CN1954871B - Discrimination method for Yanhouqing preparation for treating throat disease - Google Patents

Abstract

A Chinese medicine in the form of dripping pill, dispersing tablet, or micropill for treating thirst, painful throat, hoar seness, cough, etc is prepared from 8 Chinese- medicinal materials including Japanese peristrophe herb, coral ardisia root, thinleaf adina root, etc. Its preparing process and quality control method are also disclosed.

Description

The discrimination method of the clear preparation of throat of treatment pharyngolaryngitis

Technical field

The present invention is a kind of discrimination method for the treatment of the clear preparation of throat of pharyngolaryngitis, belongs to technical field of Chinese medicine.

Technical background

Throat is the first line of defence of human respiratory.In recent years; because the harm of atmospheric pollution and cigarette poison; the influence of occupational factor; drink; factors such as the stimulation of dust; pharyngolaryngitis is in rising trend; especially in dry winter; air pollution; dust particle rolls up; very easily bring out sphagitis, cause people's hoarseness; aphonia; dry throat is puckery; pharynx is itched; pharyngalgia; expectoration is not well; the pharynx foreign body sensation is often with the tcs response of " uttering a sound or a word; noise made in coughing or vomiting "; the sensation that these be can't get rid of; as untimely diagnosis and treatment; prevention, easily repeatedly, very obstinate; troublesome throughout one's life probably; even can cause the generation of other disease, happiness in all one's life shortcoming therefore and to some extent makes us suffering untold misery.Throat health is becoming the topic of new millennium medical expert and consumers in general's common concern.Though because what big defect pharyngo-laryngitis chronica is not, become the difficult problem of modern medicine really, generally be difficult to return to one's perfect health.For this reason, expert call: vigilant chronic pharyngitis harm, seek the throat medication of special efficacy and the healthy fashion that natural throat health products are just becoming the new millennium people.So clinical treatment must promptly and accurately be selected appropriate drug preparation and methods of treatment.The clear sheet of throat is made up of peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, menthol eight flavor medicines.Monarch drug in a prescription in peristrophe wind and heat dispersing, the removing toxicity for detumescence side of being, bicolor, japanese avens root, root of Herba Gonostegiae hirtae heat-clearing and fire-reducing, subdhing swelling and detoxicating, blood circulation and promoting silt, expelling phlegm and arresting coughing are ministerial drug altogether; Tinosporae, balloonflower root are opened a surname's lung qi, the apocenosis of eliminating the phlegm, and are adjutant in the side, peppermint, menthol wind-dispelling heat-dissipating, and the clear sharp head is for making medicine.This side's expelling wind to resolve the exterior, clearing heat and detoxicating, clearing throat are used for pharyngalgia, dry throat, hoarseness, or symptoms such as fever and aversion to wind, cough are arranged.The clear sheet of throat is determined curative effect clinically, welcome by extensive patients.But also found some problems in long-term clinical practice, fallen behind such as formulation that dose is big, onset is slow, and product quality is not ideal enough, and the formulation kind is abundant inadequately, is suitable for crowd's narrow range, takes inconvenience etc.In view of such circumstances, optimize technology, improve formulation, the control method that improves the quality becomes bastard feverfew throat clearing particle urgent problem.

Summary of the invention

The objective of the invention is to: a kind of clear preparation of throat for the treatment of pharyngolaryngitis and preparation method thereof and method of quality control are provided; The present invention is directed to prior art, formulations such as the dripping pill that provides, dispersing tablet have not only solved granule and taken inconvenience and the relatively poor problem of mouthfeel, and disintegrative are good, the bioavilability height; Preparation method provided by the present invention can effectively prepare needs preparation, guarantee that the preparation production technique obtain is scientific and reasonable; The method of quality control that is provided, the means, technical method of the index that detects, detection etc. can be provided to relevant production, testing agency, so that better control the quality of said preparation, guarantee the security of medication, can better instruct production, make controlling of production process rationally strict more, make consumer's energy full appreciation product quality.

The present invention constitutes like this: calculate according to weight, it mainly is by peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, the preparation that menthol 2.5g is made, comprise: injection comprises: parenteral solution, the powder pin, freeze-dried powder, gel, tablet, dispersing tablet, capsule, soft capsule, microcapsules, granule, pill, micropill, powder, pill, sustained release preparation, controlled release preparation, gel, oral liquid, soft extract, all acceptable formulations on the pharmacy such as extract and film.Say accurately: described preparation is dripping pill, soft capsule, dispersing tablet.

