CN108034693A - The microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers - Google Patents
The microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers Download PDFInfo
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Abstract
The invention belongs to drug measurement techniques field, and in particular to a kind of microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers.The microbial limit tests of the Bisoprolol Hemifumarate Capsule in Healthy Volunteers, step are as follows：(1) prepared by bacterium solution；(2) prepared by test liquid：Bisoprolol Hemifumarate Capsule in Healthy Volunteers test sample 10g is taken, is placed in sterile homogenizing bag, adds 45 DEG C of pH7.0 sterile NaCl peptone buffer agents of 100mL, 1min is patted with homogenizer, shakes up, as 1：10 test liquid；(3) aerobic bacteria, the measure of yeast and mold sum；(4) inspection of escherichia coli.The present invention can prevent the appearance of sample false negative in checkout procedure, reduce patient medication security risk.
Description
Technical field
The invention belongs to drug measurement techniques field, and in particular to a kind of microbial limit of Bisoprolol Hemifumarate Capsule in Healthy Volunteers
Inspection method.
Background technology
Bisoprolol fumarate, is beta-blocker, has high-affinity to the β1-receptor of bronchus and vascular smooth muscle,
So as to distend the blood vessels, blood pressure reduces.Suitable for hypertension, coronary heart disease and moderate to severe chronic stable heart failure etc.
Disease.Bisoprolol Hemifumarate Capsule in Healthy Volunteers is suitable for the treatment of essential hypertension, is more commonly used in bisoprolol fumarate's medicine
One kind.
The limitation standard in microbe of non-sterile medicine is method of administration based on medicine and potentially hazardous to patient health
And Chinese medicine particularity and work out.Microbial decolorization system check it is non-regulation sterilization preparation and its raw material, auxiliary material by
The method of microbial contamination degree.Inspection item includes bacterial population, fungi count, yeast count and Control bacteria examination.
Microbial limit tests on Bisoprolol Hemifumarate Capsule in Healthy Volunteers not yet have been reported that so far.
The content of the invention
The object of the present invention is to provide a kind of microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers, in microorganism
The phenomenon of false negative is avoided during limit test, reduces patient medication security risk.
The microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers of the present invention, step are as follows：
(1) prepared by bacterium solution
Staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, white is prepared respectively
Color candida albicans bacteria suspension and aspergillus niger spore suspension；
(2) prepared by test liquid
Bisoprolol Hemifumarate Capsule in Healthy Volunteers test sample 10g is taken, is placed in sterile homogenizing bag, adds 45 DEG C of pH7.0 of 100mL sterile
Sodium chloride-peptone buffer agent, pats 1min with homogenizer, shakes up, as 1：10 test liquid；
(3) aerobic bacteria, the measure of yeast and mold sum
1. test sample group
Take 1:10 test liquid 1mL note wares, pour into pancreas junket soy agar culture medium, and plate is inverted 30~35 DEG C and is cultivated 5 days；
Take 1:10 test liquid 1mL note wares, pour into Sabouraud glucose agar, and plate is inverted 20~25 DEG C of cultures 7
My god；
2. bacterium solution group
Take pH7.0 sterile NaCls-peptone buffer agent 9.9mL and 0.1mL bacterium solution to mix, take 1mL to note ware；
Wherein, staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, white
Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in pancreas junket soya peptone agar medium, and plate is inverted 30~35 DEG C
Culture 3 days；
Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in Sabouraud glucose agar, plate
20~25 DEG C are inverted to cultivate 5 days；
3. test group
Take 1:10 test liquid 9.9mL and 0.1mL test organisms mixes, and takes 1mL to note ware,
Wherein, staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, white
Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in pancreas junket soya peptone agar medium, and plate is inverted 30~35 DEG C
Culture 3 days；Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in Sabouraud glucose agar, plate
