【Summary】
Sulfonamides (SAs) are group of synthetic antibiotic agents that have been widely used for therapeutic and prophylactic purposes in both animals and human. They are used as feed additives as well, mainly for fattening of calves and pigs. SAs have seen an emergence in the aquaculture industry due to their effectiveness against gram-negative and gram-positive organisms.
HPLC/GC-MS method is now widely used in the detection of sulfonamides residues, but its high cost and complex operation make it unacceptable.
【Use Principles】
Sulfonamides ELISA Test Kit is an indirect competitive enzyme-labeled immunoassay. The sulfonamides antigen is precoated on the wells. The precoated antigen compete the sulfonamides antibody (antibody solution) with sulfonamides in the sample, anti- sulfonamides antibody binds to the sulfonamides-HRP enzyme conjugate. Then pipette the substrate solutions to the wells to convert the color.
The color of unknown samples is compared to the color of the standards and the sulfonamides concentration of the samples is derived.
【Cross-reactivity】
Sulfamerazine 100%
Sulfadiazine 188%
Sulfamonomethoxine 413%
Sulfaquinoxaline 44%
Sulfachlorpyridazine 75%
Sulfaclozine 68%
Sulfametoxydiazine 365%
Sulfamethazine 38%
Sulfamethoxypyridazine 42%
Sulfamethazole 33%
Sulfaethoxypyridazine 35%
Sulfadoxine 361%
Sulfadimethoxine 299%
Sulfisomidine 129%
Sulfamethizole 140%
Sulfabenzamide 39%
【Materials Required But Not Provided】
- Devices:
—- Microplate Reader (450nm/630nm filter)
—- Oscillator
—- Vortex
—- Rubber Bulb Syringe
—- Termovap Sample Concentrator
—- Homogenizer
—- Centrifuge
—- Constant Temperature Incubator
—- Centrifuge tube: 2mL, 10mL/ 7mL, 50mL
—- Balance (sensibility: 0.01g)
—- Graduated pipette: 10mL
—- Micropipette: Single channel pipette (10-100µL, 100-1000µL) Multi-channel pipette (30-300µL)
1. Reagent:
—- Deionized water
—- Neutral alumina (100-200mesh)
—- Ethyl acetate (AR)
—- Methanol (AR)
—- Acetonitrile (AR)
—- N-hexane (AR)
—- 18M Concentrated Sulfuric Acid (H2SO4)
【Provided Materials and Reagent】
Component | 96 wells |
Antigen coated plate | 96T |
5 vials of standard | 1mL each |
High concentration standard (1ppm) | 1mL each |
Enzyme Conjugate | 6mL |
Antibody Solution | 6mL |
Substrate A | 6mL |
Substrate B | 6mL |
Concentrated Wash Buffer (10×) | 40mL |
Concentrated sample diluting buffer I | 40mL |
Concentrated sample diluting buffer II | 40mL |
Concentrated sample extracting buffer | 15 mL |
Sample extractant | 1 |
Package Insert | 1 |
Testing Report | 1 |
Plate Cover Sheet | 2 |
Note: The concentrations of the standards are 0ppb, 0.5ppb, 1.5ppb, 4.5ppb and 13.5ppb.