The method for making of the clear preparation of throat of described treatment pharyngolaryngitis: get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours, filter, merge with above-mentioned aqueous solution, when being evaporated to 60 ℃ of relative densities 1.10～1.12 clear cream, adding ethanol makes and contains alcohol amount and reach 60%, left standstill 24 hours, filtrate is concentrated, dry at decompression recycling ethanol below 80 ℃, adds menthol, spray into above-mentioned volatile oil, make different preparations.Pill in the described preparation prepares like this: get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours, filter, merge with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mixes and is ground into fine powder, press extract powder: matrix=1: 1～3 add matrix PEG4000, spray into above-mentioned volatile oil behind the heating and melting, mixing is transferred to the dripping pill machine, drip 70 ℃～90 ℃ of system temperature, cooling medium is 5 ℃～30 ℃ a dimethyl silicon oil, and dripping speed is 20～40d/min, drips distance at 2～12cm, collect dripping pill and remove the dimethyl silicon oil on surface, promptly.Pill in the described preparation prepares like this: get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours, filter, merge with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mixes and is ground into fine powder, press extract powder: matrix=1: 1.5 adding matrix PEG4000, spray into above-mentioned volatile oil behind the heating and melting, mixing is transferred to the dripping pill machine, drip 85 ℃ of system temperature, cooling medium is 10 ℃～20 ℃ a dimethyl silicon oil, and dripping speed is 30d/min, drips distance at 4～8cm, collect dripping pill and remove the dimethyl silicon oil on surface, promptly.Soft capsule in the described preparation prepares like this: get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours, filtering, merge with above-mentioned aqueous solution, is 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, adding ethanol makes and contains alcohol amount and reach 60%, left standstill 24 hours, and filtered, filtrate concentrates at decompression recycling ethanol below 80 ℃, dry, add menthol, mix and be ground into fine powder, spray into above-mentioned volatile oil again, mixing, airtight 2 hours, press extract powder again: matrix=1: 1.5 adds soybean oil, the mixed-matrix of soybean lecithin, heating and melting, mixing gets soft capsule content; The preparation of glue: with gelatin: glycerine: water=1: 0.7: 1.0, getting gelatin adds an amount of distilled water and makes its imbibition, in addition the water of glycerine and remainder is put and be heated to 70～80 ℃ in the glue pot, mix, add the gelatin that expands and stir, make it to dissolve into uniform glue, in 60 ℃ of insulations 2～3 hours, leave standstill, remove the come-up foam, filter standby with cloth bag.Soft capsule content and glue are pressed into soft capsule in encapsulating machine, put in the drum dryer and finalize the design, whole ball, drying, promptly.Disket in the described preparation prepares like this: get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, and the aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours filters, and merges with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mix and be ground into fine powder, add 10% microcrystalline cellulose, 7% crospolyvinylpyrrolidone, mixing adds ethanol, the system softwood, granulate drying, the whole grain of 40 mesh sieves, compressing tablet behind the mixing, other gets menthol, adds ethanol and makes dissolving in right amount, sprays into, spray into above-mentioned volatile oil again, mixing, airtight 2 hours, promptly.

The quality control method of the clear preparation of throat of described treatment pharyngolaryngitis, its discrimination method comprise following all or part of content:

A. the thin-layered chromatography of japanese avens root is differentiated in the preparation

It is an amount of to get this product powder, adds methanol extraction, and extract evaporate to dryness, residue add water makes dissolving, adds the ethyl acetate jolting and extracts, and acetic acid ethyl fluid evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Other the Chinese waxmyrtle root control medicinal material of fetching water is an amount of, adds ethyl acetate extraction, and extract evaporate to dryness, residue add methyl alcohol makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution respectively, puts on same silica gel g thin-layer plate, and with methenyl choloride-acetone=4～6: 0.3～0.7 is developping agent, launch, take out, dry, put under the uviol lamp and inspect, or spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

B. one or both thin-layered chromatography discriminating in bicolor medicinal material, the Bergenin in the preparation

It is an amount of to get this product powder, adds the methyl alcohol ultrasonic Extraction, and extract concentrates, as need testing solution; Get the bicolor control medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Bergenin reference substance, adds methyl alcohol and makes reference substance solution; According to the thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution respectively, puts on same silica gel g thin-layer plate, with methenyl choloride-ethyl acetate-ethanol=4～6: be developping agent at 3～5: 2～4, launches, and takes out, dry, spray is to contain the developer that mixes of ferric trichloride and the potassium ferricyanide; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

