine in human plasma and other biological samples by high-performance liquid chromatography.
AuthorsTeerlink T, Nijveldt RJ, de Jong S, van Leeuwen PA
JournalAnal Biochem
PubMed ID11950212
'Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase (NOS), may be related to reduced biosynthesis of nitric oxide in diseases associated with accelerated atherosclerosis. The closely related compound symmetric dimethylarginine (SDMA) does not inhibit NOS, but may compete with arginine for cellular uptake, thereby limiting substrate availability for ... More
Appraisal of four pre-column derivatization methods for the high-performance liquid chromatographic determination of free amino acids in biological materials.
AuthorsFürst P, Pollack L, Graser TA, Godel H, Stehle P
JournalJ Chromatogr
PubMed ID2324214
'Reversed-phase high-performance liquid chromatography (RP-HPLC) is a powerful method for assaying physiological amino acid concentrations in biological fluids. Four pre-column derivatization methods, with o-phthaldialdehyde (OPA), 9-fluorenylmethyl chloroformate (FMOC-Cl), phenyl isothiocyanate (PITC) and 1-dimethylaminonaphthalene-5-sulphonyl chloride (dansyl-Cl), were assessed with respect to their applicability in biological research. The methods permit the measurement ... More
Glutathione measurement by high-performance liquid chromatography separation and fluorometric detection of the glutathione-orthophthalaldehyde adduct.
AuthorsNeuschwander-Tetri BA, Roll FJ
JournalAnal Biochem
PubMed ID2774172
'Glutathione reacts with orthophthalaldehyde to form a stable, highly fluorescent tricyclic derivative which is easily separated and quantitated by high-performance liquid chromatography. Separation of the glutathione adduct is achieved by isocratic elution over a reverse-phase column with 7.5% methanol/92.5% 0.15 M sodium acetate, pH 7.00. The adduct is detected fluorometrically ... More
Synthesis and characterization of a new fluorescent phospholipid.
AuthorsChang BC, Huang L
JournalBiochim Biophys Acta
PubMed ID476119
'A novel fluorescent phospholipid, whose structure was tentatively assigned as 1-(2''-thio-1''-hydroxyethyl)-2-(ethylphosphatidyl)isoindole), was synthesized by reacting O-phthalaldehyde and beta-mercaptoethanol with phosphatidylethanolamine. The fluorescent lipid product was purified by silicic acid chromatography. The purity was demonstrated by thin-layer chromatography. This fluorescent phospholipid could not form stable lipid vesicles. However, a mixture of ... More
Inactivation of yeast hexokinase by o-phthalaldehyde: evidence for the presence of a cysteine and a lysine at or near the active site.
AuthorsPuri RN, Bhatnagar D, Roskoski R
JournalBiochim Biophys Acta
PubMed ID3140897
'Yeast hexokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1), a homodimer, was rapidly and irreversibly inactivated by o-phthalaldehyde at 25 degrees C (pH 7.3). The reaction followed pseudo-first-order kinetics over a wide range of the inhibitor concentration. The second-order-rate constant for the inactivation of hexokinase was estimated to be 45 M-1.s-1. Hexokinase was ... More
Catalytic mechanism and molecular recognition of E. coli UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase probed by mutagenesis.
AuthorsHernick M, Fierke CA
JournalBiochemistry
PubMed ID17176046
'UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase (LpxC) is a metal-dependent deacetylase that catalyzes the hydrolysis of UDP-3-O-myristoyl-N-acetyl-glucosamine to form UDP-3-O-myristoyl-glucosamine and acetate. This is the committed step in the biosynthesis of lipid A, and therefore, LpxC is a target for the development of antimicrobial agents in the treatment of Gram-negative infections. To facilitate the ... More
Neuronal nitric oxide synthase self-inactivates by forming a ferrous-nitrosyl complex during aerobic catalysis.
AuthorsAbu-Soud HM, Wang J, Rousseau DL, Fukuto JM, Ignarro LJ, Stuehr DJ
JournalJ Biol Chem
PubMed ID7559438
'Neuronal NO synthase (NOS) is a flavin-containing hemeprotein that generates NO from L-arginine, NADPH, and O2. NO has recently been proposed to autoinhibit NOS. We have investigated whether a NOS heme-NO complex forms during aerobic steady-state catalysis. Visible and resonance Raman spectra recorded during steady-state NO synthesis by NOS showed ... More
RP-HPLC tryptic mapping of IgG1 proteins with post-column fluorescence derivatization.
AuthorsGulati D, Bongers J, Burman S
JournalJ Pharm Biomed Anal
PubMed ID10703956
'Peptide mapping is an important analytical technique widely used to study the primary structure of proteins. In quality control settings, it is employed as an identity test to probe for small changes in protein primary structure. A great challenge in peptide mapping is to minimize the detection limit for peptides ... More
Observations of the use of o-phthalaldehyde condensation for the measurement of histamine.
AuthorsSiegel PD, Lewis DM, Petersen M, Olenchock SA
JournalAnalyst
PubMed ID2256552
'The in vitro study of mast cell degranulation utilises the measurement of histamine as a quantitative marker of this process. Histamine is most commonly assayed, following organic extraction, by condensing it with o-phthalaldehyde (OPT) and thereby obtaining a highly fluorescent adduct. A number of variables that might affect the performance ... More
Evaluation of a liquid chromatographic method for the determination of fumonisins in corn, poultry feed, and Fusarium culture material.
