JPH02180707A - Production of phosphorus compound grain assemblage - Google Patents

Abstract

PURPOSE:To obtain the porous assemblage appropriate for an oxygen fixing carrier, etc., with good reproducibility without using an org. binder by charging a slurry of a calcium phosphate compd. into a vessel having an inclined rotating shaft, and concentrating the slurry while heating and rotating the vessel. CONSTITUTION:A vessel lid 18 also used as a supporting rod is provided at the top of the obliquely arranged vessel 10, the lid 18 is pivoted on a rotary bearing 20, and the lid 18 is rotatably supported on a holder 22. A trap tube 24 is branched and connected to the lid 18, and the trap tube 24 is dipped in a cooling water tank 26. The slurry 14 of a calcium phosphate compd. (e.g. hydroxyapatite) free of an org. binder is supplied into the vessel 10 through a supply pipe 12, the inside of the vessel 10 is depressurized by a vacuum pump 28 and heated above room temp. by a thermostatic bath 16, the slurry 14 is concentrated while rotating the vessel, and the assemblage of porous phosphorus compd. grains is obtained.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a method for producing a phosphoric acid compound particle aggregate. The phosphoric acid compound particle aggregate produced by the method of the present invention can be used as a chromatography filler for separating and adsorbing cells and physiologically active substances, a support for culturing animal cells, and a support or immobilization carrier for enzymes. Can be done.

[Conventional technology] Conventionally, phosphoric acid compounds such as hydroxyapatite,
It is used for the separation and adsorption of cells and physiologically active substances, as a culture support for animal cells, and as an immobilization carrier for enzymes and cells.
However, a combination of particle size, surface area, pore volume, particle shape, compressive strength, bulk specific gravity, etc. that is particularly suitable for such separation materials and supports is not known.

Furthermore, there is no known method for producing phosphoric acid compound particles having a specific combination of particle size, specific surface area, pore volume, particle shape, compressive strength, bulk specific gravity, etc. with good reproducibility. Phosphoric acid compound particles having an average particle size of about 1LL11 to about 1100U are produced by a centrifugal force method or a spray granulation method. However, large-sized phosphoric acid compound particles having an average particle size exceeding 100 μm cannot be produced by the centrifugal force method or the spray granulation method.

On the other hand, phosphoric acid compound particles having an average particle diameter of 100 μm to 2000 μm are produced by a transfer granulation method in which an organic binder is added. however.

In the conventional transfer granulation method, a firing step is essential in order to add an organic binder, and during this step, the yield may decrease due to rupture of the binder, and particle properties 1 such as pore volume and specific surface area may be controlled. There are some problems that make it difficult.

In addition, a submerged granulation method is known as a general granulation method using an inorganic slurry as a raw material. In this method, a polymer flocculant or a liquid crosslinking agent is added to an inorganic slurry, and the slurry is granulated by stirring the slurry. However, when this method is applied to the granulation of phosphate compound particles, problems similar to those described for the transfer granulation method arise due to the polymeric flocculant or liquid crosslinking agent.

In this way, in the conventional technology, the average particle size is 20 μI.
or about 5000 μl of phosphoric acid compound particle aggregate,
There is no known method for manufacturing it while controlling its pore volume, surface area, etc. with high precision.

In view of the above circumstances, the inventors of the present application have found that the problems of the prior art described above can be solved by first stirring a slurry that does not contain a polymer flocculant or a liquid crosslinking agent using a rotary blade. (Japanese Patent Application No. 63-58059). However, the porous spheres produced inside the container contain a large amount of water and have low mechanical strength.
Therefore, in this method, since the rotary blade is used, the speed of one rotation is high and the rotation time is long (I see, the generated spherical bodies are destroyed by the rotary blade, and as a result, it is difficult to generate spherical bodies with a larger particle size. The yield of spheroids decreases due to
Destroyed flakes become mixed in as irregular objects that are difficult to remove.
Problems such as this arise.

[Problems to be Solved by the Invention] Therefore, an object of the present invention is to provide a phosphoric acid compound having specific physical properties that exhibits excellent effects when used as a chromatography filler or a support for animal cells. The object of the present invention is to provide a manufacturing method that allows particle aggregates to be manufactured with good reproducibility.

[Means for Solving the Problems] As a result of intensive research, the inventors of the present application have discovered a combination of physical properties that exhibits excellent effects when used as a chromatography filler or a support for animal cells. The present invention was completed based on the discovery that these can be obtained with good reproducibility by rotating the container itself and stirring the raw material slurry.

That is, it includes a step of loading a slurry of a calcium phosphate compound containing no organic binder into a container having an inclined rotation axis, and a step of heating the container above room temperature and rotating the slurry while concentrating it. A method for producing a porous phosphate compound particle aggregate is provided.