C. the thin-layered chromatography of balloonflower root is differentiated in the preparation

It is an amount of to get this product powder, adds the mixed solution of ethanol solution of sulfuric acid and water, and reflux is put cold, add the methenyl choloride jolting and extract, methenyl choloride liquid adds water washing, discards water liquid, methenyl choloride liquid anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Other gets the balloonflower root control medicinal material, shines medicinal material solution in pairs with legal system; According to thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution, puts respectively on same silica gel g thin-layer plate, and with methenyl choloride-ether=4～5: 5～6 be developping agent, launches, and takes out, and dries, and spray is with 10% ethanol solution of sulfuric acid, be heated to spot develop the color clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color.

D. one or both thin-layered chromatography discriminating in peppermint medicinal material, the menthol in the preparation

It is an amount of to get this product powder, adds the sherwood oil jolting and extracts, and extract is as need testing solution; Other gets the peppermint medicinal material, shines medicinal material solution in pairs with legal system; Get the menthol reference substance, add sherwood oil and make reference substance solution; Test according to thin-layered chromatography, it is an amount of to draw above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-methenyl choloride=8～10: be developping agent at 1.5～2.5: 0.8～1.2, launch, take out, dry, spray is with the vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.

Say that accurately discrimination method comprises following all or part of content:

A. the thin-layered chromatography of japanese avens root is differentiated in the preparation

It is an amount of to get this product powder, adds the methyl alcohol ultrasonic Extraction, and extract evaporate to dryness, residue add water makes dissolving, adds the ethyl acetate jolting and extracts, and acetic acid ethyl fluid evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Other the Chinese waxmyrtle root control medicinal material of fetching water is an amount of, adds the ethyl acetate ultrasonic Extraction, and extract evaporate to dryness, residue add methyl alcohol makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution respectively, put on same silica gel g thin-layer plate, be developping agent with methenyl choloride-acetone=5: 0.5, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to spot develop the color clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

B. one or both thin-layered chromatography discriminating in bicolor medicinal material, the Bergenin in the preparation

It is an amount of to get this product powder, adds the methyl alcohol ultrasonic Extraction, and extract concentrates, as need testing solution; Get the bicolor control medicinal material, shine medicinal material solution in pairs with legal system; Other gets the Bergenin reference substance, adds methyl alcohol and makes reference substance solution; According to thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, and be developping agent with methenyl choloride-ethyl acetate-ethanol=5: 4: 3, launch, take out, dry, spray to contain the developer that mixes of ferric trichloride and the potassium ferricyanide; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;

C. the thin-layered chromatography of balloonflower root is differentiated in the preparation

It is an amount of to get this product powder, add the mixed solution of 7% ethanol solution of sulfuric acid-water=1: 3, reflux is put cold, adding the methenyl choloride jolting extracts, methenyl choloride liquid adds water washing, discards water liquid, methenyl choloride liquid anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Other gets the balloonflower root control medicinal material, shines medicinal material solution in pairs with legal system; According to thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with methenyl choloride-ether=4: 6, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to spot develop the color clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color.

D. one or both thin-layered chromatography discriminating in peppermint medicinal material, the menthol in the preparation

It is an amount of to get this product powder, adds the sherwood oil jolting and extracts, and extract is as need testing solution; Other gets the peppermint medicinal material, shines medicinal material solution in pairs with legal system; Get the menthol reference substance, add sherwood oil and make reference substance solution; According to the thin-layered chromatography test, it is an amount of to draw above-mentioned three kinds of solution, puts respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-methenyl choloride=9: 2: 1 was developping agent, launched, and took out, dry, spray is with the vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.

