CN109668887A - A kind of inspection method of trace protein of Chinese medicine injection - Google Patents
A kind of inspection method of trace protein of Chinese medicine injection Download PDFInfo
- Publication number
- CN109668887A CN109668887A CN201910142841.9A CN201910142841A CN109668887A CN 109668887 A CN109668887 A CN 109668887A CN 201910142841 A CN201910142841 A CN 201910142841A CN 109668887 A CN109668887 A CN 109668887A
- Authority
- CN
- China
- Prior art keywords
- chinese medicine
- inspection method
- traditional chinese
- injection
- tube
- 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.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/82—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a precipitate or turbidity
Abstract
The present invention provides a kind of inspection methods of trace protein of Chinese medicine injection, it includes following operating procedure: 1) taking traditional Chinese medicine stoste, sulfosalisylic acid solution is added to mix；2) step 1) mixed liquor is added in centrifuge tube, is centrifuged；3) observation centrifugation bottom of the tube, judging result.The inspection method of trace protein of Chinese medicine injection of the present invention increases high speed centrifugation step on the basis of " Chinese Pharmacopoeia " method, has strong anti-interference ability, can dramatically increase detection positive rate.Because the method is easy to operate, strong antijamming capability, it can be used in the safety evaluatio of traditional Chinese medicine.
Description
Technical field
The present invention relates to a kind of inspection methods of traditional Chinese medicine, and in particular to a kind of trace protein of Chinese medicine injection
Inspection method belongs to safety of Chinese medicine injection evaluation field.
Background technique
Traditional Chinese medicine results from last century the forties earliest, is started, has been widely used in by scholars such as Qian Xinzhong
It is clinical.The version Pharmacopoeia of the People's Republic of China in 2015 (referred to as " Chinese Pharmacopoeia ") one just included 'Zhichuanling, erigeron breviscapus,
Folk prescriptions and the compound injections such as ZHUSHEYONG SHUANGHUANGLIAN (freeze-drying), Breviscapine, qingkailing injections.Traditional Chinese medicine energy
It promotes drug effect, the curative effect of certain Chinese medicines is enable preferably to be played, promoting form of Chinese drug development, that expands Chinese medicine applies model
It encloses, service numerous people health etc. is performed meritorious deeds never to be obliterated really.Technologically speaking, industry has generally solved Chinese medicine note
The validity problem of agent is penetrated, safety is still industry problem.
The curative effect of traditional Chinese medicine comes from oral and external application earliest, does not have drug administration by injection mode.Most traditional Chinese medicine is ground
Hair also originates from the curative effect of oral agents, such as qingkailing injections, Shuanhuanglian injection, danshen injections and Erigeron Breviscapus Injection.
The oral traditional Chinese medicine ingredients that can be absorbed, which are substantially all, belongs to small-molecule substance, because macromolecular substances are not easy to absorb, or needs to drop
It could be absorbed after solution, and catabolite can not be reassembled into original macromolecular substances in body after absorbing.
Traditional Chinese medicine has the feature of bio-extract, may contain the macromolecular substances such as protein, tannin and resin.
Modern research shows that the big molecular impurity of unknown structure is the important material base for leading to safety of Traditional Chinese Medicine injections.To the greatest extent
The mechanism that pipe traditional Chinese medicine macromolecular substances cause safety issue is not fully understood, but its harm is gradually recognized by industry
It can.Therefore, " Chinese Pharmacopoeia " is since the macromolecular substances version in 2000 in monitoring traditional Chinese medicine, including protein, tannin
And resin.In these macromolecular substances, it is considered that it is the most serious that heterologous protein enters safety issue caused by body
Complexity, therefore the protein material in traditional Chinese medicine belongs to substance of keeping under strict control since two thousand.
Residual of the protein material from medicinal material extract in traditional Chinese medicine, current inspection method are by Chinese medicine injection
Whether agent stoste mixes with 30% sulfosalicylic acid 1:1, muddiness occur and judge whether traditional Chinese medicine contains macromolecular complex
Matter, the limitation of inspection is in 30 μ g/ml or so.Later someone improves trace protein of Chinese medicine injection inspection using absorption development process
Checking method, the sensitivity of inspection can achieve 12 μ g/ml (101865926 B of CN), 0.5 μ g/ml (101975861 B of CN),
Even 5ng/ml (102253036 B of CN).
