CN106872634A - Facial paralysis quality determining method just scattered again - Google Patents
Facial paralysis quality determining method just scattered again Download PDFInfo
- Publication number
- CN106872634A CN106872634A CN201710132017.6A CN201710132017A CN106872634A CN 106872634 A CN106872634 A CN 106872634A CN 201710132017 A CN201710132017 A CN 201710132017A CN 106872634 A CN106872634 A CN 106872634A
- Authority
- CN
- China
- Prior art keywords
- solution
- facial paralysis
- thin
- medicinal material
- prepared
- 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
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- 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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
-
- 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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/10—Enterobacteria
-
- 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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/14—Streptococcus; Staphylococcus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
Abstract
The invention discloses the quality determining method that a kind of facial paralysis is just scattered again, facial paralysis quality determining method just scattered again includes following several：(1) proterties：Khaki powder；Gas is pungent, bitter；(2) differentiate：Including pseudo-ginseng, prepared RADIX ET RHIZOMA RHEI, Chinese violet, the bark of official magnolia indentification by TLC；(3) check：Inspection including granularity, appearance uniform degree, moisture, content uniformity and microbial limit；(4) assay：Assay including ginsenoside Rg1.By above method, facial paralysis quality standard just scattered again can be made, so as to realize, to facial paralysis quality testing just scattered again, the experiment proved that, in this way for standard detection facial paralysis is just dissipating again, can be very good to ensure the stability of preparation, so as to ensure its curative effect.
Description
Technical field
The present invention relates to technical field of Chinese medicine, and in particular to facial paralysis quality determining method just scattered again.
Background technology
Facial paralysis is a kind of acute peripheral facial paralysis caused by the nonsuppurative inflammation of facial nerve.It is mainly shown as patient facial region
Expression myoparalysis, i.e. distortion of commissure, the category traditional Chinese medical science mouthful out-of-the-way (apoplex involving the channels and collaterals card).《Interior warp》Middle title mouth fish sticking its mouth out of the water, mouth it is out-of-the-way, and think facial paralysis
The mainly lesion of the stomach channel of foot-Yang-ming.It is bright《Jing Yue's complete work miliaria wind》Point out：" mouth eye fish sticking its mouth out of the water tiltedly have distinguishing for fever and chills ... but vim and vigour
Without losing, though then heat may not muscular atonia, though it is cold may not muscular contracture, also always understood by declining for vim and vigour." at present, this disease being thought middle medical circles more
By positive unsaturated vapor, interspaces of skin and muscles being loose, weakened defensive QI, collaterals being void and defenceless, ailment said due to cold or exposure is taken advantage of a weak point in opponent's defence the facial channels and collaterals of invasion and attack, facial Stomach Muscle Channel lose in
Foster, qi and blood numbness resistance is moistened, so that muscle is indulged to delay does not receive caused.But there are fever and chills not, there is face to suffer from cold factor, heat type facial paralysis more sympotoms caused by cold factors
The multifactor body overabundance of yang, cold-evil enter innerization heat, thermal resistance sun is bright, shaoyang meridians network, cause stagnant through vapour lock, and muscles along the regular meridians loses and support and send out in moistening.Facial paralysis
The treatment of early stage is very crucial, and whether early treatment is appropriate, directly affects recovery in the future.Hormone being given current early treatment more
To mitigate the compression of facial nerve oedema, slow down facial nerve denaturation.Early stage anemopyretic patient is more with infection, so antiviral, antibacterial
The use of element is also very necessary.External treatment of tcm treatment bell paralysis is evident in efficacy, and most researchers are matched somebody with somebody
Close acupuncture.
Peripheral facial paralysis acute stage, is most commonly seen in clinic, while acute stage is also when treating the gold of peripheral facial paralysis
Section, focuses on the treatment of this phase and nurses, and can play the curative effect got half the result with twice the effort.Using Chinese medicine external compound preparation, clinically merge
Acupuncture treatment, is a feasible program for treating peripheral facial paralysis.This treatment method can make the direct skin permeation of medicine, drug effect
Through focus, can promote Doppler flow mapping to set up, and mitigate local inflammation and ooze out and oedema, improve neural surrounding microcirculation, with rush
Enter regional flow, improve neurotrophic effect.With instant effect, short treating period, effect be good, advantage without side reaction.
Facial paralysis is just dissipating prescription and is being made up of pseudo-ginseng, prepared RADIX ET RHIZOMA RHEI, Chinese violet, the bark of official magnolia, the taste drug matching of borneol 5 again, is Kweiyang
The second affiliated hospital of college of traditional Chinese medicine Cai Wei deputy director doctor is formed for treating according to theory of traditional Chinese medical science and clinical experience aggegation for many years
Peripheral facial paralysis acute stage and the pure Chinese medicine compound external-use preparation of partial gall patient.Its composition medicine disappears with anti-inflammatory respectively
Effect of each side such as swollen, suppression inflammatory oozes out, antibacterial, analgesia, reduction capillary permeability, promotion organization wound repair.
Clinically merge acupuncture treatment to also indicate that, pain is effective after facial paralysis is just dissipating to improving ear again, and its possible mechanism is inflammation and pain
Being proportionate property, is by improving patient's local pain, the non-specific inflammatory symptoms of facial neuritis is effectively controlled.But mesh
The preceding quality determining method being applicable without the medicine, it is difficult to ensure the stability and curative effect of preparation.