20~25 DEG C are inverted to cultivate 5 days；
4. aerobic bacteria, the measure of yeast and mold sum are carried out after each plate culture
Test group clump count subtracts the value of test sample group clump count and the ratio of bacterium solution group clump count in the range of 0.5~2
It is as qualified；
(4) inspection of escherichia coli
1. prepared by bacterium solution：
Prepare escherichia coli bacteria suspension；
2. inspection method：
Test sample group：Take 1：10 test liquid 10mL, adds in 100mLTSB culture mediums, mixes, 30~35 DEG C of cultures 18
~72 it is small when, obtain culture；Above-mentioned culture 1mL is taken to be seeded in 100mL Mai Kangkai fluid nutrient mediums, 42~44 DEG C of cultures
24~48 it is small when；Taking the streak inoculation of Mai Kangkai liquid cultures, 30~35 DEG C are cultivated 18 on maconkey agar culture medium flat plate
~72 it is small when；If as a result judging there is no colony growth on maconkey agar culture medium flat plate, though or there is colony growth identification to tie
Fruit is feminine gender, judges that test sample group does not detect escherichia coli；
Test group：Take 1：10 test liquid 10mL and no more than 100cfu escherichia coli add 100mLTSB culture mediums
In, mix, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to be seeded to 100mL Mai Kangkai liquid
In body culture medium, when 42~44 DEG C of cultures 24~48 are small；The streak inoculation of Mai Kangkai liquid cultures is taken in maconkey agar culture
On base tablet, when 30~35 DEG C of cultures 18~72 are small；If as a result judging there is no colony growth on maconkey agar culture medium flat plate,
Or though it is feminine gender to have colony growth but qualification result, judges that test group does not detect escherichia coli；
Positive controls：Take pH7.0 sterile NaCls-peptone buffer agent 10mL and no more than 100cfu escherichia coli
Add in 100mLTSB culture mediums, mix, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to connect
Kind is into 100mL Mai Kangkai fluid nutrient mediums, when 42~44 DEG C of cultures 24~48 are small；Take Mai Kangkai liquid culture streak inoculations
In on maconkey agar culture medium flat plate, when 30~35 DEG C of cultures 18~72 are small；If as a result judging, maconkey agar culture medium is put down
There is no colony growth on plate, though or have colony growth qualification result for feminine gender, it is uncommon to judge that positive controls do not detect large intestine angstrom
Bacterium；
Negative control group：Take pH7.0 sterile NaCls-peptone buffer agent 10mL to add in 100mLTSB culture mediums, mix
It is even, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to be seeded to 100mL Mai Kangkai Liquid Cultures
In base, when 42~44 DEG C of cultures 24~48 are small；The streak inoculation of Mai Kangkai liquid cultures is taken in maconkey agar culture medium flat plate
On, when 30~35 DEG C of cultures 18~72 are small；If as a result judging there is no colony growth on maconkey agar culture medium flat plate, though or have
Colony growth but qualification result are feminine gender, judge that negative control group does not detect escherichia coli.
In step (1) preparation method of staphylococcus aureus bacteria suspension learn from else's experience 30~35 DEG C culture 18~24 it is small when
Staphylococcus aureus fresh cultured thing, it is 5000~10000cfu that every 1mL, which is made, containing bacterium number with 0.9% aseptic sodium chloride solution
Bacteria suspension.
In step (1) preparation method of pseudomonas aeruginosa bacteria suspension be learn from else's experience 30~35 DEG C culture 18~24 it is small when copper
Green pseudomonad fresh cultured thing, every 1mL is made containing the bacterium that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution
Suspension.
In step (1) preparation method of bacillus subtilis bacteria suspension be learn from else's experience 30~35 DEG C culture 18~24 it is small when it is withered
Careless bacillus fresh cultured thing, every 1mL is made containing the bacterium that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution
Suspension.
The preparation method of Candida albicans bacteria suspension is 20~25 DEG C of Candida albicans for cultivating 2~3 days of learning from else's experience in step (1)
Bacterium fresh cultured thing, every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution.