[Effects of the Invention] The present invention provides a method for producing a phosphoric acid compound particle aggregate that exhibits excellent effects when used as a separating agent and support for cells and physiologically active substances. According to the method of the present invention, a phosphoric acid compound particle aggregate having an average particle size of 20 μl to 5000 μl can be produced without using an organic binder. In the method of the present invention, since impurities such as an organic binder are not added, it is possible to use the phosphoric acid compound particles as they are as a separating agent or a carrier without baking them. Further, there is no decrease in yield due to rupture of the organic binder, and pore volume, specific surface area, etc. can be easily controlled.

The phosphoric acid compound particle aggregate produced by the method of the present invention is relatively large and substantially spherical, so when used as a separation agent, it is possible to efficiently separate large cells, etc. can greatly contribute to downstream processing in the biotechnology field.

Furthermore, the particle aggregate produced by the method of the present invention having the above-mentioned physical properties can be used as a support or carrier in the biochemical field, for example, as a support for culturing animal cells, or as a support for immobilizing enzymes or cells. It is particularly useful as

Furthermore, the particle aggregate produced by the method of the present invention is
Since it is manufactured without the use of rotary blades, it is virtually free of foreign contaminants such as particle fragments.

[Detailed Description of the Invention] The average particle size of the phosphoric acid compound particle aggregate produced by the method of the present invention is from 20 μI to 5000 μm, preferably from 2000 μI to 5000 μm. If the average particle diameter is smaller than 20 μm, it is too small to be effective as a support for cell culture, and if it exceeds 5000 μI, it is too large to be effective as a support for cell culture.

The total pore volume of the particle aggregate produced by the method of the present invention is between 0.01 m17 g and 1.0 g as measured by mercury porosimetry.
0 m17g, preferably 0.1 +++ I/g to 0.6 ■l/g. Total pore volume is 0.01 m17
If it is less than 1.0 +al/g, it will have high density (low separation ability and little cell separation effect), and if it exceeds 1.0+al/g, it will be highly porous and brittle, resulting in poor strength as a support for cell culture or a cell separation agent. is not enough.

The specific surface area of the particle aggregates produced by the method of the invention is between 5 rm2/g and 100 m''7g, preferably 4.
0 rm"7g to 80 m"7g. If the specific surface area is less than 5ra"1g, the density will be high, the separation ability will be low, and the effect as a cell separation agent will be low.
When the amount exceeds 100 g, adsorption properties become diverse and selective cell separation becomes difficult.

The compressive strength of the particle aggregate produced by the method of the present invention is 0.1 kg/mm" to 5.0 kg/++m"
, preferably 0.2 kg/mm” to 3.0 kg
/man''.If the compressive strength is less than 0.1 kg/mm'', the compressive strength will be too small and it will be easily destroyed when used as a chromatography packing material; If it exceeds "m+s", the density will inevitably increase and the separation power will decrease.

The particle aggregate produced by the method of the present invention has an average bulk specific gravity of 0.4 to 2.5, preferably 0.5 to 10.
It is. If the bulk specific gravity is less than 0.4, the porosity will increase, resulting in a decrease in mechanical strength, making it easy to break.2.
When it is larger than 5, the density increases and the separation ability decreases.

The average porosity of the particle aggregates produced by the method of the invention is between 1% and 85%, preferably between 40% and 70%. If the average porosity is less than 1%, the density increases and the separation ability decreases, and if it is more than 85%, the mechanical strength is too small and it is easy to break. This is the volume ratio occupied by pores.

The particle aggregate produced by the method of the present invention is substantially spherical and substantially free of irregularly shaped particles such as particles other than spherical, for example, particle fragments. There will be variations in resolution when used as a

The "phosphoric acid compound" in the present invention includes hydroxyapatite, calcium phosphate, and fluoroapatite.

The particle aggregate described above can be manufactured as follows.

First, a slurry of a phosphoric acid compound is prepared and placed in a container with an inclined rotation axis. Slurry of phosphoric acid compound is
Those conventionally known in this field can be used. However, in the method of the present invention, the concentration of the slurry influences the particle size of the particles produced, and the higher the concentration, the larger the particles produced. As described below, the rotation speed and rotation time of the container also affect the particle size, so the concentration of the slurry is appropriately selected depending on the desired particle size, rotation speed, and rotation time. However, if the phosphoric acid compound concentration in the slurry is too low or too high, it will be difficult to granulate the phosphoric acid compound, so the appropriate concentration of the aqueous slurry is 10% by weight to 60% by weight, especially 20% by weight. % to about 50% by weight. The slurry medium is most preferably water, but may contain extra-aquatic materials or impurities provided they do not adversely affect the performance of the particles produced.

Next, the container is rotated around the rotation axis while heating the container to a temperature equal to or higher than room temperature. The temperature of the container is preferably from room temperature to about 100° C. The container can be heated, for example, by rotating the container in a constant temperature bath. The rotation speed of the container affects the particle size of the particles, and the higher the rotation speed, the smaller the particle size.8 The rotation speed is 5.
rpm to 300 rpm, especially lorpm+ to 100 rpm+m are preferred, and the slurry can be concentrated, for example, by pulling the container under vacuum. The pressure in the vessel for slurry concentration was set to 0. O
1 atm to 1.0 atm, especially 0.05 atm to 0
．． Preferably, the pressure is 5 atm.