The content of the invention
It is an object of the invention to provide the quality determining method that facial paralysis is just scattered again, to overcome the deficiencies in the prior art.
The technical scheme is that such：
Facial paralysis quality determining method just scattered again, the facial paralysis just dissipates be prepared again：By 5~15 parts of pseudo-ginseng, ripe big
Yellow 5~15 parts, 10~20 parts of Chinese violet, 5~15 parts of the bark of official magnolia, 0.3~1 part of borneol assort, in addition to borneol, pseudo-ginseng, prepared RADIX ET RHIZOMA RHEI,
This four traditional Chinese medicine material of Chinese violet magnolia obovata was ground into powder through 60 DEG C of drying under reduced pressure 3 hours；Borneol is finely ground, with above-mentioned powder
Facing-up, sieving, mixing, packing is obtained final product；
Facial paralysis quality determining method just scattered again includes following several：
(1) proterties：Khaki powder；Gas is pungent, bitter；
(2) differentiate：Including pseudo-ginseng, prepared RADIX ET RHIZOMA RHEI, Chinese violet, the bark of official magnolia indentification by TLC；
(3) check：Inspection including granularity, appearance uniform degree, moisture, content uniformity and microbial limit；
(4) assay：Assay including ginsenoside Rg1.
Wherein, the TLC Identification of pseudo-ginseng comprises the following steps：
Need testing solution, negative control solution, control medicinal material solution and reference substance solution are prepared respectively；According to thin-layered chromatography
Experiment, draws need testing solution, negative control solution, control medicinal material solution and reference substance solution point in same silica gel g thin-layer plate
On, with chloroform-acetate-methanol-water (15：40：22：10) 10 DEG C of lower floor's solution arranged below are solvent, exhibition
Open, take out, dry, spray with ethanol solution of sulfuric acid, be heated to spot development clear；
In test sample chromatogram, on position corresponding with control medicinal material and reference substance chromatogram, show the fluorescent spot of same color
Point, on the negative sample chromatogram relevant position for lacking pseudo-ginseng, without this spot.
Further, the TLC Identification of prepared RADIX ET RHIZOMA RHEI comprises the following steps：
Need testing solution, negative control solution and control medicinal material solution are prepared as respectively；According to thin-layered chromatography experiment, difference
Control medicinal material solution, negative control solution and need testing solution are taken, is put on same silica gel g thin-layer plate, with n-hexane-acetic acid second
Ester-formic acid (27：10：2.4) for solvent launches, take out, dry, inspected under 365nm ultraviolet lamps；
In test sample chromatogram, there is same color spot with position with control medicinal material, negative control is noiseless.
Further, the TLC Identification of Chinese violet comprises the following steps：
Need testing solution, negative control solution and control medicinal material solution are prepared respectively；According to thin-layered chromatography experiment, draw and supply
Test sample solution, negative control solution and control medicinal material solution, put on same silica gel g thin-layer plate, with toluene-acetic acid second respectively
Ester-formic acid (5：3：1) upper solution is solvent, is launched, and is taken out, and is dried, and is placed under 365nm ultraviolet lamps and inspects；
In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show the fluorescence spot of same color, it is negative right
According to noiseless；
Further, the TLC Identification of the bark of official magnolia comprises the following steps：
Need testing solution, negative control solution and control medicinal material solution are prepared respectively；According to thin-layered chromatography, draw supply respectively
Test sample solution, negative control solution and control medicinal material solution, put on same silica gel g thin-layer plate, with petroleum ether (60~90 respectively
DEG C)-acetic ether-methanoic acid (8.5：1.5：0.2) it is solvent, launches, take out, dry, spray is with vanillin-sulfuric acid solution, heating
It is clear to spot development；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show same color spot, and negative control without dry
Disturb.
Further, facial paralysis limit test of microbe just scattered again is carried out using Plating.
Further, the assay of the ginsenoside Rg1 comprises the following steps：
Need testing solution is prepared with this product, with ginsenoside Rg1Reference substance solution is prepared, reference substance solution is drawn and is supplied to try
Product solution injection liquid chromatograph is measured, and chromatographic condition is：With Aglient ZORBAX SB-C8(150 × 4.6mm, 5 μ
M) it is chromatographic column；With acetonitrile as mobile phase A, with water as Mobile phase B, gradient elution is carried out；Flow velocity is 1ml/min；30 DEG C of column temperature；
Detection wavelength is 203nm；Gradient elution is provided as follows：
Mobile phase A：0~20 minute：18%；20~30 minutes：18 → 19%；
Mobile phase B：0~20 minute：82%；20~30 minutes：82 → 81%.
Technique effect of the invention is as follows：
By above method, facial paralysis quality standard just scattered again can be made, so as to realize to facial paralysis matter just scattered again
Amount detection, the experiment proved that, in this way for standard detection facial paralysis is just dissipating again, can be very good to ensure the stability of preparation, from
And ensure its curative effect.
