阅读说明：本技术 制备亲水玻璃微珠的方法及其应用 (Method for preparing hydrophilic glass beads and application thereof ) 是由 黄志刚 王秀柱 王丽 王伟权 刘金梁 于 2020-05-19 设计创作，主要内容包括：本文提供了一种制备亲水玻璃微珠的方法,其包括让所述玻璃微珠依次与氢氟酸溶液和尿素溶液接触。本文还提供了经该方法处理的玻璃微珠及其用于免疫检测的用途。本文提供的玻璃微珠处理方法可以在玻璃微珠灌装前随时进行,操作简单,成本低廉,能够解决玻璃微珠柱凝集卡生产过程中产生微小气泡的问题。(Provided herein is a method of preparing hydrophilic glass microspheres comprising sequentially contacting the glass microspheres with a hydrofluoric acid solution and a urea solution. Also provided herein are glass microspheres treated by this method and their use for immunoassays. The glass bead processing method provided by the invention can be carried out at any time before the glass beads are filled, is simple to operate and low in cost, and can solve the problem of generation of micro bubbles in the production process of the glass bead column agglutination card.)

1. A method for producing hydrophilic glass microspheres comprising contacting said glass microspheres in sequence with a solution of hydrofluoric acid and a solution of urea.

2. The method of claim 1, wherein the hydrofluoric acid solution has a concentration of 5% -15% (wt) and the urea solution has a concentration of 1 mol/L.

3. The method according to claim 2, wherein the glass micro beads have a particle size of 70 to 110 μm.

4. The method of claim 2 or 3, comprising:

1) mixing the glass beads in a ratio of not more than 1: 5 is mixed with the hydrofluoric acid solution in a volume ratio and reacts for at least 5 minutes at the temperature of 40-50 ℃;

2) washing the glass beads reacted in the step 1) with water;

3) mixing the glass beads cleaned in the step 2) in a mixing ratio of not higher than 1: 2.5 by volume with the urea solution and reacting at 80-105 ℃ for at least 1 hour; and

4) optionally washing the glass beads after the reaction of step 3) with water and drying.

5. The method of claim 4, wherein step 1) comprises mixing the glass microspheres in a ratio of 1: 10 volume percent was mixed with a 5% (wt) hydrofluoric acid solution.

6. The method of claim 4, wherein step 3) comprises applying the glass microspheres in a ratio of 3: 10 by volume is mixed with the urea solution.

7. The method of claim 4, wherein step 3) is carried out at 80 ℃ or 90 ℃.

8. Hydrophilic glass microspheres prepared according to any one of claims 1 to 7.

9. Use of the glass microspheres of claim 8 in an immunoassay.

10. The use of claim 9, wherein the immunoassay is a blood group test in a column agglutination card.

Technical Field

The invention relates to a method for preparing hydrophilic glass beads, in particular to a method for preparing hydrophilic glass beads capable of reducing bubble generation in the production of glass bead column agglutination cards.

Background

The glass beads as a silicate material have the characteristics of uniform size, high roundness, high light transmittance, chemical corrosion resistance, impact resistance and the like. Glass beads have numerous applications in the biomedical field, including use in the preparation of microcolumns in Column Agglutination (CAT) assays or in the preparation of column agglutination cards having a plurality of microcolumns. The glass beads can be used for packing the microcolumn, and can be used as a separation medium in the microcolumn. However, for example, in the production process of a column aggregation card, a large number of fine glass beads are put in a narrow space, and thus air is immersed in a liquid reagent along with the putting of the glass beads, thereby generating fine bubbles. A certain amount of micro-bubbles remain in the packed column and may have a serious quality influence on the function of the product.

Generally, increasing the hydrophilicity of the glass surface improves its adsorption of air and other impurities. At present, the treatment methods for preparing the super-hydrophilic wettability glass mainly comprise the following methods.

1. The micro-nano structure is formed on the surface of the glass through a nano technology, a photoetching technology, in-situ vapor deposition and the like. The glass beads treated by the process have the characteristics of controllable micro-nano structure and strong super-hydrophilic capability, but the formed micro-nano structure is fragile and cannot be slightly touched, and the instrument cost is high;

2. modifying the surface groups of the glass beads by methods such as molecular membranes, surface functionalization and the like. The super-hydrophilic glass beads prepared by the methods have low synthesis temperature and uniform chemical components, but also have the problems that the film is not resistant to friction and the super-hydrophilic effect is not long enough in maintenance time;

3. by coating the nano material coating, the super-hydrophilic capability is obtained on the surface of the glass microsphere. The super-hydrophilic glass beads prepared by the method have strong hydrophilicity and wide application range, but most of the nano materials used for coating do not have the transparency and light transmission of common glass.

Therefore, there is still a need for an improved hydrophilic treatment method for glass beads.

Disclosure of Invention

In one aspect, provided herein is a method of making hydrophilic glass microspheres comprising sequentially contacting the glass microspheres with a solution of hydrofluoric acid and a solution of urea.

In some embodiments, the concentration of the hydrofluoric acid solution is 5% to 15% (wt).

In some embodiments, the concentration of the urea solution is 1 mol/L.

In some embodiments, the glass microspheres have a particle size of 70 to 110 μm.

In some embodiments, the method comprises:

1) mixing the glass beads in a ratio of not more than 1: 5 is mixed with the hydrofluoric acid solution in a volume ratio and reacts for at least 5 minutes at the temperature of 40-50 ℃;

2) washing the glass beads reacted in the step 1) with water;

3) mixing the glass beads cleaned in the step 2) in a mixing ratio of not higher than 1: 2.5 by volume with the urea solution and reacting at 80-105 ℃ for at least 1 hour; and

4) optionally washing the glass beads after the reaction of step 3) with water and drying.

