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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Cow Fat

Classification: Biological tissue -> fat -> cow -> calf

Citations 1

"Development And Optimization Of A Liquid Chromatographic Method For The Determination Of Gentamicin In Calf Tissues"
Anal. Chim. Acta 1993 Volume 275, Issue 1-2 Pages 285-293
Florence Sar, Pierre Leroy and Alain Nicolas*, Philippe Archimbault

Abstract: Calf muscle, liver, kidney or fat (5 g) was deproteinated with 5% trichloroacetic acid (20 ml). After centrifugation at 8000 rpm and 5°C for 15 min, the supernatant solution was adjusted to pH 7.0 ± 0.2 and diluted to a known volume with 0.1 M Na2SO4 - 0.1 M K2HPO4 (pH 7). The solution (10 ml) were applied to CM-Sephadex C-25 cartridges (prep. described) in 1 mL fractions, the cartridges were washed with 0.2 M Na2SO4 - 1 mM EDTA (2 x 1 ml), water (1 ml) and 0.05 M NaOH (250 µL). Gentamicin (I) was eluted with 0.05 M NaOH (1 ml) and the eluates were mixed with 1 M HCl (100 µL), 0.5 M camphorsulfonate (pH 2.2; 100 µL). I was determined by HPLC on a column (12.5 cm x 4 mm) of LiChrospher 100 RP-18 (5 µm) operated at 45°C and protected by a guard column (0.4 cm x 4 mm) of the same material. The mobile phase (1.2 mL min-1) was 0.05 M Na dl-camphor-10-sulfonate solution in 0.1 mM EDTA (pH 2.2) - methanol (9:11). Post-column derivatization was carried out at 45°C in a PTFE knitted open-tubular reactor (3 m x 0.5 mm i.d.) with phthalaldehyde solution containing mercaptoethanol (0.5 mL min-1). Fluorimetric detection was at 440 nm (excitation at 340 nm). Detection limits were 25 ng g-1 of I in muscle and fat and 50 ng g-1 of I in kidney and liver. Recoveries were 68 to 98%.
Gentamicin HPLC Fluorescence Optimization Post-column derivatization Knotted reactor