Brief description of the drawings
Fig. 1 is the process chart of preparation method of the invention；
Fig. 2 is the thin layer discriminating figure of pseudo-ginseng of the invention；In figure, 1, pseudo-ginseng control medicinal material；2nd, notoginsenoside R and ginseng
Saponin(e Rg1 and ginsenoside Rb1's mixing reference substance；3~5, facial paralysis just dissipates test sample (lot number again：20160311,20160312,
20160313)；6th, negative test sample (scarce pseudo-ginseng)；
Fig. 3 is the thin layer discriminating figure of prepared RADIX ET RHIZOMA RHEI；In figure：1st, prepared RADIX ET RHIZOMA RHEI control medicinal material；2~4, facial paralysis just dissipates test sample again
(lot number：20160311,20160312,20160313)；5th, negative test sample (lacking prepared RADIX ET RHIZOMA RHEI medicinal material)；
Fig. 4 is the thin layer discriminating figure of Chinese violet；In figure, 1, Chinese violet control medicinal material；2~4, facial paralysis is just being dissipated for examination again
Product (lot number：20160311,20160312,20160313)；5th, negative test sample (lacking Chinese violet medicinal material)；
Fig. 5 is the thin layer discriminating figure of the bark of official magnolia；In figure, 1, bark of official magnolia control medicinal material；2~4, facial paralysis just dissipates test sample (lot number again：
20160311,20160312,20160313)；5th, negative test sample (scarce magnolia medicament)；
Fig. 6 is the thin layer discriminating figure one of the bark of official magnolia；In figure, 1, borneol control medicinal material；2nd, facial paralysis just dissipates test sample (lot number again：
20160501)；3rd, negative test sample (lacking borneol medicinal material)；
Fig. 7 is the thin layer discriminating figure two of the bark of official magnolia；In figure, 1, borneol control medicinal material；2nd, facial paralysis just dissipates test sample (lot number again：
20160501)；3rd, negative test sample (lacking borneol medicinal material).
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawings and examples, but protection scope of the present invention is not received
Any limitation of specific embodiment, but be defined in the claims.
Embodiment 1：
1st, prescription composition：Pseudo-ginseng 10g prepared RADIX ET RHIZOMA RHEI 10g Chinese violet 15g bark of official magnolia 10g borneols 0.5g.
2nd, preparation method：Technological process as shown in Figure 1, the above five tastes are assorted by 23 times of amounts of prescription, in addition to borneol,
The four traditional Chinese medicine such as pseudo-ginseng, prepared RADIX ET RHIZOMA RHEI material was ground into most fine powder through 60 DEG C of drying under reduced pressure 3 hours；Borneol is finely ground, match somebody with somebody with above-mentioned powder
Grind, sieve, mixing, packing is obtained final product.Loading amount standard：6g/ bags.
3rd, indication：It is clearing heat for detumescence, promoting blood circulation and removing obstruction in channels.For peripheral facial paralysis acute stage and partial gall patient.
4th, usage and dosage：External application, according to the state of an illness, external application, apply part acupuncture point, per cave is mixed well from vaseline, grease etc.
One patch, once a day, January is 1 course for the treatment of.Needed that the course for the treatment of can be repeated according to the state of an illness.
5th, check：According under powder pertinent regulations (《Chinese Pharmacopoeia》Four general rules 0115 of version in 2015) examined
Look into.Granularity, appearance uniform degree, moisture, content uniformity, microbial limit are checked respectively for, regulation should be met.
5.1st, granularity：(general rule 0115) is determined under powder " granularity " item, should meet regulation.
5.2nd, appearance uniform degree is determined according to (general rule 0115) under powder " appearance uniform degree " item, should meet regulation.
5.3rd, moisture：According to lower second method of aquametry (《Chinese Pharmacopoeia》Four general rules 0832 of version in 2015) carry out
Determine, moisture must not cross 9.0%.
5.4th, content uniformity：10 bags of this product is taken, (general rule 0115) checks that loading amount is poor in accordance with the law under powder " content uniformity " item
Different limit should must not be more than 2 bags, and must not have 1 bag of 1 times of overrun within ± 7% beyond content uniformity limit.
Experimental example 1：
Facial paralysis quality determining method just scattered again：
1 proterties：Three batches of samples of pilot scale are khaki powder；Gas is pungent, bitter.
2 thin layers differentiate：
The thin-layer chromatography research of 2.1 pseudo-ginseng
This product 2g, plus 5ml water are taken, is stirred evenly, add water the n-butanol 50ml of saturation, ultrasonically treated 1 hour, filtration, filtrate
Extracted with the water of 120ml n-butanol saturations, divide and take n-butanol layer, water bath method, residue 1ml methyl alcohol dissolves, used as test sample
Solution.The negative simulation preparation 2g of pseudo-ginseng is taken, negative control solution is prepared with method.Separately take pseudo-ginseng powder 0.5g, same to legal system
Into control medicinal material solution.Ginsenoside Rg is weighed again1Reference substance, ginsenoside Rb1Reference substance and Panax Notoginseng saponin R1Reference substance is fitted
Amount, is made solution of every 1ml containing 0.5mg, as reference substance solution of methyl alcohol respectively.According to thin-layered chromatography (《Chinese Pharmacopoeia》
Four general rules 0502 of version in 2015) experiment, above-mentioned each 5 μ l of four kinds of solution are drawn, put respectively on same silica gel g thin-layer plate, with
Chloroform-acetate-methanol-water (15：40：22：10) 10 DEG C of lower floor's solution arranged below are solvent, are launched, and are taken
Go out, dry, spray with 10% ethanol solution of sulfuric acid, be heated to spot development at 105 DEG C clear.In test sample chromatogram, with it is right
According on medicinal material and the corresponding position of reference substance chromatogram, show the spot of same color, lack the negative sample chromatogram phase of pseudo-ginseng
Answer on position, without this spot (see Fig. 2).
