Simple and rugged SPE method for the determination of tetracycline antibiotics in serum by HPLC using a volatile mobile phase

Summary

A simple and rugged SPE method for the determination of tetracycline (TC), minocycline (MC) and demeclocycline (DCC) in porcine serum by high performance liquid chromatography (HPLC) was developed. The spiked serum sample was pretreated with 2% phosphoric acid followed by a simple and rugged solid-phase extraction procedure using the Oasis^TM HLB extraction cartridges. High and reproducible recoveries were obtained even though the cartridges were run dry. The extracted sample analytes were injected onto a Waters SymmetryShield^TM RP₈ column. The mobile phase was a simple volatile solution containing 0.1% TFA, 2% methanol and 7% acetonitrile in Water. The antibiotics were detected at 350 nm. The calibration curves were linear from 2.0 to 25.0 μg mL⁻¹ of TC and MC with DCC as the internal standard at a concentration of 25.0 μg mL⁻¹. For six replicate analyses, the average recoveries of TC and MC from porcine serum sample fortified at the level of 2.5 μg mL⁻¹ were 96.1% with 1.3% RSD and 101% with 0.54% RSD; at level of 0.5 μg mL⁻¹ the average recoveries were 88% with 1.6% RSD and 97.8% with 1.4% RSD.