1. Principle

This test kit is based on the competitive enzyme immunoassay for the detection of Florfenicol in the tissue (chicken, pork), fish, shrimp and honey. The coupling antigens are pre-coated on the micro-well stripes. The Florfenicol in the sample and the conjugate antigens pre-coated on the micro-well stripes compete for the anti-Florfenicol antibodies. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value has a negative correlation with the Florfenicol concentration in the sample. This value is compared to the standard curve and the Florfenicol concentration is subsequently obtained.

2. Technical specifications

Sensitivity : 0.5 ppb

Detection limit:

Honey........................................................................................... 1.5 ppb

Meat, liver, shrimp, fish.................................................................. 0.5 ppb

Recovery rate:

Honey......................................................................................... 70 ±10%

Meat, liver, shrimp, fish................................................................ 80 ±10%

Cross-reaction rate:

Florfenicol........................................................................................ 100%

Florfenicol Amine............................................................................... 11%

Thiamphenicol................................................................................ < 0.1%

Chloramphenicol............................................................................ < 0.1%

3. Components

1）   Micro-well strips: 12 strips with 8 removable wells each

2）   6× standard solution (1 mL each): 0 ppb, 0.5 ppb, 1.5 ppb, 4.5 ppb, 13.5 ppb and 40.5 ppb

3）   Enzyme conjugate (12 mL)............................................................. red cap

4）   Concentrated Antibody working solution (1 mL).............................. blue cap

5）   Substrate A solution (7 mL).......................................................... white cap

6）   Substrate B solution (7 mL)......................................................... black cap

7）   Stop solution (7 mL).................................................................. yellow cap

8）   20× concentrated washing buffer (40 mL)..................................... white cap

9）   2× concentrated redissolving solution (50 mL)..................... transparent cap

4. Materials required but not provided

1)        Equipments: microplate reader, printer, mixer or stomacher, nitrogen-drying device, oscillator, centrifuge, measuring pipets, balance( a reciprocal sensibility of 0.01 g)

2)        Micropipettors: single-channel 20-200µL and 100-1000 µL, and multi-channel 250 µL;

3)        Reagents: ethyl acetate, N-hexane