General information:

The kit is used to determine the antibiotic chloramphenicol in animal MEAT, including poultry, offal, seafood (fish, shrimp, shellfish, oysters) and eggs.

Storage conditions:

Test kits should be stored at 2-8°C.

Shelf life: 18 months. The batch number and expiration date are indicated on the packaging.

Operating principle:

The reagent kit uses an immunochromatographic method using colloidal gold particles. The analysis consists of two stages: sample preparation and detection of chloramphenicol. The prepared sample is added to the well with antibodies, if antibiotics are present in the sample , they will bind to the antibodies, thus preventing subsequent binding of antibodies to antigens applied to the nitrocellulose membrane of the test strip. The result of the reaction is the coloring of the strips, which is taken into account later.

Detection limit of the test kit: 

Set composition:

6 tubes, each containing 1 strip with 8 wells with a reagent containing antibodies and 8 test strips (48 tests in total);

1a vial of Sample Dilution Buffer;

Instructions;

Materials required:

Reading device (if necessary);

Scales , vortex, homogenizer , centrifuge , 15 ml or 50 ml centrifuge tubes, evaporator and 7 ml tubes, dispenser (100-1000 µl);

Ethyl acetate, n-hexane;

Pre-treatment reagent for eggs;

Preparation of egg pretreatment reagent (900 µl)

Place a clean 1 L flask on a magnetic stirrer and add 800 ml of distilled water. Slowly pour 100 ml of concentrated sulfuric acid into the water along the side of the flask, stirring continuously. Mix well and cool to room temperature before use.

Sample preparation:

Samples for analysis are taken in accordance with STB 1036, GOST 34668 or current TNPA. The samples taken can be stored in a place protected from light at a temperature of plus 2°C to plus 4°C for 2 days or frozen at a temperature not exceeding minus 20°C for 14 days. Before sample preparation, frozen samples must be defrosted at a temperature of plus 2°C to plus 4°C.

Bring the temperature of the samples from plus 20°C to plus 25°C, keeping them at room temperature, then homogenize them using a homogenizer. The samples under study must be uniform.

From the homogenized sample, take a sample weighing  3.0 + 0.1 g.  Place the sample in a 15 ml or 50 ml centrifuge tube (for the egg sample, add 400 μl of the prepared egg pretreatment reagent, vortex well) and add  5 ml of ethyl acetate . Vortex the contents of the tube for 3 minutes (for shrimp samples, shake the tube vertically in your hands). And then centrifuge the tube with the sample at the following mode: 4000 rpm for 5 minutes.

After centrifugation, take  3 ml  of the supernatant from the test tube and transfer it to a 7 ml glass test tube. Evaporate the ethyl acetate from the test tube using an evaporator at a temperature of plus 60 ± 5°C until there is no longer any smell of ethyl acetate.

Add 2 ml n-hexane  to the dry residue in the test tube  to dissolve the precipitate. Then add  0.6 ml  sample dilution buffer and vortex for 1 minute.

Transfer the contents into a centrifuge tube and centrifuge at 4000 rpm for 1 minute.

After centrifugation, remove the upper organic layer from the test tube using a dispenser.

The bottom layer is used for further analysis and represents the test sample.

Analysis procedure:

All components of the kit should be at room temperature (20-25°C). Prepare the required number of wells with reagent and test strips. Unused reagents and components of the kit should be put in the refrigerator.

Place the well with the reagent in the well plate. Add  150 µl of the test sample to the well with the reagent.  Mix the sample with the reagent thoroughly by five to six times by pouring the liquid into the well using the dispenser until uniform pink coloring is achieved and incubate the mixture for  2 minutes .

Then place the test strip from the kit into the well with the reagent and sample and incubate for another 5 minutes.

After incubation, remove the test strip from the well with the reagent and sample, remove the porous sponge and interpret the result within 3 minutes.