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: Cell -> leucocyte -> human

Citations 2

"Method For Determination Of Free And Total Glutathione And γ-glutamylcysteine Concentrations In Human Leucocytes And Plasma"
J. Chromatogr. B 1987 Volume 420, Issue 1 Pages 152-157
Johannes Mårtensson

Abstract: Leucocytes and plasma were prepared as previously described for glutathione (Metabolism, 1986, 35, 118) except that cell lysis was performed for total glutathione(I) and γ-glutamylcysteine(II) analysis before incubation with dithiothreitol. After incubation with sulfosalicylic acid, dithiothreitol was removed from total I samples by ethyl acetate extraction. All solution were applied to a p-acetoxymercurianiline-Sepharose 4-B affinity column before elution with 25 mM mercaptoacetic acid in 50 mM HNO3 containing 1 mM Na2EDTA. HPLC was performed on a column (25 cm x 4.6 mm) of Spherisorb C18 (5 µm) with a guard column (25 cm x 4.6 mm) of Polygosil 60 C12 (25 to 40 µm). The mobile phase (1.2 mL min-1) was 5 mM Na heptanesulfonate (pH adjusted to 1.8 with H3PO4) containing 11% of methanol. Post-column derivatization with phthalaldehyde was performed before detection by measuring fluorescence at 420 nm (excitation at 350 nm). EDTA was added to all solution to prevent oxidation of the compounds. Recoveries were 98 and 96% for reduced and total I, respectively, and 96 and 92% for reduced and total II, respectively. The calibration graph was rectilinear from 20 pmol to 2 nmol of I and the detection limit was 4.5 pmol. The inter-assay coefficient of variation (n = 5) were 6.4 and 10.1% for I and II, respectively.
Glutathione γ-Glutamylcysteine HPLC Fluorescence Post-column derivatization

"A Modified Flow Injection Analysis Method To Quantify DNA Strand-breaks"
Med. Sci. Res. 1993 Volume 21, Issue 14 Pages 535-538
Saravanan, K.;Venkatesan, P.V.;Nagarajan, B.

Abstract: In this paper the authors describe the application of flow injection analysis for the quantitation of DNA strand-breaks in human leukocytes following exposure to MNNG and catechol in vitro and in rat stomach mucosal cells in vivo using flow fluorescence detector of high performance liquid chromatograph. This was compared against the conventional spectrofluorometric data.
DNA Fluorescence Method comparison