The thin-layer chromatography research of 2.2 prepared RADIX ET RHIZOMA RHEIs
This product 2g, plus methyl alcohol 15ml are taken, ultrasonic extraction 20min is filtered, and filtrate is concentrated into 1ml, used as need testing solution.
The negative simulation preparation 2g of scarce prepared RADIX ET RHIZOMA RHEI is taken, negative control solution is prepared with method.Prepared RADIX ET RHIZOMA RHEI control medicinal material powder 5g separately is taken, plus
The straight fire refluxing extraction 1h of 40ml water, filtrate water-bath Back stroke is done, and residue adds methyl alcohol 10ml to be transferred in conical flask, ultrasonic extraction 10min,
Filtrate is concentrated into 1ml, used as control medicinal material solution.According to thin-layered chromatography (《Chinese Pharmacopoeia》Four general rules 0502 of version in 2015) examination
Test, the μ l of control medicinal material solution 3, negative control solution, each 2 μ l of compound test solution are taken respectively, put in same silica gel g thin-layer plate
On, with n-hexane-ethyl acetate-formic acid (27：10：2.4) for solvent launches, take out, dry, under 365nm ultraviolet lamps
Inspect.In test sample chromatogram, there is same color spot with position with control medicinal material, negative control is noiseless (see Fig. 3).
The thin-layer chromatography research of 2.3 Chinese violets
This product 2g, plus methyl alcohol 20ml dissolvings are taken, ultrasonically treated 30min, filtration, filtrate is evaporated, and residue adds pure water 10ml, stirs
Mixing makes dissolving, filtration, and filtrate is evaporated, and residue adds the ethyl acetate 1ml to make dissolving, used as need testing solution.Take scarce Chinese violet
Feminine gender simulation preparation 2g, negative control solution is prepared with method.Chinese violet control medicinal material 1.0g separately is taken, control medicinal material is made in the same way of
Solution.According to thin-layered chromatography (《Chinese Pharmacopoeia》Four general rules 0502 of version in 2015) experiment, above-mentioned each 3 μ l of three kinds of solution are drawn,
Put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-formic acid (5：3：1) upper solution is solvent, is launched,
Take out, dry, be placed under uviol lamp (365nm) and inspect.In test sample chromatogram, on position corresponding with control medicinal material chromatogram,
The fluorescence spot of aobvious same color, negative control is noiseless (see Fig. 4).
The thin-layer chromatography research of 2.4 barks of official magnolia
Test sample 2g is taken, the negative control sample 2g prepared by formula rate is taken, methyl alcohol 15ml, ultrasonic 30min is added respectively,
Filtration, filtrate is respectively as need testing solution and negative control solution.Bark of official magnolia control medicinal material powder 0.5g, plus 5ml methyl alcohol separately are taken,
It is made in the same way of control medicinal material solution；According to thin-layered chromatography (《Chinese Pharmacopoeia》Four general rules 0502 of version in 2015) experiment, inhale respectively
Each 2 μ l of above-mentioned need testing solution, negative control solution, the μ l of control medicinal material solution 5 are taken, are put respectively on same silica gel g thin-layer plate,
With petroleum ether (60~90 DEG C)-acetic ether-methanoic acid (8.5：1.5：0.2) it is solvent, launches, take out, dry, sprays with 1%
Vanillin-sulfuric acid solution, is heated to spot development clear at 100 DEG C.In test sample chromatogram, in position corresponding with reference substance chromatogram
Put, show same color spot, and negative control is noiseless (see Fig. 5).
The thin-layer chromatography research of 2.5 borneols
With reference to " borneol in TLC, GC method detection storax capsule for treating coronary heart disease "[14]In to borneol thin-layer chromatography study：Take this product
1g, adds ethyl acetate 10ml, and extraction takes acetic acid ethyl fluid, as need testing solution.The negative simulation reagent 1g of scarce borneol is taken,
Negative controls are prepared with method.It is another to take borneol control medicinal material 1g, plus it is the solution of 2mg/ml that ethyl acetate is made concentration, as right
According to product solution.According to thin-layered chromatography (《Chinese Pharmacopoeia》Four general rules 0502 of version in 2015) experiment, draw above-mentioned three kinds of solution each 2
μ l, put on silica G thin web, with petroleum ether (30~60 DEG C)-ethyl acetate (9 respectively：1) it is solvent, launches, takes out,
Dry, spray with phosphomolybdic acid test solution, it is clear that hot blast is blown to spot development.In test sample chromatogram, corresponding with control medicinal material chromatogram
On position, without corresponding spot (see Fig. 6).To borneol in reference " compound inflammation-diminishing dissipates the indentification by TLC of middle borneol and turmeric "
Thin-layer chromatography is studied：This product 1g is taken, chloroform 10ml extractions are added, chloroform liquid is taken, as need testing solution.Take scarce ice
The negative simulation reagent 1g of piece, negative controls are prepared with method.It is another to take borneol control medicinal material, plus it is 1mg/ that ethyl acetate is made concentration
The solution of ml, as reference substance solution.According to thin-layered chromatography (《Chinese Pharmacopoeia》Four general rules 0502 of version in 2015) experiment, draw
Above-mentioned each 2 μ l of three kinds of solution, put on silica gel g thin-layer plate, with cyclohexane-ethyl acetate (17 respectively：3) it is solvent, launches,
Take out, dry, spray with phosphomolybdic acid test solution, it is clear that hot blast is blown to spot development.In test sample chromatogram with control medicinal material chromatogram phase
On the position answered, without corresponding spot (see Fig. 7).Therefore the Qualitive test of borneol excludes quality determining method of the invention.