AuthorsRice LG, Ross PF, Dejong J, Plattner RD, Coats JR
JournalJ AOAC Int
PubMed ID7580311
'The performance of a liquid chromatographic method for determining fumonisins in corn, animal feeds, and culture material was evaluated. Efficiencies of extractions with the following solvent systems were determined: acetonitrile-water (50 + 50, v/v), methanol-water (75 + 25, v/v), and 100% water. The acetonitrile solvent gave both higher extraction efficiencies ... More
Liquid chromatographic determination of D- and L-amino acids by derivatization with o-phthaldialdehyde and N-isobutyryl-L-cysteine. Applications with reference to the analysis of peptidic antibiotics, toxins, drugs and pharmaceutically used amino acids.
AuthorsBrückner H, Westhauser T, Godel H
JournalJ Chromatogr A
PubMed ID7496491
'In order to evaluate and extend the applicability of an analytical method that enables the quantitative and simultaneous high-performance liquid chromatographic determination of D- and L-amino acids (DL-AAs) by automated precolumn derivatization with o-phthaldialdehyde together with the chiral thiol N-isobutyryl-L-cysteine [J. Chromatogr., 666 (1994) 259] selected natural and synthetic bioactive ... More
Hydrogen peroxide-supported oxidation of NG-hydroxy-L-arginine by nitric oxide synthase.
AuthorsPufahl RA, Wishnok JS, Marletta MA
JournalBiochemistry
PubMed ID7531495
'The ability of murine macrophage nitric oxide synthase (NOS) to utilize peroxides in place of O2 and NADPH was investigated using hydrogen peroxide (H2O2), tert-butylhydroperoxide, and cumene hydroperoxide with both L-arginine and NG-hydroxy-L-arginine (L-NHA) as substrates. Of the three peroxides examined, only H2O2 was able to support product formation using ... More
Rapid method for micro-analysis of endogenous amino acid neurotransmitters in brain perfusates in the rat by isocratic HPLC-EC.
AuthorsPeinado JM, McManus KT, Myers RD
JournalJ Neurosci Methods
PubMed ID2879077
'A simplified method is described for the isocratic analysis of endogenous amino acid neurotransmitters contained in brain perfusates by high performance liquid chromatography (HPLC) with electrochemical detection (EC). Pre-column o-phthalaldehyde (OPA) tert-butylthiol derivatives of the amino acids were injected into a C18 3 microns column. After linear concentration curves for ... More
Quantitative determination of amino acid levels in neutral and glucosamine-containing carbohydrate polymers.
AuthorsPalace GP, Phoebe CH
JournalAnal Biochem
PubMed ID9025957
'Low-level (subanomole) determination of amino acids in samples of naturally derived polymeric carbohydrates has been demonstrated using vapor-phase acid hydrolysis and subsequent precolumn derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Application has been demonstrated for neutral polysaccharide polymers such as laminarin (beta-1,3; branched), curdland (beta-1,3; unbranched), pullulan (alpha-1, 6-maltotriose), glycogen (alpha-1,4-glucan), and inulin ... More
Formation of N delta-cyanoornithine from NG-hydroxy-L-arginine and hydrogen peroxide by neuronal nitric oxide synthase: implications for mechanism.
AuthorsClague MJ, Wishnok JS, Marletta MA
JournalBiochemistry
PubMed ID9398165
'Neuronal nitric oxide synthase (nNOS) catalyzes the oxidation of NG-hydroxy-L-arginine (NHA) by hydrogen peroxide. The amino acid products were characterized by high-performance liquid chromatography/mass spectrometry of the o-phthalaldehyde/2-mercaptoethanol derivatives and identified as citrulline and N delta-cyanoornithine (CN-orn). The assignment of CN-orn was confirmed by independent chemical synthesis and comparison of ... More
Stability of o-phthalaldehyde-sulfite derivatives of amino acids and their methyl esters: electrochemical and chromatographic properties.
AuthorsTuriák G, Volicer L
JournalJ Chromatogr A
PubMed ID8032487
'RP-HPLC coupled with 16-channel coulometric electrode array detection was used to monitor the decomposition of five amino acid o-phthalaldehyde (OPA)-sulfite derivatives (Ala, Arg, Glu, Ser, Tyr) and their methyl ester derivatives as well. At fixed OPA and sulfite concentrations inclusion of methanol and EDTA in the derivatization media has increased ... More
Influences of metal ions on the reaction of amino and imino acids with fluorogenic reagents.
AuthorsMiyano H, Toyo'oka T, Imai K, Nakajima T
JournalAnal Biochem
PubMed ID3936376
'When a twofold excess of metal ions (Cu2+, Ni2+, Zn2+, Mg2+, Ca2+, Fe3+, and Al3+) was added to a reaction medium of amino acids (1-50 microM each) with the fluorogenic reagents NBD-F (4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole), DNS-Cl (5-N,N-dimethylaminonaphthalenesulfonyl chloride), OPA (o-phthalaldehyde), and fluorescamine (4-phenylspiro[furan-2(3H),1''-phthalan]-3,3''-dione), a reduction of derivatization was observed. The effect was ... More
Determination of reduced disulfide groups in monoclonal antibodies.
AuthorsThakur ML, DeFulvio JD
JournalBiotechniques
PubMed ID2357374
'Reduction of disulfide bonds to sulfhydryl groups for direct radiolabeling of antibodies for immunoscintigraphic and therapeutic applications continues to be of considerable interest. Sensitive spectrophotometric methods have been evaluated that will enable investigators to determine submicrogram quantities of cysteine units produced, for the assurance of controlled reduction. One method, which ... More
Inactivation of guanosine cyclic 3',5'-monophosphate dependent protein kinase from bovine lung by o-phthalaldehyde.
AuthorsPuri RN, Bhatnagar D, Glass DB, Roskoski R
JournalBiochemistry
PubMed ID3002444
'Guanosine cyclic 3'',5''-monophosphate (cGMP) dependent protein kinase is inactivated by o-phthalaldehyde. The loss of phosphotransferase activity following treatment with o-phthalaldehyde was rapid, and the second-order rate constant at 25 degrees C and pH 7.3 was 35 M-1 s-1. The inactivation reaction did not follow saturation kinetics. The cGMP-dependent protein kinase ... More
Flow cytometric assays of anticancer drug resistance.
