Fluoroquinolone test kit instructions

Note: Please read the manual carefully before use (please refer to the English version, Chinese is for reference only)
Be applicable
The Abraxis fluoroquinolone test kit is suitable for the detection of fluoroquinolone in food products.
principle
The principle of the Abraxis fluoroquinolone test kit is to compete for an enzyme-linked immunoreactivity. The fluoroquinolone was extracted from the sample by an oscillating extraction using an extract. It is then diluted, filtered, and tested. The fluoroquinolone-HRP enzyme conjugate is added to the solid support microplate, then the standard and treated samples are added, and then the fluoroquinolone antibody is added to initiate the reaction. After incubation for 10 minutes, the fluoroquinolone and the enzyme-labeled fluoroquinolone in the sample competed with the fluoroquinolone antibody reaction, and the fluoroquinolone antibody and micropores were combined. The plate was washed to remove unreacted reagents. Adding a substrate developer, a colorless developer to a blue product; after 30 minutes of incubation, the reaction is terminated by adding a reaction stop solution, the color changes from blue to yellow; and the detection is performed at a wavelength of 450 nm.
The concentration of fluoroquinolone in the sample is inversely proportional to the intensity of the absorbed light.
Specificity
The Abraxis fluoroquinolone test kit does not distinguish between different types of fluoroquinolone, and the detection of different fluoroquinolone crossover rates is also different. The following is a cross-reaction listing of the kit for different fluoroquinolones.
Detection of limited meat, fish and shrimp: 1 ppb
Honey: 2 ppb

The kit consists of a kit that is stored at 2-8 degrees Celsius. Can be used during the validity period
1, 96-well ELISA plate: 1 block (12 × 8).
2, fluoroquinolone standard stock solution (Standard): 6 bottles (1ml / bottle), the concentration is 0, 0.2, 0.8, 3.2 6.4 and 16 μg / L (ppb)
3. Fluoroquinolone HRP enzyme label: 1 bottle (6ml)
4, rabbit anti-fluoroquinolone antibody: 1 bottle (6ml)
5, 5X concentrated lotion: 1 bottle (100ml)
6, substrate coloring solution: 1 bottle (12ml)
7. Stop solution: 1 bottle (12ml, 1N HCl).
8, the instructions for use
1. It is best to use the supplied reagents, do not mix them with Other companies' products, and do not mix different batches of products.
2. Dilution of the sample or the presence of impurities is likely to have an impact on the accuracy of the results.
3. Do not use expired products.
4. Return all reagents to room temperature 20-28oC before use. Do not leave at room temperature for more than 24 hours.
5. Fluoroquinolone is an antibiotic and should be treated with care.
6. Stop solution is 1N hydrochloric acid to avoid contact with skin. If contact, please rinse with plenty of water.
7. The lotion is 5X concentrated. It should be diluted with non-ionized water (such as 100ml lotion plus 400ml of non-ionized water), and the material not provided by the diluted solution washing kit is used.
1. Distilled or non-ionized water
2, measuring cylinder 100ml
3, beaker (sample extraction and collection)
4, centrifuge
5, pipette 50 μL
6, 50 and 100 μL 8-channel pipettes
7, absorbent paper
8, 450nm microplate reader
9, the timer
10. Mixer
11. SPE C18 Column 200mg/3mL
12. Acetonitrile
13. Acetonitrile
14. Washing bottles
15. Acetonitrile
16. HCl
17. Acetonitrile
18. 10 mM PBS, [0.31 g NaH2PO4-2H2O + 2.87 g Na2HPO4-12H2O + 9 g NaCl, add 1 liter of non-ionized water.
Sample processing meat sample
1. Homogenize the sample.
2. Add 5 g of homogenized sample to a 50 ml centrifuge tube and add 10 ml of acetonitrile.
3. Stunning for 5 minutes
4. Centrifuge for 5 minutes at room temperature 3000 rpm
5. Take 2 ml of the supernatant into a clean glass tube and blow dry with nitrogen.
6. Then add 2 ml of hexane, shake vigorously for 1 minute, and add 2 mL of 10 mM PBS to mix.
7. Centrifuge at room temperature for 3000 minutes at 3000 rpm.
8. Remove the upper layer of liquid.
9. Transfer the lower layer of liquid to a new tube for testing.
Honey sample (1:2 dilution)
1. Add 1g of honey to a 15ml glass bottle with a screw cap.
2. Add 5 ml of 1 M HCl and mix vigorously.
3. Centrifuge at room temperature for 3000 minutes at 3000 rpm.
4. Pre-treat the C18 column with 6 ml of methanol and 6 ml of water.
5. Pass the clear sample extract from the third step through the column (flow rate: 15 drops/min).
6. Wash the column with 3 ml water and 3 ml 5% methanol/water solution.
7. Remove the residual water sample by blowing with a weak nitrogen stream for 2 min.
8. Elute with 5% acetic acid/methanol solution (flow rate: 15 drops/min)
9. Dry the eluate with nitrogen.
10. Dissolve the dried extract in 2 ml of 10 mM PBS and mix vigorously.
11. To be tested, multiply the test result by the dilution factor of 2.
Steps to note: Standard and sample parallel can increase precision and accuracy
1. Remove the kit and sample preparation from the refrigerator before use to reach room temperature.
2. Take out the required number of microporous strips from the aluminum foil bag, place the desiccant and reseal the bag to prevent the microporous strip from getting wet.
3. Add 50 μL of 1:10 diluted standard and sample to the corresponding microwell to ensure the tip is clean.
4. Add 50 μL of enzyme conjugate to each test well.
5. Add 50 μL of antibody solution to each test well.
6, incubate for 30 minutes
7. After the incubation is completed, pour the solution in the micropore into the water tank, fill the micropores with 1X washing solution and then pour off, repeating three times, and wash the plate four times in total.
8. Clap the plate after the last wash and remove the remaining wash solution.
9. Add 100 μL of substrate to each test well.
10, incubate for 30 minutes
10. Add 100 μL of Stop Solution to each test well.
11. Read the absorbance value (OD) of each well at 450 nm.
Result analysis
1. The determination of the semi-quantitative result can be determined according to the absorbance value of the sample and the standard absorbance value. The absorbance value of the sample is lower than the absorbance value of a certain standard, indicating that the fluoroquinolone content of the sample is greater than the concentration of the standard, and the absorbance value of the sample A absorbance value above a certain standard indicates that the sample has a fluoroquinolone content less than this standard.
2. For the quantitative result determination, the standard absorbance value is required to be the X-axis, the logarithm of the standard concentration is plotted on the Y-axis, and the point of the standard concentration absorbance value is connected by a straight line, and the absorbance value of the sample is also located on the straight line, according to the sample. The absorbance value can be found on the Y-axis. If the absorbance value of the sample is greater than the minimum standard absorbance value or less than the maximum standard absorbance value, the fluoroquinolone content in this sample is less than 0.2 ppb or greater than 16 ppb. .

Black Garlic Paste is made of Peeled Black Garlic without supplements. The paste appears brown to black and is without any impurities. It has a rich garlic odor and it tastes sour with a hint of sweet.The paste is convenient and very easy to eat. It is usable directly after opening, eaten directly or spreaded on bread slices and biscuits.