In the quality control method of the clear preparation of throat of described treatment pharyngolaryngitis, its content assaying method comprises following all or part of content:

A. the high performance liquid chromatography assay of Bergenin in the preparation

It is an amount of to get this product powder, accurate claims surely, puts in the tool plug conical flask, and the accurate methyl alcohol that adds claims decide weight, and sonicated is put coldly, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filtration is got subsequent filtrate, as need testing solution; Accurate title Bergenin reference substance is an amount of, adds methyl alcohol and makes reference substance solution; According to high performance liquid chromatography test, be filling agent with 18 silylation bonded silica gels, with methanol-water=15～21: 85～79 is moving phase; The detection wavelength is 270～280nm; Accurate respectively absorption reference substance solution and need testing solution are an amount of, inject liquid chromatograph, measure; Calculate with one point external standard method or calibration curve method, this product contains the purple bergenia herb element with dosage and must not be less than 1.5mg every day;

B. the vapor-phase chromatography assay of menthol in the preparation

It is an amount of to get this product powder, and accurate the title decides, and puts in the tool plug conical flask, the accurate absolute ethyl alcohol that adds, sonicated is placed to room temperature, claim again to decide weight, supply the weight that subtracts mistake, shake up with absolute ethyl alcohol, centrifugal, it is an amount of that precision is measured supernatant, and the accurate again inner mark solution that adds is an amount of, adds absolute ethyl alcohol and is settled to debita spissitudo, shake up, as need testing solution; It is an amount of that naphthalene decided in accurate title, adds anhydrous alcohol solution and make inner mark solution; Other gets the menthol reference substance, and accurate the title decides, and the accurate inner mark solution that adds is an amount of, adds absolute ethyl alcohol and makes reference substance solution; According to the vapor-phase chromatography test, be stationary phase with polyglycol-20M, coating concentration is 8～12%; Column temperature is 110～150 ℃; Draw an amount of inject gas chromatograph of reference substance solution, calculate than positive divisor; The absorption need testing solution is an amount of, and inject gas chromatograph is measured; This product contains menthol with dosage and must not be less than 15mg every day.

Say that accurately content assaying method comprises following all or part of content:

A. the high performance liquid chromatography assay of Bergenin in the preparation

Get this product, porphyrize is got about 0.2g, accurate claims surely, puts in the tool plug conical flask, and the accurate methyl alcohol 25ml that adds claims decide weight, and sonicated 1 hour is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filtration is got subsequent filtrate, as need testing solution; Accurate title Bergenin reference substance is an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.05mg, in contrast product solution; According to high performance liquid chromatography test, be filling agent with 18 silylation bonded silica gels, be moving phase with methanol-water=18: 82; The detection wavelength is 275nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; Calculate with one point external standard method, this product contains the purple bergenia herb element with dosage and must not be less than 3.0mg every day;

B. the vapor-phase chromatography assay of menthol in the preparation

It is an amount of to get this product, and porphyrize is got 1g, and accurate the title decides, put in the tool plug conical flask, the accurate absolute ethyl alcohol 20ml that adds claims to decide weight, ice-bath ultrasonic was handled 30 minutes, was placed to room temperature, claimed to decide weight again, supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, centrifugal, precision is measured supernatant 5ml, puts in the 10ml measuring bottle, the accurate inner mark solution 2ml that adds, add absolute ethyl alcohol to scale, shake up, as need testing solution; Get naphthalene 150mg, the accurate title, decide, and puts in the 100ml measuring bottle, adds anhydrous alcohol solution and be diluted to scale, as inner mark solution; Other gets menthol reference substance 10mg, and accurate the title decides, and puts in the 10ml measuring bottle, and the accurate inner mark solution 2ml that adds adds absolute ethyl alcohol to scale, shakes up, in contrast product solution; According to the vapor-phase chromatography test, be stationary phase with polyglycol-20M, coating concentration is 10%; With Chromosorb W (AW-DMCS) (60-80 order) is carrier, and column temperature is 140 ℃; Draw reference substance solution 1 μ l, inject gas chromatograph calculates than positive divisor; Draw need testing solution 1 μ l, inject gas chromatograph is measured; This product contains menthol with dosage and must not be less than 20mg every day.

Compare with technology with existing formulation, the invention solves formulation and be suitable for crowd's narrow range, take inconvenience, the unfavorable problem of medicine stability.Its preparation formulation taking convenience, bioavilability height, good stability, easy to carry, good mouthfeel, absorption be fast, it is wide to be suitable for the crowd; The preparation method who is provided can effectively prepare needs preparation, guarantee that the preparation variety effect obtain is remarkable, production technology is scientific and reasonable, has overcome the problem that existing product exists; The method of quality control that is provided can more fully be controlled the quality of said preparation; Reached purpose of the present invention.

