A kind of method and product suitable for detecting the content of oquindox in veterinary drug preparations
A technology for olaquindox and reagents, which is applied in the field of detecting the content of olaquindox in veterinary drug preparations, can solve the problems of complicated processing process, poor purification effect, and much waste of organic solvents, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0030] Embodiment 1, the preparation of immunogen and coating former
[0031] One, the preparation of olaquindox hapten
[0032] Rinse the 50ml round bottom flask, dry it with ethanol, fix it on the stirrer, and add a stirring bar. Weigh 500mg (1.9mmol) of the compound shown in formula II, dissolve it in 5ml of pyridine, stir at RT for 5min, add 155mg (1.55mmol) of succinic anhydride, heat to 80°C, react, and monitor by pointing the plate for 1h / time.
[0033] It was found by TLC that the reaction was complete, treated, adjusted the pH value to neutral with 2M HCl, extracted with 10ml of ethyl acetate water*3, dried with anhydrous sodium sulfate and concentrated, added 1.5g of 100 mesh silica gel to mix and mix the sample, and used 10 g of 200-mesh silica gel was packed into a column and subjected to column chromatography. Gradient elution was performed with ethyl acetate:petroleum ether 10:1. The required solution was collected, spin-dried, and collected to obtain the compou...
Embodiment 2
[0039] Embodiment 2, preparation and structural identification of olaquindox artificial antigen
[0040] One, the preparation of olaquindox artificial antigen
[0041] 1. Synthesis of immunogen
[0042] (1) Dissolve 18.4mg hapten in 1.5ml DMF, stir at 200rpm for 10min, add 7.25mg of CDI to dissolve, stir at room temperature (500rpm) and activate for 2-3h.
[0043] (2) Weigh 50mg of BSA and dissolve it in 3.5ml of CB solution, stir at 200rpm for 10min to fully dissolve, cool down in an ice bath to 0-4°C, stir at 1000rpm, add the reaction solution in step 1 dropwise (1ml / min), 500rpm stirring reaction 24h.
[0044] (3) Put the reaction product into distilled water and rinse the dialysis bag (10cm), dialyze with 1L of 0.01M PBS (1×, pH7.2) at 4°C (100rpm) for 3 days, change the solution 3 times a day (once in the morning, once in the evening) , changing the medium 9 times in total, centrifuge the dialysis product at 5000rpm for 6min, aliquot 1.5ml / tube, number the antigen, and...
Embodiment 3
[0051] Embodiment 3, olaquindox artificial antigen immunization animal prepares monoclonal antibody
[0052] 1. Animal immunity
[0053] Use 100 μg of the immunogen (olaquindox-BSA) prepared in Example 2, dissolve the immunogen in physiological saline and mix it with Freund's complete adjuvant in equal volume, and inject subcutaneously on the back of the neck to immunize 6-8 weeks old Balb / c On the 7th, 14th, and 28th day after the initial immunization, the female mice were mixed with equal volumes of the immunogen and Freund's incomplete adjuvant, and each additional immunization was given once, and the immune complex was 100 μg / rat 3 days before the fusion, without Freund's adjuvant A booster immunization was given again.
[0054] 2. Cell fusion and cloning
[0055] According to the conventional method, the splenocytes of the immunized mice were mixed with the mouse myeloma cells (SP2 / 0) in the logarithmic growth phase, and then the preheated fusion agent (PEG4000) was slowl...
PUM
| Property | Measurement | Unit |
|---|---|---|
| reaction rate constant | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More