CN107019710B - Method for increasing passing rate of bacteria-removing and filtering effective components of nocardia rubra cell wall skeleton emulsion - Google Patents
Method for increasing passing rate of bacteria-removing and filtering effective components of nocardia rubra cell wall skeleton emulsion Download PDFInfo
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- 238000001914 filtration Methods 0.000 title claims abstract description 47
- 239000000839 emulsion Substances 0.000 title claims abstract description 27
- 108010039939 Cell Wall Skeleton Proteins 0.000 title claims abstract description 17
- 241000187563 Rhodococcus ruber Species 0.000 title claims abstract description 17
- 210000004520 cell wall skeleton Anatomy 0.000 title claims abstract description 17
- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000000227 grinding Methods 0.000 claims abstract description 99
- 239000007788 liquid Substances 0.000 claims abstract description 88
- 238000003756 stirring Methods 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000008215 water for injection Substances 0.000 claims abstract description 23
- 239000000843 powder Substances 0.000 claims abstract description 21
- 239000000463 material Substances 0.000 claims abstract description 17
- 238000002156 mixing Methods 0.000 claims abstract description 17
- 239000000243 solution Substances 0.000 claims abstract description 15
- 230000001954 sterilising effect Effects 0.000 claims abstract description 15
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 13
- 238000005406 washing Methods 0.000 claims abstract description 11
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 9
- 238000005086 pumping Methods 0.000 claims abstract description 4
- 239000004480 active ingredient Substances 0.000 claims abstract description 3
- 239000002994 raw material Substances 0.000 claims abstract description 3
- 239000003814 drug Substances 0.000 claims description 39
- 239000011324 bead Substances 0.000 claims description 28
- 238000003801 milling Methods 0.000 claims description 19
- 238000000265 homogenisation Methods 0.000 claims description 14
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 claims description 13
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims description 13
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims description 13
- 238000011049 filling Methods 0.000 claims description 12
- 238000002347 injection Methods 0.000 claims description 12
- 229940090044 injection Drugs 0.000 claims description 12
- 239000007924 injection Substances 0.000 claims description 12
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 claims description 9
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 claims description 9
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 claims description 9
- 229940031439 squalene Drugs 0.000 claims description 9
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 claims description 9
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 claims description 8
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 8
- 229920000053 polysorbate 80 Polymers 0.000 claims description 8
- 229940068968 polysorbate 80 Drugs 0.000 claims description 8
- 229940080526 mannitol injection Drugs 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 5
- 238000007599 discharging Methods 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- 239000004570 mortar (masonry) Substances 0.000 claims description 5
- 238000005070 sampling Methods 0.000 claims description 5
- 238000011068 loading method Methods 0.000 claims description 4
- 239000011148 porous material Substances 0.000 claims description 2
- 239000002002 slurry Substances 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims 1
- 229950008882 polysorbate Drugs 0.000 claims 1
- 108010073243 Nocardia cell wall skeleton Proteins 0.000 abstract description 12
- 239000002245 particle Substances 0.000 abstract description 2
- 241000894006 Bacteria Species 0.000 abstract 1
- 238000010008 shearing Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 8
- 239000006185 dispersion Substances 0.000 description 6
- 238000011045 prefiltration Methods 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 238000004945 emulsification Methods 0.000 description 5
- 206010028980 Neoplasm Diseases 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- 229930195725 Mannitol Natural products 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 239000013589 supplement Substances 0.000 description 3
- 239000003082 abrasive agent Substances 0.000 description 2
- 239000010431 corundum Substances 0.000 description 2
- 229910052593 corundum Inorganic materials 0.000 description 2
- 239000010432 diamond Substances 0.000 description 2
- 229910003460 diamond Inorganic materials 0.000 description 2
- 238000011146 sterile filtration Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000002223 garnet Substances 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- HBMJWWWQQXIZIP-UHFFFAOYSA-N silicon carbide Chemical compound [Si+]#[C-] HBMJWWWQQXIZIP-UHFFFAOYSA-N 0.000 description 1
- 229910010271 silicon carbide Inorganic materials 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
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- Health & Medical Sciences (AREA)
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Abstract
