CN110484596A - A kind of gentamicine sulphate injection Sterility Test Methods - Google Patents
A kind of gentamicine sulphate injection Sterility Test Methods Download PDFInfo
- Publication number
- CN110484596A CN110484596A CN201910827523.6A CN201910827523A CN110484596A CN 110484596 A CN110484596 A CN 110484596A CN 201910827523 A CN201910827523 A CN 201910827523A CN 110484596 A CN110484596 A CN 110484596A
- Authority
- CN
- China
- Prior art keywords
- gentamicin sulfate
- culture medium
- sterility
- sulfate injection
- injection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000002347 injection Methods 0.000 title claims abstract description 123
- 239000007924 injection Substances 0.000 title claims abstract description 123
- 230000036512 infertility Effects 0.000 title claims abstract description 89
- 238000010998 test method Methods 0.000 title abstract description 17
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 title abstract description 11
- CEAZRRDELHUEMR-UHFFFAOYSA-N gentamicin Chemical compound O1C(C(C)NC)CCC(N)C1OC1C(O)C(OC2C(C(NC)C(C)(O)CO2)O)C(N)CC1N CEAZRRDELHUEMR-UHFFFAOYSA-N 0.000 title abstract 5
- 229910021653 sulphate ion Inorganic materials 0.000 title abstract 5
- 238000000034 method Methods 0.000 claims abstract description 107
- 239000001963 growth medium Substances 0.000 claims abstract description 79
- 238000012360 testing method Methods 0.000 claims abstract description 53
- 238000002360 preparation method Methods 0.000 claims abstract description 38
- 241000894006 Bacteria Species 0.000 claims abstract description 36
- 238000001914 filtration Methods 0.000 claims abstract description 25
- 239000013641 positive control Substances 0.000 claims abstract description 23
- 239000000872 buffer Substances 0.000 claims abstract description 17
- 238000010790 dilution Methods 0.000 claims abstract description 16
- 239000012895 dilution Substances 0.000 claims abstract description 16
- 238000005374 membrane filtration Methods 0.000 claims abstract description 15
- 241000588724 Escherichia coli Species 0.000 claims abstract description 14
- RDEIXVOBVLKYNT-VQBXQJRRSA-N (2r,3r,4r,5r)-2-[(1s,2s,3r,4s,6r)-4,6-diamino-3-[(2r,3r,6s)-3-amino-6-(1-aminoethyl)oxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-5-methyl-4-(methylamino)oxane-3,5-diol;(2r,3r,4r,5r)-2-[(1s,2s,3r,4s,6r)-4,6-diamino-3-[(2r,3r,6s)-3-amino-6-(aminomethyl)oxan-2-yl]o Chemical compound OS(O)(=O)=O.O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H](CC[C@@H](CN)O2)N)[C@@H](N)C[C@H]1N.O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H](CC[C@H](O2)C(C)N)N)[C@@H](N)C[C@H]1N.O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N RDEIXVOBVLKYNT-VQBXQJRRSA-N 0.000 claims description 118
- 239000012528 membrane Substances 0.000 claims description 62
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 54
- 239000007853 buffer solution Substances 0.000 claims description 53
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 27
- 229930003268 Vitamin C Natural products 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 27
- 235000019154 vitamin C Nutrition 0.000 claims description 27
- 239000011718 vitamin C Substances 0.000 claims description 27
- 239000003085 diluting agent Substances 0.000 claims description 25
- 108010050327 trypticase-soy broth Proteins 0.000 claims description 24
- 239000002609 medium Substances 0.000 claims description 23
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 19
- 239000012530 fluid Substances 0.000 claims description 15
- CWERGRDVMFNCDR-UHFFFAOYSA-M thioglycolate(1-) Chemical compound [O-]C(=O)CS CWERGRDVMFNCDR-UHFFFAOYSA-M 0.000 claims description 15
- 102000002260 Alkaline Phosphatase Human genes 0.000 claims description 14
- 108020004774 Alkaline Phosphatase Proteins 0.000 claims description 14
- 238000009630 liquid culture Methods 0.000 claims description 14
- 230000001954 sterilising effect Effects 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 12
- 239000001888 Peptone Substances 0.000 claims description 9
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical group C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000011010 flushing procedure Methods 0.000 claims description 8
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 8
- 229920000053 polysorbate 80 Polymers 0.000 claims description 8
- 239000011148 porous material Substances 0.000 claims description 8
- 229910052708 sodium Inorganic materials 0.000 claims description 8
- 239000011734 sodium Substances 0.000 claims description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims description 7
- 238000001471 micro-filtration Methods 0.000 claims description 5
- 239000013642 negative control Substances 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 230000035899 viability Effects 0.000 claims description 3
- 230000008569 process Effects 0.000 abstract description 12
- RDEIXVOBVLKYNT-HDZPSJEVSA-N (2r,3r,4r,5r)-2-[(1s,2s,3r,4s,6r)-4,6-diamino-3-[(2r,3r,6s)-3-amino-6-[(1r)-1-aminoethyl]oxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-5-methyl-4-(methylamino)oxane-3,5-diol;(2r,3r,4r,5r)-2-[(1s,2s,3r,4s,6r)-4,6-diamino-3-[(2r,3r,6s)-3-amino-6-(aminomethyl)oxan-2 Chemical compound OS(O)(=O)=O.O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H](CC[C@@H](CN)O2)N)[C@@H](N)C[C@H]1N.O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H](CC[C@H](O2)[C@@H](C)N)N)[C@@H](N)C[C@H]1N.O1[C@H]([C@@H](C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N RDEIXVOBVLKYNT-HDZPSJEVSA-N 0.000 abstract 1
- 206010015856 Extrasystoles Diseases 0.000 abstract 1
- 230000003042 antagnostic effect Effects 0.000 abstract 1
- 238000007689 inspection Methods 0.000 description 19
- 238000005406 washing Methods 0.000 description 18
- 230000003385 bacteriostatic effect Effects 0.000 description 14
- 230000000694 effects Effects 0.000 description 14
- 102000004190 Enzymes Human genes 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 13
- 241000228245 Aspergillus niger Species 0.000 description 11
- 238000012795 verification Methods 0.000 description 11
- 241000222122 Candida albicans Species 0.000 description 8
- 241000193470 Clostridium sporogenes Species 0.000 description 8
- 230000009286 beneficial effect Effects 0.000 description 8
- 229940095731 candida albicans Drugs 0.000 description 8
- 229940071127 thioglycolate Drugs 0.000 description 8
- 238000005352 clarification Methods 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 239000000725 suspension Substances 0.000 description 7
- 229930182566 Gentamicin Natural products 0.000 description 6
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 6
- 239000011550 stock solution Substances 0.000 description 6
- 244000063299 Bacillus subtilis Species 0.000 description 5
- 235000014469 Bacillus subtilis Nutrition 0.000 description 5
- 241000191967 Staphylococcus aureus Species 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000013190 sterility testing Methods 0.000 description 3
