Development and Fast Screening of Salbutamol Residues in Swine Serum by an Enzyme-Linked Immunosorbent Assay in Taiwan

Abstract

The analysis of salbutamol in swine serum is the more practical basis for large scale surveillance programs in Taiwan. Objectives of the study were to develop a new assay and to compare with a commercially available kit in field test screens. A simple and reliable enzyme-linked immunosorbent assay (ELISA) to monitor the presence of beta-agonist, salbutamol, in 1,358 field samples of swine serum that were collected from local meat markets was described. The method proved to be suitable and sensitive for the detection of beta-agonist residues caused by growth promoting dosage. The limit of detection of the developed ELISA directly performed on diluted serum was 0.25 ng/mL. The application and the results of two ELISA kits (homemade and commercially available) for the screening of salbutamol were presented. For further confirmation, all samples that showed to be ELISA positive for salbutamol residues were analyzed by GC-MS. Adopting 1 ng/mL salbutamol as a cutoff value, the commercial beta-agonist ELISA had a sensitivity of 89.2% and a specificity of 86.7% versus GC-MS at a cutoff of 1 ng/mL. The homemade salbutamol ELISA had a sensitivity of 81.1% and a specificity of 98.6% and gave a low proportion of false-positive rate results. The reliability of the developed kit in terms of the percentage of false-positive rate results is evaluated. In conclusion, a sensitive, specific salbutamol ELISA has been developed that could serve as a rapid screening assay, and the detection of positive samples at the place of sampling can result in more effective control of the illegal use of beta-agonists.

Screening of salbutamol residues in swine meat and animal feed by an enzyme immunoassay in Taiwan

“…To minimize matrix effects, two common routes are often taken, dilution or sample cleanup such as liquid or solid phase extraction. Our recent report has shown that the dilution of serum decreased and eliminated the matrix effect [10]. Obviously, the more the meat extract is diluted the smaller the matrix effect one would expect, but with concomitant lowering of the assay sensitivity.…”

“…The value of immunoassays for salbutamol residue analysis in a variety of matrices has been widely reported [7][8][9][10]. Effective surveillance for the illicit use of ␤-agonists requires sensitive and satisfactory analytical techniques for their detection in serum and tissues from treated animals, and even includes the animal feed detection, with strengthening management of feed resource and realizing the security limit of drug residue.…”

“…The productions of polyclonal antibodies specific for salbutamol have been fully described previously [10]. Briefly, a derivative between succinic anhydride and salbutamol was prepared by stirring in dry ethanol.…”

“…The typical salbutamol standard curve obtained using the homemade ELISA method has been reported previously [10] and the parameters were presented in Table 3. As a result of the linear detection range (the concentration corresponding to 16.5-79.8% inhibition of binding) was 0.05-1.0 g kg −1 .…”

Development of a Competitive ELISA for the Detection of a Furaltadone Marker Residue, 3-Amino-5-Morpholinomethyl-2-Oxazolidinone (AMOZ), in Cultured Fish Samples

“…The ELISA was carried out using the methodology described previously [18]. One hundred microliters of anti-AMOZ polyclonal antibody (2.5 µg/ml) in a coating buffer (50 mM carbonate/bicarbonate buffer, pH 9.6) was added to each well of a split-type microtiter plate (12 strips of 8 wells each) with a µFill microplate dispenser (Bio-Tek), and incubation was performed overnight at 4°C.…”