However, absorption development process has the advantage of high sensitivity, but method is related to the operations such as absorption, dyeing and decoloration,
Step is complex, and influence factor as a result is more, is unfavorable for for the technology being standardized popularization.
Although " Chinese Pharmacopoeia " has protein inspection method (sulfosalicylic acid turbidimetry), traditional Chinese medicine is generally all
There is certain color to may interfere with result judgement.Therefore, a kind of easy to operate, strong antijamming capability trace protein is developed
Inspection method still makes progress meaning.
Summary of the invention
To solve the above problems, the present invention provides a kind of inspection method of trace protein of Chinese medicine injection, it includes
Following operating procedure:
1) traditional Chinese medicine stoste is taken, adds sulfosalisylic acid solution to mix, obtains mixed liquor；
2) step 1) mixed liquor is added in centrifuge tube, is centrifuged；
3) observation centrifugation bottom of the tube, judging result.
Further, step 1) the traditional Chinese medicine stoste is traditional Chinese medicine injection or injection traditional Chinese medicine by appended
The solution of operation instructions preparation；And/or the sulfosalisylic acid solutions are 30% (g/ml).
Further, the step 1) stoste and the volume ratio of sulfosalisylic acid solution are 1:9~9:1.
Further, the stoste and the volume ratio of sulfosalisylic acid solution are 1:1.
Further, the volume of the step 2) mixed liquor is 0.5~10mL.
Further, the volume of the mixed liquor is 2mL.
Further, the step 2) centrifuge tube is conical centrifuge tube.
Further, the step 2) centrifugal speed is greater than or equal to 5000 revs/min, and centrifugation time is 1~10 minute.
Further, 10000 revs/min of the centrifugal speed, centrifugation time are 5 minutes.
Further, step 3) the observation centrifugation bottom of the tube is observation bottom with the presence or absence of precipitating.
Further, the step 3) judging result is that centrifugation bottom of the tube has precipitating to show to check that the positive, no precipitating show
It checks negative.
The inspection method of trace protein of Chinese medicine injection of the present invention increases high speed on the basis of " Chinese Pharmacopoeia " method
Centrifugation step has strong anti-interference ability, and can dramatically increase detection positive rate.Because the method is easy to operate, anti-interference ability
By force, it can be used in the safety evaluatio of traditional Chinese medicine.
Obviously, above content according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific embodiment of form by the following examples remakes further specifically above content of the invention
It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on above content of the present invention
The technology realized all belongs to the scope of the present invention.
Specific embodiment
Raw material used in the specific embodiment of the invention is obtained by purchase commercial product.
The trace protein of Chinese medicine injection inspection of the present invention of embodiment 1
1) traditional Chinese medicine stoste 1ml is taken, adds 30% sulfosalicylic acid of 1ml to mix, obtains mixed liquor；
2) 2ml mixed liquor is added in conical centrifuge tube, 10000 revs/min of centrifugation 5min centrifugations；
3) whether observation centrifugation bottom of the tube has precipitating to generate, and centrifugation bottom of the tube has precipitating to show to check positive, no precipitating table
Bright inspection is negative.
The trace protein of Chinese medicine injection inspection of the present invention of embodiment 2
1) traditional Chinese medicine stoste 1ml is taken, adds 30% sulfosalicylic acid of 1ml to mix, obtains mixed liquor；
2) 0.5ml mixed liquor is added in conical centrifuge tube, 10000 revs/min of centrifugation 1min；
3) whether observation centrifugation bottom of the tube has precipitating to generate, and centrifugation bottom of the tube has precipitating to show to check positive, no precipitating table
Bright inspection is negative.
The trace protein of Chinese medicine injection inspection of the present invention of embodiment 3
1) traditional Chinese medicine stoste 1ml is taken, adds 30% sulfosalicylic acid of 9ml to mix, obtains mixed liquor；
2) 10ml mixed liquor is added in conical centrifuge tube, 10000 revs/min of centrifugation 10min centrifugations；
3) whether observation centrifugation bottom of the tube has precipitating to generate, and centrifugation bottom of the tube has precipitating to show to check positive, no precipitating table
Bright inspection is negative.
The trace protein of Chinese medicine injection inspection of the present invention of embodiment 4
1) traditional Chinese medicine stoste 9ml is taken, adds 30% sulfosalicylic acid of 1ml to mix, obtains mixed liquor；
2) 10ml mixed liquor is added in conical centrifuge tube, 10000 revs/min of centrifugation 10min centrifugations；
3) whether observation centrifugation bottom of the tube has precipitating to generate, and centrifugation bottom of the tube has precipitating to show to check positive, no precipitating table
Bright inspection is negative.