- 239000012137 tryptone Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000012982 microporous membrane Substances 0.000 description 2
- 108010077805 Bacterial Proteins Proteins 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 206010024971 Lower respiratory tract infections Diseases 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229940126574 aminoglycoside antibiotic Drugs 0.000 description 1
- 239000002647 aminoglycoside antibiotic agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000002977 hyperthermial effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000004708 ribosome subunit Anatomy 0.000 description 1
- 208000013223 septicemia Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/22—Testing for sterility conditions
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Public Health (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention discloses a kind of gentamicine sulphate injection Sterility Test Methods, using membrane-filter procedure, the preparation step of culture includes carrying out membrane filtration using bigeminy closed film filter, the amount that buffer rinses is every film 100-800mL, and the additional amount of culture medium is every film 100mL；Check groups preparation step, which includes that gentamicine sulphate injection is added in dilution, to be diluted, and the volume of dilution is 0-10 times of injection volume；The amount of injection filtering is that every film filtering potency is 800,000 units；Positive controls preparation step includes that, using escherichia coli as Positive contrast bacteria, Positive contrast bacteria is added after culture medium is added.Gentamicine sulphate injection Sterility Test Methods provided by the invention adjust the elements such as sterility test process, culture medium, buffer for gentamicin sulphate antagonistic property, accurately and reliably can carry out sterility test to gentamicine sulphate injection.
Description
Technical Field
The invention relates to the field of medicine detection, in particular to a method for checking the sterility of gentamicin sulfate injection.
Background
Gentamicin sulfate is an aminoglycoside antibiotic, has good antibacterial effect on various gram-negative bacteria and gram-positive bacteria, and has an action mechanism of combining with a bacterial ribosome subunit to inhibit the synthesis of bacterial protein. Gentamicin sulfate is suitable for treating serious infection caused by gram-negative bacteria, such as septicemia, lower respiratory tract infection, intestinal tract infection, etc. The sanitary index of the gentamicin sulfate injection is required to be aseptic, and in the production process, although the gentamicin sulfate injection is sterilized, part of microorganisms in a low-activity or dormant state can be not killed or removed.
The sterility test method is a test method for establishing sterility reliability of sterile drugs and the like, and whether the method is accurate or not is very important for controlling the safety of the drugs. The sterility test needs to be carried out on the injection specified in the Chinese pharmacopoeia, and the sterility test method is specifically required in the 2015 edition of the Chinese pharmacopoeia, but the gentamicin sulfate injection has an obvious inhibition effect on bacteria, influences the reliability of the sterility test, and needs to eliminate the bacteriostasis of the medicine through the study on the applicability of the sterility test methodology. For the injection with bacteriostatic activity, a membrane filtration method is generally selected for sterility test, a plurality of methodological applicability tests are carried out, and the final sterility test method is determined by eliminating bacteriostatic activity through diluting the medicine, washing the membrane and the like. There is a need to identify an accurate method for sterility testing of gentamicin sulfate injection.
Disclosure of Invention
The invention aims to provide a method for checking the sterility of gentamicin sulfate injection, which aims at the bacteriostatic property of gentamicin sulfate, adjusts the factors such as sterility checking process, culture medium, buffer solution and the like, and can accurately and reliably check the sterility of the gentamicin sulfate injection.
In order to realize the aim, the invention provides a method for checking the sterility of gentamicin sulfate injection, which adopts a membrane filtration method, cultures comprise a test group, a positive control group and a negative control group,
the preparation steps of each group of cultures comprise membrane filtration by using a duplex closed membrane filter, wherein the flushing amount of a buffer solution is 100mL per membrane and 800mL per membrane, and the adding amount of a culture medium is 100mL per membrane;
the preparation method of the test group comprises the steps of adding gentamicin sulfate injection into diluent for dilution, wherein the volume of the diluent is 0-10 times of that of the gentamicin sulfate injection; the filtration quantity of the gentamicin sulfate injection is that the filtration titer of each membrane is 80 ten thousand units;
the preparation method of the positive control group comprises the steps of adopting Escherichia coli as a positive control bacterium, and adding the positive control bacterium after adding a culture medium;
wherein,
the diluent is a sodium chloride-peptone buffer solution with pH7.0 or a sterile sodium chloride solution with 0.9 percent; the buffer solution is a sodium chloride-peptone buffer solution with pH 7.0; the culture medium comprises one of a thioglycollate fluid culture medium or a trypticase soytone liquid culture medium.
By adopting the scheme, as the gentamicin sulfate has an antibacterial effect, the injection stock solution is diluted by adding the diluent, and the antibacterial effect is reduced by washing the buffer solution for multiple times after the membrane filtration, so that the sterility test is accurately carried out. The inventor determines that the injection stock solution is not diluted or the dilution range, the proper membrane filter and the flushing amount after membrane filtration in the process of carrying out the sterile inspection on the gentamicin sulfate injection by referring to a sterile inspection method in 'Chinese pharmacopoeia' 2015 edition, researching the applicability of the sterile inspection method of the gentamicin sulfate injection and carrying out method applicability tests for multiple times; the Escherichia coli is determined to be sensitive bacteria of gentamicin sulfate and is suitable to be used as positive control bacteria. The aseptic inspection operation process generally requires that the aseptic process and the aseptic process are carried out separately in order to prevent pollution.