AuthorsHedley DW
JournalAnn N Y Acad Sci
PubMed ID8098593
Derivatization of thiol-containing compounds.
AuthorsShimada K, Mitamura K
JournalJ Chromatogr B Biomed Appl
PubMed ID7820279
'The determination of thiol-containing compounds in biological fluids is important in biochemistry and clinical chemistry. In this paper, derivatization reagents for thiols are reviewed with respect to their reactivity, selectivity, spectroscopic characteristics and their applicability especially to high-performance liquid chromatography. Derivatization used in ultraviolet and electrochemical detection. The derivatization reagents ... More
Effect of an ectokinase inhibitor, K252b, on degranulation and Ca2+ signals of RBL-2H3 cells and human basophils.
AuthorsTeshima R, Saito Y, Ikebuchi H, Rajiva De Silva N, Morita Y, Nakanishi M, Sawada J, Kitani S
JournalJ Immunol
PubMed ID9218617
'We examined the effects of K252b, an ectoprotein kinase inhibitor of microbial origin, on the activation process of RBL-2H3 cells by cross-linking of IgE receptors by the endoplasmic reticulum Ca2+-ATPase inhibitor 2,5-di(tert-butyl)-1,4-hydroquinone or by the Ca2+ ionophore A23187. Analysis of phosphorylation of ectoproteins following IgE receptor cross-linking revealed that K252b ... More
A fluorometric microassay for histamine release from human gingival mast cells.
AuthorsZhao ZZ, Sugerman PB, Walsh LJ, Savage NW
JournalJ Periodontal Res
PubMed ID11519696
'Mast cells are important effector cells of the immune system. We describe a rapid and inexpensive microassay to determine histamine release from human gingival mast cells. The assay is based on the coupling of histamine with o-phthalaldehyde (OPT) at a highly alkaline pH to form a fluorescent product. Using this ... More
Cigarette smoke and its formaldehyde component inhibit bradykinin-induced calcium increase in pig aortic endothelial cells.
AuthorsMazák I, Wittmann I, Wagner L, Wagner Z, Degrell P, Vas T, Molnár GA, Nagy J
JournalEndothelium
PubMed ID12200957
'Bradykinin-induced increase in the intracellular concentration of free calcium evokes an activation of the endothelial nitric oxide synthase (eNOS) enzyme, producing nitric oxide (NO). Cigarette smoke inhibits the eNOS-NO-cGMP signaling pathway. The pathomechanism of this deleterious effect of smoke on NO production is unknown. The aim of this study was ... More
o-phthaldehyde fluorescence microassay for the determination of antibody concentration.
AuthorsLi X, Abdi K, Mentzer SJ
JournalJ Immunol Methods
PubMed ID8034969
'o-Phthaldehyde (OPA) reagent reacts with primary amines in aqueous solution at room temperature. When the reaction occurs in the presence of 2-mercaptoethanol, a bright blue fluorescence is produced. We investigated the application of the OPA reagent in the routine determination of monoclonal antibody concentration. The OPA reagent produced maximal fluorescence ... More
High-performance liquid chromatography of histamine and 1-methylhistamine with on-column fluorescence derivatization.
AuthorsSaito K, Horie M, Nose N, Nakagomi K, Nakazawa H
JournalJ Chromatogr
PubMed ID1374421
'An on-column fluorometric derivatization method was developed for the determination of histamine and 1-methylhistamine (HMs) by high-performance liquid chromatography. The system for the derivatization consisted only of a commercially available single-plunger pump and a reversed-phase C18 column supported on synthetic polymer with a mobile phase of acetonitrile and alkaline borate ... More
A fluorescent assay for the determination of UDP-GlcNAc: Gal beta 1,3GalNAc-R (GlcNAc to GalNAc) beta 1,6-N-acetylglucosaminyltransferase activity.
AuthorsPalmerini CA, Datti A, Alunni S, VanderElst IE, Orlacchio A
JournalAnal Biochem
PubMed ID7762797
'A fluorescent assay for UDP-GlcNAc: Gal beta 1,3Gal-NAc-R beta 1,6-N-acetylglucosaminyltransferase (core 2 GlcNAc-T) activity has been developed involving dansylation of the enzyme reaction product. Core 2 GlcNAc-T detection was performed using unlabeled UDP-GlcNAc as the donor and Gal beta 1,3GalNAc alpha-pAp as the acceptor. The product, Gal beta 1,3(GlcNAc beta ... More
HPLC with o-phthalaldehyde precolumn derivatization to measure total, oxidized, and protein-bound glutathione in blood, plasma, and tissue.
AuthorsParoni R, De Vecchi E, Cighetti G, Arcelloni C, Fermo I, Grossi A, Bonini P
JournalClin Chem
PubMed ID7882521
'This HPLC assay with o-phthalaldehyde precolumn derivatization is used to measure the total, oxidized, and protein-bound forms of glutathione in human blood, plasma, and rat tissue. Total glutathione (i.e., sum of reduced, oxidized, and protein-bound fractions) was determined after reduction with dithiothreitol and protein precipitation with perchloric acid (PCA). A ... More
Realities of automating OPA HPLC amino acid analyses.
AuthorsKrok KA, Seaver SS
JournalBiotechniques
PubMed ID1910784
'The levels of free amino acids in culture medium can easily be monitored by reversed-phase HPLC chromatography on a C18 column following pre-column derivatization with o-pthaladehyde. The method does not require the prior removal of proteins. A guard column does that. When this method was automated, problems started to occur ... More
Kinetic centrifugal analyzer and manual determination of serum urea nitrogen, with use of o-phthaldialdehyde reagent.