The stirring time also influences the particle size of the resulting particles; the longer the stirring time, the larger the resulting particles (thus, the stirring time depends on the desired particle size and slurry concentration and the stirring speed). The choice will be made as appropriate depending on the balance.However,
If the stirring time is too short, the yield of granulation will be poor, so the stirring time is preferably one hour or more.The stirring time can be shortened by heating. Furthermore, if the stirring time is 20 hours or more, the growth of particle size will almost stop, so from the economic point of view, the stirring time is preferably 20 hours or less.

By the above method, the porous phosphoric acid compound particle aggregate described above can be efficiently and reproducibly obtained in the container.

The porous phosphoric acid compound particle aggregate obtained by the above method can be used as it is as a separating agent or a support for cell culture, but it can also be further calcined. By firing, the pore size distribution can be easily controlled and mechanical strength can be improved. Firing conditions are not particularly limited, but 200°C to 1200°C, particularly 30°C.
1 hour to 50 hours at 0℃ to 900℃, especially 3
It is preferable to carry out the treatment for 10 hours to 10 hours.

The method of the present invention described above can be carried out using, for example, an apparatus as schematically shown in FIG. A phosphate compound slurry supply pipe 12 is introduced into the container 10, and the end of the supply pipe 12 is connected to a phosphate compound slurry supply pump 13. Slurry is supplied from a slurry tank (not shown) to the container via the supply pipe 12. In FIG. 6, 14 indicates the phosphoric acid compound slurry supplied in this way.At least the portion of the container IO containing the slurry 14 is immersed in a constant temperature bath 16. A container lid 18 is provided at the top of the container IO, and the container lid 18 also functions as a support rod. The container lid 18 is pivoted on a rotation bearing 20 and rotates in the direction of the arrow while being supported on a support base 22.

Further, a trap pipe 24 is branched from the container lid 18, and a portion of the trap pipe 24 is immersed in a cooling water tank 26. A vacuum pump 28 is connected to the end of the trap pipe 24, and when the vacuum pump 28 is operated to reduce the pressure inside the container 10, the slurry trapped in the trap pipe 24 is reused.

Hereinafter, the present invention will be explained based on Examples; however, the present invention is not limited to the Examples.

[Example] Using the apparatus shown in Figure A, spherical particle aggregates were produced in 11 dishes under the production conditions shown in Table 1. The phosphoric acid compound slurry used was a hydroxyapatite aqueous slurry, and the hydroxyapatite concentration was as shown in Table 1. After production, one particle was dried at 50°C for 10 hours. The average particle diameter of the obtained particles was measured. The results are shown in Table 1. The average particle diameter was measured by taking an optical microscope photograph. The microscope used had an objective lens of 0.7 to 4 times magnification and an eyepiece lens of approximately 10 times magnification.

! The sample obtained in Example 4 was heated at 300°C, 500°C or 70°C.
It was baked at 0° C. for 3 hours, and the physical properties shown in Table 2 were measured.

The results are shown in Table 2.

In addition, the specific measurement method of each physical property value was as follows.

Specific product Carlo Erba [5por manufactured by Carlo Erba1]
It was calculated by measuring the adsorption/desorption isotherm of nitrogen gas using t+5aticSeries 1800.

L Mouse Yu 11 Based on the Mercury Seitai method, Shimadzu Corporation's mercury intrusion porosimeter rMicromeritics Auropor
Measured using e92001.

Shimadzu Corporation's mercury intrusion porosimeter rMicroa+eritics Auropo based on the Mercury Seitai method
It was determined from a pore size distribution diagram using re9200J. This pore size distribution map was obtained using "Bore Plot System J" (a software program manufactured by Takasugi Seisakusho).

L1 Technique 1 The dry weight of 10 ml of the particle aggregate was measured, and the weight per unit volume (c+a''l) was calculated.

5YU1J This is the ratio occupied by all the pores present in the particle aggregate, calculated from the pore volume and the theoretical density of hydroxyapatite.

The measurement was carried out under the following measurement conditions using a universal testing machine Autograph AG-8 manufactured by Tamayu Pressure 1 Rin Takashi Seisakusho.

(Test speed 0.5 tars 1 minute, load cell 10
0gf1

[Brief explanation of the drawing]

The figure is a diagram schematically showing one embodiment of a manufacturing apparatus for manufacturing a phosphate compound particle aggregate of the present invention. 10... Container, 12... Slurry supply pipe, 13...
・Slurry supply pump, 14...Slurry, 16...
・Thermostatic chamber, 18... Container lid, 20... Rotating bearing, 2
2... Support stand, 24... Trap pipe, 26... Water tank, 28... Vacuum pump patent applicant Toa Fuel Industries Co., Ltd.

Claims (1)

[Claims] A porous method comprising the steps of: loading a slurry of a calcium phosphate compound without an organic binder into a container having an inclined axis of rotation; warming the container above room temperature; and rotating the slurry while concentrating it. A method for producing a phosphoric acid compound particle aggregate.