According to another specific embodiment, the invention discloses a method for checking the sterility of gentamicin sulfate injection, wherein the buffer washing amount is 300mL per membrane; the gentamicin sulfate injection is added into the diluent for dilution, and the volume of the diluent is 10 times of that of the gentamicin sulfate injection.
By adopting the scheme, the method applicability test confirms that the Escherichia coli, the staphylococcus aureus, the bacillus subtilis, the clostridium sporogenes, the candida albicans and the aspergillus niger all grow after being cultured for 24 hours under the condition, and the method is suitable for the sterility test of the gentamicin sulfate injection.
According to another embodiment of the invention, the method for aseptically examining gentamicin sulfate injection disclosed in the embodiment of the invention is characterized in that the pH value of trypticase soytone liquid medium at 25 ℃ is 5.5-6.5.
By adopting the scheme, after the pH value of the trypticase soy peptone liquid medium is reduced to 5.5-6.5, the growth of clostridium sporogenes is not obviously influenced, the growth effects of candida albicans and aspergillus niger are better, the influence of the gentamicin sulfate on bacteriostasis is weakened, and after 600mL buffer solution is added for flushing, three bacteria all obviously grow after 72 hours of culture; therefore, it was determined that lowering the pH of trypticase Soy peptone broth was beneficial for gentamicin sulfate injection sterility testing.
According to another embodiment of the present invention, the method for examining the sterility of gentamicin sulfate injection disclosed in the embodiments of the present invention comprises culturing sterile alkaline phosphatase with a viability unit of 800-.
By adopting the scheme, the alkaline phosphatase has no obvious influence on the growth of the thalli, but the influence of the bacteriostatic action of the gentamicin sulfate is weakened; therefore, it was determined that the addition of alkaline phosphatase is advantageous for the sterility test of gentamicin sulfate injection.
According to another embodiment of the present invention, the method for aseptically examining gentamicin sulfate injection disclosed in the embodiment of the present invention comprises aseptically treating alkaline phosphatase, and the treating step comprises preparing a solution, and filtering and sterilizing the solution with a microfiltration membrane filter having a pore size of 0.22 μm.
By adopting the scheme, the enzyme preparation is filtered and sterilized by the microporous filter membrane, and then the sterilized culture medium is added, so that the sterility and the safety of the culture medium are high under the condition of not influencing the enzyme activity.
According to another specific embodiment of the invention, the culture medium comprises 0.05-0.1% by mass of sterile vitamin C.
By adopting the scheme, the vitamin C has no obvious influence on the growth of bacteria in the vitamin C, but the influence of the bacteriostatic action of the gentamicin sulfate is weakened; therefore, the addition of vitamin C is determined to be beneficial to the sterility test of gentamicin sulfate injection.
According to another embodiment of the invention, the method for aseptically inspecting gentamicin sulfate injection disclosed by the embodiment of the invention comprises the step of aseptically processing vitamin C, wherein the processing step comprises filtering and sterilizing the vitamin C injection by using a microfiltration membrane filter with the pore diameter of 0.22 μm.
By adopting the scheme, the sterilized culture medium is added after the vitamin C is filtered and sterilized by the microporous filter membrane, and the sterility and the safety of the culture medium are high under the condition of not influencing the enzyme activity.
According to another embodiment of the present invention, the method for examining the sterility of gentamicin sulfate injection disclosed by the embodiment of the present invention further comprises a culture medium control group; media control preparation a positive control was prepared, with the addition of media as standard media.
By adopting the scheme, the culture medium control group is added, and compared with the positive control group, the growth time and the thallus growth concentration difference are not obvious, so that the condition that the thallus growth is not influenced by adding the enzyme preparation, the vitamin C and the pH value change is determined.
According to another specific embodiment of the present invention, the method for examining the sterility of gentamicin sulfate injection disclosed in the embodiment of the present invention comprises 0.1% by mass of tween-80 in the buffer.
By adopting the scheme, the buffer solution washes the filtered film, so that the influence of the gentamicin sulfate on the bacteriostasis is weakened, and the growth of bacteria is not obviously influenced; therefore, it was determined that the addition of tween-80 to the buffer facilitates the sterility test of gentamicin sulfate injection.
The invention provides a sterile inspection method suitable for gentamicin sulfate injection through the research on the applicability of the sterile inspection method, and the method is used for determining the dilution of the stock solution of the injection, the selection of instruments for membrane filtration, the membrane flushing process after filtration, positive control bacteria and the like in the sterile inspection process; aiming at the bacteriostatic property of the gentamicin sulfate, an adjusted culture medium is also determined, the adjusted culture medium does not have obvious influence on the growth of bacteria, the bacteriostatic action of the gentamicin sulfate can be reduced, and the sterility test of the gentamicin sulfate injection is facilitated; the adjusted buffer solution is determined, and the sterility check of the gentamicin sulfate injection is facilitated. Tests prove that the sterility test method provided by the invention can accurately and reliably carry out sterility test on the gentamicin sulfate injection.
Detailed Description
For purposes of the following detailed description, it is to be understood that the application may assume various alternative variations and step sequences, except where expressly specified to the contrary. Moreover, other than in any operating examples, or where otherwise indicated, all numbers expressing, for example, quantities of ingredients used in the specification and claims are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties to be obtained by the present application. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.
The terminology used in the present application is for the purpose of describing particular embodiments only and is not intended to be limiting. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items. The expression "at least one" of an expression, for example, modifies an entire list of elements when preceding or following the list of elements, without modifying individual elements of the list.
Further, the terms "comprises" or "comprising," when used in this specification, specify the presence of stated features, regions, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, regions, integers, steps, operations, elements, components, and/or groups thereof.
As used herein, "about" or "approximately" includes the recited value and means, for example, within an acceptable range of deviation for the particular value as determined by one of ordinary skill in the art in view of the measurement in question and the error associated with measurement of the particular quantity (i.e., limitations of the measurement system). All ratios of components refer to weight percent (wt.%), unless otherwise specified; unless otherwise indicated, all parameter ranges disclosed include the endpoints and all values therebetween.