AuthorsLevinson SS
JournalClin Chem
PubMed ID719870
'o-Phthaldialdehdye reagent was used in assaying urea with a miniature centrifugal analyzer. Seventeen samples can be assayed kinetically in 2 min at a current cost of less than a penny per test. The reaction is initiated in the instrument upon simultaneous mixing of the o-phthaldialdehyde reagent containing 10 microliter of ... More
Difference between histidine and histamine in the mechanistic pathway of the fluorescence reaction with ortho-phthalaldehyde.
AuthorsYoshimura T, Kamataki T, Miura T
JournalAnal Biochem
PubMed ID2221352
'Histamine reacts with ortho-phthalaldehyde (OPA) in an alkaline medium to form an unstable fluorescent adduct (Fbase-Hm). Acidification of the solution to pH 2-4 gives a stable and highly fluorescent adduct (Facid-Hm). In contrast, histidine develops a relatively stable fluorescent adduct (Fbase-Hd) with OPA in an alkaline medium. Upon acidification of ... More
Separation strategies for o-phthalaldehyde-mercaptoethanol derivatives of amino acids for reversed-phase high-performance liquid chromatography.
AuthorsSchmidt J, McClain CJ
JournalJ Chromatogr
PubMed ID3667769
'Twenty-three o-phthalaldehyde-mercaptoethanol derivatives of primary amino acids in serum were separated with Waters C18 5-microns Radial-Pak Resolve columns, using aqueous phosphate-methanol mobile phase with acetate and tetrahydrofuran as modifiers. Resolution is critical in this system for glutamine/histidine, citrulline/glycine/threonine/3-methylhistidine, and tryptophan/methionine derivatives, and is affected by small changes in column properties ... More
Fluorescence properties of o-phthaldialdehyde derivatives of amino acids.
AuthorsChen RF, Scott C, Trepman E
JournalBiochim Biophys Acta
PubMed ID427201
'The fluorescence properties of the products formed by reaction of o-phthaldialdehyde with amino acids and their derivatives, in the presence of thiol compounds, have been studied. The emission spectra, quantum yields, and lifetimes depend on the primary amine and thiol compound used; the observations confirm the report (Simsons, S.S., Jr. ... More
Determination of sisomicin in eluate from dried blood spot on filter paper disc for monitoring of blood level in rat, by reversed-phase high-performance liquid chromatography after pre-column fluorimetric derivatization.
AuthorsFujimoto T, Tawa R, Hirose S
JournalChem Pharm Bull (Tokyo)
PubMed ID2720847
'About 20 microliters of whole blood obtained by venipuncture from rat tail vein was spotted onto a filter paper and the blood spot was punched out (5 mm diameter). Sisomicin (SISO) in the dried blood spot (DBS) was extracted effectively into 0.5 M Na2HPO4 solution by ultrasonication and determined by ... More
Simple and sensitive determination of plasma N tau-methylhistidine by high-performance liquid chromatography using pre-column derivative formation with o-phthalaldehyde-2-mercaptoethanol.
AuthorsNagasawa T, Sakai T, Onodera R
JournalJ Chromatogr
PubMed ID1885715
'A simple, rapid and sensitive assay method for plasma N tau-methylhistidine by isocratic high-performance liquid chromatography has been developed. The deproteinized plasma was treated with o-phthalaldehyde-2-mercaptoethanol. The derivatives were separated on a LiChrospher 100 RP-18 column within 10 min. The detection limit for N tau-methylhistidine was 0.5 pmol. The plasma ... More
High-performance liquid chromatographic determination of N-nitroso-N-alkylureas by pre-column fluorescence derivatization and application to blood analysis.
AuthorsSano A, Ohashi M, Kodama M, Takitani S
JournalJ Chromatogr
PubMed ID8294537
'A method for the derivatization and separation of N-nitroso-N-alkylureas [alkyl = methyl (NMU), ethyl (NEU), and n-butyl (NBU)] has been developed. Fluorescent derivatives were formed with sodium sulphide, taurine and o-phthalaldehyde and separated by reversed-phase high-performance liquid chromatography. The limits of detection of standard NMU, NEU and NBU were 0.25, ... More
Determination of homocysteine thiolactone and homocysteine in cell cultures using high-performance liquid chromatography with fluorescence detection.
AuthorsMukai Y, Togawa T, Suzuki T, Ohata K, Tanabe S
JournalJ Chromatogr B Analyt Technol Biomed Life Sci
PubMed ID11885855
'A sensitive and simple method utilising fluorometric detection for the simultaneous routine monitoring of homocysteine thiolactone (HTL) and homocysteine (Hcy) in biological samples has been developed. Separation relies on isocratic ion-pairing and reversed-phase chromatography while the principle of the detection is that the lactone ring in HTL molecule is cleaved ... More
Protein-heparin interactions measured by BIAcore 2000 are affected by the method of heparin immobilization.
AuthorsOsmond RI, Kett WC, Skett SE, Coombe DR
JournalAnal Biochem
PubMed ID12423639
'Surface plasmon resonance (SPR) biosensors such as the BIAcore 2000 are a useful tool for the analysis of protein-heparin interactions. Generally, biotinylated heparin is captured on a streptavidin-coated surface to create heparinized surfaces for subsequent binding analyses. In this study we investigated three commonly used techniques for the biotinylation of ... More
Water-soluble carbodiimide for the fluorescent measurement of the carboxyl group produced by enzyme reactions.