The invention discloses a method for improving the passing rate of sterilization and filtration active ingredients of nocardia rubra cell wall skeleton emulsion, which comprises the following steps: 1. premixing and dispersing raw materials and auxiliary materials: grinding and homogenizing red nocardia cell wall skeleton raw powder and auxiliary materials; 2. stirring, grinding and grinding: grinding rotating speed: 3600-3800 rpm, setting the highest temperature of an equipment outlet: 40 ℃; and the grinding efficiency is improved by adopting graded grinding. First grinding to obtain (solution A). Washing the grinding cavity and the pipeline (solution B) with a proper amount of water for injection, and uniformly mixing the solution A and the solution B to obtain solution AB; dividing the AB liquid into two parts, and grinding each sample for 45 min; wherein the pumping speed of the peristaltic pump is 10% for the first 10min and 20% for the second 10min, and finally the grinding is finished at the pumping speed of 25%. The invention obviously grinds the emulsion to superfine particle size, greatly reduces the interception of the bacteria removing filter to the red nocardia cell wall skeleton raw powder, improves the passing rate of effective components, and then improves the yield of finished products under the conditions of ensuring the product quality and meeting the preset application of the products.
Description
Technical Field
The invention relates to a method for improving the passing rate of effective components of sterilization and filtration of an emulsion which is an intermediate product of a red nocardia rubra cell wall skeleton in the technical field of microbial pharmacy.
Background
The effective component of Ikejia is a red nocardia cell wall skeleton, the specification of the red nocardia cell wall skeleton (N-CWS) for injection is 200 mu g/bottle, the arabinose content is 66-104 mu g/bottle, and the N-CWS content is calculated by measuring the arabinose content. Therefore, the loading of the cell wall skeleton of the nocardia rubra for injection depends on the arabinose content of the emulsion after filtration. Under the condition of unchanged feeding amount, if the arabinose content of the filtered emulsion is high, the corresponding per-bottle loading is reduced, and the yield (batch) is increased.
An important factor influencing the high and low arabinose content is the passing rate of the effective components after the liquid medicine is sterilized and filtered. The less the active ingredient is retained during the sterile filtration process, the higher the sugar content correspondingly.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method which not only maintains the activity and performance of the original substances, but also improves the maximum passing rate of the effective components in the sterilization and filtration process of the materials, namely, improves the product yield under the conditions of ensuring the product quality and meeting the preset application of the product; but also improve the method for quickly and fully releasing the medicine in the human body. The process is stable and reliable and has uniform quality.
The invention realizes the above purposes by the following technical scheme:
a method for improving the passing rate of effective components of nocardia rubra cell wall skeleton emulsion in sterilization and filtration comprises the following steps:
1) premixing and dispersing raw materials and auxiliary materials:
grinding raw and auxiliary materials: placing the weighed raw powder into a mortar of a grinder, adding a mixed solution of squalene, polysorbate 80, a proper amount of 20% mannitol injection and water for injection according to the prescription amount, and grinding for 15-25 min;
homogenizing: adding the ground liquid medicine into a sample cup of a homogenizer for low-pressure homogenization, and homogenizing at 900-1100 bar for one time; homogenizing under high pressure for 1700-1900 bar three times; collecting the homogenized solution in a container, cleaning the cavity of the homogenizer with appropriate amount of water for injection, washing out the residual medicinal liquid, and mixing with the previous homogenized solution; the solid-liquid ratio is 1 g: 500-700 ml;
2) stirring, grinding and grinding:
firstly, grinding: grinding beads of 0.08-0.2mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 85-95%; solid-liquid ratio: 1 g: 500-700 ml;
a. pouring the homogenized liquid medicine into a circulating hopper; setting the rotating speed of a stirring mill rotor at 3600-3800 rpm, and setting the highest outlet temperature: 40 ℃;
b. starting a stirring mill, wherein the whole milling time is 85-95min, the first 10min of the pump speed of the peristaltic pump is 8-12%, the second 10min of the peristaltic pump is 15-25%, and finally, the milling is finished at the pump speed of 20-30%; discharging the liquid medicine to obtain liquid medicine A;
c. washing the grinding cavity and the pipeline with a proper amount of water for injection to obtain liquid B, and uniformly mixing the liquid A and the liquid B to obtain liquid AB, wherein the solid-liquid ratio is 1 g: 800-;
secondly, grinding: grinding beads of 0.02-0.07mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 85-95%;
a. unloading the grinding beads with the diameter of 0.08-0.2mm, and loading the grinding beads with the diameter of 0.02-0.07 mm;
dividing the AB liquid into two parts, pouring the two parts into a circulating cup of an assembled bead mill in batches, setting the stirring mill rotation speed to be 3600rpm-3800rpm, and grinding for 40-50 minutes; the pumping speed of the peristaltic pump is 8-12% for the first 10min and 20% for the second 15-25min, and then the grinding is finished at the flow rate of 20-30%;
d. adding the injection water to the full amount according to the formula amount, starting the stirring slurry, and stirring at 400-600rpm for 8-12 min.