Experimental example 2：Microbial decolorization
1st, the preparation of test liquid：This product 10ml is taken, is added and is no more than 45 DEG C of PH7.0 sterile NaCls-peptone dilutions
It is stirring while adding to 100ml, test sample is fully mixed, obtain final product 1:10 test liquid.
2nd, microorganism count method：The total counting number of aerobic bacteria：Take 1:10 1 ware of test liquid 1ml dispensings, 1ml/ wares are pressed《China
Pharmacopeia》2015 editions four non-sterile product limit test of microbe：Plating is carried out under microorganism count method (general rule 1105) item
Colony counting.
The total counting number of yeast and mold：Take 1:10 2 wares of test liquid 2ml dispensings, 1ml/ wares are pressed《Chinese Pharmacopoeia》2015
Four non-sterile product limit test of microbe of version：Plating in microorganism count method (general rule 1105) carries out colony counting.
3rd, Control bacteria examination：
Staphylococcus aureus：Take 1:10 test liquids according to《Chinese Pharmacopoeia》2015 editions four non-sterile product microorganism limits
Degree is checked：Corresponding Control bacteria examination method is carried out under Control bacteria examination method (general rule 1106).
Pseudomonas aeruginosa：Take 1:10 test liquids according to《Chinese Pharmacopoeia》2015 editions four non-sterile product microbial limits
Check：Corresponding Control bacteria examination method is carried out under Control bacteria examination method (general rule 1106).
Microbial limit test of drugs method applicability test report
Culture medium, diluent and source：
0.9% sterile NaCl-peptone buffer agent, pancreas junket soya peptone agar, Sabouraud's dextrose agar, pancreas junket soya peptone
Fluid nutrient medium, Mai Kangkai fluid nutrient mediums, maconkey agar culture medium etc..(Shanghai Sheng Si biochemical technologies Co., Ltd produces,
Press《Chinese Pharmacopoeia》2015 editions relevant regulations are prepared and sterilized).
Strain and source：
Above strain is National Institute for Food and Drugs Control and derives from strain.
Laboratory apparatus：Autoclave sterilizer, electronic balance (1/10th), toilet's clean work station, colony counting
Device etc..
As a result：Applicability is recorded and result is as follows：
1st, aerobic bacteria, mould and saccharomycete sum method of counting Selection experiment the results are shown in Table 1；
2nd, aerobic bacteria, mould and saccharomycete sum method of counting checking recovery test are shown in Table 2-4 (subordinate list records 1-3)；
3rd, bacterium --- the result of staphylococcus aureus inspection method see the table below 5 for control；
4th, bacterium --- the result of pseudomonas aeruginosa inspection method see the table below 6 for control；
The aerobic bacteria of table 1, mould and saccharomycete sum method of counting Selection experiment result
Remarks：" --- " represents and does not do this.
Conclusion：
1、1:10 test liquid uses Plating (1ml/ wares) EHEC, Candida albicans, the rate of recovery of aspergillus niger
More than 70% can be reached；1:10 test liquid can reach 70% using the rate of recovery of Plating (0.5ml/ wares) bacillus subtilis
More than, 1:10 test liquid can reach more than 70% using the rate of recovery of Plating (0.2ml/ wares) staphylococcus aureus.
2nd, facial paralysis just dissipates again has bacteriostatic active ingredients (such as：Chinese violet, bark of official magnolia etc.), therefore carrying out microorganism checking
When, there are obvious bacteriostasis, especially the inhibition to staphylococcus aureus is obvious, next to that to withered grass gemma
Bacillus has certain inhibitory action, so carried out using Plating, and effect of the medicine to bacillus subtilis is not it is obvious that
Take conventional method.According to the result, verification operation step is considered, it is final to determine to use Plating (1:10 for examination
Liquid 1ml, 5 plates of dispensing) limit test of microbe just scattered again to carry out facial paralysis.
The aerobic bacteria of table 2, mould and saccharomycete sum method of counting checking recovery test record 1
Note：Test group：Test liquid+experiment bacterium solution；Test sample control group：Test liquid+dilution；Bacterium solution group：Dilution+examination
Test bacterium solution.
The aerobic bacteria of table 3, mould and saccharomycete sum method of counting checking recovery test record 2
The aerobic bacteria of table 4, mould and saccharomycete sum method of counting checking recovery test record 3
The checking test record of the staphylococcus aureus inspection method of table 5
Remarks：In mannitol sodium chloride plates："+" is labeled with colonies typical growth；"-" indicates asepsis growth.
In coagulase test of blood plasma："+" is labeled with the clotting of plasma；"-" sign blood plasma does not solidify.
" --- " represents and does not do this.
The checking test record of the pseudomonas aeruginosa inspection method of table 6
Remarks：In mannitol sodium chloride plates："+" is labeled with colonies typical growth；"-" indicates asepsis growth.
In oxidase test：There is the change of pink-aubergine in culture on "+" sign paper；Trained on "-" sign paper
Support thing nondiscolouring or only show pink and change.
In pyo experiment：There is pink discolouration reaction in "+" sign；"-" sign occurs without pink.
" --- " represents and does not do this.
