Fluoroquinolone ELISA Kit

Sample

Meat, milk, tissues, serum, urine

Species Reactivity

N/A

Detection Method

cELISA

Intended Use

This product can be used for the qualitative and quantitative detection of Fluoroquinolones residues in: Tissues, Aquatic Products, Chicken By-products, Duck By-products, milk, and milk powder.

Contents of Kit

1. 96-well Microtiter Plate × 1 (pre-coated with coupled antigen)
2. Standards × 6 vials (1 mL/vial): (1 μg/L = 1 ppb = 1 μg/kg) 0 ppb, 0.1 ppb, 0.3 ppb, 0.9 ppb, 2.7 ppb, 8.1 ppb
3. High Concentration Standard: (1 mL/vial) 100 ppb
4. Antibody Working Solution: 7 mL (Green Cap)
5. Enzyme-Labeled Secondary Antibody Concentrate: 1 mL (Red Cap)
6. Substrate Solution A: 7 mL (White Cap)
7. Substrate Solution B: 7 mL (Red Cap)
8. Stop Solution: 7 mL (Yellow Cap)
9. 20× Concentrated Wash Buffer: 40 mL (Transparent Cap)
10. 2× Concentrated Reconstitution Solution: 50 mL (Blue Cap)

Storage

Storage Condition: Store at 2~8°C.
Shelf Life: 12 months.

Precision

Intra-assay and Inter-assay coefficients of variation (CV) are both < 10%.

Detection Limit

Detection Limits (LOD) for Specific Drugs:
Norfloxacin: 0.2 ppb
Difloxacin: 0.2 ppb
Enrofloxacin: 0.2 ppb
Flumequine: 0.2 ppb
Sarafloxacin: 0.2 ppb
Danofloxacin: 0.2 ppb
Pefloxacin: 0.2 ppb
Ciprofloxacin: 0.2 ppb
Enoxacin: 0.3 ppb
Ofloxacin (Racemate): 0.3 ppb

Lower Detection Limits (LOD) for Sample Matrices:
Tissue: 0.3 ppb
Aquatic Products: 0.3 ppb
Chicken By-products: 0.3 ppb
Duck By-products: 0.3 ppb
Bovine Serum: 0.3 ppb
Milk: 3 ppb
Milk Powder: 6 ppb

Sensitivity

0.1 ppb

General Description

Fluoroquinolones are a class of highly important broad-spectrum antibiotics that have developed rapidly over the past 20 years. They inhibit bacterial DNA gyrase and are known for their broad antibacterial spectrum, high efficacy, low toxicity, and strong tissue penetration. They have become one of the most important anti-infective drugs in veterinary medicine and aquaculture, widely used for treatment, prevention, and growth promotion. Their resistance potential and possible carcinogenicity have raised widespread concern. In tissues, the marker residues for Enrofloxacin are Enrofloxacin and its metabolite Ciprofloxacin, with the highest residue concentrations found in liver and kidney tissues, followed by muscle and skin tissue attached to fat. Its metabolite, Ciprofloxacin (CIF), still possesses biological activity. Ofloxacin (OFL) primarily exists in tissues in its parent form. The metabolic rate of Pefloxacin (PEF) in the body is close to 100%, and its metabolite is Norfloxacin (NOR).

This kit is a new generation of drug residue detection product developed using ELISA technology. Compared with instrumental analytical techniques such as High-Performance Liquid Chromatography (HPLC), it features rapidity, simplicity, accuracy, and high sensitivity. The total operation time is only 45 minutes, which can minimize operational errors and workload.

Standard Curve

Two methods can be used. Method 1 for rough estimation, Method 2 for quantitative determination. Note: Sample absorbance value is inversely proportional to Fluoroquinolones concentration.
1. Qualitative / Semi-quantitative Determination:
Compare the average absorbance values of samples with those of the standards to estimate the concentration range.
Example Standard Absorbance Values:
0 ppb = 1.788; 0.1 ppb = 1.321; 0.3 ppb = 0.857; 0.9 ppb = 0.404; 2.7 ppb = 0.192; 8.1 ppb = 0.093.

If Sample 1 OD = 0.258: Concentration is between 0.9 - 2.7 ppb.

If Sample 2 OD = 0.956: Concentration is between 0.1 - 0.3 ppb.
Multiply the obtained concentration by the appropriate sample dilution factor to get the residue concentration in the original sample.

2. Quantitative Determination:

Calculation of Percentage Absorbance (%):
% Absorbance = (B / B0) × 100%

B = Average absorbance of the standard or sample solution

B0 = Average absorbance of the 0 ppb standard solution

Plotting Standard Curve and Calculation:
Plot the standard curve with the % Absorbance (y-axis) of the standards against the logarithm of the standard concentration (ppb, x-axis). Determine the concentration of the sample from the standard curve based on its %Absorbance. Multiply by the appropriate sample dilution factor to obtain the actual concentration of Fluoroquinolones in the original sample.
Sample Dilution Factors:
Tissue: 2
Aquatic 2
Chicken By-products: 2
Duck By-products: 2
Bovine Serum: 2
Milk: 20
Milk Powder: 50