3) Pre-filtering: pre-filtering the medicinal liquid with 0.4-0.5 μm filter;
4) and (3) degerming and filtering: filtering the medicinal liquid with a filter of 0.4-0.5 μm with a sterilizing filter.
In a preferred embodiment of the present invention, the step 4) further comprises the step of sampling and measuring the content of the arabinose which is an effective component after the aseptic filtration.
In a preferred embodiment of the present invention, the pore size of the sterile filtration in step 4) is 0.22 μm. .
The material of the grinding bead of the invention can be natural abrasive materials, including diamond, corundum, garnet, quartz, and the like. Or synthetic abrasives including diamond-based, carbide-based (e.g., silicon carbide), corundum-based, and the like.
The invention has the beneficial effects that: 1. optimizing and designing an operation process: the effective pretreatment enables the material to be micronized, and the proper rheological property of the suspension is manufactured, so that a good material concentration and particle foundation is provided for the stirring and grinding effect; 2. high-energy grinding is carried out at low temperature, so that thermal stability is realized, and the activity and performance of the original substance are maintained. 3. The stirring mill selects grinding beads with different specifications for graded grinding, effectively improves the crushing effect, improves the passing rate of effective components of the product under the conditions of ensuring the product quality and meeting the preset application of the product, and then improves the yield. 3. The tumor inhibition rate is basically kept unchanged while the product yield is improved.
Detailed Description
Influence of different emulsification methods on the passing rate of effective components in sterilization and filtration of emulsion
Example 1:
1. the red nocardia cell wall skeleton raw powder and the auxiliary materials are accurately weighed and prepared according to the prescription amount.
Prescription (prior art):
nocardia rubra cell wall skeleton raw powder 1g
Proper amount of medium
The medium formula comprises:
2. processing parameters:
grinding agate: mixing the raw powder, squalene, polysorbate 80, 20% mannitol injection and water for injection, and grinding for 20 min;
homogenizing: adding the ground liquid medicine into a sample cup of a homogenizer for low-pressure homogenization, wherein the liquid medicine is homogenized once at 1000bar and is homogenized for three times at 1800 bar; collecting the homogenized solution in a container, cleaning the cavity of the homogenizer with appropriate amount of water for injection, washing out the residual medicinal liquid, and mixing with the previous homogenized solution; the solid-liquid ratio is 1 g: 600ml (concentrated).
Stirring, grinding and grinding:
a. primary grinding: grinding beads of 0.1mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 90%; solid-liquid ratio: 1 g: the rotating speed of a 600ml stirring mill rotor is 3600 rpm;
starting the stirring mill, wherein the whole milling time is 90min, the first 10min of the peristaltic pump is 10%, the second 10min is 20%, and finally, the milling is finished at the pump speed of 25%. Discharging the liquid medicine to obtain liquid medicine A;
washing the grinding cavity and the pipeline (liquid B) with a proper amount of water for injection, and uniformly mixing the liquid A and the liquid B to obtain (liquid AB) so that the solid-liquid ratio is 1 g: 900ml (concentrated);
b. secondary grinding: grinding beads of 0.05mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 90%; solid-liquid ratio: 1 g: the rotating speed of a 900ml mixing and grinding rotor is 3700 rpm; starting the stirring mill, wherein the whole milling time is 90min, the first 10min of the peristaltic pump is 10%, the second 10min is 20%, and finally, the milling is finished at the pump speed of 25%. Adding water for injection to full dose according to the formula amount. The stirring paddle was turned on and stirred at 500rpm for 10 min.