In step (1) preparation method of aspergillus niger spore suspension be learn from else's experience 20~25 DEG C culture 5~7 days aspergillus niger it is new
Fresh culture, adds 3~5mL, 0.9% aseptic sodium chloride solutions, spore is eluted, and suctions out in spore suspension to sterile test tube, uses
Every 1mL is made containing the spore suspension that spore count is 5000~10000cfu in 0.9% aseptic sodium chloride solution.
In step (4) preparation method of escherichia coli bacteria suspension be learn from else's experience 30~35 DEG C culture 18~24 it is small when large intestine
Angstrom uncommon bacterium fresh cultured thing, is made every 1mL with 0.9% aseptic sodium chloride solution and is hanged containing the bacterium that bacterium number is 5000~10000cfu
Liquid.
Compared with prior art, the present invention have the advantages that：
The present invention can prevent the appearance of sample false negative in checkout procedure, reduce patient medication security risk.
Embodiment
The present invention is described further with reference to embodiments.
Embodiment 1
(1) prepared by bacterium solution
1. staphylococcus aureus, pseudomonas aeruginosa, bacillus subtilis
Pseudomonas aeruginosa, staphylococcus aureus, bacillus subtilis when 30~35 DEG C of cultures 18~24 of learning from else's experience are small
Fresh cultured thing, every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution.
2. Candida albicans
Learn from else's experience 20~25 DEG C of cultures Candida albicans fresh cultured things of 2~3 days, with 0.9% aseptic sodium chloride solution system
Into every 1mL containing the bacteria suspension that bacterium number is 5000~10000cfu.
3. aspergillus niger
Learn from else's experience 20~25 DEG C culture 5~7 days aspergillus niger fresh cultured thing, add 3~5mL, 0.9% sterile NaCls molten
Liquid, spore is eluted, and suctions out in spore suspension to sterile test tube, every 1mL is made with 0.9% aseptic sodium chloride solution and contains spore count
For the spore suspension of 5000~10000cfu.
(2) prepared by test liquid
Bisoprolol Hemifumarate Capsule in Healthy Volunteers test sample 10g is taken, is placed in sterile homogenizing bag, adds 45 DEG C of pH7.0 of 100mL sterile
Sodium chloride-peptone buffer agent, pats 1min with homogenizer, shakes up, as 1：10 test liquid.
(3) aerobic bacteria, the measure of yeast and mold sum
1. test sample group
Take 1:10 test liquid 1mL note wares, pour into pancreas junket soy agar culture medium, and plate is inverted 30~35 DEG C and is cultivated 5 days；
Take 1:10 test liquid 1mL note wares, pour into Sabouraud glucose agar, and plate is inverted 20~25 DEG C of cultures 7
My god；
Count results are shown in Table 1.
2. bacterium solution group
Take pH7.0 sterile NaCls-peptone buffer agent 9.9mL and 0.1mL bacterium solution to mix, take 1mL to note ware；
Wherein, staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, white
Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in pancreas junket soya peptone agar medium, and plate is inverted 30~35 DEG C
Culture 3 days；Count results are shown in Table 2；
Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in Sabouraud glucose agar, plate
20~25 DEG C are inverted to cultivate 5 days；Count results are shown in Table 3.
3. test group
Take 1:10 test liquid 9.9mL and 0.1mL test organisms mixes, and takes 1mL to note ware,
Wherein, staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, white
Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in pancreas junket soya peptone agar medium, and plate is inverted 30~35 DEG C
Culture 3 days, count results are shown in Table 4；Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately to Sabouraud dextrose fine jade
In fat culture medium, plate is inverted 20~25 DEG C and is cultivated 5 days, and count results are shown in Table 5.
4. aerobic bacteria, the measure of yeast and mold sum are carried out after each plate culture
Test group clump count subtracts the value of test sample group clump count and the ratio of bacterium solution group clump count in the range of 0.5~2
It is as qualified；Measurement result is shown in Table 6, table 7.Aerobic bacteria, the ratio of yeast and mold are in the range of 0.5~2, and detection is for examination
Product are qualified.