Conclusion：
Through this verification experimental verification, facial paralysis Aerobic Count just scattered again can be pressed《Chinese Pharmacopoeia》2015 editions four general rules 1105
Lower Plating (1:10 test liquids, 0.2ml/ wares) counted；Mould and the total counting number of saccharomycete can be pressed《Chinese Pharmacopoeia》2015
Version 1105 lower Platings of general rule (take 1:10 2 wares of test liquid 2ml dispensings, 1.0ml/ wares) count；Golden yellow Portugal in control bacterium
The inspection of grape coccus and pseudomonas aeruginosa can be pressed《Chinese Pharmacopoeia》The conventional method specified under 2015 editions general rules 1106 is examined
Look into.
Experimental example 3：Assay
Ginsenoside Rg1 according to high performance liquid chromatography (《Chinese Pharmacopoeia》Four general rules 0512 of version in 2015) determine.
1. instrument and reagent
Instrument：The high performance liquid chromatographs of Agilent 1260；Plum Teller-support benefit AE electronic analytical balances (METTLER
The a ten thousandths of TOLEDO ten).
Reagent：Acetonitrile [chromatographically pure, the U.S. world (TEDIA) Co., Ltd]；Water (Wahaha drinking pure water, Kweiyang baby
Heartily Food Co., Ltd's manufacture).
Reference substance：(for assay, in terms of 91.7%, lot number is content ginsenoside Rg1's reference substance：110703-
201530), purchased from Chinese food drug assay research institute.
Sample：Facial paralysis just dissipates (lot number again：20160501 20160502 20160503).
2. prepared by reference substance solution
The preparation of reference substance solution：Precision weighs ginsenoside Rg1 reference substance 4.536mg, plus methyl alcohol is made every 1ml containing people
The solution of ginseng saponin(e Rg1 0.416mg, obtains final product.
3. the selection of chromatographic condition：
Carrying out ginsenoside Rg1During chromatography condition, bibliography has carried out chromatographic column, mobile phase and column temperature
Screening.
The selection of 3.1 chromatographic columns
Different chromatographic columns is separated to chemical composition and had a certain impact, and Diamonsil Plus C18 colors have been investigated herein
Spectrum post (250 × 4.6mm, 5 μm)；Ultimate XB-C18 chromatographic columns (250 × 4.6mm, 5 μm)；Aglient ZORBAX SB-
C18(250 × 4.6mm, 5 μm) and Aglient ZORBAX SB-C8(150 × 4.6mm, 5 μm) four kinds of chromatographic column, the results are shown in Table 7.
The ginsenoside Rg of table 71Chromatogram column condition selects table
It is compared by chromatogram under the conditions of different chromatographic columns, as a result shows Ultimate XB-C18Before chromatographic column peak
Prolong, symmetry is poor, separating degree is low；Diamonsil plus C18 chromatographic column peak stretchings, peak shape is poor；Agilent Zorbax SB-
C18Chromatographic column peak trails, inferior separating effect；Agilent Zorbax SB-C8Preferably, separating degree is higher for chromatographic column peak shape, therefore
Determine Agilent Zorbax SB-C8(150 × 4.6mm, 5 μm) chromatographic column is experiment post.
The investigation of 3.2 mobile phases
Three kinds of different elution systems are investigated, 8-10 is the results are shown in Table.
The elution system one of table 8
The elution system two of table 9
The elution system three of table 10
By comparing the chromatographic peak of three of the above difference elution system, the wherein ginsenoside Rg of elution system one1With other
Component peaks are not kept completely separate, inferior separating effect；The chromatographic peak of elution system two is separated preferably, and peak shape is sharper；The ginseng of elution system three
Saponin(e Rg1It is not kept completely separate with other components peak, and feminine gender has interference, inferior separating effect.It is thus determined that elution system two is stream
Dynamic phase condition.
The investigation of 3.3 column temperatures
Investigated respectively 25 DEG C, 30 DEG C, 35 DEG C when chromatographic isolation effect.
Result shows, when column temperature is 25 DEG C, ginsenoside Rg1It is not kept completely separate with other components peak, inferior separating effect；Post
Temperature is 30 DEG C, 35 DEG C it is unobvious on the influence of the separating effect of chromatographic peak, it is contemplated that situations such as room temperature, selection column temperature is 30 DEG C.
To sum up, ginsenoside Rg is determined1Chromatographic condition be Agilent Zorbax SB-C8(150 × 4.6mm, 5 μm) color
Spectrum post；Acetonitrile-water gradient：0~20min (18:82), 21~30min (19:81)；Flow velocity：1ml/min；Column temperature：30℃；
Detection wavelength：203nm；Analysis time：30min；Sample size：10μl.Number of theoretical plate presses ginsenoside Rg1Calculating should be not less than
4000。
Certainly, it is more than the concrete application example of invention, the present invention also has other implementation methods, all use is equally replaced
Change or equivalent transformation formed technical scheme, all fall within protection domain of the presently claimed invention.