Fourthly, after the liquid medicine is treated, transferring the emulsion into a filtering system for filtering, transferring the emulsion into the filtering system for pre-filtering, wherein the aperture of a filter element used for pre-filtering is 0.45 mu m; the aperture of the sterilizing filter element is 0.22 μm.
Sampling and measuring the content of the effective component arabinose.
And tumor inhibition test: according to the third supplement of 'red nocardia rubra cell wall skeleton for injection' in Chinese pharmacopoeia 2010 edition.
Example 2:
1. the red nocardia cell wall skeleton raw powder and the auxiliary materials are accurately weighed and prepared according to the prescription amount.
Prescription (prior art):
nocardia rubra cell wall skeleton raw powder 1g
Proper amount of medium
The medium formula comprises:
2. processing parameters:
grinding agate: mixing the raw powder, squalene, polysorbate 80, 20% mannitol injection and water for injection, and grinding for 18 min;
homogenizing: adding the ground liquid medicine into a sample cup of a homogenizer for low-pressure homogenization, wherein the liquid medicine is homogenized once at 1000bar and is homogenized for 1700bar three times at high pressure; collecting the homogenized solution in a container, cleaning the cavity of the homogenizer with appropriate amount of water for injection, washing out the residual medicinal liquid, and mixing with the previous homogenized solution; the solid-liquid ratio is 1 g: 700ml (concentrated).
Stirring, grinding and grinding:
a. primary grinding: grinding beads of 0.2mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 85%; solid-liquid ratio: 1 g: the rotating speed of a 700ml stirring mill rotor is 3700 rpm;
starting the stirring mill, wherein the whole milling time is 85min, the first 10min of the peristaltic pump is 10%, the second 10min is 20%, and finally, the milling is finished at the pump speed of 25%. Discharging the liquid medicine to obtain liquid medicine A;
washing the grinding cavity and the pipeline (liquid B) with a proper amount of water for injection, and uniformly mixing the liquid A and the liquid B to obtain (liquid AB) so that the solid-liquid ratio is 1 g: 800ml (concentrated);
b. secondary grinding: grinding beads with the diameter of 0.1mm are stirred, ground and emulsified, and the filling degree of the grinding beads is 85 percent; solid-liquid ratio: 1 g: the rotating speed of a 700ml mixing and grinding rotor is 3700 rpm; starting the stirring mill, wherein the whole milling time is 85min, the first 10min of the peristaltic pump is 10%, the second 10min is 20%, and finally, the milling is finished at the pump speed of 25%. Adding water for injection to full dose according to the formula amount. The stirring paddle was turned on and stirred at 500rpm for 10 min.
Fourthly, after the liquid medicine is treated, transferring the emulsion into a filtering system for filtering, transferring the emulsion into the filtering system for pre-filtering, wherein the aperture of a filter element used for pre-filtering is 0.45 mu m; the aperture of the sterilizing filter element is 0.22 μm.
Sampling and measuring the content of the effective component arabinose.
And tumor inhibition test: according to the third supplement of 'red nocardia rubra cell wall skeleton for injection' in Chinese pharmacopoeia 2010 edition.
Example 3:
1. the red nocardia cell wall skeleton raw powder and the auxiliary materials are accurately weighed and prepared according to the prescription amount.
