A kind of agarose gel microsphere containing glucosamine group and preparation method thereof
A technology of agarose gel and agarose microspheres, applied in the field of agarose gel microspheres containing glucosamine group and its preparation, can solve the problem of poor mechanical strength and pore size uniformity of microspheres, complex preparation process, and easy influence on activity and other problems, to achieve the effect of low cost, high purification efficiency and large adsorption capacity
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Embodiment 1
[0060] Weigh 40g of agarose solid, add 1000mL of ultrapure water, mix well to make an agarose solution, then slowly add NaBH 4 0.0004g, then continue to use the pump to add 20wt% NaOH solution at a flow rate of 0.5mL / min, mechanically stir and mix, the stirring speed is 80rpm; under stirring conditions, continue to use the pump to add epoxy chlorine at a flow rate of 0.3mL / min Propane, react at 50°C for 1 hour, gradually reduce the flow rate of epichlorohydrin to 0.03mL / min when the reaction reaches 0.3h, and reduce the flow rate of NaOH solution to 0.05mL / min; Add 10wt% glucosamine sulfate solution to the solution at a flow rate of / min, stop adding epichlorohydrin and NaOH solution, and react at 55°C for 1h; filter the resulting solution with a 4040 ultrafiltration membrane, concentrate and dry to obtain amino Glucose-Sepharose.
[0061] figure 2 It is the electrophoresis comparison diagram of nucleic acid separated by the prepared glucosamine-based agarose (A) and unmodi...
Embodiment 2
[0065] Weigh 80g of agarose solid, add 1000mL of ultrapure water, mix well to make an agarose solution, slowly add NaBH 4 0.008g, then continue to add 30wt% NaOH solution with a pump at a flow rate of 1mL / min, mechanically stir and mix, and the stirring speed is 100rpm; Glyceryl ether, react at 55°C for 2 hours, gradually reduce the flow rate of ethylene glycol diglycidyl ether to 0.05mL / min when the reaction reaches 0.5h, and reduce the flow rate of NaOH solution to 0.1mL / min; Add 20wt% glucosamine hydrochloride solution to the solution with the pump at a flow rate of 0.02mL / min, stop adding ethylene glycol diglycidyl ether and NaOH solution, and react at 65°C for 1 hour; filter the resulting solution with an 8040 ultrafiltration membrane, Concentrate and dry to obtain glucosamine-based agarose.
[0066] Figure 4 It is the electrophoresis comparison diagram of nucleic acid separated by the prepared glucosamine-based agarose (A) and unmodified agarose (B). It can be seen f...
Embodiment 3
[0070] Weigh 60g of agarose microspheres, wash with water and drain, add 1000mL of ultrapure water and mix evenly to prepare agarose microsphere solution, slowly add NaBH 4 0.0012g, then continue to add 40wt% NaOH solution with a pump at a flow rate of 0.8mL / min, and mix evenly with low-frequency ultrasonic vibration, and the ultrasonic frequency is 25KHz; Alcohol diglycidyl ether, react at 40°C for 8 hours, gradually reduce the flow rate of ethylene glycol diglycidyl ether to 0.03mL / min when the reaction reaches 4h, and reduce the flow rate of NaOH solution to 0.06mL / min; Add 15wt% glucosamine hydrochloride solution to the solution with a pump at a flow rate of 0.05mL / min, stop adding ethylene glycol diglycidyl ether and NaOH solution, and react at 45°C for 1.5h; The solution is filtered with a filter cloth and washed to prepare agarose gel microspheres containing glucosamine base, and the obtained microspheres are stored in 20 wt% ethanol solution. The glucosamine group co...
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