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Cleaning method of agarose microspheres

A technology of agarose microspheres and agarose heat, applied in separation methods, microsphere preparation, chemical instruments and methods, etc., can solve the problems of high recycling costs, environmental pollution, etc., achieve stable yields, easy cleaning process, The effect of high yield

Active Publication Date: 2022-03-18
WUHAN RUIFA MEDICAL DEVICES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cleaning method of the above-mentioned microspheres will use a large amount of organic reagents in actual production, the recovery cost in the use process is high, and the production process is easy to cause environmental pollution

Method used

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  • Cleaning method of agarose microspheres
  • Cleaning method of agarose microspheres

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] S1: Preparation of agarose microspheres: Weigh 4.0g of agarose powder in 100mL of purified water, heat it to dissolve completely, which is the water phase; weigh 12g of Span 80 in 300mL of liquid paraffin, stir well and control the temperature at 60~ 80°C, as the oil phase. Drop the water phase into the oil phase, stir at 60-80°C for 30-60 minutes, cool at 2-5°C / min to below 20°C, and let it solidify for 1 hour to form microspheres.

[0042] S2: After cooling and solidifying the emulsion into balls, let it stand at room temperature for 16-24 hours. After standing and stratifying, the upper layer of the emulsion is a clear oil phase, about 200mL, and the lower layer is an emulsified layer; suck out the upper oil phase to obtain Emulsion layer of microspheres.

[0043] S3: After adding an equal volume of purified water to the obtained emulsified layer, after stirring at a constant speed of 150 rpm for 10 minutes, let it stand for 15 minutes to separate the water phase fr...

Embodiment 2

[0048] Compared with embodiment 1, embodiment 2 only increases the number of repetitions of step S3. The number of repetitions of step S3 in step S4 in embodiment 2 is 4 times, and the rest of the steps are the same as in embodiment 1, and will not be repeated here. Table 2 records the mass of microspheres collected after step S3 each time.

[0049] Table 2 Effect of the number of operations in step S3 on the quality of the obtained microspheres

[0050] Operations 1 2 3 4 5 Mass of each microsphere / g 79.64 10.81 1.37 0.52 0.03 Total mass of microspheres / g 79.64 90.45 91.82 92.34 92.37 % of total mass 86.22％ 97.92％ 99.40％ 99.97％ ——

[0051] As can be seen from Table 2, when the number of operations of step S3 is the 4th time, almost all microspheres have been obtained, that is, when the number of times of repeating step S3 in step S4 is 3 times, all microspheres in the emulsified layer can be separate out. And step S3 needs t...

Embodiment 3～6

[0053] Compared with Example 1, Examples 3-6 are different in that the amount of purified water added and the stirring rate involved in step S3 are changed, and the rest of the steps are the same as in Example 1, and will not be repeated here. The specific parameters are shown in Table 3.

[0054] Table 3 Parameter changes of Examples 3 to 6 and the quality of the obtained microspheres

[0055] Example Volume ratio of purified water to emulsified layer Stirring rate / rpm Microsphere mass / g 3 0.5:1 150 70.73 4 3:1 150 92.17 5 1:1 50 91.35 6 1:1 200 91.78

[0056] Compared with Example 1, Examples 3 and 4 only changed the volume ratio of the purified water added in step S3 to the emulsified layer. It can be seen from Table 3 that the volume ratio of purified water to the emulsified layer will affect the quality of the finally obtained microspheres, and if the volume ratio is small, the quality of the obtained microspheres will decr...

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Abstract

The invention provides a method for cleaning agarose microspheres, which comprises the following steps: standing and layering an agarose microsphere emulsion obtained by an emulsification-cooling method, and separating an upper oil phase to obtain a lower agarose microsphere emulsion layer; adding water into the agarose microsphere emulsion layer, standing for layering, separating to obtain an upper emulsion layer and a lower agarose microsphere water-phase layer, and separating from the lower water-phase layer to obtain agarose microspheres; and repeatedly adding water into the upper emulsion layer for layering separation, mixing the agarose microspheres obtained by each time of separation, and washing with water to obtain the high-purity agarose microspheres. According to the method, firstly, the emulsion is subjected to standing, so that most of oil phases are separated out, and the volume of an emulsion layer to be cleaned is greatly reduced, so that the maximum cleaning amount which can be realized at a time can be increased, and the running cost of equipment is saved; meanwhile, reduction of the volume of the emulsion layer must reduce the dosage of the cleaning liquid, the cleaning cost is further reduced, and the method is suitable for industrial production; the obtained microspheres have good physicochemical properties after the cross-linking reaction is completed.

Description

technical field [0001] The invention relates to the technical field of solid-liquid separation, in particular to a method for cleaning agarose microspheres. Background technique [0002] Agarose microspheres have extremely high added value and are mainly used for the separation and purification of macromolecular biologically active substances such as viruses, proteins, and enzymes, and are widely used in industries such as biomedicine, blood products, and vaccines. As a natural polysaccharide biofiller, agarose microspheres have the advantages of good biocompatibility, porosity, hydrophilicity and no charged groups, and are widely used in the field of separation chromatography. At the same time, the surface of agarose microspheres contains a large number of hydroxyl groups, which are easy to chemically react with many groups, so it is easier to modify the surface and bond groups with different functions, making it suitable for different separation objects and separation requ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D17/025B01J13/02
CPCB01D17/0208B01J13/02
Inventor 余曾成高双双庄晓慧张磊王业富
Owner WUHAN RUIFA MEDICAL DEVICES CO LTD
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