On-line metal chelate affinity chromatography clean-up for the high-performance liquid chromatographic determination of tetracycline antibiotics in animal tissues.
An on-line high-performance liquid chromatographic (HPLC) method for the determination of tetracycline, oxytetracycline, chlortetracycline and demeclocycline using metal chelate affinity chromatography-reversed-phase HPLC has been developed. The drugs were extracted with succinate buffer and the extract diluted with EDTA-pentanesulphonate buffer. Diluted extract was then absorbed onto a C8 or XAD-2 solid-phase extraction (SPE) cartridge and eluted with methanol. The eluate was then injected onto a TSKgel chelate column which had been preloaded with copper(II). The tetracyclines were eluted from this column onto the analytical column (Polymer Labs. PLRP-S) with an EDTA-containing buffer. Elution of the analytical column was via a methanol-acetonitrile gradient and detection was by UV at 350 nm. Average recoveries at the 10, 20, 50 and 300 micrograms kg-1 levels were 50-80%. The limit of detection (LOD) was 10 micrograms kg-1 for oxytetracycline and tetracycline and 20 micrograms kg-1 for chlortetracycline and demeclocycline. The method was validated for sheep liver and cattle kidney.
Stubbings
G
G
Ministry of Agriculture, Fisheries and Food, CSL Food Science Laboratory, Colney, Norwich, UK.
Tarbin
J A
JA
Shearer
G
G
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D002417:Cattle; D002614:Chelating Agents / Q000737:chemistry*; D002751:Chlortetracycline / Q000032:analysis; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D003707:Demeclocycline / Q000032:analysis; D004350:Drug Residues / Q000032:analysis*; D004492:Edetic Acid / Q000737:chemistry*; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D010118:Oxytetracycline / Q000032:analysis; D015203:Reproducibility of Results; D012756:Sheep; D013752:Tetracycline / Q000032:analysis
D000900:Anti-Bacterial Agents; D002614:Chelating Agents; D003707:Demeclocycline; D004492:Edetic Acid; D013752:Tetracycline; D002751:Chlortetracycline; D010118:Oxytetracycline