Beneficial effects of the present invention are further illustrated below by way of test example.
The following test of 1 standard protein solution of test example
Precision weighing 0.1g bovine serum albumin(BSA) is dissolved in the standard egg for being configured to that concentration is 10mg/ml in 10ml distilled water
Then white solution takes the 1ml standard protein solution to be diluted to the standard protein solution that 10ml concentration is 1mg/ml.Then 5ml is taken
Concentration is that the standard protein solution of 1mg/ml is successively diluted 7 times with 2 times of dilution methods again, it can thus be concluded that concentration be followed successively by 1000,
500,8 kinds of standard protein solution of 250,125,62.5,31.25,15.63 and 7.81 μ g/ml.Above-mentioned 8 kinds of concentration is taken respectively
Standard protein solution 1ml is mixed in equal volume with the 30% sulfosalisylic acid solution 1ml newly prepared, is placed 5 minutes and is seen whether out
Existing muddiness, the results are shown in Table 1.
The standard protein solution 1ml for separately taking above-mentioned 8 kinds of concentration, with the 30% sulfosalisylic acid solution 1ml that newly prepares in point
Bottom centrifuge tube is placed in 10000 revs/min of supercentrifuge after mixing in equal volume and is centrifuged 5 minutes, and whether observation centrifugation bottom of the tube goes out
It now precipitates, the results are shown in Table 1.
Table 1 the result shows that: after mixing observe (CP method) and centrifugation after observe (this law) result it is consistent.
The inspection of 1 standard protein of table
The protein inspection of 2 traditional Chinese medicine stoste of test example
Take 6 kinds of traditional Chinese medicine stostes (Sofflower injection, batch number: 1031504213；Danshen injections, product batch
Number: 1005104；Qingkailing injections, batch number: 1003272；Shuanhuanglian injection, batch number: 2010032437；It relaxes
Xuening injection, batch number: 1041611272；Erigeron Breviscapus Injection, batch number: 20130533；It is pharmaceutical factory listing
Qualified products) each 1ml, observe and mix without adding the 30% sulfosalisylic acid solution that 1ml is newly prepared after muddiness, place
5min has seen whether that muddy or precipitating generates.If there is muddy or precipitating by injection with water for injection carry out one time 2 times
Such as method inspection after dilution, until checking negative.It the results are shown in Table 2.
Separately take this 6 kinds of each 1ml of traditional Chinese medicine stoste in sharp bottom test tube, after 10000 revs/min of centrifugation 5min have no precipitating
The 30% sulfosalisylic acid solution that 1ml is newly prepared is added, then 10000 revs/min of centrifugation 5min again after mixing have been seen whether
Precipitating generates.If any precipitating, such as method inspection after injection is carried out 2 times of dilutions with water for injection, until inspection feminine gender is
Only.It the results are shown in Table 2.
Table 2 is as it can be seen that the trace protein of 6 kinds of traditional Chinese medicine stostes of observation (CP method) inspection is feminine gender after mixing.
However, observation (this law) checks discovery after centrifugation, danshen injections and qingkailing injections are the positive；Even danshen injections
8 times of dilution, qingkailing injections also check for the positive after diluting 4 times.Show that this law is sensitiveer in terms of traditional Chinese medicine inspection.
26 kinds of traditional Chinese medicine stock protein quality inspections of table are looked into
Test example 3 goes the protein inspection of macromolecular traditional Chinese medicine injection
Take 6 kinds of traditional Chinese medicine stostes (Sofflower injection, batch number: 1031504213；Danshen injections, product batch
Number: 1005104；Qingkailing injections, batch number: 1003272；Shuanhuanglian injection, batch number: 2010032437；It relaxes
Xuening injection, batch number: 1041611272；Erigeron Breviscapus Injection, batch number: 20130533；It is pharmaceutical factory listing
Qualified products) through 10K molecular sieve processing after be made remove macromolecular injection.It takes and respectively removes macromolecular injection 1ml, observe without muddy
The 30% sulfosalisylic acid solution that 1ml is newly prepared is added after turbid to mix, places 5min, has seen whether that muddy or precipitating produces
It is raw.Injection carried out with water for injection if there is muddy or precipitating after 2 times of dilutions such as method inspection, until checking feminine gender
Until.It the results are shown in Table 3.