AuthorsKobayashi M, Chiba Y
JournalAnal Biochem
PubMed ID8080075
'Fluorescent measurement of the carboxyl group was achieved using water-soluble carbodiimide, EDC (1-ethyl-3-(3- dimethylaminopropyl)carbodiimide). Detection of the active intermediate from the glucuronic acid-EDC reaction mixture using the OPA (o-phthalaldehyde) reagent described in a previous paper was greatly improved to about 4 ng/tube (17 pmol) glucuronic acid owing to a modification ... More
Direct detection of endogenous histamine in rat peritoneal mast cells by in-capillary derivatization high-performance capillary electrophoresis.
AuthorsOguri S, Ohta Y, Suzuki C
JournalJ Chromatogr B Biomed Sci Appl
PubMed ID10677007
'A simple method for the detection of endogenous histamine in rat peritoneal mast cells was evaluated using on-line mode in-capillary derivatization high-performance capillary electrophoretic (ICD-HPCE) techniques, which were previously developed by our group [S. Oguri et al., J. Chromatogr. A, 787 (1997) 253-260]. The method involves a suspension of peritoneal ... More
An automated fluorescence assay for subnanogram quantities of protein in the presence of interfering material.
AuthorsHammer KD, Nagel K
JournalAnal Biochem
PubMed ID3728981
'An automated assay for measuring nanogram and subnanogram quantities of protein in microliter samples was developed with the fluorometric reagent omicron-phthaldialdehyde/mercaptoethanol. Low molecular weight interfering substances were separated within the analysis by gel filtration. The technique allowed measurement of biological fluids without any sample pretreatment. The method presented proved to ... More
Fluorimetric determination of pantothenic acid in foods by liquid chromatography with post-column derivatization.
AuthorsPakin C, Bergaentzlé M, Hubscher V, Aoudé-Werner D, Hasselmann C
JournalJ Chromatogr A
PubMed ID15117078
'A method to determine the content of free pantothenic acid in various foods by reverse phase liquid chromatography-fluorimetry is reported. It includes a purification of the samples by successive passages through anion and cation exchange cartridges and a post-column derivatization of pantothenic acid as the fluorescent 1-alkylthio-2-alkylisoindole (reaction of beta-alanin, ... More
An automated high-performance liquid chromatography procedure for the quantitation of L- and D-amino acids by means of stepwise precolumn derivatization.
AuthorsHardy GP, Van Hemert FJ, Meijer AJ, Goudsmit J
JournalAnal Biochem
PubMed ID11401304
A convenient spectrophotometric method for the kinetic analysis of the enzymatic hydrolysis of N-acyl peptides using phthaldialdehyde.
AuthorsRowlett R, Murphy J
JournalAnal Biochem
PubMed ID6789706
A fluorimetric assay for available lysine in proteins.
AuthorsGoodno CC, Swaisgood HE, Catignani GL
JournalAnal Biochem
PubMed ID6795961
Quantitative determination of phosphatidylethanolamine and phosphatidylserine in liposomes and on HPTLC plates with o-phthalaldehyde.
AuthorsZellmer S, Lasch J
JournalAnal Biochem
PubMed ID8053560
Fluorimetric determination of rat growth hormone in pituitary cell culture by high-performance liquid chromatography with post-column derivatization.
AuthorsNakazawa H
JournalJ Chromatogr
PubMed ID3654890
Fluorometric determination of sisomicin, an aminoglycoside antibiotic, in dried blood spots on filter paper by reversed-phase high-performance liquid chromatography with pre-column derivatization.
AuthorsFujimoto T, Tawa R, Hirose S
JournalChem Pharm Bull (Tokyo)
PubMed ID3416375
Fluorometric analysis of amino sugars and derivatized neutral sugars.
AuthorsPerini F, Peters BP
JournalAnal Biochem
PubMed ID6181711
Fluorescence analysis of picoliter samples.
AuthorsMroz EA, Lechene C
JournalAnal Biochem
PubMed ID7356167
Reverse-phase liquid chromatographic analysis of amino acids after reaction with o-phthalaldehyde.
AuthorsGardner WS, Miller WH
JournalAnal Biochem
PubMed ID7356137
Simultaneous determination of radioactivity and o-phthalaldehyde derived fluorescence in low levels of amino acids.
AuthorsBeecher GR, Dohm GL, Peacock I
JournalJ Chromatogr
PubMed ID546906
Fluorescent chemoaffinity labeling. Potential application of a new affinity labeling technique to glucocorticoid receptors.
AuthorsSimons SS, Thompson EB, Johnson DF
JournalBiochemistry
PubMed ID508723
o-Phthalaldehyde and the fluorogenic detection of peptides.
AuthorsJoys TM, Kim H
JournalAnal Biochem
PubMed ID464307
Amino acid analysis with o-phthalaldehyde detection: effects of reaction temperature and thiol on fluorescence yields.
AuthorsCronin JR, Pizzarello S, Gandy WE
JournalAnal Biochem
PubMed ID434461
Fluorimetric assay for prolinase and partial characterisation in cultured skin fibroblasts.
AuthorsButterworth J, Priestman D
JournalClin Chim Acta
PubMed ID7094345
Automated fluorometric amino acid analysis: the determination of nonprotein cyclic imino acids.
AuthorsBleecker AB, Romeo JT
JournalAnal Biochem
PubMed ID7103060
A rapid fluorometric assay for measurement of peptidase activity.
AuthorsPorter DH, Swaisgood HE, Catignani GL
JournalAnal Biochem
PubMed ID7051897
Fluorometric analysis of polyamines, histamine, and 1-methylhistamine.
AuthorsPerini F, Sadow JB, Hixson CV
JournalAnal Biochem
PubMed ID37783
Fluorometric quantitation of amino sugars in the picomole range.
AuthorsCarroll SF, Nelson DR
JournalAnal Biochem
PubMed ID44436
o-phthalaldehyde for the fluorometric assay of nonprotein amino compounds.