In the description of the present invention, unless otherwise specified, terms have the same meaning as those generally understood by those skilled in the art, but if different, the definition of the present invention shall control; unless otherwise specified, the test methods are all conventional methods; unless otherwise specified, the raw materials and test materials used in the present invention are all available commercially; unless otherwise specified, the percentages (%) in the present invention are mass percentages (% by mass).
In the present invention, the term "membrane" refers to a filter having a pore size of not more than 0.45. mu.m.
In the present invention, the test strain should be passaged no more than 5 generations (0 th generation of a dried strain obtained from a strain collection center) and preserved by using an appropriate strain preservation technique.
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention are described in detail below.
The invention provides a gentamicin sulfate injection sterility test method, which adopts a membrane filtration method, wherein cultures comprise a test group, a positive control group and a negative control group, the preparation steps of each group of cultures comprise membrane filtration by using a duplex closed membrane filter, the flushing amount of a buffer solution is 800mL per membrane, and the adding amount of a culture medium is 100mL per membrane;
the preparation method of the test group comprises the steps of adding gentamicin sulfate injection into diluent for dilution, wherein the volume of the diluent is 0-10 times of that of the gentamicin sulfate injection; the filtration quantity of the gentamicin sulfate injection is that the filtration titer of each membrane is 80 ten thousand units;
the preparation method of the positive control group comprises the steps of adopting Escherichia coli as a positive control bacterium, and adding the positive control bacterium after adding a culture medium;
wherein the diluent is a sodium chloride-peptone buffer solution with pH7.0 or a 0.9% sterile sodium chloride solution; the buffer solution is a sodium chloride-peptone buffer solution with pH 7.0; the culture medium comprises one of a thioglycollate fluid culture medium or a trypticase soytone liquid culture medium.
The membrane filtration method is a common detection method for the sterility test of the medicine, is mature, has higher accuracy, is particularly suitable for the medicine with antibacterial property by adopting the membrane filtration method, and can reduce the bacteriostatic action by washing away with a buffer solution. The invention carries out the sterility test on the gentamicin sulfate injection, and the liquid state is easy to filter through a film. As gentamicin sulfate has an antibacterial effect, the injection stock solution can be diluted by adding a diluent, and the antibacterial effect can be reduced by washing the buffer solution for multiple times after the membrane filtration, so that the sterility test can be accurately carried out. The inventor determines that the gentamycin sulfate injection is subjected to sterility test, the stock solution of the injection is not diluted or the dilution range, a suitable membrane filter and the flushing amount after membrane filtration by referring to a sterility test method in 'Chinese pharmacopoeia' 2015 edition, researching the applicability of the gentamycin sulfate injection sterility test and carrying out the method applicability test for multiple times; the Escherichia coli is determined to be sensitive bacteria of gentamicin sulfate and is suitable to be used as positive control bacteria. The aseptic inspection operation process generally requires that the aseptic process and the aseptic process are carried out separately in order to prevent pollution.
According to another specific embodiment, the invention discloses a method for checking the sterility of gentamicin sulfate injection, wherein the buffer washing amount is 300mL per membrane; the gentamicin sulfate injection is added into the diluent for dilution, and the volume of the diluent is 10 times of that of the gentamicin sulfate injection
The inventor determines, by studying the applicability of the aseptic examination method for gentamicin sulfate injection, that the examination method comprises diluting the injection with 10-fold volume of diluent, and washing each membrane with 300mL of pH7.0 NaCl-peptone buffer solution after filtration. The method is subjected to a method applicability test, and the method is confirmed to be suitable for the sterility check of gentamicin sulfate injection by culturing escherichia coli, staphylococcus aureus, bacillus subtilis, clostridium sporogenes, candida albicans and aspergillus niger for 24 hours under the condition of the method.
According to another embodiment of the invention, the method for aseptically examining gentamicin sulfate injection disclosed in the embodiment of the invention is characterized in that the pH value of trypticase soytone liquid medium at 25 ℃ is 5.5-6.5.
Standard tryptic soy peptone broth was provided in the Chinese pharmacopoeia, and the pH at 25 ℃ after sterilization was 7.3. + -. 0.2. The inventor finds in experiments that in a standard trypticase soytone liquid medium, clostridium sporogenes, candida albicans and aspergillus niger cultured in the medium are all affected by the bacteriostatic action of gentamicin sulfate, and particularly, after the aspergillus niger is added into 500mL of buffer solution for washing, people do not grow after the aspergillus niger is cultured for 72 hours; after the pH value of a trypticase soy peptone liquid culture medium is reduced to 5.5-6.5, the growth of clostridium sporogenes is not obviously influenced, the growth effects of candida albicans and aspergillus niger are better, the influence of the gentamicin sulfate on bacteriostasis is weakened, and after 600mL of buffer solution is added for washing, three bacteria all obviously grow after being cultured for 72 hours; therefore, it was determined that lowering the pH of trypticase Soy peptone broth was beneficial for gentamicin sulfate injection sterility testing.
According to another embodiment of the present invention, the method for examining the sterility of gentamicin sulfate injection disclosed in the embodiments of the present invention comprises culturing sterile alkaline phosphatase with a viability unit of 800-.
The inventor finds in experiments that the addition of alkaline phosphatase to the culture medium has no significant effect on the growth of the bacteria cultured therein, but has a reduced effect on the gentamicin sulfate bacteriostasis; therefore, it was determined that the addition of alkaline phosphatase is advantageous for the sterility test of gentamicin sulfate injection.
According to another embodiment of the invention, the method for checking the sterility of gentamicin sulfate injection disclosed by the embodiment of the invention,alkaline phosphataseThe method comprises preparing solution, and filtering with microporous membrane filter with pore diameter of 0.22 μm for sterilization.
