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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Plasma Rat

Classification: Biological fluid -> blood -> plasma -> rat

Citations 6

"Determination Of The Fluorescent Drugs Dipyridamole And Benzydamine In Rat Plasma By Liquid Chromatography With Peroxyoxalate Chemiluminescence Detection"
Anal. Chim. Acta 1991 Volume 251, Issue 1-2 Pages 247-253
Atsuhiko Nishitani, Yukie Tsukamoto, Susumu Kanda and Kazuhiro Imai*

Abstract: Plasma (10 µL) or, for dipyridamole (I) determination, plasma diluted 10-fold with 150 mM imidazole buffer solution (pH 6.0) was mixed with imidazole buffer solution (40 µL; as above), internal standard solution (20 µL) and acetonitrile (130 µL) and the mixture was centrifuged at 2500 g for 5 min. The internal standards were 5-(NN-dimethylaminonaphthalene)-1-sulfonyl-L-phenylalanine for I and I for benzydamine hydrochloride (II). Sample solution was injected into a flow system (described with diagram) containing a column (15 cm x 4.6 mm) of TSK ODS 8OTm (5 µm) operated at 40°C with 50 mM imidazole buffer (pH 6.0) - acetonitrile (1:1) as eluent. The eluate was mixed with 0.25 mM bis-[4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl]oxalate and 12.5 mM H2O2 solution in acetonitrile - ethyl acetate (1:1) before detection of chemiluminescence. Calibration graphs were rectilinear from 2.5 to 200 nM-I and 2.5 to 100 µM-II. Detection limits were 345 pM-I and 147 nM-II.
Benzydamine Dipyridamole Chemiluminescence LC Buffer Column Heated reaction Internal standard

"Sulfur-selective Detector For Liquid Chromatography Based On Sulfur Monoxide-ozone Chemiluminescence"
Anal. Chem. 1994 Volume 66, Issue 18 Pages 2841-2851
Thomas B. Ryerson, Andrew J. Dunham, Robert M. Barkley, and Robert E. Sievers

Abstract: The design and performance of a S-selective detector that can be used at liquid flow rates characteristic of HPLC, ion chromatography and FIA is described. It converts S-containing compounds in the liquid phase into SO under pressure and at elevated temperature, typically 2000 psig and 300°C, respectively. The SO is allowed to permeate across a flat Teflon membrane into a He stream and is swept to a reaction cell where O3 is added. The photoemission resulting from the SO + O3 reaction is monitored with a photomultiplier tube. The parameters most affecting the detector response are the reaction capillary temperature and pressure, the composition of the liquid mobile phase and the oxidation state of S in the analyte. The mobile phase/reaction medium chosen was He-sparged water adjusted to pH 3 with H3PO4. The detector had a linear response of 3 orders of magnitude of analyte and a selectivity of 106 for S compounds over non-S-containing species. It was applied to the reversed-phase HPLC analysis of S-containing pesticides, proteins and blood thiols and FIA of acid-soluble thiols in rat plasma. The detection limits were in the order of 10 ppb S in aqueous solution.
Sulfur compounds HPLC HPIC Chemiluminescence Teflon membrane

"Determination Of Thiamine And Its Phosphate Esters In Human And Rat Blood By High Performance Liquid Chromatography With Post-column Derivatization"
J. Chromatogr. A 1985 Volume 332, Issue 1 Pages 181-188
Mieko Kimura and Yoshinori Itokawa

Abstract: Blood, erythrocytes and plasma were deproteinized with aqueous 10% trichloroacetic acid. After centrifugation, the supernatant solution were analyzed by HPLC on a µBondapak C18 column (25 cm x 4 mm) with 0.2 M NaH2PO4 in aqueous 0.3% acetonitrile as the mobile phase (1.0 mL min-1); the eluates were subjected to post-column derivatization with 0.1% of K3Fe(CN)6 in aqueous 15% NaOH and spectrofluorimetric detection at 450 nm (excitation at 375 nm). The detection limit for thiamine and its phosphate esters was 30 fmol and recoveries were 95.0 to 100.2%. The calibration graphs were rectilinear for 50 to 200 fmol.
Thiamine Thiamine triphosphate Thiamine monophosphate HPLC Fluorescence Post-column derivatization

"Quantification Of Physiological Amino Acids By Gradient Ion-exchange High Performance Liquid Chromatography"
J. Chromatogr. B 1993 Volume 613, Issue 2 Pages 223-230
Svend Erik Møller

Abstract: Human or rat plasma, human cerebrospinal fluid or 0.4 M HClO4 extracts of rat brain were mixed with sulfosalicylic acid and α-aminoadipic acid (internal standard) at 4°C, then filtered before analysis on a column (15 cm x 3 mm) of 5 µm Li cation-exchange material (sulfonated divinylbenzene-styrene copolymer) equipped with a guard column (2 cm x 3 mm) and maintained at 50°C. The mobile phase (0.2 and 0.3 ml/min) comprised a gradient (details given) of 0.64 M lithium citrate buffer (pH 7.50) in 0.24 M lithium citrate buffer (pH 2.27) and detection was by post-column phthalaldehyde derivatization and fluorimetry at 448 nm (excitation at 340 nm). Intra-assay RSD for 28 amino-acids present in human plasma were 3.4-17.6% and the inter-assay RSD were 2.7-39.0%. The response was rectilinear for 50-5000 pmol for 32 amino-acids, and up to 40 compounds could be determined in a 3-h run. A single-column gradient lithium ion-exchange chromatographic method with post-column derivatization and fluorimetric detection for the quantification of physiological amino acids is described. The method runs automatically, requires a minimum of sample preparation, separates all amino acids in plasma and cerebrospinal fluid, and most compounds in brain extract, in addition to some amino acids used therapeutically and in pharmacological studies. About 40 compounds can be quantitated within a run time of 3 h. The within-assay and between-assay coefficients of variations for principal amino acids in plasma samples are satisfactory. The system has performed conveniently and with high stability in the daily routine work and is cost-saving based on laboratory-prepared buffers.
Amino Acids HPLC Fluorescence Post-column derivatization

"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

"Determination Of Immunostimulative Acylpeptide (FK 565) [N-(1-oxoheptyl)-D-γ-glutamyl-L-erythro-α,e-diaminopimelyl-D-alanine] In Plasma By HPLC With Fluorogenic Post-column Derivatization"
Bunseki Kagaku 1987 Volume 36, Issue 11 Pages 787-791
Tashiro, Y.;Suzuki, A.;Noda, K.;Noguchi, H.

Abstract: Rat or mouse plasma was deproteinized with 6% trichloroacetic acid solution and the supernatant solution was applied to a Bond-Elut C18 cartridge. The retained FK 565 was eluted with mobile phase [20 mM phosphate buffer (pH 2.5) - acetonitrile (79:21)] at 0.5 mL min-1 and was determined on a TSK-gel LS-410 ODS Sil column (15 cm x 4 mm) with fluorimetric detection by using post-column derivatization with phthalaldehyde in the presence of 2-mercaptoethanol. The coefficient of variation were <2.3%. The limits of determination were 50 and 20 ng mL-1 of FK 565 in rat and mouse plasma, respectively.
HPLC Fluorescence Post-column derivatization