The applicant finds in development process, is the assurance product quality, the screening of auxiliary material, process conditions, and the screening of all conditions of method of quality control is most important.The applicant has carried out a series of experiments, with method and parameter of the supplementary product kind of the preparation technology that selects pharmaceutical preparation provided by the invention, use and consumption and ratio, quality control etc.; To guarantee science, rationality, the feasibility of invention.

Experimental example 1: Study on extraction

1. volatile oil extraction conditions screening

(1) factor selective volatilization oil extraction effect is subjected to the influence of factors such as amount of water, extraction time.Therefore choose amount of water and extraction time as factor, the varying level of high spot reviews factor is to the influence of volatile oil extraction effect.

(2) to determine to select oil mass be evaluation index to index, and assay method is as follows:

Take by weighing respectively three parts of peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g respectively, volatile oil is collected in boiling simultaneously, receives oil mass every 1 hour record.

(3) test: test arrangement and the results are shown in following table.

Volatile oil extracts investigates table as a result

As seen from the above table, amount of water is put forward oil mass for 10 times and 12 times and is more or less the same, and extracts after 6 hours volatile oil and carries substantially to the greatest extent, from saving time and the angle of the energy, selects to add 10 times of water and extracts 6 hours.And carry out demonstration test according to these process conditions.

2. the screening of water boiling and extraction condition

(1) factor is selected: the decocting for Chinese herbal medicine extraction effect is subjected to the influence of factors such as amount of water, extraction time, extraction time.Though prior art is investigated extraction time, extraction time, amount of water is not done any research as key factor, and the varying level of the applicant's high spot reviews factor is to decocting the influence of extraction effect.Take all factors into consideration the selection factor level in conjunction with aspects such as production cost, the energy.

(2) index is determined:

1. select the medicinal extract recovery rate as evaluation index.Medicinal extract is the material base of solid pharmaceutical preparation performance curative effect, and its yield height directly influences preparation process, is reasonable, effective control device so be chosen as the extraction index.

2. Bergenin content: extract yield height can not reflect fully that effective ingredient extracts situation, so select in the prescription the contained principal ingredient Bergenin of bicolor content as the decoction screening index simultaneously; With reference to relevant document, adopt reversed phase high efficiency liquid phase method to measure Bergenin content.

(3) decoct test: take by weighing respectively three parts of peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g respectively, add 10 times of water respectively and extract volatile oil, aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours, filter, filtrate and above-mentioned aqueous solution merge, being evaporated to relative density is the clear cream of 1.10～1.12 (60 ℃), adds ethanol and makes and contain alcohol amount and reach 60%, leaves standstill 24 hours, filter, filtrate decompression concentrates, drying, and it is heavy to claim to decide cream, calculate paste-forming rate, test findings sees the following form.

Amount of water is investigated table as a result

Tested number amount of water (doubly) medicinal extract recovery rate (%) Bergenin content (mg/g)

1 6 3.31 12.28

2 8 4.48 17.68

3 10 4.52 17.52

As seen from the above table: medicinal extract recovery rate and Bergenin content were higher when amount of water was 10 times and 8 times amount, and not significantly difference between the two, guaranteeing under the sufficient prerequisite of extracts active ingredients,, determining that extracting amount of water is 8 times of amounts in order to save cost and to shorten man-hour.

(4) confirmatory experiment carries out confirmatory experiment by above extraction process condition, the experimental result tabulation:

Decoct amount of water confirmatory experiment result

Scheme tested number medicinal extract recovery rate (%) Bergenin content (mg/g)

1 4.68 17.45

Decoct each 8 times of amounts 2 4.35 18.02 1 time

3 5.02 17.23

Remarks: investigation amount 500g

As seen decoct the optimal combination condition by the result of confirmatory experiment and extract that medicinal extract yield and the fluctuation of Bergenin content are little as a result, as seen this extraction process condition is reasonable, feasible and stable.

In sum, peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint add 10 times of water extraction volatile oil in the side, aqueous solution after distillation device is in addition collected, and dregs of a decoction boiling 1.5 hours filters, filtrate and distillate are evaporated to the clear cream that relative density is 1.10～1.12 (60 ℃) after merging, add ethanol and make and contain alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate decompression concentrates, drying.

Experimental example 2 reclaims ethanol research

For the ease of production operation control with avoid loss of effective components, take decompression recycling ethanol, and concentrating under reduced pressure, cream, standby.The condition of alcohol extracting soup concentrating under reduced pressure is: temperature is 60 ℃, and vacuum tightness is 0.08～0.1Mpa.

