Active substances: Amoxicillin
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However, the recovery values were not improved to a satisfying rate assuming that the analytes get lost during extraction and clean-up procedure.
Another explanation for the low recovery rates could be different polarities of the compounds, assuming that the analytes partially migrated into the aqueous phase after separation. Solid-Phase Extraction To improve ionization of the analytes due to better matrix removal solid-phase extraction SPE was tested for sample clean-up.
Acetonitrile was used as extraction solvent in order to get a clean supernatant preventing clogging of the cartridges.
After conditioning the cartridge with methanol and water, the diluted sample extract was applied to the SPE column, followed by washing the cartridge with water and elution of the analytes with methanol.
With this step, recovery rates for tetracyclines were three times higher than with acetonitrile, but unfortunately, the number of detected analytes did not increase.
For a better explanation of the low recovery rates, the samples were quantified with matrix-matched calibration resulting in better recoveries for nearly all substances, except in case of tetracyclines and macrolides.
In addition, possible losses of analytes during washing step were assessed and the elution step was conducted twice to evaluate if the volume of elution solvent was sufficient. Therefore, the washing solution was analyzed and it was noticed that sulfadiazine was partially getting lost during washing step.
Tetracyclines were also found after second elution indicating that tetracyclines retained in the stationary phase maybe explaining the low recoveries.
Due to the fact that not all substances were detectable and that SPE might be more time consuming, another sample preparation was examined.
Based on already published studies Lehotay et al. First results demonstrated that nearly all substances could be detected, except penicillin V, amoxicillin, and ceftiofur assuming that these substances are getting rapidly degraded.
Therefore, matrix effects were evaluated by quantifying with matrix-matched calibration fortifying a blank milk samples at five different concentration levels 0.
The recovery rates demonstrated increasing as well as decreasing adjustments which let us to assume that the matrix is not the most important influence factor, subsequently performing quantification by procedure matched calibration PMC to eliminate possible influences from the matrix and sample preparation.
For this, five blank milk samples were fortified with antibiotic standard mix at different concentrations 0. In accordance with this quantification, acceptable recovery rates were obtained for all substances Fig.
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