AuthorsReeder DJ, Sniegoski LT, Schaffer R
JournalAnal Biochem
PubMed ID26286
Measurement of protein thiol groups and glutathione in plasma.
AuthorsHu ML
JournalMethods Enzymol
PubMed ID8015473
The fluorescent staining of heparin in mast cells using berberine sulfate: compatibility with paraformaldehyde or o-phthalaldehyde induced fluorescence and metachromasia.
AuthorsDimlich RV, Meineke HA, Reilly FD, McCuskey RS
JournalStain Technol
PubMed ID6160648
Automated precolumn derivatization of amino acids, small peptides, brain amines and drugs with primary amino groups for reversed-phase high-performance liquid chromatography using naphthalenedialdehyde as the fluorogenic label.
AuthorsKoning H, Wolf H, Venema K, Korf J
JournalJ Chromatogr
PubMed ID2127935
A rapid and sensitive fluorescence assay for angiotensin-converting enzyme.
AuthorsConroy JM, Lai CY
JournalAnal Biochem
PubMed ID210690
Sensitive fluorescent determination of trypsin-like proteases.
AuthorsBrown F, Freedman ML, Troll W
JournalBiochem Biophys Res Commun
PubMed ID4795362
O-phthalaldehyde: fluorogenic detection of primary amines in the picomole range. Comparison with fluorescamine and ninhydrin.
AuthorsBenson JR, Hare PE
JournalProc Natl Acad Sci U S A
PubMed ID1054843
O-Phthalaldehyde, in the presence of 2-mercaptoethanol, reacts with primary amines to form highly fluorescent products. Picomole quantities of amino acids, peptides, and proteins can be detected easily. o-Phthalaldehyde is five to ten times more sensitive than fluorescamine and is soluble and stable in aqueous buffers. ... More
A comparison of fluorescamine and o-phthaldialdehyde as effective blocking reagents in protein sequence analyses by the Beckman sequencer.
AuthorsBhown AS, Cornelius TW, Volanakis JE, Bennett JC
JournalAnal Biochem
PubMed ID6614468
Use of o-phthaldialdehyde to chemically reduce the newly generated amino termini responsible for the progressively increasing background during an extended amino acid sequence analysis in a liquid phase sequencer has been described. The results have been compared with Fluram blocking using apomyoglobin and rabbit C-reactive protein as standard and unknown ... More
Derivatization of posttranslationally modified amino acids.
AuthorsTeerlink T
JournalJ Chromatogr B Biomed Appl
PubMed ID7820276
After a brief overview of posttranslational modifications of protein amino acids, the use of various derivatizing reagents for amino acid analysis is discussed. Derivatization and chromatographic separation of hydroxyproline, methylhistidine, and phosphorylated amino acids are discussed in detail to illustrate some of the strategies that can be applied to the ... More
Identification of the lysine residue involved in the inactivation of lamb liver 6-phosphogluconate dehydrogenase by fluorescein 5'-isothiocyanate.
AuthorsHanau S, Dallocchio F, Rippa M
JournalBiochem Int
PubMed ID1815497
Fluorescein 5'-isothiocyanate binds almost selectively at the active site of lamb liver NADP-dependent 6-phosphogluconate dehydrogenase causing the inactivation of the enzyme. The substrate and the coenzyme protect against the loss of catalytic activity. The enzyme derivative was digested with trypsin, the labelled peptide was isolated by h.p.l.c. and its amino ... More
Investigation of spectrin binding to phospholipid vesicles using isoindole fluorescent probe. Thermal properties of the bound and unbound protein.
AuthorsMichalak K, Bobrowska M, Sikorski AF
JournalGen Physiol Biophys
PubMed ID2079201
Fluorescence of isoindole probe covalently bound to spectrin from pig erythrocytes, and fluorescence of tryptophanyl residues were used to study spectrin interaction with phospholipid bilayers. Evidence would be provided for conformational changes of spectrin occurring upon its binding to lipid bilayers. Fluorescence quenching experiments allowed to determine thermal stability of ... More
Formation and instability of o-phthalaldehyde derivatives of amino acids.
AuthorsGarcía Alvarez-Coque MC, Medina Hernández MJ, Villanueva Camañas RM, Mongay Fernández C
JournalAnal Biochem
PubMed ID2658682
o-Phthalaldehyde reacts with amino acids in the presence of a thiol to give highly fluorescent 1-alkyl-thio-2-alkyl-substituted isoindoles. However, the instability of the derivatives limits the general utility of the reaction. Mechanistic descriptions of isoindole formation and degradation proposed in the last years, which permit a better understanding of the factors ... More
Recent advances in capillary electrophoresis of proteins, peptides and amino acids.
AuthorsNovotny MV, Cobb KA, Liu JP
JournalElectrophoresis
PubMed ID2257844
The current status of high-performance capillary electrophoresis as an analytical separation method for proteins, peptides and amino acids is assessed. Recent advances in suppressing the effects of electroosmotic flow and irreversible adsorption of proteins at the capillary wall are reviewed, together with procedures for optimal separations of peptides and amino ... More
Flow-cytometric determination of glutathione alterations in vital cells by o-phthaldialdehyde (OPT) staining.
AuthorsTreumer J, Valet G
JournalExp Cell Res
PubMed ID2420623
A flow-cytometric method for the detection of changes of cellular glutathione content in vital cells is described. The reaction is based on formation of a fluorescent product between o-phthaldialdehyde (OPT) and reduced glutathione (GSH). OPT is a more GSH-specific dye than other thiol-specific dyes (e.g., bromobimanes), because it forms a ... More
Determination of glutamate by high-performance liquid chromatography with fluorescence detection in superfusates of rat brain preparations.