Generally, the culture medium required for sterility test is prepared and then sterilized at high temperature and high pressure, but the structure of the enzyme preparation is easily damaged and loses activity at high temperature, so that the inhibition effect on the bacteriostatic action of gentamicin sulfate can be reduced. The enzyme preparation can be prepared by purchasing sterile enzyme preparation, sterilizing other components of the culture medium at high temperature, and adding; the enzyme preparation purchased may also be subjected to non-hyperthermic sterilization methods, such as irradiation, filtration, etc. The invention adopts the microporous filter membrane to filter and sterilize the enzyme preparation, and then adds the sterilized culture medium, under the condition of not influencing the enzyme activity, the culture medium has high sterile safety.
According to another specific embodiment of the invention, the culture medium comprises 0.05-0.1% by mass of sterile vitamin C.
The inventor finds in experiments that the addition of 0.05-0.1% of sterilized vitamin C to the culture medium has no significant effect on the growth of the bacteria cultured therein, but weakens the bacteriostatic effect of gentamicin sulfate; therefore, the addition of vitamin C is determined to be beneficial to the sterility test of gentamicin sulfate injection.
According to another embodiment of the invention, the method for aseptically inspecting gentamicin sulfate injection disclosed by the embodiment of the invention comprises the step of aseptically processing vitamin C, wherein the processing step comprises filtering and sterilizing the vitamin C injection by using a microfiltration membrane filter with the pore diameter of 0.22 μm.
The culture medium is sterilized at high temperature and high pressure after being prepared, but the structure of the vitamin C is easy to damage under the high-temperature state, and the inhibition effect on the bacteriostatic action of the gentamicin sulfate can be reduced. Optionally, the sterile vitamin C injection can be added after high-temperature sterilization of other components in the culture medium; the non-high temperature sterilization method can also be carried out on the vitamin C injection. The invention adopts the microporous filter membrane to filter and sterilize the vitamin C, and then adds the sterilized culture medium, under the condition of not influencing the enzyme activity, the culture medium has high sterility and safety.
According to another embodiment of the present invention, the method for examining the sterility of gentamicin sulfate injection disclosed by the embodiment of the present invention further comprises a culture medium control group; media control preparation a positive control was prepared, with the addition of media as standard media.
The term "standard medium" refers to a medium prepared by a method of preparing a trypticase Soy peptone liquid medium or a thioglycolate fluid medium according to the sterility test method of "Chinese pharmacopoeia" 2015 edition, and does not include an enzyme preparation, vitamin C, a change in pH, and the like. And adding a culture medium control group, and comparing with the positive control group, wherein the growth time and the thallus growth concentration difference are not obvious, so that the addition of the enzyme preparation, the vitamin C and the change of the pH value have no influence on the thallus growth.
According to another embodiment of the present invention, the method for sterility test of gentamicin sulfate injection disclosed in the embodiment of the present invention comprises 0.1% tween-80 in the buffer.
The inventor finds in experiments that 0.1% of tween-80 is added into the buffer solution, and the buffer solution washes the filtered film, so that the influence of the gentamicin sulfate on bacteriostasis is weakened, and the growth of bacteria is not obviously influenced; therefore, it was determined that the addition of tween-80 to the buffer facilitates the sterility test of gentamicin sulfate injection.
The following will describe the embodiments of the present invention in further detail by using specific examples, in which gentamicin sulfate injection with a titer of 8 ten thousand units and a specification of 2mL is selected. And the method applicability verification test is carried out on the sterility test method of the embodiment according to the specified method of Chinese pharmacopoeia 2015 edition, and whether the method is applicable or not is verified.
Example 1
1. A method for checking the sterility of gentamicin sulfate injection comprises the following steps:
1.1 preparing a culture medium, a bacterial liquid, a diluent and a buffer solution: preparing a thioglycollate fluid culture medium and a trypticase soytone liquid culture medium; preparing Escherichia coli suspension with bacteria number less than 100 cfu/mL; a sodium chloride-peptone buffer of buffer pH7.0 was prepared.
1.2 Experimental group preparation: after a group of duplex closed membrane filters are wetted by buffer solution, 10 bottles of gentamicin sulfate injection samples are filtered and passed through, each membrane is washed by 800mL of buffer solution, each time by 100mL, and 2 filter cylinders containing samples are obtained; 100mL of thioglycolate fluid medium and 100mL of trypticase Soy peptone broth were added to each of the two cartridges, and cultured and observed day by day.
1.3 Positive control preparation: after a group of duplex closed membrane filters are wetted by buffer solution, washing each membrane by 800mL of buffer solution, and washing each membrane by 100mL each time; 100mL of thioglycollate fluid medium was added to each cartridge, followed by 1mL of E.coli suspension, and the cartridges were incubated and observed day by day.
1.4 negative control preparation: after a group of duplex closed membrane filters are wetted by buffer solution, washing each membrane by 800mL of buffer solution, and washing each membrane by 100mL each time; after 100mL of thioglycolate fluid medium and tryptone Soy peptone broth medium were added to the cartridges, respectively, they were cultured and observed day by day.
2. The method of this example was subjected to a method suitability verification test, comprising the following steps:
2.1 preparing a culture medium, a bacterial liquid, a diluent and a buffer solution: preparing a thioglycollate fluid culture medium and a trypticase soytone liquid culture medium; respectively preparing staphylococcus aureus, bacillus subtilis, clostridium sporogenes, candida albicans and aspergillus niger suspension with the bacteria number less than 100 cfu/mL; a sodium chloride-peptone buffer of buffer pH7.0 was prepared.
2.2 Experimental group preparation: wetting three groups of duplex closed membrane filters by using buffer solution, filtering gentamicin sulfate injection samples by 30 bottles, and then washing each membrane by 800mL of buffer solution, wherein each time is 100mL, so as to obtain 6 filter cartridges containing samples; wherein, 100mL of thioglycolate fluid culture medium is respectively added into each cylinder of the 3 filter cylinders, and 1mL of staphylococcus aureus, escherichia coli and clostridium sporogenes suspension is respectively added into each cylinder; and adding 100mL of trypticase soy peptone liquid culture medium into each of the other 3 filter cylinders, adding 1mL of bacillus subtilis, candida albicans and aspergillus niger liquid, culturing and observing day by day.
