C/N: EE 108    Sensitivity: 0.5 ppb    Test Range: 0.5~50 ppb

1. BACKGROUND 
    Sulfonamides are widely used as feed additives, mainly for fattening of calves and pigs. Combined with inhibitors of dihydrofolate reductase such as trimethoprim, tetroxoprim, or pyrimethamine sulfonamides are also used in veterinary medicine for the treatment of intestinal infections, mastitis, pulmonitis and other (systemic) diseases. Sulfonamide residues may therefore occur in food of animal origin such as meat and milk. Particularly the transmission of the carcinogenic sulfamethazine represents a threat to human health. According to the EU-Law a  maximum residue limit for all substances of the sulfonamide-group of 100 ppb in muscle, fat, liver and kidney and of 100 ppb in milk is valid.

2. PURPOSE
    This handbook establishes procedures for determining sulfamethazine in grain and processed grain products. The test is a competitive direct ELISA that provides exact concentrations in parts per billion (ppb). Free toxin in the sample and controls competes with enzyme-labeled toxin (conjugate) for the antibody binding sites. After a wash step, substrate reacts with the bound enzyme conjugate to produce blue color.

3. KIT CONTENTS
1)Microtiter plate (8 wells×12) precoated with antibodies to mouse IgG.
2)6×sulfamethazine standard solutions (1ml/each):0ng/ml, 0.5ng/ml, 2.0ng/ml, 5.0ng/ml, 20.0ng/ml, 50.0ng/ml
Controls provided: 0ppb, 0.5ppb, 2.0ppb, 5.0ppb, 20.0ppb, and 50.0ppb
3)Antibody (100×): sulfamethazine              (0.2ml)
4)Enzyme Conjugate(100×)                    (0.2ml)
5)Washing buffer(10×)                        (50ml)
6)Antibody and Enzyme diluent buffer            (30ml)
7)Diluent buffer A                            (40ml)
8)Diluent buffer B                            (40ml)
9)Diluent buffer C                            (40ml)
10)TMB                                              (18ml)
11)Stop solution                               (7ml)
12)Product Manual