1 test sample group count results (unit of table：cfu)
| Aerobic bacteria | Yeast and mold |
| 0 | 0 |
2 aerobic bacteria bacterium solution group count results (unit of table：cfu)
| Staphylococcus aureus | Pseudomonas aeruginosa | Bacillus subtilis | Candida albicans | Aspergillus niger |
| 78 | 73 | 80 | 65 | 65 |
3 yeast and mold bacterium solution group bacterium solution group count results (unit of table：cfu)
| Candida albicans | Aspergillus niger |
| 85 | 62 |
4 aerobic bacteria test group count results (unit of table：cfu)
| Staphylococcus aureus | Pseudomonas aeruginosa | Bacillus subtilis | Candida albicans | Aspergillus niger |
| 67 | 70 | 80 | 57 | 62 |
5 yeast and mold test group count results (unit of table：cfu)
| Candida albicans | Aspergillus niger |
| 83 | 57 |
6 aerobic bacteria ratio of table
| Staphylococcus aureus | Pseudomonas aeruginosa | Bacillus subtilis | Candida albicans | Aspergillus niger |
| 0.86 | 0.96 | 1.00 | 0.88 | 0.95 |
7 yeast and mold ratio of table
| Candida albicans | Aspergillus niger |
| 0.98 | 0.92 |
(4) Control bacteria examination method applicability is tested：The inspection of escherichia coli
1. prepared by bacterium solution：
Escherichia coli fresh cultured thing when 30~35 DEG C of cultures 18~24 of learning from else's experience are small, it is molten with 0.9% sterile NaCl
Every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu in liquid.
2. inspection method：
Test sample group：Take 1：10 test liquid 10mL, adds in 100mLTSB culture mediums, mixes, 30~35 DEG C of cultures 18
~72 it is small when, obtain culture；Above-mentioned culture 1mL is taken to be seeded in 100mL Mai Kangkai fluid nutrient mediums, 42~44 DEG C of cultures
24~48 it is small when；Taking the streak inoculation of Mai Kangkai liquid cultures, 30~35 DEG C are cultivated 18 on maconkey agar culture medium flat plate
~72 it is small when；If as a result judging there is no colony growth on maconkey agar culture medium flat plate, though or there is colony growth identification to tie
Fruit is feminine gender, judges that test sample group does not detect escherichia coli；
Test group：Take 1：10 test liquid 10mL and no more than 100cfu escherichia coli add 100mLTSB culture mediums
In, mix, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to be seeded to 100mL Mai Kangkai liquid
In body culture medium, when 42~44 DEG C of cultures 24~48 are small；The streak inoculation of Mai Kangkai liquid cultures is taken in maconkey agar culture
On base tablet, when 30~35 DEG C of cultures 18~72 are small；If as a result judging there is no colony growth on maconkey agar culture medium flat plate,
Or though it is feminine gender to have colony growth but qualification result, judges that test group does not detect escherichia coli；
Positive controls：Take pH7.0 sterile NaCls-peptone buffer agent 10mL and no more than 100cfu escherichia coli
Add in 100mLTSB culture mediums, mix, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to connect
Kind is into 100mL Mai Kangkai fluid nutrient mediums, when 42~44 DEG C of cultures 24~48 are small；Take Mai Kangkai liquid culture streak inoculations
In on maconkey agar culture medium flat plate, when 30~35 DEG C of cultures 18~72 are small；If as a result judging, maconkey agar culture medium is put down
There is no colony growth on plate, though or have colony growth qualification result for feminine gender, it is uncommon to judge that positive controls do not detect large intestine angstrom
Bacterium；
Negative control group：Take pH7.0 sterile NaCls-peptone buffer agent 10mL to add in 100mLTSB culture mediums, mix
It is even, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to be seeded to 100mL Mai Kangkai Liquid Cultures
In base, when 42~44 DEG C of cultures 24~48 are small；The streak inoculation of Mai Kangkai liquid cultures is taken in maconkey agar culture medium flat plate
On, when 30~35 DEG C of cultures 18~72 are small；If as a result judging there is no colony growth on maconkey agar culture medium flat plate, though or have
Colony growth but qualification result are feminine gender, judge that negative control group does not detect escherichia coli.