2.3 preparation of a bacterial liquid control group: after the three groups of duplex closed membrane filters are wetted by buffer solution, each membrane is washed by 800mL of buffer solution, and each time, 100mL of buffer solution is used; wherein, 100mL of thioglycolate fluid culture medium is respectively added into each cylinder of the 3 filter cylinders, and 1mL of staphylococcus aureus, escherichia coli and clostridium sporogenes suspension is respectively added into each cylinder; and adding 100mL of trypticase soy peptone liquid culture medium into each of the other 3 filter cylinders, adding 1mL of bacillus subtilis, candida albicans and aspergillus niger liquid, culturing and observing day by day.
2.4 negative control preparation: after a group of duplex closed membrane filters are wetted by buffer solution, washing each membrane by 800mL of buffer solution, and washing each membrane by 100mL each time; after 100mL of thioglycolate fluid medium and tryptone Soy peptone broth medium were added to the cartridges, respectively, they were cultured and observed day by day.
2.5 sample set preparation: after a group of duplex closed membrane filters are wetted by buffer solution, 10 bottles of gentamicin sulfate injection samples are filtered and passed through, each membrane is washed by 800mL of buffer solution, each time by 100mL, and 2 filter cylinders containing samples are obtained; 100mL of thioglycolate fluid medium and 100mL of trypticase Soy peptone broth were added to each of the two cartridges, and cultured and observed day by day.
3. The method applicability verification test results of the present example are as follows:
TABLE 1
Note: + means liquid turbidity; denotes clarification of the liquid
The results in table 1 show that 6 experimental bacteria all grew after 72 hours of culture, indicating that the examination method of example 1 is suitable for the sterility examination of gentamicin sulfate injection.
Example 2
The method for checking the sterility of gentamicin sulfate injection and the method for verifying the applicability refer to example 1, except that: also prepared is a 0.9% sterile sodium chloride solution of the diluent; adding 100mL of diluent into each 10 bottles of gentamycin sulfate injection samples for dilution before filtering; each membrane was washed with 500mL of buffer, 100mL each time.
The method applicability verification test results of the present example are as follows:
TABLE 2
Note: + means liquid turbidity; denotes clarification of the liquid
The results in Table 2 show that 6 experimental bacteria all grew after 72 hours of culture, indicating that the examination method of example 2 is suitable for the sterility examination of gentamicin sulfate injection.
Example 3
The method for checking the sterility of gentamicin sulfate injection and the method for verifying the applicability refer to example 1, except that: adding 200mL of buffer solution into every 10 bottles of gentamycin sulfate injection samples before filtering to be used as diluent for dilution; each membrane was washed with 300mL of buffer, 100mL each time.
The method applicability verification test results of the present example are as follows:
TABLE 3
Note: + means liquid turbidity; denotes clarification of the liquid
The results in Table 3 show that 6 experimental bacteria all grew after 48 hours of culture, indicating that the examination method of example 3 is suitable for the sterility examination of gentamicin sulfate injection.
Example 4
The method for checking the sterility of gentamicin sulfate injection and the method for verifying the applicability refer to example 1, except that: preparing trypticase soy peptone liquid culture medium, and making the pH value of the trypticase soy peptone liquid culture medium at 25 ℃ to be 6.5 after the trypticase soy peptone liquid culture medium is sterilized; each membrane was washed with 700mL of buffer, 100mL each time.
The method applicability verification test results of the present example are as follows:
TABLE 4
Note: + means liquid turbidity; denotes clarification of the liquid
The results in Table 4 show that 6 experimental bacteria all grow after being cultured for 72 hours, which indicates that the inspection method in example 4 is suitable for the sterility inspection of gentamicin sulfate injection, and the results in example 1 show that the Aspergillus niger growth is obviously facilitated after the pH value of the tryptone soy peptone liquid medium is adjusted.
Example 5
The method for checking the sterility of gentamicin sulfate injection and the method for verifying the applicability refer to example 1, except that: the preparation process of the culture medium comprises adding 1000U of alkaline phosphatase into every 100mL of sterilized solution, filtering and sterilizing the solution by a microporous membrane filter with the pore diameter of 0.22 μm; adding 100mL of diluent into each 10 bottles of gentamycin sulfate injection samples for dilution before filtering; each membrane is washed by 500mL of buffer solution, and each time, 100mL is carried out; the sterility test method further comprises a culture medium control group preparation step:
1.5 Medium control preparation: preparing a standard thioglycollate fluid medium without the addition of alkaline phosphatase; after a group of duplex closed membrane filters are wetted by buffer solution, washing each membrane by 500mL of buffer solution, and washing each membrane by 100mL each time; 100mL of standard culture medium and 1mL of E.coli suspension were added to each cartridge, incubated and observed day by day.
The method applicability verification test results of the present example are as follows:
TABLE 5
Note: + means liquid turbidity; denotes clarification of the liquid
The results in Table 5 show that 6 experimental bacteria all grow after 48 hours of culture, which indicates that the inspection method in example 5 is suitable for the sterility inspection of gentamicin sulfate injection, and the results in example 2 indicate that the addition of alkaline phosphatase to the culture medium is obviously beneficial to the sterility inspection of gentamicin sulfate injection.
Example 6
The method for checking the sterility of gentamicin sulfate injection and the method for verifying the applicability refer to example 1, except that: the preparation process of the culture medium comprises the steps of adding 0.1 mass percent of vitamin C into the sterilized culture medium filtered and sterilized by a microporous filter membrane filter with the aperture of 0.22 mu m; adding 200mL of diluent into every 10 bottles of gentamycin sulfate injection samples for dilution before filtering; each membrane is washed by 300mL of buffer solution, each time 100 mL; the sterility test method further comprises a culture medium control group preparation step:
1.5 Medium control preparation: preparing a standard thioglycolate fluid medium without vitamin C; after a group of duplex closed membrane filters are wetted by buffer solution, each membrane is washed by 300mL of buffer solution, and each time, 100mL of buffer solution is used; 100mL of standard culture medium and 1mL of E.coli suspension were added to each cartridge, incubated and observed day by day.