Prescription (prior art):
nocardia rubra cell wall skeleton raw powder 1g
Proper amount of medium
The medium formula comprises:
2. processing parameters:
grinding agate: mixing the raw powder, squalene, polysorbate 80, 20% mannitol injection and water for injection, and grinding for 25 min;
homogenizing: adding the ground liquid medicine into a sample cup of a homogenizer for low-pressure homogenization, wherein the liquid medicine is homogenized once at 1000bar and is homogenized for 1900bar three times at high pressure; collecting the homogenized solution in a container, cleaning the cavity of the homogenizer with appropriate amount of water for injection, washing out the residual medicinal liquid, and mixing with the previous homogenized solution; the solid-liquid ratio is 1 g: 500ml (concentrated).
Stirring, grinding and grinding:
a. primary grinding: grinding beads of 0.1mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 90%; solid-liquid ratio: 1 g: the rotating speed of a 600ml stirring mill rotor is 3600 rpm;
starting the stirring mill, wherein the whole milling time is 90min, the first 10min of the peristaltic pump is 10%, the second 10min is 20%, and finally, the milling is finished at the pump speed of 25%. Discharging the liquid medicine to obtain liquid medicine A;
washing the grinding cavity and the pipeline (liquid B) with a proper amount of water for injection, and uniformly mixing the liquid A and the liquid B to obtain (liquid AB) so that the solid-liquid ratio is 1 g: 900ml (concentrated);
b. secondary grinding: grinding beads of 0.02mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 90%; solid-liquid ratio: 1 g: the rotating speed of a 900ml mixing and grinding rotor is 3700 rpm; starting the stirring mill, wherein the whole milling time is 95min, the first 10min of the pump speed of the peristaltic pump is 10%, the second 10min is 20%, and finally, the milling is finished at the pump speed of 25%. Adding water for injection to full dose according to the formula amount. The stirring paddle was turned on and stirred at 500rpm for 10 min.
Fourthly, after the liquid medicine is treated, transferring the emulsion into a filtering system for filtering, transferring the emulsion into the filtering system for pre-filtering, wherein the aperture of a filter element used for pre-filtering is 0.45 mu m; the aperture of the sterilizing filter element is 0.22 μm.
Sampling and measuring the content of the effective component arabinose.
And tumor inhibition test: according to the third supplement of 'red nocardia rubra cell wall skeleton for injection' in Chinese pharmacopoeia 2010 edition.
Comparative example 1 (prior art) milling, shearing, homogenizing;
1. the red nocardia cell wall skeleton raw powder and the auxiliary materials are accurately weighed and prepared according to the prescription amount (the prescription is the same as the example 1).
2. Processing parameters
Grinding: putting the weighed raw powder into a mortar of a grinder, and grinding for 40 min; then adding squalene with the formula amount, and grinding for 20 min; adding polysorbate 80 according to the formula amount, grinding for 10min, adding appropriate amount of mixed solution of mannitol and water for injection, and grinding for 10 min.
Secondly, the dispersion head of the high-speed shearing machine is inserted into the liquid medicine to the bottom of about 1cm, and a power supply is turned on to carry out shearing dispersion. The rotation speed was adjusted to 17000rpm and the shear was carried out for 20 min.
Thirdly, transferring the cut liquid medicine into a sample injection cup of a high-pressure homogenizer, starting the high-pressure homogenizer for homogenization, adding the ground liquid medicine into the sample injection cup of the homogenizer for low-pressure homogenization at 1100bar for one time, and high-pressure homogenization at 1900bar for three times;
and fourthly, homogenizing the liquid medicine to obtain uniform emulsion. The emulsion was transferred to a filtration system for filtration. The aperture of the filter element used in the first pre-filtration is 1.0 μm, the aperture of the filter element used in the second pre-filtration is 0.65 μm, and the aperture of the filter element used in the third degerming filtration is 0.22 μm.
Comparative example 2 (prior art) milling, shearing, homogenizing;
1. the red nocardia cell wall skeleton raw powder and the auxiliary materials are accurately weighed and prepared according to the prescription amount (the prescription is the same as the example 1).
2. Processing parameters
Grinding: putting the weighed raw powder into a mortar of a grinder, and grinding for 30 min; then adding squalene with the formula amount, and grinding for 15 min; adding polysorbate 80 according to the formula amount, grinding for 15min, adding appropriate amount of mixed solution of mannitol and water for injection, and grinding for 20 min.