Claims (7)
1. the quality determining method that facial paralysis is just dissipating again, it is characterised in that：The facial paralysis just dissipates be prepared again：By pseudo-ginseng 5~
15 parts, 5~15 parts of prepared RADIX ET RHIZOMA RHEI, 10~20 parts of Chinese violet, 5~15 parts of the bark of official magnolia, 0.3~1 part of borneol assort, in addition to borneol, three
7th, this four traditional Chinese medicine material of prepared RADIX ET RHIZOMA RHEI, Chinese violet magnolia obovata was ground into powder through 60 DEG C of drying under reduced pressure 3 hours；Borneol is finely ground,
With above-mentioned powder facing-up, sieve, mixing, packing is obtained final product；
Facial paralysis quality determining method just scattered again includes following several：
(1) proterties：Khaki powder；Gas is pungent, bitter；
(2) differentiate：Including pseudo-ginseng, prepared RADIX ET RHIZOMA RHEI, Chinese violet, the bark of official magnolia indentification by TLC；
(3) check：Inspection including granularity, appearance uniform degree, moisture, content uniformity and microbial limit；
(4) assay：Assay including ginsenoside Rg1.
2. the quality determining method that facial paralysis according to claim 1 is just dissipating again, it is characterised in that：The thin-layer chromatography mirror of pseudo-ginseng
Other method comprises the following steps：
Need testing solution, negative control solution, control medicinal material solution and reference substance solution are prepared respectively；According to thin-layered chromatography examination
Test, draw need testing solution, negative control solution, control medicinal material solution and reference substance solution point on same silica gel g thin-layer plate,
With chloroform-acetate-methanol-water (15：40：22：10) 10 DEG C of lower floor's solution arranged below are solvent, are launched,
Take out, dry, spray with ethanol solution of sulfuric acid, be heated to spot development clear；
In test sample chromatogram, on position corresponding with control medicinal material and reference substance chromatogram, show the fluorescence spot of same color,
Lack on the negative sample chromatogram relevant position of pseudo-ginseng, without this spot.
3. the quality determining method that facial paralysis according to claim 1 is just dissipating again, it is characterised in that：The thin-layer chromatography of prepared RADIX ET RHIZOMA RHEI
Discrimination method comprises the following steps：
Need testing solution, negative control solution and control medicinal material solution are prepared as respectively；According to thin-layered chromatography experiment, it is right to take respectively
According to medicinal material solution, negative control solution and need testing solution, put on same silica gel g thin-layer plate, with n-hexane-ethyl acetate-
Formic acid (27：10：2.4) for solvent launches, take out, dry, inspected under 365nm ultraviolet lamps；
In test sample chromatogram, there is same color spot with position with control medicinal material, negative control is noiseless.
4. the quality determining method that facial paralysis according to claim 1 is just dissipating again, it is characterised in that：The thin layer color of Chinese violet
Spectrum discrimination method comprises the following steps：
Need testing solution, negative control solution and control medicinal material solution are prepared respectively；According to thin-layered chromatography experiment, test sample is drawn
Solution, negative control solution and control medicinal material solution, put on same silica gel g thin-layer plate, with toluene-ethyl acetate-first respectively
Acid (5：3：1) upper solution is solvent, is launched, and is taken out, and is dried, and is placed under 365nm ultraviolet lamps and inspects；
In test sample chromatogram, on position corresponding with control medicinal material chromatogram, show same color fluorescence spot, negative control without
Interference.
5. the quality determining method that facial paralysis according to claim 1 is just dissipating again, it is characterised in that：The thin-layer chromatography mirror of the bark of official magnolia
Other method comprises the following steps：
Need testing solution, negative control solution and control medicinal material solution are prepared respectively；According to thin-layered chromatography, test sample is drawn respectively
Solution, negative control solution and control medicinal material solution, put on same silica gel g thin-layer plate respectively, with petroleum ether (60~90 DEG C)-
Acetic ether-methanoic acid (8.5：1.5：0.2) it is solvent, launches, take out, dry, spray with vanillin-sulfuric acid solution, is heated to spot
Point colour developing is clear；
In test sample chromatogram, on position corresponding with reference substance chromatogram, show same color spot, and negative control is noiseless.
6. the quality determining method that facial paralysis according to claim 1 is just dissipating again, it is characterised in that：Carried out using Plating
Facial paralysis limit test of microbe just scattered again.