AuthorsIoannou N, Georgopoulos A, Liapi C, Palaiologos G
JournalAnal Biochem
PubMed ID9784191
A method is described for the determination of glutamate in superfusates of cortical or hippocampal rat brain slices. This method is based on the precolumn derivatization of amino acids with orthopthaldialdehyde and mercaptopropionic acid, separation by high-performance liquid chromatography, and detection by fluorescence. By adjusting the concentrations of reagents and ... More
Fluorescent method for detection of cleaved collagens using O-phthaldialdehyde (OPA).
AuthorsGo K, Horikawa Y, Garcia R, Villarreal FJ,
JournalJ Biochem Biophys Methods
PubMed ID17573117
Analysis of collagen degradation remains an important but cumbersome task. Traditional methods with dansyl chloride derivatization of collagen have been used to quantify collagen damage. Fluorescent labeling reagents have been developed that offer advantages such as greater solubility in water and low background emission. One such reagent is o-phthalaldehyde (OPA). ... More
A high-performance liquid chromatographic assay for acid ceramidase activity in cultured fibroblasts from patients with Farber's disease and from controls.
AuthorsMitsuo K, Kobayashi T, Shinnoh N, Goto I
JournalClin Chim Acta
PubMed ID3383428
The high-performance liquid chromatographic (HPLC) method we devised for assay of acid ceramidase activity involves coupling of a fluorescent probe to the enzymatically released sphingosine in the reaction mixture and detection of the fluorescent sphingosine derivative by reverse-phase HPLC. Using the method, acid ceramidase activity in fibroblast homogenates was accurately ... More
High-performance liquid chromatographic method for the evaluation of possible interferences in basophil-histamine release measurements.
AuthorsSiegel PD, Lewis DM, Olenchock SA
JournalAnal Biochem
PubMed ID1699455
Basophil activation studies commonly rely on the measurement of histamine following extraction and condensation to o-phthalaldehyde (OPT) as a quantitative measurement of degranulation. Specificity has long been recognized as a problem with this method. We have described a new high-performance liquid chromatographic method that allows for both a qualitative and ... More
Quantitative determination of thiolipids in organic solution, in membranes, and on HPTLC plates.
AuthorsZellmer S, Lasch J, Rothe U
JournalJ Lipid Res
PubMed ID9392435
In the presence of ortho-phthalaldehyde and glucosamine, thiolipids form fluorescent isoindole derivatives. This reaction can be used to quantify single- and double-chain mercaptans in membranes (liposomes) and micellar solutions. The lower detection limit is 100 pmol. In addition, the assay allows the detection of 1.9 nmol thiolipids on HPTLC plates ... More
Separation of tetrodotoxin and paralytic shellfish poisons by high-performance liquid chromatography with a fluorometric detection using o-phthalaldehyde.
AuthorsOnoue Y, Noguchi T, Nagashima Y, Hashimoto K, Kanoh S, Ito M, Tsukada K
JournalJ Chromatogr
PubMed ID6853629
Tetrodotoxin (TTX) and a variety of paralytic shellfish poisons (PSPs) were extracted from toxic specimens of puffer and scallop, and quantitated by high-performance liquid chromatography with a fluorometric detection using o-phthalaldehyde. Fluorescence spectra for the TTX- and PSP-fluorophors in 0.05 M borate buffer (pH 10) showed maxima at 453 nm ... More
Microplate assay for measurement of histamine release from mast cells.
AuthorsPrice JA
JournalBiotechniques
PubMed ID9149882
A microplate-format assay for measurement of histamine release by activated rat peritoneal mast cells, with accompanying microplate-format spectrofluorometric assay of histamine, is demonstrated. Reproducibility and linearity of the histamine assay are comparable to the historical large format assay. Transfer of samples in the 96-well format, from histamine release through spectrofluorometric ... More
Use of water-soluble 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide for the fluorescent determination of uronic acids and carboxylic acids.
AuthorsKobayashi M, Ichishima E
JournalAnal Biochem
PubMed ID2278381
Reaction between glucuronic acid and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was monitored by the o-phthalaldehyde (OPA) method, which was developed for the fluorescent assay of compounds containing an amino group. About 1 nmol of glucuronic acid was detected by this method. This EDC-OPA method was effective in detecting not only acidic sugar but ... More
High-performance liquid chromatographic method with fluorescence detection for the determination of total homocyst(e)ine in plasma.
AuthorsFermo I, Arcelloni C, De Vecchi E, Viganò S, Paroni R
JournalJ Chromatogr
PubMed ID1639902
A high-performance liquid chromatographic method for the determination of total plasma homocyst(e)ine [H(e)] after reduction with sodium tetrahydroborate and precolumn derivatization with o-phthaldialdehyde is described. The analyses, carried out on a reversed-phase C18 column, were based on spectrofluorimetric detection. The sensitivity was 1 pmol per injection and the intra- and ... More
Simultaneous quantification of cycloserine and its prodrug acetylacetonylcycloserine in plasma and urine by high-performance liquid chromatography using ultraviolet absorbance and fluorescence after post-column derivatization.
D-Cycloserine is a broad-spectrum antibiotic used with other antibiotics to treat various forms of tuberculosis. Its prodrug sodium (R)-4-[(1-methyl-3-oxo-1-butenyl)amino]-3-isoxazolidinone hemihydrate, developed for better aqueous stability and solubility, is combined with another broad-spectrum antibiotic, fludalanine. An ion-pair, reversed-phase high-performance liquid chromatographic assay has been developed to simultaneously detect cycloserine and its ... More
Quantitative analysis of glutathione in rat central nervous system: comparison of GSH in infant brain with that in adult brain.