Secondly, the dispersion head of the high-speed shearing machine is inserted into the liquid medicine to the bottom of about 1.5cm, and a power supply is turned on to carry out shearing dispersion. The rotation speed was adjusted to 17000rpm and the shear was carried out for 20 min.
Thirdly, transferring the cut liquid medicine into a sample cup of a high-pressure homogenizer, starting the high-pressure homogenizer for homogenization, adding the ground liquid medicine into the sample cup of the homogenizer for low-pressure homogenization at 1000bar) for one time, and performing high-pressure homogenization at 1800bar) for three times;
and fourthly, homogenizing the liquid medicine to obtain uniform emulsion. The emulsion was transferred to a filtration system for filtration. The aperture of the filter element used in the first pre-filtration is 1.0 μm, the aperture of the filter element used in the second pre-filtration is 0.65 μm, and the aperture of the filter element used in the third degerming filtration is 0.22 μm.
Comparative example 3 (prior art) milling, shearing, homogenizing;
1. the red nocardia cell wall skeleton raw powder and the auxiliary materials are accurately weighed and prepared according to the prescription amount (the prescription is the same as the example 1).
2. Processing parameters
Grinding: putting the weighed raw powder into a mortar of a grinder, and grinding for 20 min; then adding squalene with the formula amount, and grinding for 10 min; adding polysorbate 80 according to the formula amount, grinding for 20min, adding appropriate amount of mixed solution of mannitol and water for injection, and grinding for 10 min.
Secondly, the dispersion head of the high-speed shearing machine is inserted into the liquid medicine to the bottom of about 2cm, and a power supply is turned on to carry out shearing dispersion. The rotation speed was adjusted to 17000rpm and the shear was carried out for 20 min.
Transferring the cut liquid medicine to a sample injection cup of a high-pressure homogenizer, starting the high-pressure homogenizer for homogenization, adding the ground liquid medicine into the sample injection cup of the homogenizer for low-pressure homogenization at 900bar) for one time, and high-pressure homogenization at 1700bar) for three times;
and fourthly, homogenizing the liquid medicine to obtain uniform emulsion. The emulsion was transferred to a filtration system for filtration. The aperture of the filter element used in the first pre-filtration is 1.0 μm, the aperture of the filter element used in the second pre-filtration is 0.65 μm, and the aperture of the filter element used in the third degerming filtration is 0.22 μm.
And (3) comparing the emulsification results:
from the above experiments it can be seen that example 1/2/3 has a clear advantage over comparative example 1/2/3 in using homogeneous emulsification: A. the arabinose loss of the filtered liquid medicine is obviously reduced, which shows that the efficiency is obviously improved by stirring, grinding, emulsifying and grinding, the superfine grain size emulsion can be effectively prepared, and the passing rate of degerming and filtering after the emulsification of the raw powder is improved; B. and (4) increasing the batch. The filling amount of the cell wall skeleton of the red nocardia rubra for injection depends on the arabinose content of the filtered emulsion, and under the condition that the feeding amount is not changed, the arabinose content of the filtered emulsion is improved due to the addition of stirring and grinding for emulsification, the filling amount per bottle under the same concentration is reduced, the yield (batch) is increased, and the batch yield is increased; C. the finished product has stable quality and meets the standard; D. the emulsified liquid medicine is stirred and ground, and only needs to be filtered in a second stage and passes through a filter element with the diameter of 0.22 mu m; compared with the three-stage filtration of the comparative example 1, the operation process is reduced, and the material cost of the filter element is saved.
