University of North Florida
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Stuart Chalk, Ph.D.
Department of Chemistry
University of North Florida
Phone: 1-904-620-1938
Fax: 1-904-620-3535
Website: @unf

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Classification: Biological fluid -> urine -> rat

Citations 5

"Rapid Separation And Measurement Of Rat Urinary Kallikrein By High Performance Liquid Chromatography With A Continuous-flow Enzyme Detector"
J. Chromatogr. A 1983 Volume 264, Issue 2 Pages 249-257
Yoshihiko Funae, Hiroko Akiyama, Susumu Imaoka, Masanori Takaoka and Shiro Morimoto

Abstract: A prepared urine sample or the partially purified urinary enzyme was subjected to anion-exchange HPLC at 20°C to 25°C on a column (30 cm x 4 mm) of IEX-540 DEAE SIL (5±1 µm) with gradient elution by 1 M Na acetate in 0.02 M Tris - acetate (pH 7.5) at 0.7 mL min-1. The eluate was collected in a PTFE helical coil, and either the peptidase activity was continuously measured with use of N-(4-methylcoumarin-7-yl)prolylphenylalanylargininamide or the esterase activity was continuously measured with use of N2-tosyl-L-arginine methyl ester as substrate and fluorimetric detection. Gel-permeation HPLC on two columns of TSK-GEL G3000SW, with 0.05 M sodium phosphate buffer (pH 7.5) - 0.1 M NaCl as mobile phase (0.5 mL min-1) was also used to separate kallikrein before its activity was measured; confirmation of identity was by the inhibitory effect of aprotinin. The detection of peptidase activity was more sensitive than that of esterase activity.
Kallikrein HPIC Apparatus Reactor

"Improved High Performance Liquid Chromatographic Method For The Determination Of 6-N,N,N-trimethyl-lysine In Plasma And Urine: Biomedical Application Of Chromatographic Figures Of Merit And Amine Mobile Phase Modifiers"
J. Chromatogr. B 1986 Volume 380, Issue 2 Pages 285-299
Paul E. Minkler, Elizabeth A. Erdos, Stephen T. Ingalls, Ronda L. Griffin and Charles L. Hoppel

Abstract: Samples of blood and urine were collected from rats and human subjects; the urine samples were hydrolyzed with concentrated HCl at 105°C for 18 h. The hydrolysate or blood plasma samples were purified on successive columns (3.5 cm x 5 mm) of Dowex 50-X8 (NH4+ form) and Dowex 1-X8 (OH- form), respectively; elution was effected with aqueous 2.2 M NH3. The eluate was analyzed by HPLC on a cartridge (10 cm x 5 mm) of Radial-Pak C18 (10 µm) equipped with a pre-column (5 cm x 4 mm) of Co:Pell ODS; the mobile phase consisted of 2.5 mM Na dodecyl sulfate - 30 mM NaH2PO4 - 20 mM 3-(dimethylamino)propane-1,2-diol in acetonitrile. Post-column derivatization was carried out by treating the eluate with phthalaldehyde - 2-mercaptoethanol in 0.5 M H3BO3. Detection was by fluorimetry at 418 nm (excitation at 240 nm). The detection limit was 0.2 nmol mL-1 of analyte. The effect of pH of the mobile phase on the performance of the system was investigated.
6-N,N,N-Trimethyllysine HPLC Fluorescence Optimization Post-column derivatization

"Flow Injection Proton Nuclear-magnetic-resonance Spectroscopy Combined With Pattern-recognition Methods Implications For Rapid Structural Studies And High-throughput Biochemical Screening"
Anal. Commun. 1997 Volume 34, Issue 11 Pages 339-341
Manfred Spraul, Martin Hofmann, Michael Ackermann, Andrew W. Nicholls, Stephen J. P. Damment, John N. Haselden, John P. Shockcor, Jeremy K. Nicholson and John C. Lindon

Abstract: The applicability of novel NMR flow probe technology has been tested by the measurement of 300 MHz H-1 NMR spectra of a series of rat urine samples, Compared with conventional automatic operation, the method resulted in a significantly increased rate of sample throughput, required minimal spectrometer optimization before each measurement and avoided the need for expensive and fragile NMR sample tubes, The NMR approach has been coupled with computer methods for spectral data reduction and classification using, in this case, principal components analysis, The flow probe NMR approach offers distinct advantages in situations where large numbers of samples require NMR analysis in a short period of time, These could include routine samples from high throughput chemical synthesis, biofluid samples for drug toxicity monitoring as shown here, samples for clinical diagnosis or real-time analysis in chemical production facilities. 7 References
Drugs Nuclear magnetic resonance Principal component analysis Optimization

"Improved Flow Injection Analysis Method For Determining Selenium In Biological Samples, And The Effect Of Captopril Administration On Selenium Levels And Glutathione Peroxidase Activity In Rat"
Biol. Pharm. Bull. 1993 Volume 16, Issue 11 Pages 1069-1072
Nobuyuki SUZUKI, Eriko FUJITA, Hiroyuki YASUI, Eiji AOYAMA, Hisashi TANAKA, Terumichi NAKAGAWA

Abstract: An improved flow injection analysis (FIA) method has been developed for the determination of trace selenium in biological samples, and this method has been applied to investigate the effect of captopril, an antihypertensive drug having a thiol group, on selenium concentrations in the rat blood, liver and urine. After oral administration of captopril, selenium levels in the blood decreased, while those in the liver increased significantly. However, no pronounced effect was observed on the urinary excretion rate. The glutathione peroxidase activities in the blood and the liver were comparable to the changes in the selenium levels.
Selenium Captopril

"Simultaneous Determination Of Phencyclidine And Its Metabolites In Rat Urine By High-Performance Liquid Chromatography"
Anal. Sci. 1998 Volume 14, Issue 5 Pages 979-982
Ryoko Chiba, Megumi Fukushi And Akira Tanaka

Abstract: A high-performance liquid chromatography with chemiluminescence detection using tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)32+] for the simultaneous determination of phencyclidine and its major metabolites in rat urine was developed. The urine sample was applied onto a disposable cartridge column (Bond Elut Certify) of silica solid-phase extraction. The charged sample was then washed with an acidic solution and methanol prior to its elution with 2% ammonium hydroxide in methanol, and the eluate was chromatographed. In this method, a TSKgel ODS 80Ts column and a mobile phase containing 60% methanol, 50 mM phosphate buffer (pH 7.0) and 0.12 mM tris(2,2'- bipyridyl)chloride hexahydrate in 10 mM sulfuric acid as a reagent solution were used. The calibration curves for phencyclidine and its metabolites showed good linearity over the range 25 - 400 ng mL-1 per 20 µl injection. As a result, the method was successfully applied to a simultaneous determination of phencyclidine and its three major metabolites in actual rat urine.
Phencyclidine 4-Phenyl-4-piperidinocyclohexanol 1-(1-Phenylcyclohexyl)-4-hydroxypiperidine 1-(1-Phenyl-4-hydroxycyclohexyl)-4-hydroxypiperidine HPLC Chemiluminescence Post-column derivatization