This 6 kinds are separately taken to go each 1ml of macromolecular traditional Chinese medicine injection in sharp bottom test tube, 10000 revs/min of centrifugation 5min have no heavy
The 30% sulfosalisylic acid solution that 1ml is newly prepared is added behind shallow lake, 10000 revs/min of centrifugation 5min, then observation are again after mixing
It is no to there is precipitating to generate.It is negative until checking such as method inspection after injection is carried out 2 times of dilutions with water for injection if any precipitating
Until property.It the results are shown in Table 3.
Table 3 is as it can be seen that the trace protein of 6 kinds of traditional Chinese medicine stostes of observation (CP method) inspection is feminine gender after mixing.
However, observation (this law) checks discovery after centrifugation, danshen injections and qingkailing injections are the positive；Even after 2 times of dilution
Check the also positive.Show that this law is sensitiveer in terms of traditional Chinese medicine inspection.
Table 3 goes the inspection of protein in six kinds of traditional Chinese medicines of macromolecular
To sum up: the inspection method of trace protein of Chinese medicine injection of the present invention, though it not can increase inspection sensitivity, because
It has strong anti-interference ability, detection positive rate can be dramatically increased, compared with " Chinese Pharmacopoeia " method, anti-interference ability is stronger, quasi-
Exactness is higher, can be used in the safety evaluatio of traditional Chinese medicine.
Claims (10)
1. a kind of inspection method of trace protein of Chinese medicine injection, it is characterised in that: it includes following operating procedure:
1) traditional Chinese medicine stoste is taken, adds sulfosalisylic acid solution to mix, obtains mixed liquor；
2) step 1) mixed liquor is added in centrifuge tube, is centrifuged；
3) observation centrifugation bottom of the tube, judging result.
2. inspection method according to claim 1, it is characterised in that: step 1) the traditional Chinese medicine stoste is Chinese medicine note
Penetrate the solution that liquid or injection traditional Chinese medicine are prepared by appended operation instructions；And/or the sulfosalisylic acid solutions
For 30% (g/ml).
3. inspection method according to claim 1, it is characterised in that: the step 1) stoste and sulfosalisylic acid solution
Volume ratio is 1:9~9:1.
4. inspection method according to claim 3, it is characterised in that: the volume ratio of the stoste and sulfosalisylic acid solution
For 1:1.
5. inspection method according to claim 1, it is characterised in that: the volume of the step 2) mixed liquor be 0.5~
10mL。
6. inspection method according to claim 5, it is characterised in that: the volume of the mixed liquor is 2mL.
7. inspection method according to claim 1, it is characterised in that: the step 2) centrifuge tube is conical centrifuge tube, institute
Centrifugal speed is stated more than or equal to 5000 revs/min, centrifugation time is 1~10 minute.
8. inspection method according to claim 7, it is characterised in that: 10000 revs/min of the centrifugal speed, centrifugation time
It is 5 minutes.
9. inspection method according to claim 1, it is characterised in that: step 3) the observation centrifugation bottom of the tube is observation bottom
Portion is with the presence or absence of precipitating.
10. inspection method according to claim 1, it is characterised in that: the step 3) judging result is centrifugation bottom of the tube
There is precipitating to show to check that the positive, no precipitating show to check negative.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910142841.9A CN109668887A (en) | 2019-02-26 | 2019-02-26 | A kind of inspection method of trace protein of Chinese medicine injection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910142841.9A CN109668887A (en) | 2019-02-26 | 2019-02-26 | A kind of inspection method of trace protein of Chinese medicine injection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109668887A true CN109668887A (en) | 2019-04-23 |
Family
ID=66151779
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910142841.9A Pending CN109668887A (en) | 2019-02-26 | 2019-02-26 | A kind of inspection method of trace protein of Chinese medicine injection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109668887A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112964709A (en) * | 2021-02-18 | 2021-06-15 | 济南尚诚医药科技有限公司 | Method for detecting protein in sample |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1739598A (en) * | 2005-08-23 | 2006-03-01 | 咸阳步长医药科技发展有限公司 | Chinese medicine injection for treating cancer and its prepn process and quality control method |