Experimental example 3 separates, concentrates and drying process research

Separating technology research: for the ease of production operation control, extract adopts 200 order filter clothes to filter.

Concentration technology research: concentrate and adopt the triple effect concentration tank to concentrate, extract is concentrated into the thick paste that relative density is 1.10～1.12 (60 ℃); Recovery ethanol concentrates, and the relative density of concentrate is that the concentrated condition of 1.25～1.30 (70 ℃) is: temperature is 84 ℃ of effects, two 80 ℃ of effects, 70 ℃ of triple effects, and vacuum tightness is an effect-0.025Mpa, two effect-0.045Mpa, triple effect-0.065Mpa.

Drying process research: the vacuum drying condition is: 60～70 ℃, and-0.08Mpa.

Experimental example 4: Study on Forming

4.1 Disket Study on Forming

Dispersing tablet meet water rapidly disintegration form the water dispersion tablet of uniform sticky suspension, it is poor to have solved former formulation disintegrative, stripping is shortcoming slowly, and the dispersing tablet that the applicant makes is fully disintegration in the 3min in 19 ℃～21 ℃ water, and suspension ability is good, bioavilability is high, be uniformly dispersed.

Check disintegration time limited: adopting changes the basket method, and the lift disintegration tester is got 6, observes the situation by screen cloth, and percent of pass height then disintegrative is good, more pleasant bulk absorption.

Group microcrystalline cellulose % crospolyvinylpyrrolidone % disintegration time s

1 10 3 110

2 10 5 95

3 10 7 72

4 15 3 105

5 15 5 136

6 15 7 89

7 18 3 213

8 18 5 114

9 18 7 101

The result shows that optimum process condition is for adding 10% microcrystalline cellulose, 7% crospolyvinylpyrrolidone, and mixing adds ethanol, the system softwood.

4.2 soft capsule Study on Forming

4.2.1 the adsorbing base rate is investigated

Medicinal powder: matrix suspension situation

1: 1.0 inhomogeneous suspension

1: 1.5 even suspension

1: 2.0 even suspension

4.2.2 auxiliary material is to the influence of composition:

Group Bergenin (mg/g)

Medicinal powder 0.51

Add after the matrix 0.49

The result shows that optimum process condition is for pressing medicinal powder: matrix=1: 1.5, Bergenin content did not have significant change after medicinal powder added matrix.

4.3 dripping pill Study on Forming

4.3.1 the initial option of matrix

Dripping pill requires preparation to bring into play curative effect rapidly, and medicine contains volatile ingredient in addition, so select fusing point low, the PEG4000 with fine dispersion power and big cohesion satisfies the requirement of clinical treatment and effective component character as matrix.Primary Study shows that when making matrix with PEG4000, the hardness of dripping pill, flowability are all good, the results are shown in following table.

Matrix screening experiment result

The hangover of matrix species hardness roundness

PEG4000 ++++++tail do not held in the palm

PEG6000 ++++ a little holds in the palm tail

Annotate: +++show fine; ++ show better; + show general

4.3.2 medicine and substrate composition

Mix the back situation with matrix and drip the system complexity with medicine, establish the proportioning of medicine and matrix.See the following form.The result shows that the ratio of medicine and Macrogol 4000 is at 1: 1.5 o'clock, and medicine and matrix amalgamation are better, and hardness, roundness suit, and is easy to the system of dripping.

The proportioning test result of medicine and matrix

The proportioning hardness roundness hangover of medicine and matrix

1: 1+-obviously

1: 1.2 ++-obviously

1: 1.5 ++++++do not trail

1: 2 +++++ do not trail

1: 3 ++++ do not trail

Annotate: +++show fine; ++ show better; + show general;-differential

4.3.3 cooling medium is selected

Earlier with medicine: the ratio of Macrogol 4000=1: 1.5, from methyl-silicone oil, liquid paraffin, soybean oil, peanut oil, rape seed oil, select suitable condensing agent.Decline rate, moulding situation with dripping pill are index, with the investigation result of each index by good to poorly using " +++" successively, " ++ ", "+", "-" expression the results are shown in following table.

The selection experimental result of cooling medium

Condensing agent decline rate moulding situation

Dimethyl silicon oil ++++++

Liquid paraffin-++

Soybean oil ++-

Rape seed oil +++

Annotate: +++show fine; ++ show better; + show general

As seen from the above table, cooling medium selection dimethyl silicon oil is good.