AuthorsKudo H, Kokunai T, Kondoh T, Tamaki N, Matsumoto S
JournalBrain Res
PubMed ID2334850
Reduced glutathione (GSH) plays an important role in cellular protection against damage from free radicals and other electrophiles. GSH in the rat central nervous system was quantitatively analyzed. In adult brain, tissue from the cerebellum and cerebrum had higher GSH levels than that from the brainstem and spinal cord. Infant ... More
Determination of aminoglycosides in pharmaceutical formulations--II. High-performance liquid chromatography.
AuthorsFabre H, Sekkat M, Blanchin MD, Mandrou B
JournalJ Pharm Biomed Anal
PubMed ID2490559
A post-column derivatization procedure using OPA and fluorescence detection has been used for the determination of seven aminoglycosides (dibekacin, framycetin, kanamycin, netilmicin, sisomicin, tobramycin and gentamicin) in commercial pharmaceutical formulations. The linearity, precision and detection limits were satisfactory. Recoveries from eye drops, ointments, injections and capsules were comparable (P = ... More
Glutathione localization by a novel o-phthalaldehyde histofluorescence method.
AuthorsMurray GI, Burke MD, Ewen SW
JournalHistochem J
PubMed ID3536805
Glutathione in tissues forms an intense fluorophore with a solution of o-phthalaldehyde at room temperature. We have studied the loss of glutathione from tissue sections and find that it is not measurable from thick sections. The fluorescence spectra of the induced fluorophore between glutathione and o-phthalaldehyde are identical in model ... More
Glutathione content of cultured cells and rodent brain regions: a specific fluorometric assay.
AuthorsMokrasch LC, Teschke EJ
JournalAnal Biochem
PubMed ID6486436
The glutathione contents of cultured cells and rodent brain were determined by a fluorometric procedure which eliminates the interference of endogenous histidine-containing compounds. Cultured neonatal rat and hamster astrocytes, human dermal fibroblasts, and mouse and rat brain regions were assayed. o-Phthalaldehyde forms a fluorescent complex with glutathione, oxidized glutathione, and ... More
Sensitive high-performance liquid chromatographic determination of catecholamines in rat brain using a laser fluorimetric detection system.
AuthorsTodoriki H, Hayashi T, Naruse H, Hirakawa AY
JournalJ Chromatogr
PubMed ID6672024
A sensitive high-performance liquid chromatographic method for the determination of catecholamines in rat brains has been developed using a fluorescence detector equipped with a continuous wave laser as an excitation light source. A new pre-purification and derivatization method was established and confirmed to be useful for the determination of catecholamines ... More
Determination of amino sugars and amino acids in glycoconjugates using precolumn derivatization with o-phthalaldehyde.
AuthorsAltmann F
JournalAnal Biochem
PubMed ID1381156
This report examines the RP-HPLC separation of o-phthalaldehyde derivatives of amino acids, amino sugars, and amino sugar alcohols using either 2-mercaptoethanol or 3-mercaptopropionic acid. A method with pmol sensitivity for the analysis of N-acetylamino sugars of glycoconjugates was elaborated. Upon hydrolysis, amino sugars are reduced with borohydride. Automated precolumn derivatization ... More
Measurement of total plasma and cerebrospinal fluid homocysteine by fluorescence following high-performance liquid chromatography and precolumn derivatization with o-phthaldialdehyde.
AuthorsHyland K, Bottiglieri T
JournalJ Chromatogr
PubMed ID1447351
Precolumn derivatization of amino acids with o-phthaldialdehyde followed by high-performance liquid chromatographic separation and fluorescence detection is used in many clinical and experimental laboratories for the measurement of primary amino acids. This technique was adapted for the measurement of homocysteine in plasma and cerebrospinal fluid (CSF) following alkylation of the ... More
Derivatization reactions for neurotransmitters and their automation.
AuthorsBovingdon ME, Webster RA
JournalJ Chromatogr B Biomed Appl
PubMed ID7820275
Many reagents suitable for the derivatization of neurotransmitters are selective for the amino function. Others, however, are selective for the carboxyl-, thiol- and hydroxyl function, and recently, reagents selective for more than one function have been produced. Interest persists in the established reagents, with their well understood behaviour which assists ... More
A model system for convenient fluorescent labeling of sugar chain in taka-amylase A.
AuthorsKobayashi M, Sasaki Y, Chiba Y
JournalBiosci Biotechnol Biochem
PubMed ID9404062
A convenient detection of sugar chains in Taka-amylase A (TAA) was done by using 40 micrograms of enzyme, where a decrease in the UV absorption of NaIO4 during the periodate oxidation reaction was monitored. The periodate-oxidized sugar chain was labeled with a fluorescent reagent, N-1-ethylenediaminonaphthalene (EDAN), by incubation at pH ... More
Incorporation of norvaline at leucine positions in recombinant human hemoglobin expressed in Escherichia coli.
AuthorsApostol I, Levine J, Lippincott J, Leach J, Hess E, Glascock CB, Weickert MJ, Blackmore R
JournalJ Biol Chem
PubMed ID9360970
We report here a novel finding that norvaline can be incorporated in place of leucine in recombinant human hemoglobin expressed in Escherichia coli. The presence of the norvaline was confirmed by several analytical methods such as amino acid analysis, peptide mapping, electrospray mass spectrometry, and Edman protein sequencing. It appears ... More
A rapid and sensitive fluorometric assay method for the determination of nitrilase activity.
AuthorsBanerjee A, Sharma R, Banerjee UC
JournalBiotechnol Appl Biochem
PubMed ID12549980
A rapid, simple and sensitive fluorometric assay method for the determination of nitrilase activity is described. 3-Cyanopyridine was hydrolysed to nicotinic acid by Rhodococcus rhodochrous and the liberated NH(3) was allowed to react with buffered o -phthaldialdehyde-2-mercaptoethanol solution (pH 7.4) to form a fluorochrome. The fluorescence intensity was found to ... More