COMPARISON OF EFFICIENCY OF TWO TYPES SPE COLUMNS FOR DETECTION OF BETA AGONISTS IN LIVER WITH LC-MS/MS METHOD
Date Issued
2024-09-22
Author(s)
Jovanov, Stefan
Angeleska, Aleksandra
Abstract
Beta-agonists are a large group of chemical compounds that can be used as drugs to treat bronchial
asthma and other pulmonary diseases, due to their spasmolytic properties. Moreover, they are use as
tocolytics and heart tonics. Beta-agonists can be illegally used in meat producing animals to accelerate
growth and increase muscle mass, thereby increasing total red meat yield. Consumption of meat from
treated animals incur a risk to human health, because the residues from beta agonists accumulate and
persist in animal tissues. Due to adverse side effects in human, the Commission of European
Communities has banned the use of these compounds in slaughter animals. Тhere are various extraction
methods for the analysis of beta agonists. The aim of this study was to compare the efficiency of elution
procedure of ten beta agonists from liver with two types of solid-phase extraction (SPE) columns. The
sample preparation procedure includes homogenization of liver samples, double extraction with
phosphate buffer, enzymatic hydrolysis with β-glucoronidasa/aril sulfatase from Helix pomatia and
solid phase extraction with two types of SPE columns: Discovery® DSC-MCAX (300 mg, 6 ml) and
Strata Screen-C (500 mg, 6 ml). The analyses were performed with liquid chromatography tandem mass
spectrometry (LC-MS/MS). The validation of the method was done according to the criteria established
in Commission Decision 2002/657. Accuracy (recovery) and precision of the method were used for
comparison of the efficiency of the proposed SPE columns. The recovery of the method using DSC
MCAX columns ranges from 75.20 % to 109.4 %. A higher recovery can be observed with Strata
Screen-C for some β-agonists. For clenbuterol at spiking levels of 0.25 and 0.75 ng/g, the recoveries
were 129.0 % and 130.61 %, respectively, while for terbutaline at the spiking levels of 0.25 and 0.5
ng/g, the recoveries were 244.80 % and 275.76 %, respectively. Zilpaterol was not detected using Strata
Screen-C columns. The coefficient of variation ranges from 3.43 % to 15.73 % (DSC-MCAX columns)
and from 4.07% to 33.57 % (Strata Screen-C). The difference in the efficiency of the two types of
columns is a result of the phases they contain. DSC-MCAX columns contain C8 (octyl), while Strata
Screen-C columns contain C18 (octadecyl) phase. In the elution of samples, the C8 phase does not
retain all analytes of interest. According to the obtained results DSC-MCAX columns fulfil the criteria,
while Strata Screen-C columns don’t meet the criteria established in the 2002/657/EC.
asthma and other pulmonary diseases, due to their spasmolytic properties. Moreover, they are use as
tocolytics and heart tonics. Beta-agonists can be illegally used in meat producing animals to accelerate
growth and increase muscle mass, thereby increasing total red meat yield. Consumption of meat from
treated animals incur a risk to human health, because the residues from beta agonists accumulate and
persist in animal tissues. Due to adverse side effects in human, the Commission of European
Communities has banned the use of these compounds in slaughter animals. Тhere are various extraction
methods for the analysis of beta agonists. The aim of this study was to compare the efficiency of elution
procedure of ten beta agonists from liver with two types of solid-phase extraction (SPE) columns. The
sample preparation procedure includes homogenization of liver samples, double extraction with
phosphate buffer, enzymatic hydrolysis with β-glucoronidasa/aril sulfatase from Helix pomatia and
solid phase extraction with two types of SPE columns: Discovery® DSC-MCAX (300 mg, 6 ml) and
Strata Screen-C (500 mg, 6 ml). The analyses were performed with liquid chromatography tandem mass
spectrometry (LC-MS/MS). The validation of the method was done according to the criteria established
in Commission Decision 2002/657. Accuracy (recovery) and precision of the method were used for
comparison of the efficiency of the proposed SPE columns. The recovery of the method using DSC
MCAX columns ranges from 75.20 % to 109.4 %. A higher recovery can be observed with Strata
Screen-C for some β-agonists. For clenbuterol at spiking levels of 0.25 and 0.75 ng/g, the recoveries
were 129.0 % and 130.61 %, respectively, while for terbutaline at the spiking levels of 0.25 and 0.5
ng/g, the recoveries were 244.80 % and 275.76 %, respectively. Zilpaterol was not detected using Strata
Screen-C columns. The coefficient of variation ranges from 3.43 % to 15.73 % (DSC-MCAX columns)
and from 4.07% to 33.57 % (Strata Screen-C). The difference in the efficiency of the two types of
columns is a result of the phases they contain. DSC-MCAX columns contain C8 (octyl), while Strata
Screen-C columns contain C18 (octadecyl) phase. In the elution of samples, the C8 phase does not
retain all analytes of interest. According to the obtained results DSC-MCAX columns fulfil the criteria,
while Strata Screen-C columns don’t meet the criteria established in the 2002/657/EC.
Subjects
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