7. the quality determining method that facial paralysis according to claim 1 is just dissipating again, it is characterised in that：The ginsenoside Rg1
Assay comprise the following steps：
Need testing solution is prepared with this product, with ginsenoside Rg1Reference substance solution is prepared, reference substance solution is drawn molten with test sample
Liquid injection liquid chromatograph is measured, and chromatographic condition is：With Aglient ZORBAX SB-C8(150 × 4.6mm, 5 μm) is
Chromatographic column；With acetonitrile as mobile phase A, with water as Mobile phase B, gradient elution is carried out；Flow velocity is 1ml/min；30 DEG C of column temperature；Detection
Wavelength is 203nm；Gradient elution is provided as follows：
Mobile phase A：0~20 minute：18%；20~30 minutes：18 → 19%；
Mobile phase B：0~20 minute：82%；20~30 minutes：82 → 81%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710132017.6A CN106872634A (en) | 2017-03-07 | 2017-03-07 | Facial paralysis quality determining method just scattered again |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710132017.6A CN106872634A (en) | 2017-03-07 | 2017-03-07 | Facial paralysis quality determining method just scattered again |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106872634A true CN106872634A (en) | 2017-06-20 |
Family
ID=59170638
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710132017.6A Pending CN106872634A (en) | 2017-03-07 | 2017-03-07 | Facial paralysis quality determining method just scattered again |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106872634A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824244A (en) * | 2005-12-29 | 2006-08-30 | 贵州益佰制药股份有限公司 | Quelity control method of Chinese medicinal preparation for hepatatitis toxin resolving |
| CN101115493A (en) * | 2005-08-12 | 2008-01-30 | 河乃建仁 | Agent for hair growth |
| CN101698034A (en) * | 2009-11-05 | 2010-04-28 | 广州中一药业有限公司 | Method for controlling quality of modified inspirex extract |
| CN102068504A (en) * | 2009-11-19 | 2011-05-25 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Quality control method of phlegm-eliminating pill for treating pyrophlegm stagnation and dry feces |
| CN105548406A (en) * | 2016-01-20 | 2016-05-04 | 李兴惠 | Quality control method for Huju hepatitis B preparation and preparation |
-
2017
- 2017-03-07 CN CN201710132017.6A patent/CN106872634A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101115493A (en) * | 2005-08-12 | 2008-01-30 | 河乃建仁 | Agent for hair growth |
| CN1824244A (en) * | 2005-12-29 | 2006-08-30 | 贵州益佰制药股份有限公司 | Quelity control method of Chinese medicinal preparation for hepatatitis toxin resolving |
| CN101698034A (en) * | 2009-11-05 | 2010-04-28 | 广州中一药业有限公司 | Method for controlling quality of modified inspirex extract |
| CN102068504A (en) * | 2009-11-19 | 2011-05-25 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Quality control method of phlegm-eliminating pill for treating pyrophlegm stagnation and dry feces |
| CN105548406A (en) * | 2016-01-20 | 2016-05-04 | 李兴惠 | Quality control method for Huju hepatitis B preparation and preparation |
Non-Patent Citations (4)
| Title |
|---|
| 国家药典委员会 编: "《中华人民共和国药典 2010年版 一部》", 31 January 2010 * |
| 张振秋 等主编: "《药物分析》", 31 January 2016 * |
| 张瑞 等: "泡沫浮选-固相提取分离富集三七提取液中的人参皂苷", 《分析化学》 * |
| 蔡伟 等: "面瘫复正贴膏与针刺并用对周围性面瘫（急性期）患者临床疗效的观察", 《中医临床研究》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhong et al. | Therapeutic use of traditional Chinese herbal medications for chronic kidney diseases | |
| Liu et al. | Potential antidepressant properties of Radix polygalae (Yuan Zhi) | |
| Lau et al. | Synergistic interaction between Astragali Radix and Rehmanniae Radix in a Chinese herbal formula to promote diabetic wound healing | |
| CN105477006A (en) | Stomatitis clearing active component group and fingerprint characteristic spectrum constructing and quality detecting method for same | |
| CN101259172B (en) | Chinese medicinal composition for treating acne, preparation and application thereof | |
| CN105477166B (en) | A kind of preparation method and its method of quality control of duhuo jisheng decoction granule | |
| Chen et al. | Saikosaponin a and saikosaponin d inhibit proliferation and migratory activity of rat HSC-T6 cells | |
| Qiu et al. | The effect of Chaihu-Shugan-San and its components on the expression of ERK5 in the hippocampus of depressed rats | |
| Yue et al. | Screening for anti‐inflammatory and bronchorelaxant activities of 12 commonly used Chinese herbal medicines | |
| CN101708274B (en) | Quality control method of pill preparation for treating chest impediment and cardialgia | |
| Zheng et al. | Effects of herbal drugs in Mahuang decoction and their main components on intestinal transport characteristics of Ephedra alkaloids evaluated by a Caco-2 cell monolayer model | |
| Van Le et al. | Effects of steaming on saponin compositions and antiproliferative activity of Vietnamese ginseng | |
| Zhan et al. | The sulfur-fumigation reduces chemical composition and biological properties of Angelicae sinensis radix | |
| Lee et al. | Astragaloside II promotes intestinal epithelial repair by enhancing L-arginine uptake and activating the mTOR pathway | |
| CN104042824B (en) | Preparation and detecting methods of traditional Chinese medicine composition for treating wind-cold-wetness evil | |
| Bekara et al. | Evaluating the antidepressant Effect of Verbena officinalis L.(Vervain) aqueous extract in adult rats | |
| CN102353735B (en) | Quality detection method for Tongmai Tangyanming capsule | |
| Yan et al. | Chemical characterization and neuroprotective properties of copper nanoparticles green-synthesized by nigella sativa L. seed aqueous extract against methadone-induced cell death in adrenal phaeochromocytoma (PC12) cell line | |
| Ma et al. | Effects of traditional Chinese medicinal plants on anti-insulin resistance bioactivity of DXMS-induced insulin resistant HepG2 cells | |
| CN101890087A (en) | Composition containing coptis root, rhubarb and baikal skullcap root | |
| CN107252465B (en) | Analysis method of traditional Chinese medicine composition for regulating allergic constitution | |
| Jäger et al. | Activity of a traditional South African epilepsy remedy in the GABA-benzodiazepine receptor assay | |
| Nikolaou et al. | Accidental poisoning after ingestion of “aphrodisiac” berries: diagnosis by analytical toxicology | |
| Gryszczynska et al. | Bioactive compounds determination in the callus and hydroalcoholic extracts from Salvia miltiorrhiza and Salvia przewalskii–Preliminary study on their anti-alcoholic activity effects | |
| CN106872634A (en) | Facial paralysis quality determining method just scattered again |
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: 20170620 |
|
| RJ01 | Rejection of invention patent application after publication |