The method applicability verification test results of the present example are as follows:
TABLE 6
The results in table 6 show that 6 experimental bacteria all grow after 24 hours of culture, which indicates that the inspection method in example 6 is suitable for the sterility inspection of gentamicin sulfate injection, and the results in example 3 indicate that the addition of vitamin C to the culture medium is obviously beneficial to the sterility inspection of gentamicin sulfate injection.
Example 7
The method for checking the sterility of gentamicin sulfate injection and the method for verifying the applicability refer to example 6, except that: adding tween-80 with the mass percent of 0.1 percent in the buffer solution preparation process; the gentamicin sulfate injection sample is not diluted before being filtered; the amount of vitamin C added to the culture medium is 0.05 percent of the mass of the culture medium, and each membrane is washed by 500mL of buffer solution, 100mL each time.
The method applicability verification test results of the present example are as follows:
TABLE 7
Note: + means liquid turbidity; denotes clarification of the liquid
The results in Table 7 show that 6 experimental bacteria all grew after 72 hours of culture, indicating that the examination method of example 7 is suitable for the sterility examination of gentamicin sulfate injection. And the results of the comparison with the example 1 show that the culture medium is added with vitamin C, and the buffer solution 1 is added with Tween-80, which is obviously beneficial to the sterile inspection of gentamicin sulfate injection.
Example 8
The method for checking the sterility of gentamicin sulfate injection and the method for verifying the applicability refer to example 5, except that: preparing trypticase soy peptone liquid culture medium, and making the pH value of the sterilized trypticase soy peptone liquid culture medium at 25 ℃ be 5.5; the amount of alkaline phosphatase added to the medium was 800U per 100 mL.
The method applicability verification test results of the present example are as follows:
TABLE 8
Note: + means liquid turbidity; denotes clarification of the liquid
The results in Table 8 show that 6 experimental bacteria all grew after 72 hours of culture, indicating that the examination method of example 8 is suitable for the sterility examination of gentamicin sulfate injection. And the result of the comparison with the example 2 shows that the addition of alkaline phosphatase to the culture medium and the adjustment of the pH of the trypticase soytone liquid culture medium are obviously beneficial to the aseptic examination of gentamicin sulfate injection.
The sterility test methods of examples 1-8, by the suitability verification test, indicate suitability for the sterility test of gentamicin sulfate injection. The sterile inspection method suitable for gentamicin sulfate injection is determined through the embodiments, and the methods comprise the steps of determining dilution of injection stock solution, selection of instruments for membrane filtration, membrane flushing after filtration, positive control bacteria and the like in the sterile inspection process; aiming at the bacteriostatic property of the gentamicin sulfate, an adjusted culture medium is also determined, the adjusted culture medium does not have obvious influence on the growth of bacteria, the bacteriostatic action of the gentamicin sulfate can be reduced, and the sterility test of the gentamicin sulfate injection is facilitated; the adjusted buffer solution is determined, and the sterility check of the gentamicin sulfate injection is facilitated. Tests prove that the sterility test method provided by the invention can accurately and reliably carry out sterility test on the gentamicin sulfate injection.
While the invention has been described with reference to certain preferred embodiments thereof, it will be understood by those skilled in the art that the foregoing is a more particular description of the invention than is possible with reference to the specific embodiments, which are not to be construed as limiting the invention. Various changes in form and detail, including simple deductions or substitutions, may be made by those skilled in the art without departing from the spirit and scope of the invention.
Claims (9)
1. A method for checking the sterility of gentamicin sulfate injection by membrane filtering method features that the culture contains test group, positive control group and negative control group,
the preparation step of each group of the culture comprises the steps of performing membrane filtration by using a duplex closed membrane filter, wherein the flushing amount of a buffer solution is 100mL per membrane, and the adding amount of a culture medium is 100mL per membrane;
the preparation step of the test group comprises the steps of adding the gentamicin sulfate injection into a diluent for dilution, wherein the volume of the diluent is 0-10 times that of the gentamicin sulfate injection; the filtration quantity of the gentamicin sulfate injection is that the filtration titer of each membrane is 80 ten thousand units;
the preparation method of the positive control group comprises the steps of adopting Escherichia coli as a positive control bacterium, and adding the positive control bacterium after the culture medium is added;
wherein,
the diluent is a sodium chloride-peptone buffer solution with pH7.0 or a sterile sodium chloride solution with 0.9 percent; the buffer solution is a sodium chloride-peptone buffer solution with pH 7.0; the culture medium comprises one of a thioglycollate fluid culture medium or a trypticase soytone liquid culture medium.
2. The method for checking the sterility of gentamicin sulfate injection according to claim 1, wherein the buffer is washed in an amount of 300mL per membrane; the gentamicin sulfate injection is added into a diluent for dilution, and the volume of the diluent is 10 times of that of the gentamicin sulfate injection.
3. The method for examining the sterility of gentamicin sulfate injection according to claim 1, wherein the pH of the trypticase Soy peptone liquid medium at 25 ℃ is 5.5-6.5.
4. The method for sterility test of gentamicin sulfate injection as claimed in claim 1, wherein the culture medium contains sterile alkaline phosphatase with viability unit 800-1000U per 100 mL.
5. The method for examining the sterility of gentamicin sulfate injection as claimed in claim 4, wherein said alkaline phosphatase is sterilized by a sterilization treatment comprising preparing a solution and sterilizing the solution by filtration through a microfiltration membrane filter having a pore size of 0.22 μm.
6. The method for checking the sterility of gentamicin sulfate injection according to claim 1, wherein the culture medium comprises 0.05-0.1% by mass of sterile vitamin C.
7. The method for examining the sterility of gentamicin sulfate injection as claimed in claim 6, wherein the vitamin C is sterilized, and the step of treating comprises filtering the vitamin C injection with a microfiltration membrane filter having a pore size of 0.22 μm for sterilization.
8. The method for checking the sterility of gentamicin sulfate injection according to any one of claims 3-7, further comprising a culture medium control group; the preparation of the culture medium control group is prepared according to the positive control group, and the added culture medium is the standard culture medium.
