Development of a rapid screening test for veterinary sedatives and the beta-blocker carazolol in porcine kidney by ELISA

Abstract

Sedatives and tranquillisers are frequently used to reduce stress during the transportation of food producing animals. The most widely used classes of sedatives include the butyrophenone azaperone, the phenothiazines acepromazine, propionylpromazine, chlorpromazine and the [small beta]-blocker, carazolol. For regulatory control purposes, tolerances for azaperone and carazolol have been set by the European Union as 100 and 25 [micro sign]g kg(-1), respectively. Furthermore, the use of the phenothiazines is prohibited and therefore has a zero tolerance. A method for the detection of residues of five tranquillisers and one [small beta]-blocker using a single ELISA plate has been developed. Kidney samples (2.5 g) were extracted with dichloromethane and applied to a competitive enzyme immunoassay using three polyclonal antibodies raised in rabbits against azaperol, propionylpromazine and carazolol conjugates. In sample matrix, the azaperol antibody cross-reacted 28.0% with azaperone and the propionylpromazine antibody cross-reacted 24.9% with acepromazine and 11.7% with chlorpromazine. In the ELISA, the detection capabilities of the six sedatives, azaperol, azaperone, carazolol, acepromazine, chlorpromazine, and propionylpromazine are 5, 15, 5, 5, 20 and 5 [micro sign]g kg(-1), respectively. The proposed method is a sensitive and rapid multi-residue technique that offers a cost effective alternative to current published procedures, without any concession on the ability to detect sedative misuse.

Preparation of a monoclonal antibody and development of an indirect competitive ELISA for the detection of chlorpromazine residue in chicken and swine liver

“…3, which showed that the IC 50 value of chlorpromazine was 0.73 ppb, indicating excellent sensitivity. The MAbs raised in our study showed more than 20 times higher sensitivity for chlorpromazine than that raised in published research, 7 in which chlorpromazine was determined by an ELISA procedure using the polyclonal antibody obtained by immunisation of a propionylpromazine-protein conjugate in rabbit, with the detection capability of 20 ppb. The monoclonal antibody demonstrates remarkable specificity toward chlorpromazine.…”

Rapid method for the determination of tranquilizers and a beta-blocker in porcine and bovine kidney by liquid chromatography with tandem mass spectrometry

“…They include enzyme-linked immunosorbent assay (ELISA) [5], thin layer chromatography (TLC) [6,7], liquid chromatography (LC) coupled with ultraviolet [7,8], fluorescence [8,9] and electrochemical detection [10]. Several methods using liquid and gas chromatography with mass spectrometry (MS) have been also published [11][12][13][14].…”

Comparison of the analysis of β-blockers by different techniques

“…Different techniques have been used to determine ␤-blockers in urine, including methods based on spectrofluorimetry [6][7][8], ELISA [9,10], and chromatography [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26]. Most of the chromatographic methods are based on gas chromatography/mass spectrometry coupling (GC/MS), requiring the derivatization of polar groups (aminopropanol chain) of compounds [11,14,[17][18][19][21][22][23]25,26].…”