Measurement result is shown in Table 8.
8 escherichia coli employment and suitability test (E ＆ ST) result of table
| Test sample | Test group | Positive bacteria control group | Negative control group |
| - | + | + | - |
Claims (7)
1. a kind of microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers, it is characterised in that step is as follows：
(1) prepared by bacterium solution
Staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, white is prepared respectively to read
Pearl bacterium bacteria suspension and aspergillus niger spore suspension；
(2) prepared by test liquid
Bisoprolol Hemifumarate Capsule in Healthy Volunteers test sample 10g is taken, is placed in sterile homogenizing bag, adds 45 DEG C of sterile chlorinations of pH7.0 of 100mL
Sodium-peptone buffer agent, pats 1min with homogenizer, shakes up, as 1：10 test liquid；
(3) aerobic bacteria, the measure of yeast and mold sum
1. test sample group
Take 1:10 test liquid 1mL note wares, pour into pancreas junket soy agar culture medium, and plate is inverted 30~35 DEG C and is cultivated 5 days；
Take 1:10 test liquid 1mL note wares, pour into Sabouraud glucose agar, and plate is inverted 20~25 DEG C and is cultivated 7 days；
2. bacterium solution group
Take pH7.0 sterile NaCls-peptone buffer agent 9.9mL and 0.1mL bacterium solution to mix, take 1mL to note ware；
Wherein, staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, Candida albicans
Bacterium bacteria suspension and aspergillus niger spore suspension are added separately in pancreas junket soya peptone agar medium, and plate is inverted 30~35 DEG C of cultures
3 days；
Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in Sabouraud glucose agar, and plate is inverted
20~25 DEG C are cultivated 5 days；
3. test group
Take 1:10 test liquid 9.9mL and 0.1mL test organisms mixes, and takes 1mL to note ware,
Wherein, staphylococcus aureus bacteria suspension, pseudomonas aeruginosa bacteria suspension, bacillus subtilis bacteria suspension, Candida albicans
Bacterium bacteria suspension and aspergillus niger spore suspension are added separately in pancreas junket soya peptone agar medium, and plate is inverted 30~35 DEG C of cultures
3 days；Candida albicans bacteria suspension and aspergillus niger spore suspension are added separately in Sabouraud glucose agar, and plate is inverted
20~25 DEG C are cultivated 5 days；
4. aerobic bacteria, the measure of yeast and mold sum are carried out after each plate culture
Test group clump count subtracts the value of test sample group clump count
It is qualified；
(4) inspection of escherichia coli
1. prepared by bacterium solution：
Prepare escherichia coli bacteria suspension；
2. inspection method：
Test sample group：Take 1：10 test liquid 10mL, adds in 100mLTSB culture mediums, mixes, 30~35 DEG C of cultures 18~72
Hour, obtain culture；Above-mentioned culture 1mL is taken to be seeded in 100mL Mai Kangkai fluid nutrient mediums, 42~44 DEG C of cultures 24~
48 it is small when；Taking the streak inoculation of Mai Kangkai liquid cultures, 30~35 DEG C are cultivated 18~72 on maconkey agar culture medium flat plate
Hour；If as a result judging there is no colony growth on maconkey agar culture medium flat plate, though or there is the colony growth qualification result to be
Feminine gender, judges that test sample group does not detect escherichia coli；
Test group：Take 1：10 test liquid 10mL and no more than 100cfu escherichia coli add 100mLTSB culture mediums in, mix
It is even, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to be seeded to 100mL Mai Kangkai Liquid Cultures
In base, when 42~44 DEG C of cultures 24~48 are small；The streak inoculation of Mai Kangkai liquid cultures is taken in maconkey agar culture medium flat plate
On, when 30~35 DEG C of cultures 18~72 are small；If as a result judging there is no colony growth on maconkey agar culture medium flat plate, though or have
Colony growth but qualification result are feminine gender, judge that test group does not detect escherichia coli；
Positive controls：Take pH7.0 sterile NaCls-peptone buffer agent 10mL and added no more than 100cfu escherichia coli
In 100mLTSB culture mediums, mix, when 30~35 DEG C of cultures 18~72 are small, obtain culture；Above-mentioned culture 1mL is taken to be seeded to
In 100mL Mai Kangkai fluid nutrient mediums, when 42~44 DEG C of cultures 24~48 are small；Mai Kangkai liquid culture streak inoculations are taken in wheat
On Kang Kai agar medium tablets, when 30~35 DEG C of cultures 18~72 are small；If as a result judge on maconkey agar culture medium flat plate
There is no colony growth, though or have colony growth qualification result for feminine gender, judge positive controls do not detect escherichia coli；
Negative control group：Take pH7.0 sterile NaCls-peptone buffer agent 10mL to add in 100mLTSB culture mediums, mix, 30
When~35 DEG C of cultures 18~72 are small, culture is obtained；Above-mentioned culture 1mL is taken to be seeded in 100mL Mai Kangkai fluid nutrient mediums,
When 42~44 DEG C of cultures 24~48 are small；The streak inoculation of Mai Kangkai liquid cultures is taken on maconkey agar culture medium flat plate, 30
When~35 DEG C of cultures 18~72 are small；If as a result judging there is no colony growth on maconkey agar culture medium flat plate, though or there is a bacterium colony
Growth but qualification result are feminine gender, judge that negative control group does not detect escherichia coli.
2. the microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers according to claim 1, it is characterised in that step
Suddenly in (1) preparation method of staphylococcus aureus bacteria suspension be learn from else's experience 30~35 DEG C culture 18~24 it is small when golden yellow grape
Coccus fresh cultured thing, every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution.
3. the microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers according to claim 1, it is characterised in that step
Suddenly in (1) preparation method of pseudomonas aeruginosa bacteria suspension be learn from else's experience 30~35 DEG C culture 18~24 it is small when pseudomonas aeruginosa
Fresh cultured thing, every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution.
4. the microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers according to claim 1, it is characterised in that step
Suddenly in (1) preparation method of bacillus subtilis bacteria suspension be learn from else's experience 30~35 DEG C culture 18~24 it is small when bacillus subtilis
Fresh cultured thing, every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution.
5. the microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers according to claim 1, it is characterised in that step
Suddenly the preparation method of Candida albicans bacteria suspension is 20~25 DEG C of Candida albicans fresh cultureds for cultivating 2~3 days of learning from else's experience in (1)
Thing, every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution.
6. the microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers according to claim 1, it is characterised in that step
Suddenly in (1) preparation method of aspergillus niger spore suspension be learn from else's experience 20~25 DEG C culture 5~7 days aspergillus niger fresh cultured thing,
Add 3~5mL, 0.9% aseptic sodium chloride solutions, spore is eluted, suction out in spore suspension to sterile test tube, it is sterile with 0.9%
Every 1mL is made containing the spore suspension that spore count is 5000~10000cfu in sodium chloride solution.
7. the microbial limit tests of Bisoprolol Hemifumarate Capsule in Healthy Volunteers according to claim 1, it is characterised in that step
Suddenly in (4) preparation method of escherichia coli bacteria suspension be learn from else's experience 30~35 DEG C culture 18~24 it is small when escherichia coli it is fresh
Culture, every 1mL is made containing the bacteria suspension that bacterium number is 5000~10000cfu with 0.9% aseptic sodium chloride solution.
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| CN112980918B (en) * | 2019-12-17 | 2022-12-30 | 杭州远大生物制药有限公司 | Culture medium for detecting mixed bacteria in live bacteria preparation |
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