9. The method for checking the sterility of gentamicin sulfate injection according to any one of claims 1 to 7, wherein tween-80 is included in the buffer in an amount of 0.1% by mass.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910827523.6A CN110484596B (en) | 2019-09-03 | 2019-09-03 | Method for checking sterility of gentamicin sulfate injection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910827523.6A CN110484596B (en) | 2019-09-03 | 2019-09-03 | Method for checking sterility of gentamicin sulfate injection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110484596A true CN110484596A (en) | 2019-11-22 |
| CN110484596B CN110484596B (en) | 2023-04-04 |
Family
ID=68556342
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910827523.6A Active CN110484596B (en) | 2019-09-03 | 2019-09-03 | Method for checking sterility of gentamicin sulfate injection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110484596B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111500671A (en) * | 2020-04-13 | 2020-08-07 | 苏州市药品检验检测研究中心 | Method for checking sterility of docetaxel micelle for injection |
| CN113388663A (en) * | 2021-06-09 | 2021-09-14 | 深圳善康医疗健康产业有限公司 | Indissolvable implant sterility inspection method and sterility filtering device |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130041241A1 (en) * | 2009-07-02 | 2013-02-14 | John T. Felts | Pecvd coating methods for capped syringes, cartridges and other articles |
| CN107287276A (en) * | 2017-07-03 | 2017-10-24 | 中国大冢制药有限公司 | Compound sodium acetate electrolyte injection Sterility Test Methods |
| CN108148888A (en) * | 2018-03-09 | 2018-06-12 | 金花企业(集团)股份有限公司西安金花制药厂 | Yeast and mold microbial limit detection method |
-
2019
- 2019-09-03 CN CN201910827523.6A patent/CN110484596B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130041241A1 (en) * | 2009-07-02 | 2013-02-14 | John T. Felts | Pecvd coating methods for capped syringes, cartridges and other articles |
| CN107287276A (en) * | 2017-07-03 | 2017-10-24 | 中国大冢制药有限公司 | Compound sodium acetate electrolyte injection Sterility Test Methods |
| CN108148888A (en) * | 2018-03-09 | 2018-06-12 | 金花企业(集团)股份有限公司西安金花制药厂 | Yeast and mold microbial limit detection method |
Non-Patent Citations (3)
| Title |
|---|
| 周自永等: "《新编常用药物手册》", 31 May 1992, 金盾出版社 * |
| 杨晓莉等: ""2015年版《中国药典》无菌检查法解读"", 《中国药业》 * |
| 符春娜: ""硫酸庆大霉素注射液无菌检查方法学验证"", 《企业科技与发展》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111500671A (en) * | 2020-04-13 | 2020-08-07 | 苏州市药品检验检测研究中心 | Method for checking sterility of docetaxel micelle for injection |
| CN113388663A (en) * | 2021-06-09 | 2021-09-14 | 深圳善康医疗健康产业有限公司 | Indissolvable implant sterility inspection method and sterility filtering device |
| CN113388663B (en) * | 2021-06-09 | 2024-02-20 | 深圳善康医药科技股份有限公司 | Sterile inspection method and sterile filter device for insoluble implant |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110484596B (en) | 2023-04-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Evans et al. | Susceptibility of bacterial biofilms to tobramycin: role of specific growth rate and phase in the division cycle | |
| Gad et al. | Detection of icaA, icaD genes and biofilm production by Staphylococcus aureus and Staphylococcus epidermidis isolated from urinary tract catheterized patients | |
| CN110484596B (en) | Method for checking sterility of gentamicin sulfate injection | |
| CN113491692B (en) | Application of C-JUN N-terminal kinase inhibitor SU3327 | |
| Dawei et al. | In vitro biofilm forming potential of Streptococcus suis isolated from human and swine in China | |
| Baldassarri et al. | Effect of iron limitation on slime production by Staphylococcus aureus | |
| CN111500671A (en) | Method for checking sterility of docetaxel micelle for injection | |
| Serichantalergs et al. | Emerging fluoroquinolone and macrolide resistance of Campylobacter jejuni and Campylobacter coli isolates and their serotypes in Thai children from 1991 to 2000 | |
| CN111850086A (en) | Sterile detection method of voriconazole for injection | |
| CN109402219B (en) | Sterility inspection method of azithromycin eye drops | |
| Oche et al. | Isolation of biofilm producing methicillin-resistant Staphylococcus aureus from hospitalized orthopaedic patients in Kano State, Nigeria | |
| Blasetti et al. | A protocol for microbiologically safe preparation, storage, and use of autologous serum eye-drops in low-income countries | |
| CN113293194A (en) | Sterility test method for moxifloxacin hydrochloride eye drops | |
| CN110804646B (en) | Antibacterial activity removal method for rifampicin capsule microorganism limit control bacteria inspection | |
| CN112831539A (en) | In-vitro detection kit for aerobic microorganisms | |
| CN112899341A (en) | Selective enrichment medium for yersinia enterocolitica and preparation method thereof | |
| Clark | Antibiotic susceptibilities of the Vibrionaceae to meropenem and other antimicrobial agents | |
| Mashaly et al. | Adhesins encoding genes and biofilm formation as virulence determinants in methicillin resistant Staphylococcus aureus causing hospital acquired infections | |
| Blondeau et al. | Evaluation of aerobic Bactec 6A non-resin-and 16A resin-containing media for the recovery of microorganisms causing peritonitis | |
| Becherucci et al. | Validation of rapid sterility test with BacT/ALERT 3D on human mesenchymal stem cells as cell therapy product inside an hospital cell factory | |
| Hadke et al. | Anti-Microbial Activity of Antibiotic Produced From Fungi by Fermentation Process. | |
| CN114958956A (en) | Sterility inspection method for biological product containing mercuride | |
| MUNAWARA et al. | Isolation, characterization and identification of contaminant bacteria from sugarcane (Saccharum officinarum L.) in vitro culture. | |
| CN109988733A (en) | A kind of streptococcus specific solid medium, preparation method and applications | |
| Edwar et al. | Biofilm And Hemolysis Profile Index in Bacteria Isolated from Pre-Cesarean Surgery and Post Cesarean Infections |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |