CN108403783A - A kind of humulus green pepper oral solution and preparation method thereof and detection method - Google Patents
A kind of humulus green pepper oral solution and preparation method thereof and detection method Download PDFInfo
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Abstract
The invention discloses a kind of humulus green pepper oral solution and preparation method thereof and detection methods, in parts by weight, take 10 20 parts of humulus grass, 0.6 1.5 parts of white pepper；Humulus grass adds alcohol reflux to extract and is concentrated to give humulus grass concentrate；White pepper beats powder, and after sieving plus water is evaporated under reduced pressure, and obtains pepper aqua aromatica；Remaining Aqueous extracts are discarded, addition alcohol reflux extraction, concentration is stood, and takes supernatant as pepper alkali extracting solution；The supernatant of humulus grass concentrate is mixed with pepper alkali extracting solution, centrifuging and taking supernatant adds pepper aqua aromatica；It is diluted with water, dispenses and humulus green pepper oral solution is made, detection method includes：Using thin-layer chromatography to humulus grass therein and white pepper Qualitive test；The active constituent content of humulus grass is measured using HPLC methods.The present invention is by humulus green pepper oral solution made from specific preparation method, and active constituent content is high, stable quality, while establishing detection method, and easy to operate, specificity is strong, as a result accurately, can be used for the quality control of humulus green pepper oral solution.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, especially a kind of humulus green pepper oral solution and preparation method thereof and detection method.
Background technology
Rotavirus enteritis is the acute infectious disease of the digestive tract caused by rotavirus, infant is occurred mainly in, often by A
Group rotavirus causes, and it is even dead that rotavirus infection can cause serious gastroenteritis, diarrhea, de- water and rock-soil coupling, shock
It dies, can alleviate clinical symptoms caused by rotavirus infection in spite of some drugs at present, but treat rotavirus not yet
The specific medicament of diarrhea, humulus green pepper side are on the basis of clinical experience for many years, for the baby diarrhea exploitation caused by rotavirus
Prescription, and arrange to obtain in conjunction with folk tradition proved recipe, there is good therapeutic effect to the baby diarrhea caused by rotavirus.But mesh
Preceding preparation method is irregular, also lacks unified detection method to control product quality.
Invention content
The purpose of the present invention is to overcome above-mentioned existing technical deficiency, a kind of humulus green pepper oral solution is provided, to colyliform disease
Baby diarrhea caused by poison has good therapeutic effect.
It is a further object to provide the preparation methods of above-mentioned humulus green pepper oral solution.
It is a still further object of the present invention to provide the detection methods of above-mentioned humulus green pepper oral solution so that the quality of humulus green pepper oral solution
Controllably, curative effect is stablized.
To achieve the above object, the present invention uses following technical proposals：
A kind of preparation method of humulus green pepper oral solution, it is characterised in that include the following steps：In parts by weight, humulus grass is taken
10-20 parts, 0.6-1.5 parts of white pepper；Humulus grass adds 8-12 times to measure 75% ethyl alcohol, and flow back 1.2-2h, and more than extraction once, merging carries
Liquid is taken, then concentration is the humulus grass concentrate that 1mL concentrates are equivalent to 1g crude drugs, stand for standby use；White pepper beats powder, adds after sieving
4-6 times is measured water, is evaporated under reduced pressure, and when the moisture rate of outflow is less than 2mL/min, is stopped distillation, is obtained pepper aqua aromatica, 1mL
Pepper aqua aromatica is equivalent to 0.5g crude drugs；Remaining Aqueous extracts are discarded, 75% ethyl alcohol, refluxing extraction one are measured in 5-7 times of addition
More than secondary, each 1.2-2h merges extracting solution, is concentrated into 1mL extracting solutions and is equivalent to 0.3g crude drugs, filters, and filtrate stands, and takes
Supernatant is as pepper alkali extracting solution；The supernatant of humulus grass concentrate and pepper alkali extracting solution are pressed 4:0.8-1.2 is mixed, then from
The heart takes supernatant, adds the amount of above-mentioned pepper aqua aromatica 70%-90%；It is finally diluted with water 8-12 times, trichlorine sugarcane is added
Sugar, packing.
A kind of humulus green pepper oral solution, it is characterised in that be made by above-mentioned preparation method.
The humulus green pepper oral solution is not less than 0.80mg per the 1mL gauge of rutin containing general flavone；The humulus green pepper oral solution is every
1mL gauge containing cosmosiin is not less than 0.10mg.
The detection method of above-mentioned humulus green pepper oral solution, it is characterised in that including：Using thin-layer chromatography to humulus grass therein and in vain
Pepper Qualitive test；The active constituent content of humulus grass is measured using HPLC methods.
The present invention is by humulus green pepper oral solution made from specific preparation method, and active constituent content is high, stable quality, simultaneously
It is promulgated according to Chinese Pharmacopoeia Commission《<Chinese Pharmacopoeia>Quality standards in Chinese drugs studies and defines technology requirement》Establish the present invention
Special formulation product detection method, in preparation humulus green pepper and white pepper carry out indentification by TLC, to the humulus in preparation
Grass carries out assay, and the method for foundation is easy to operate, and specificity is strong, as a result accurately, can be used for the quality control of humulus green pepper oral solution
System.
Description of the drawings
Fig. 1 is humulus green pepper oral solution (humulus grass) TLC chromatograms of the embodiment of the present invention 1, wherein：1,2,3 be respectively sample 1,
Sample 2,3 test sample of sample, 4 be humulus grass control medicinal material, and 5 be cyanidenon reference substance, and 6 be negative control.
Fig. 2 is humulus green pepper oral solution (pepper) TLC chromatograms of the embodiment of the present invention 1, wherein：1,2,3 be respectively sample 21,
Sample 2,3 test sample of sample, 4 be pepper control medicinal material, and 5 be pipering reference substance, and 6 be negative control.
Fig. 3 is that control substance of Rutin solution scans absorption curve in 400-600nm wave-length coverages.
Specific implementation mode
The present invention is a kind of preparation method of humulus green pepper oral solution, is included the following steps：
In parts by weight, 10-20 parts of humulus grass, 0.6-1.5 parts of white pepper are taken.Preferably, 20 parts of humulus grass, white pepper
1.5 part.
Humulus grass adds 8-12 times to measure 75% ethyl alcohol, and flow back 1.2-2h, extracts primary above (preferably twice), merges extraction
Liquid, then concentration is the humulus grass concentrate that 1mL concentrates are equivalent to 1g crude drugs, stand for standby use.Preferably, the humulus grass exists
80-90 DEG C of temperature flows back.
White pepper beats powder, and after sieving plus 4-6 times is measured water, is evaporated under reduced pressure, when the moisture rate of outflow is less than 2mL/min
When, stop distillation, obtains pepper aqua aromatica, 1mL pepper aqua aromaticas are equivalent to 0.5g crude drugs；Remaining Aqueous extracts are discarded, are added
5-7 times is added to measure 75% ethyl alcohol, refluxing extraction is primary above (preferably twice), and each 1.2-2h merges extracting solution, is concentrated into
1mL extracting solutions are equivalent to 0.3g crude drugs, and filtering, filtrate stands, and takes supernatant as pepper alkali extracting solution.Preferably, described
White pepper beat powder after, cross 120 mesh sieve.
The supernatant of humulus grass concentrate and pepper alkali extracting solution are pressed 4:0.8-1.2 is mixed, then is centrifuged, and supernatant is taken, then
The amount of above-mentioned pepper aqua aromatica 70%-90% is added；It is finally diluted with water 8-12 times, Sucralose is added, it is oral to obtain humulus green pepper
Liquid, packing.Preferably, after the supernatant by humulus grass concentrate is mixed with pepper alkali extracting solution, in rotating speed 3400-
Centrifugation 15-25min is carried out under 3800rpm.Preferably, in the humulus green pepper oral solution, a concentration of 0.25- of Sucralose
0.35%.The pH value 5.0-6.0 of humulus green pepper oral solution, relative density are not less than 1.02.Preferably, every 10mL is pressed in the packing
It is dispensed.
The humulus green pepper oral solution made from above-mentioned preparation method is in light brown transparency liquid, clarifies free from admixture, and nothing is heavy after standing
It forms sediment and occurs.In prescription, humulus grass has the effect of clearing heat and detoxicating, inducing diuresis for treating strangurtia, clearing away deficiency-heat, is monarch drug in a prescription；It is equipped with the temperature of white pepper
Middle cold dispelling, lower gas dissolving phlegm are ministerial drug, and two medicine phases are closed, and play the work(of the puckery antidiarrheal of temperature altogether.
Preferably, it is not less than 0.80mg in terms of rutin containing general flavone per 1mL；Preferably, humulus green pepper oral solution contains big wave per 1mL
This chrysanthemum glycosides gauge is not less than 0.10mg.
Humulus green pepper oral solution is made of humulus grass and white pepper, therefore humulus grass and white pepper are required for carrying out Qualitive test, and humulus
Grass is monarch drug in a prescription, should generally have the assay, detection method to include：Using thin-layer chromatography to humulus grass therein and the qualitative mirror of white pepper
Not；The active constituent content of humulus grass is measured using HPLC methods.
Preferably, the Qualitive test of the humulus grass, includes the following steps：
It takes humulus green pepper oral solution 20mL in separatory funnel, is extracted with 8-12mL ethyl acetate, lower layer depth is released after being layered
Brown liquid pours out upper layer of extraction liquid；Lower layer's dark brown liquid is poured into separatory funnel, then is extracted with 8-12mL ethyl acetate
Once, merge extract liquor twice, 50-70 DEG C of water bath method, residue adds appropriate ethyl acetate to dissolve, and filtering takes filtrate to be settled to
2mL, as test solution；Humulus grass control medicinal material coarse powder 1.4-1.6g is taken, ethyl acetate 28-32mL is added, is carried out at 50 DEG C
Ultrasonic 40min, filtering take 50-70 DEG C of water bath method of filtrate, residue that appropriate ethyl acetate is added to dissolve, and filtering takes filtrate to be settled to
1mL, as a contrast medicinal material solution；Precision weighs cyanidenon reference substance 0.00101g, adds appropriate ethyl acetate to dissolve, and constant volume
To 10mL, product solution as a contrast；It is tested according to thin-layered chromatography, draws test solution 15 μ L, 5 μ L of reference substance solution, right
According to 5 μ L of medicinal material solution, puts respectively on the silica gel g thin-layer plate in same using sodium carboxymethylcellulose as binder, be with volume ratio
16:4:1.2:0.8 toluene-ethyl acetate-methyl alcohol-formic acid is solvent, is unfolded, and takes out, dries, and is sprayed with 5% alchlor
Ethanol, it is clear to be heated to spot development, sets and is inspected under ultraviolet lamp, and test sample is with control medicinal material, reference substance in chromatography identical bits
It sets, shows the fluorescence spot of same color.
Preferably, the Qualitive test of the white pepper, includes the following steps：
It takes humulus green pepper oral solution 10mL, 60-80 DEG C of water bath method, residue that absolute ethyl alcohol 1mL is added to make dissolving, filters, take filtrate
It is settled to 1mL, as test solution；Powder 1.4-1.6g in pepper control medicinal material is taken, 75% ethyl alcohol 8-10mL, refluxing extraction are added
25-35min filters off the dregs of a decoction, takes filtrate to be settled to 5mL, above-mentioned solution 2mL, 50-70 DEG C of water bath method, residue is taken to add anhydrous second
Alcohol 1mL makes dissolving, and filtering takes filtrate to be settled to 1mL, as a contrast medicinal material solution；Pipering reference substance 0.00423g is taken, palm fibre is set
In colo(u)r specification bottle, absolute ethyl alcohol is added to dissolve, is settled to 1mL, as a contrast product solution；It tests, is drawn for examination according to thin-layered chromatography
10 μ L of product solution, 5 μ L of reference substance solution, 10 μ L of control medicinal material solution are put respectively in same with sodium carboxymethylcellulose as bonding
On the silica gel g thin-layer plate of agent, with volume ratio for 7:2:1 toluene-ethyl acetate-acetone is solvent, is unfolded, and takes out, dries,
With 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development, sets and is inspected under ultraviolet lamp for spray, test sample and control medicinal material, right
According to product in chromatography same position, the fluorescence spot of same color is shown.
Preferably, the active constituent content measuring of the humulus grass, according to ultraviolet spectrophotometry and high performance liquid chromatography
Method is tested, and steps are as follows：
General flavone rutin measures：(1) prepared by reference substance solution：Precision is weighed through the dry rutin standard items to permanent quality
0.01025g is placed in 50mL volumetric flasks, adds 80% ethyl alcohol to scale, shakes up product storing solution as a contrast.Precision measures deposit
Liquid 5.0mL, is placed in 10mL volumetric flasks and 5% sodium nitrite solution 0.5mL is added and shake up, and places 6min, adds 10% tri-chlorination
Aluminum solutions 0.5mL shakes up, and places 6min, then 4% sodium hydroxide solution 4.0mL is added to shake up, is then settled to quarter with 80% ethyl alcohol
Spend line, shake up, place 15min to get；(2) prepared by test solution：Precision measures humulus green pepper oral solution 2.0mL, and 80 DEG C of water-baths are steamed
Dry, residue adds appropriate 80% ethyl alcohol to make dissolving, filtering, takes filtrate, then be settled to 5mL with 80% ethyl alcohol, and precision measures 1.0mL,
10mL volumetric flasks are placed in, and 5% sodium nitrite solution 0.5mL is added and shakes up, 6min is placed, adds 10% liquor alumini chloridi 0.5mL
It shakes up, places 6min, then 4% sodium hydroxide solution 4.0mL is added to shake up, be then settled to graduation mark with 80% ethyl alcohol, shake up, put
Set 15min to get；(3) wavelength selection：It prepares not plus the solution of color developing agent is only added as placebo solution in reference substance.It takes
Control substance of Rutin solution scans absorption curve in 400-600nm wave-length coverages, determines that Detection wavelength is 506nm；(4) it measures
Method：Test solution is taken, absorbance is measured at 506nm, humulus green pepper oral solution is per 1mL rutins containing general flavone (C27H30O16) gauge,
0.80mg must not be less than；
Cosmosiin measures：(1) chromatographic condition and system suitability：Chromatographic column Agilent TC-C18250 ×
4.6mm；Mobile phase is -0.2% phosphoric acid (22 of second eyeball:78)；Column temperature is 25 DEG C；Detection wavelength is 350nm；Flow velocity is 1.0mL/
min；10 μ L of sample size.Number of theoretical plate is calculated by cosmosiin should be not less than 5000；(2) prepared by test solution：Precision is inhaled
It takes humulus green pepper oral solution 10.0mL in 60mL separatory funnels, 20mL ethyl acetate is added, lower layer depth is released in shaking out after standing
Brown liquid pours out upper layer acetic acid ethyl acetate extract.Lower layer's dark brown liquid is refunded into separatory funnel, again with 20mL ethyl acetate
Extraction is primary.Merging extract liquor, 60 DEG C of water bath methods, residue twice adds methanol to make dissolving, filters, filtrate is taken to be settled to 10mL,
To obtain the final product；(3) prepared by reference substance solution：Precision weighs cosmosiin reference substance 0.00104g, constant volume after adding proper amount of methanol to dissolve
To 10mL to get；(4) measuring method：It is accurate respectively to draw reference substance solution and each 10 μ L of test solution, inject liquid chromatogram
Instrument measures, and humulus green pepper oral solution is per 1mL (C containing cosmosiin21H20O10) gauge, 0.10mg must not be less than.
With reference to example, the present invention will be further elaborated, it should which explanation, following the description is merely to solution
The present invention is released, its content is not defined.
Embodiment 1
Humulus green pepper oral solution prescription is：Humulus grass 20g, white pepper 1.5g.Preparation method：The each component of recipe ratio, humulus grass is taken to add 10 times
75% ethyl alcohol is measured, flow back 1.5h under 85 DEG C of constant temperature, and extraction twice, merges extracting solution, and then water-bath concentration is that 1mL concentrates are suitable
In 1g crude drugs, stood in shady place, it is spare；White pepper beats powder, crosses 120 mesh sieve, adds 5 times of amount water, is evaporated under reduced pressure, works as moisture
When the rate of outflow is less than 2mL/min, stops distillation, obtain pepper aqua aromatica, 1mL pepper aqua aromaticas are equivalent to 0.5g crude drugs；It will
Remaining Aqueous extracts discard, and add 6 times of 75% ethyl alcohol of amount, and refluxing extraction 2 times, each 1.5h merges extracting solution, is concentrated into 1mL
Extracting solution is equivalent to 0.3g crude drugs, and filtering, filtrate stands overnight, and takes supernatant as pepper alkali extracting solution.Humulus grass is concentrated
Liquid supernatant presses 4 with pepper alkali extracting solution:1 mixing, then (3600rpm, 20min) is centrifuged, supernatant is taken, adds above-mentioned 4/5
Deal pepper aqua aromatica is diluted with water 10 times, and Sucralose (a concentration of 0.3%) in humulus green pepper oral solution, humulus green pepper mouthful is added
Liquid pH value 5.0-6.0 is taken, relative density is not less than 1.02, dispenses, every 10mL.Character：In light brown transparency liquid, nothing is clarified
Impurity.
Humulus green pepper oral solution made from the above method (totally 3 batches, sample 1~3) is taken, it is as follows according to the detection method of the present invention：
1) indentification by TLC of humulus grass：It takes this product 20mL in separatory funnel, is extracted with 10mL ethyl acetate, wait being layered
Lower layer's dark brown liquid is released afterwards, pours out upper layer of extraction liquid.Lower layer's dark brown liquid is poured into separatory funnel, then with 10mL second
Acetoacetic ester extraction is primary, merges extract liquor twice, 60 DEG C of water bath methods, and residue adds appropriate ethyl acetate to dissolve, and filtering takes filtrate
It is settled to 2mL, as test solution；It takes humulus grass control medicinal material coarse powder 1.5g, is added ethyl acetate 30mL, ultrasound (50 DEG C,
100KHz) 40min, filtering take 60 DEG C of water bath methods of filtrate, residue that appropriate ethyl acetate is added to dissolve, and filtering takes filtrate to be settled to
1mL, as a contrast medicinal material solution；Precision weighs cyanidenon reference substance (National Institute for Food and Drugs Control, lot number：
111520-201605) 0.00101g adds appropriate ethyl acetate to dissolve, and is settled to 10mL, as a contrast product solution；It takes and lacks
The sample 20mL of humulus grass ingredient is handled by test solution method, as negative sample solution.According to thin-layered chromatography (middle traditional Chinese medicines
2015 editions four general rules 0502 of allusion quotation) experiment, draw 15 μ L of test solution, 5 μ L of reference substance solution, control medicinal material solution and feminine gender
Each 10 μ L of sample solution, put on the silica gel g thin-layer plate in same using sodium carboxymethylcellulose as binder, respectively with toluene-second
Acetoacetic ester-methyl alcohol-formic acid (16:4:1.2:0.8) it is solvent, is unfolded, take out, dry, sprays with 5% alchlor ethanol,
It is clear to be heated to spot development, sets and is inspected under ultraviolet lamp (365nm), test sample is with control medicinal material, reference substance in chromatography identical bits
It sets, shows the fluorescence spot of same color, as shown in Figure 1.
2) indentification by TLC of white pepper：This product 10mL is taken, 70 DEG C of water bath methods, it is molten that residue adds absolute ethyl alcohol 1mL to make
Solution, filtering, takes filtrate to be settled to 1mL, as test solution；Take pepper control medicinal material (National Institute for Food and Drugs Control,
Lot number：Powder 1.5g in 121441-200401) adds 75% ethyl alcohol 9mL, refluxing extraction 30min, filters off the dregs of a decoction, filtrate is taken to be settled to
5mL takes above-mentioned solution 2mL, 60 DEG C of water bath methods, residue that absolute ethyl alcohol 1mL is added to make dissolving, and filtering takes filtrate to be settled to 1mL,
Medicinal material solution as a contrast；Take pipering reference substance (National Institute for Food and Drugs Control, lot number：110775-201405)
0.00423g is set in brown measuring bottle, and absolute ethyl alcohol is added to dissolve, and is settled to 1mL, as a contrast product solution；Take lack white pepper at
The sample 10mL divided is handled by test solution method, as negative sample solution.According to thin-layered chromatography (Chinese Pharmacopoeia 2015
Four general rules 0502 of version) experiment, draw 10 μ L of test solution, 5 μ L of reference substance solution, 10 μ L of control medicinal material solution, negative sample
10 μ L of product solution, put respectively on the silica gel g thin-layer plate in same using sodium carboxymethylcellulose as binder, with toluene-acetic acid second
Ester-acetone (7:2:1) it is solvent, is unfolded, take out, dry, sprays with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development
It is clear, it sets and is inspected under ultraviolet lamp (365nm), test sample in chromatography same position, shows identical face with control medicinal material, reference substance
The fluorescence spot of color, as shown in Figure 2.
3) the general flavone Assaying of Rutin of humulus grass, according to ultraviolet spectrophotometry (2015 editions four general rules of Chinese Pharmacopoeia
0401) it tests：
(1) prepared by reference substance solution：Precision is weighed through dry rutin standard items (the Chinese food drug assay to permanent quality
Research institute, lot number：100080-200707) 0.01025g is placed in 50mL volumetric flasks, is added 80% ethyl alcohol to scale, is shaken up conduct
Reference substance storing solution.Precision measures storing solution 5.0mL, is placed in 10mL volumetric flasks and 5% sodium nitrite solution 0.5mL is added and shake
It is even, 6min is placed, 10% aluminum trichloride solution 0.5mL is added to shake up, places 6min, then 4% sodium hydroxide solution 4.0mL is added to shake
It is even, be then settled to graduation mark with 80% ethyl alcohol, shake up, place 15min to get；
(2) prepared by test solution：Precision measures humulus green pepper oral solution 2.0mL, 80 DEG C of water bath methods, and residue adds appropriate 80%
Ethyl alcohol makes dissolving, filtering, takes filtrate, then be settled to 5mL with 80% ethyl alcohol, and precision measures 1.0mL, is placed in 10mL volumetric flasks, and
5% sodium nitrite solution 0.5mL is added to shake up, places 6min, 10% liquor alumini chloridi 0.5mL is added to shake up, places 6min, then add
4% sodium hydroxide solution 4.0mL shakes up, and is then settled to graduation mark with 80% ethyl alcohol, shakes up, place 15min to get；
(3) wavelength selection:It prepares not plus the solution of color developing agent is only added as placebo solution in reference substance.Take rutin pair
Absorption curve is scanned in 400-600nm wave-length coverages according to product solution, determines that Detection wavelength is 506nm, as shown in Figure 3.
(4) measuring method：Test solution is taken, absorbance is measured at 506nm, this product is per 1mL containing general flavone with rutin
(C27H30O16) gauge must not be less than 0.80mg.
The determination of total flavonoids of 3 sample humulus green pepper oral solutions the results are shown in Table 1, as a result meet regulation.
1 sample size measurement result (n=3) of table
The cosmosiin assay of humulus grass, according to high performance liquid chromatography (2015 editions four general rules 0512 of Chinese Pharmacopoeia)
Experiment：
(1) chromatographic condition and system suitability：250 × 4.6mm of chromatographic column Agilent TC-C18；Mobile phase is
- 0.2% phosphoric acid (22 of second eyeball:78)；Column temperature is 25 DEG C；Detection wavelength is 350nm；Flow velocity is 1.0mL/min；10 μ L of sample size.
Number of theoretical plate is calculated by cosmosiin should be not less than 5000；
(2) prepared by test solution：Precision draws humulus green pepper oral solution 10.0mL in 60mL separatory funnels, and 20mL second is added
Acetoacetic ester, shaking out release lower layer's dark brown liquid after standing, pour out upper layer acetic acid ethyl acetate extract.By lower layer's dark-brown
Liquid refunds separatory funnel, is extracted again once with 20mL ethyl acetate.Extract liquor, 60 DEG C of water bath methods, residue add twice for merging
Methanol makes dissolving, filtering, take filtrate be settled to 10mL to get；
(3) prepared by reference substance solution：Precision weigh cosmosiin reference substance (Chengdu Inst. of Biology, Chinese Academy of Sciences,
Lot number：MUST-1310300) 0.00104g, add proper amount of methanol dissolve after be settled to 10mL to get；
(4) measuring method：It is accurate respectively to draw reference substance solution and each 10 μ L of test solution, liquid chromatograph is injected, is surveyed
Fixed, this product is per 1mL (C containing cosmosiin21H20O10) gauge, 0.10mg must not be less than；
The cosmosiin assay of 3 sample humulus green pepper oral solutions the results are shown in Table 2, as a result meet regulation.
2 sample size measurement result (n=3) of table
Although the above-mentioned specific implementation mode to the present invention is described, not to the limit of the scope of the present invention
System, based on the technical solutions of the present invention, those skilled in the art need not make the creative labor can make it is each
Kind modification or deformation are still within protection scope of the present invention.
Claims (10)
1. a kind of preparation method of humulus green pepper oral solution, it is characterised in that include the following steps：In parts by weight, humulus grass is taken
10-20 parts, 0.6-1.5 parts of white pepper；Humulus grass adds 8-12 times to measure 75% ethyl alcohol, and flow back 1.2-2h, and more than extraction once, merging carries
Liquid is taken, then concentration is the humulus grass concentrate that 1mL concentrates are equivalent to 1g crude drugs, stand for standby use；White pepper beats powder, adds after sieving
4-6 times is measured water, is evaporated under reduced pressure, and when the moisture rate of outflow is less than 2mL/min, is stopped distillation, is obtained pepper aqua aromatica, 1mL
Pepper aqua aromatica is equivalent to 0.5g crude drugs；Remaining Aqueous extracts are discarded, 75% ethyl alcohol, refluxing extraction one are measured in 5-7 times of addition
More than secondary, each 1.2-2h merges extracting solution, is concentrated into 1mL extracting solutions and is equivalent to 0.3g crude drugs, filters, and filtrate stands, and takes
Supernatant is as pepper alkali extracting solution；The supernatant of humulus grass concentrate and pepper alkali extracting solution are pressed 4:0.8-1.2 is mixed, then from
The heart takes supernatant, adds the amount of above-mentioned pepper aqua aromatica 70%-90%；It is finally diluted with water 8-12 times, trichlorine sugarcane is added
Sugar finally obtains humulus green pepper oral solution.
2. the preparation method of humulus green pepper oral solution according to claim 1, it is characterised in that：The humulus grass is at 80-90 DEG C
It flows back.
3. the preparation method of humulus green pepper oral solution according to claim 1, it is characterised in that：It is described by humulus grass concentrate
After supernatant is mixed with pepper alkali extracting solution, centrifugation 15-25min is carried out at rotating speed 3400-3800rpm.
4. the preparation method of humulus green pepper oral solution according to claim 1, it is characterised in that：The pH of the humulus green pepper oral solution
Value 5.0-6.0, relative density are not less than 1.02.
5. a kind of humulus green pepper oral solution, it is characterised in that：Preparation method is made described in claim 1-4 any claims.
6. humulus green pepper oral solution according to claim 5, it is characterised in that：The humulus green pepper oral solution contains general flavone per 1mL
Rutin gauge is not less than 0.80mg；The humulus green pepper oral solution is not less than 0.10mg per 1mL gauge containing cosmosiin.
7. the detection method of humulus green pepper oral solution described in claim 5 or 6, it is characterised in that including：Using thin-layer chromatography to it
In humulus grass and white pepper Qualitive test；The active constituent content of humulus grass is measured using HPLC methods.
8. the detection method of humulus green pepper oral solution according to claim 7, it is characterised in that：The qualitative mirror of the humulus grass
Not, include the following steps：
It takes humulus green pepper oral solution 20mL in separatory funnel, is extracted with 8-12mL ethyl acetate, lower layer's dark-brown is released after being layered
Liquid pours out upper layer of extraction liquid；Lower layer's dark brown liquid is poured into separatory funnel, then one is extracted with 8-12mL ethyl acetate
It is secondary, merge extract liquor twice, 50-70 DEG C of water bath method, residue adds appropriate ethyl acetate to dissolve, and filtering takes filtrate to be settled to
2mL, as test solution；Humulus grass control medicinal material coarse powder 1.4-1.6g is taken, ethyl acetate 28-32mL is added, is carried out at 50 DEG C
Ultrasonic 40min, filtering take 50-70 DEG C of water bath method of filtrate, residue that appropriate ethyl acetate is added to dissolve, and filtering takes filtrate to be settled to
1mL, as a contrast medicinal material solution；Precision weighs cyanidenon reference substance 0.00101g, adds appropriate ethyl acetate to dissolve, and constant volume
To 10mL, product solution as a contrast；It is tested according to thin-layered chromatography, draws test solution 15 μ L, 5 μ L of reference substance solution, right
According to 5 μ L of medicinal material solution, puts respectively on the silica gel g thin-layer plate in same using sodium carboxymethylcellulose as binder, be with volume ratio
16:4:1.2:0.8 toluene-ethyl acetate-methyl alcohol-formic acid is solvent, is unfolded, and takes out, dries, and is sprayed with 5% alchlor
Ethanol, it is clear to be heated to spot development, sets and is inspected under ultraviolet lamp, and test sample is with control medicinal material, reference substance in chromatography identical bits
It sets, shows the fluorescence spot of same color.
9. the detection method of humulus green pepper oral solution according to claim 7, it is characterised in that：The qualitative mirror of the white pepper
Not, include the following steps：
It takes humulus green pepper oral solution 10mL, 60-80 DEG C of water bath method, residue that absolute ethyl alcohol 1mL is added to make dissolving, filters, take filtrate constant volume
To 1mL, as test solution；Powder 1.4-1.6g in pepper control medicinal material is taken, 75% ethyl alcohol 8-10mL, refluxing extraction 25- are added
35min filters off the dregs of a decoction, takes filtrate to be settled to 5mL, above-mentioned solution 2mL, 50-70 DEG C of water bath method, residue is taken to add absolute ethyl alcohol
1mL makes dissolving, and filtering takes filtrate to be settled to 1mL, as a contrast medicinal material solution；Pipering reference substance 0.00423g is taken, brown is set
In measuring bottle, absolute ethyl alcohol is added to dissolve, is settled to 1mL, as a contrast product solution；It is tested according to thin-layered chromatography, draws test sample
10 μ L of solution, 5 μ L of reference substance solution, 10 μ L of control medicinal material solution, put respectively in same using sodium carboxymethylcellulose as binder
Silica gel g thin-layer plate on, with volume ratio be 7:2:1 toluene-ethyl acetate-acetone is solvent, is unfolded, and takes out, dries, and is sprayed
With 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development, sets and is inspected under ultraviolet lamp, and test sample and control medicinal material compare
Product show the fluorescence spot of same color in chromatography same position.
10. the detection method of humulus green pepper oral solution according to claim 7, it is characterised in that：The humulus grass it is effective at
Divide assay, is tested according to ultraviolet spectrophotometry and high performance liquid chromatography, steps are as follows：
General flavone rutin measures：(1) prepared by reference substance solution：Precision is weighed through the dry rutin standard items to permanent quality
0.01025g is placed in 50mL volumetric flasks, adds 80% ethyl alcohol to scale, shakes up product storing solution as a contrast.Precision measures deposit
Liquid 5.0mL, is placed in 10mL volumetric flasks and 5% sodium nitrite solution 0.5mL is added and shake up, and places 6min, adds 10% tri-chlorination
Aluminum solutions 0.5mL shakes up, and places 6min, then 4% sodium hydroxide solution 4.0mL is added to shake up, is then settled to quarter with 80% ethyl alcohol
Spend line, shake up, place 15min to get；(2) prepared by test solution：Precision measures humulus green pepper oral solution 2.0mL, and 80 DEG C of water-baths are steamed
Dry, residue adds appropriate 80% ethyl alcohol to make dissolving, filtering, takes filtrate, then be settled to 5mL with 80% ethyl alcohol, and precision measures 1.0mL,
10mL volumetric flasks are placed in, and 5% sodium nitrite solution 0.5mL is added and shakes up, 6min is placed, adds 10% liquor alumini chloridi 0.5mL
It shakes up, places 6min, then 4% sodium hydroxide solution 4.0mL is added to shake up, be then settled to graduation mark with 80% ethyl alcohol, shake up, put
Set 15min to get；(3) wavelength selection：It prepares not plus the solution of color developing agent is only added as placebo solution in reference substance.It takes
Control substance of Rutin solution scans absorption curve in 400-600nm wave-length coverages, determines that Detection wavelength is 506nm；(4) it measures
Method：Test solution is taken, absorbance is measured at 506nm, humulus green pepper oral solution is per 1mL rutins containing general flavone (C27H30O16) gauge,
0.80mg must not be less than；
Cosmosiin measures：(1) chromatographic condition and system suitability：Chromatographic column Agilent TC-C18 250 ×
4.6mm；Mobile phase is -0.2% phosphoric acid (22 of second eyeball:78)；Column temperature is 25 DEG C；Detection wavelength is 350nm；Flow velocity is 1.0mL/
min；10 μ L of sample size.Number of theoretical plate is calculated by cosmosiin should be not less than 5000；(2) prepared by test solution：Precision is inhaled
It takes humulus green pepper oral solution 10.0mL in 60mL separatory funnels, 20mL ethyl acetate is added, lower layer depth is released in shaking out after standing
Brown liquid pours out upper layer acetic acid ethyl acetate extract.Lower layer's dark brown liquid is refunded into separatory funnel, again with 20mL ethyl acetate
Extraction is primary.Merging extract liquor, 60 DEG C of water bath methods, residue twice adds methanol to make dissolving, filters, filtrate is taken to be settled to 10mL,
To obtain the final product；(3) prepared by reference substance solution：Precision weighs cosmosiin reference substance 0.00104g, constant volume after adding proper amount of methanol to dissolve
To 10mL to get；(4) measuring method：It is accurate respectively to draw reference substance solution and each 10 μ L of test solution, inject liquid chromatogram
Instrument measures, and humulus green pepper oral solution is per 1mL (C containing cosmosiin21H20O10) gauge, 0.10mg must not be less than.
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| CN104491011A (en) * | 2014-12-18 | 2015-04-08 | 南方医科大学中西医结合医院 | Traditional Chinese medicine composition for treating infantile diarrhea caused by rotavirus and preparation method thereof |
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| CN104491011A (en) * | 2014-12-18 | 2015-04-08 | 南方医科大学中西医结合医院 | Traditional Chinese medicine composition for treating infantile diarrhea caused by rotavirus and preparation method thereof |
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Application publication date: 20180817 |