Claims (3)
1. A method for improving the passing rate of effective components of nocardia rubra cell wall skeleton emulsion in sterilization and filtration comprises the following steps:
1) premixing and dispersing raw materials and auxiliary materials:
grinding raw and auxiliary materials: placing the weighed raw powder into a mortar of a grinder, adding a mixed solution of squalene, polysorbate 80, a proper amount of 20% mannitol injection and water for injection according to the prescription amount, and grinding for 15-25 min; wherein, the prescription is as follows:
nocardia rubra cell wall skeleton raw powder 1g
Proper amount of medium
The medium formula comprises:
20% mannitol injection 250ml
3ml of squalene
Polysorbate 802 ml
1500ml of water for injection;
homogenizing: adding the ground liquid medicine into a sample cup of a homogenizer for low-pressure homogenization, and homogenizing at 900-1100 bar for one time; homogenizing under high pressure for 1700-1900 bar three times; collecting the homogenized solution in a container, cleaning the cavity of the homogenizer with appropriate amount of water for injection, washing out the residual medicinal liquid, and mixing with the previous homogenized solution; the solid-liquid ratio is 1 g: 500-700 ml;
2) stirring, grinding and grinding:
firstly, grinding: grinding beads of 0.08-0.2mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 85-95%; solid-liquid ratio: 1 g: 500-700 ml;
a. pouring the homogenized liquid medicine into a circulating hopper; setting the rotating speed of a stirring mill rotor at 3600-3800 rpm, and setting the highest outlet temperature: 40 ℃;
b. starting a stirring mill, wherein the whole milling time is 85-95min, the first 10min of the pump speed of the peristaltic pump is 8-12%, the second 10min of the peristaltic pump is 15-25%, and finally, the milling is finished at the pump speed of 20-30%; discharging the liquid medicine to obtain liquid medicine A;
c. washing the grinding cavity and the pipeline with a proper amount of water for injection to obtain liquid B, and uniformly mixing the liquid A and the liquid B to obtain liquid AB, wherein the solid-liquid ratio is 1 g: 800-;
secondly, grinding: grinding beads of 0.02-0.07mm are stirred, milled and emulsified, and the filling degree of the grinding beads is 85-95%;
a. unloading the grinding beads with the diameter of 0.08-0.2mm, and loading the grinding beads with the diameter of 0.02-0.07 mm;
b. dividing the AB liquid into two parts, pouring the two parts into a circulating cup of an assembled bead mill in several times, setting the stirring mill rotation speed to be 3600rpm-3800rpm, and grinding for 40-50 minutes; the pumping speed of the peristaltic pump is 8-12% for the first 10min and 20% for the second 15-25min, and then the grinding is finished at the flow rate of 20-30%;
d. adding injection water to full amount according to the formula amount, starting the stirring slurry, and stirring at 400-600rpm for 8-12 min;
3) pre-filtering: pre-filtering the medicinal liquid with 0.4-0.5 μm filter;
4) and (3) degerming and filtering: filtering the medicinal liquid with a filter of 0.4-0.5 μm with a sterilizing filter.
2. The effective component for improving the sterilization and filtration of nocardia rubra cell wall skeleton emulsion as claimed in claim 1
The method for determining the passing rate is characterized by further comprising the step of sampling and measuring the content of the effective component arabinose after the step 4) of sterilizing and filtering.
3. The method for increasing the passing rate of the sterilization filtration active ingredients of nocardia rubra cell wall skeleton emulsion according to claim 1, wherein the method comprises the following steps: the pore diameter of the sterilization filter in the step 4) is 0.22 μm.
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| CN104083407A (en) * | 2014-07-08 | 2014-10-08 | 福建省山河药业有限公司 | Method for improving production of nocardia rubra frame emulsion |
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Non-Patent Citations (1)
| Title |
|---|
| Novel Glycosidic Linkage between Arabinogalactan and Peptidoglycan in the Cell Wall Skeleton of Nocardia rubra AN-115;Mamoru Fujioka et al.;《Journal of General Microbiology》;19851231;1323-1329 * |
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Denomination of invention: A Method to Improve the Passing Rate of Effective Components in the Sterilization and Filtration of Red Nocardia Cell Wall Skeleton Emulsion Granted publication date: 20200811 Pledgee: Bank of Xiamen Limited by Share Ltd. Putian branch Pledgor: FUJIAN SHANHE PHARMACEUTICAL Co.,Ltd. Registration number: Y2024980011713 |
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