In some embodiments, step 1) comprises coupling the glass microspheres in a ratio of 1: 10 volume percent was mixed with a 5% (wt) hydrofluoric acid solution.

In some embodiments, step 3) comprises coupling the glass microspheres in a ratio of 3: 10 by volume is mixed with the urea solution.

In some embodiments, the reaction in step 3) is at 80 ℃ or 90 ℃.

In another aspect, provided herein are hydrophilic glass microspheres made using the above-described method.

In another aspect, provided herein is the use of the above-described glass beads in an immunoassay.

In some embodiments, the immunoassay is a blood group test in a column agglutination card.

The glass bead processing method provided by the invention can be carried out at any time before the glass beads are filled, is simple to operate and low in cost, and can solve the problem of generation of micro bubbles in the production process of the glass bead column agglutination card.

Detailed Description

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

"glass" is a common amorphous inorganic material, generally made of many inorganic minerals (such as quartz sand, borax, boric acid, heavy mineral)Spar, barium carbonate, limestone, feldspar, soda ash and the like) as main raw materials, and a small amount of auxiliary raw materials are added. Its main components are silicon dioxide and other oxides (such as Na)₂O, CaO, etc.). In some embodiments, the glass employed herein is a silicate glass, such as soda lime glass, quartz glass, and the like. The "glass microspheres" are generally spherical and may have a particle size of between 1 μm and 1mm, for example, between 20 μm and 500 μm, or between 50 μm and 200 μm. Of course, the methods provided herein can also handle glass microspheres outside this size range.

"blood typing" refers to the detection of antigens on blood cells and/or corresponding antibodies in plasma. Common blood group tests include the ABO and Rh detection of red blood cells and the detection of platelet and related antibodies.

The preparation method of the hydrophilic glass beads comprises the step of reacting the glass beads with a hydrofluoric acid solution and a urea (or called carbamide) solution sequentially under a heating condition.

In some embodiments, glass microspheres are reacted with a hydrofluoric acid solution in the following manner: mixing the glass beads with hydrofluoric acid solution with the concentration of 5-15% (wt) under the condition of heating, wherein the mixing volume ratio of the glass beads to the hydrofluoric acid solution is 1: 5 or less, e.g., 1: 10,1: 15,1: 20, etc. The reaction temperature is usually maintained above 40 deg.C, for example 40-50 deg.C, for example around 45 deg.C. The reaction time can be adjusted depending on the hydrofluoric acid concentration and the reaction temperature, and the reaction time can be usually 5 minutes or more, for example, 10 minutes, 20 minutes, or the like. After the reaction, the glass beads were washed with clean water to remove the hydrofluoric acid solution until the water after washing was detected to be approximately neutral (pH 6-8).

In some embodiments, hydrofluoric acid treated glass microspheres are reacted with an aqueous urea solution in the following manner: mixing glass beads with 1mol/L of urea aqueous solution, wherein the volume ratio of the glass beads to the urea aqueous solution is not more than 1: 2.5 (e.g., volume ratio of 1: 3, 1: 4, 1: 5, etc.). The glass beads were allowed to react with urea with heating for 1 hour or more. The reaction temperature is preferably 80 to 105 ℃ such as 82, 85, 88, 90, 95, 100 ℃ and the like. The solution after the reaction may be alkaline or weakly alkaline, followed by washing the glass beads with water until the washed water is detected to be approximately neutral (pH 6-8).

In some more specific embodiments, the methods of making hydrophilic glass microspheres provided herein can comprise:

1) preparing 5-15% (wt) hydrofluoric acid solution with distilled water or deionized water, and heating to 40-50 deg.C;

2) adding glass beads into a hydrofluoric acid solution, wherein the total amount of the glass beads is not more than 20% (vol) of the hydrofluoric acid solution;

3) reacting for 5 minutes or more under heating, and keeping constant stirring during the reaction;

4) after the reaction is finished, standing, discharging the upper-layer acid solution after the glass beads are precipitated, and washing the glass beads with water until the washed water is neutral (pH 6-8);

5) adding the cleaned glass beads into a pre-prepared 1mol/L urea aqueous solution, wherein the total amount of the glass beads is not more than 40% (vol) of the urea aqueous solution;

6) heating the mixture of the glass beads and the urea solution, and reacting at the temperature of 80-90 ℃ for 1 hour or more;

7) after the reaction is finished, discharging the upper urea solution after the glass beads are precipitated;

8) washing the glass beads with water until the washed water is neutral (pH 6-8);

9) and (5) drying the glass beads.

We have found that by improving the hydrophilicity of the surfaces of glass beads, the time for the infiltration process of the glass beads in the production process of the glass bead column agglutination card can be reduced, and the number of bubbles generated in the process of applying the glass bead column agglutination card to the production process of the glass bead column agglutination card can be greatly reduced.

The glass beads made by the methods herein can be used in a variety of applications for detection using column agglutination methods. For example, it can be used for various blood type test cards, including the filling of positively-typed and negatively-typed microcolumns. After the glass beads are filled into the microcolumn, gaps among the glass beads play a role of a molecular sieve and are used for blocking agglutinated red blood cells. During the centrifugation of the blood type card, most of agglutinated red blood cells are positioned above the glass microsphere column, and unagglutinated red blood cells can reach the bottom of the microsphere column through pores among the glass microspheres under the action of centrifugal force. Therefore, the objective of distinguishing between positive and negative experimental results is achieved by separating agglutinated versus non-agglutinated erythrocytes.

Specific examples are provided below for further detailed description.