| CN101007072A (en) * | 2007-01-18 | 2007-08-01 | 天津红日药业股份有限公司 | Quality-control method of a traditional Chinese medicine 'Xuebijing' injection |
| CN101865926A (en) * | 2010-06-13 | 2010-10-20 | 云南中医学院 | Detection method for trace protein of Chinese medicinal injection |
| CN103940929A (en) * | 2014-05-05 | 2014-07-23 | 山东丹红制药有限公司 | Method for detecting traditional Chinese medicine injection for treating cardiovascular and cerebrovascular diseases |
| CN104950101B (en) * | 2015-07-20 | 2016-08-24 | 石药银湖制药有限公司 | A kind of inspection method of Chinese medicine trace resin |
-
2019
- 2019-02-26 CN CN201910142841.9A patent/CN109668887A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1739598A (en) * | 2005-08-23 | 2006-03-01 | 咸阳步长医药科技发展有限公司 | Chinese medicine injection for treating cancer and its prepn process and quality control method |
| CN101007072A (en) * | 2007-01-18 | 2007-08-01 | 天津红日药业股份有限公司 | Quality-control method of a traditional Chinese medicine 'Xuebijing' injection |
| CN101865926A (en) * | 2010-06-13 | 2010-10-20 | 云南中医学院 | Detection method for trace protein of Chinese medicinal injection |
| CN103940929A (en) * | 2014-05-05 | 2014-07-23 | 山东丹红制药有限公司 | Method for detecting traditional Chinese medicine injection for treating cardiovascular and cerebrovascular diseases |
| CN104950101B (en) * | 2015-07-20 | 2016-08-24 | 石药银湖制药有限公司 | A kind of inspection method of Chinese medicine trace resin |
Non-Patent Citations (1)
| Title |
|---|
| 蒋云霞: "《分析化学》", 28 February 2015, 中国环境科学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112964709A (en) * | 2021-02-18 | 2021-06-15 | 济南尚诚医药科技有限公司 | Method for detecting protein in sample |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104458367B (en) | A kind of d-dimer and the compound quality-control product of FDP and preparation method thereof | |
| Martini et al. | Neurohumoral control of the release of adrenocorticotrophic hormone | |
| CN101317852B (en) | Powder injection of compound glycyrrhizic acid glycosides and preparation method thereof | |
| EP3336537B1 (en) | Method for identifying animal skin/hide | |
| WO2017032277A1 (en) | Bovine lung enoxaparin sodium, preparation method therefor, and application thereof | |
| CN109668887A (en) | A kind of inspection method of trace protein of Chinese medicine injection | |
| CN103777022B (en) | A kind of method for detecting Vaccinum Encephalitis B viral antigen content | |
| CN106645481A (en) | Method for testing dissolution curve of quick release dosage form drug-amoxicillin and clavulanate potassium dispersible tablet | |
| CN109668886A (en) | A kind of inspection method of the micro tannin of traditional Chinese medicine | |
| CN103675297A (en) | Medicament-induced hemolytic anemia detection kit and detection method thereof | |
| CN103616376A (en) | Method for detecting steroidal anti-inflammation drugs in Chinese patent medicines and health-care products | |
| CN102475687B (en) | Decitabine freeze-dry powder injection solution | |
| CN103142991B (en) | Cerebroprotein hydrolysate and lyophilized powder thereof for injection | |
| CN104198669A (en) | Method for determining immune activity of tracheitis vaccine by using trace hemolysis spectrophotometry | |
| CN101975861A (en) | Method for detecting trace protein of Chinese medicine injection | |
| Ma et al. | Ultrasound-assisted extraction of arabinogalactan and dihydroquercetin simultaneously from Larix gmelinii as a pretreatment for pulping and papermaking | |
| CN104721153B (en) | Injection aminomethylbenzoic acid freeze-drying powder-injection pharmaceutical composition | |
| CN107281317A (en) | A kind of oral liquid for improving body immunity and preparation method thereof | |
| CN113876704A (en) | Epirubicin hydrochloride injection and preparation method thereof | |
| CN102680642B (en) | A kind of method detecting macromolecular substances in ginseng and astragalus injection for strengthening body | |
| Jiang et al. | Effects of raw and sulfur-fumigated Rhizoma Dioscoreae on rat hippocampal neurotransmitter concentrations | |
| CN108498531A (en) | A kind of chitosan oral solution and preparation method thereof | |
| CN107303385B (en) | Preparation method of cerebroprotein hydrolysate solution | |
| Pei et al. | Detection of bacterial endotoxin in paclitaxel liposome. | |
| CN102253036B (en) | Filtration interception method for detecting trace amounts of proteins in traditional Chinese medicine injection |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190423 |
|
| RJ01 | Rejection of invention patent application after publication |