4.3.4 dripping the system temperature selects

The too high meeting of polyethylene glycols heating temperature color and luster occurs and deepens to be difficult for solidifying.Drip the system temperature as can be known when 80 ℃ of left and right sides by prerun test, it is very slow to drip speed, and moulding is bad.Select to drip the system temperature about 85 ℃, a speed is moderate, and moulding is good.By preferred medicine and substrate composition, medicinal extract and matrix to be put in the water-bath heat, fusion adds in the dripping pill machine, and the different temperatures insulation is spent so that pill shaped circle is whole, and the state of oozing is an index, the results are shown in following table.

Drip the selection experimental result of system temperature

Drip the system temperature (℃) roundness oozes state

70-can not drip

80+slower

85 +++easily ooze

90+too is fast

Annotate: +++show fine; ++ show better; + show general;-differential

4.3.5 coolant temperature is investigated

By preferred medicine and substrate composition, mix, be heated to 80-90 ℃, treat whole fusions after, get in right amount, splash into respectively in the dimethyl-silicon oil coolant of different temperatures, observe dripping pill moulding situation, the results are shown in following table.

Coolant temperature is selected

Coolant temperature drips apart from dripping the warm dripping pill moulding situation of speed material

85 ℃ of roundness of 10 ℃ of 6cm 30～40d/min are good, forming

85 ℃ of roundness of 20 ℃ of 6cm 30～40d/min are good, forming

85 ℃ of roundness of gradient cooling 6cm 30～40d/min are good, forming

Annotate: the gradient cooling means is: top is 10～20 ℃, and the bottom is 5～10 ℃.

Last table shows that under above-mentioned three kinds of chilling temperatures, the mouldability of this product is all good, is easy operation, is 10～20 ℃ so select coolant temperature.

4.3.6. resitting an exam, ball examines

By above optimum condition, select the dropper of different bores, drip and make ball, with the whole degree of pill shaped circle, hardness, hangover the results are shown in following table for index.

Ball is resit an exam and is examined experimental result

Heavy (mg) roundness hardness hangover of bore (inside/outside mm/mm) ball

3.0/4.0 40 ++++ tail do not held in the palm

4.0/5.0 60 ++++++tail do not held in the palm

5.0/6.0 70-+the holder tail

Annotate: +++show fine; ++ show better; + show general;-differential

The above results shows, the water dropper bore is that appearance index such as the dripping pill roundness of the water dropper of 4.0/5.0 (inside/outside mm/mm) system of dripping and hardness are better, so selection water dropper bore is 4.0/5.0 (inside/outside mm/mm).

4.3.7 dripping speed investigates

By above optimum condition, select the different speed of dripping, drip and make ball, with the whole degree of pill shaped circle, hardness, trailing is index, the results are shown in following table.

Drip speed and investigate experimental result

Drip the hangover of speed (d/min) roundness hardness

20 ++++ do not trail

30 ++++++do not trail

40-+++hangover

Annotate: +++show fine; ++ show better; + show general;-differential

So can determine that by last table dripping speed is 30d/min.

4.3.8 drip apart from investigating

By above optimum condition, select the different distances of dripping, drip and make ball, with the whole degree of pill shaped circle, hardness, hangover the results are shown in following table for index.

Drip apart from investigating experimental result

Drip apart from (cm) weight differential dripping pill outward appearance

2--adhesion of----dripping pill, roundness is poor

4 6% dripping pill outward appearance roundings, smooth surface

8 8% dripping pill outward appearance roundings, smooth surface

12 17% dripping pill outward appearance roundings, smooth surface

Last table shows, when dripping apart from the time at 4～8cm, and dripping pill outward appearance rounding, smooth surface, weight differential is little, is 4～8cm so select to drip a distance.

Experimental example 5: the research of antiinflammatory action

5.1 the influence of P-xylene induced mice otitis

Get 50 of mouse, the male and female dual-purpose, body weight 18～22g is divided at random: control group, the clear sheet group of throat, preparation group of the present invention, 10 every group.Clear sheet group of throat and formulation components of the present invention are not pressed the dosage gastric infusion in the table 4, control group is given isopyknic physiological saline, 1h after the administration, every mouse auris dextra is coated with the dimethylbenzene with 0.3ml, cause scorching back 4 hours, take off the auricle of ears same position, weigh with the 8mm card punch, with two ear weight differences is the swelling index, the results are shown in following table.

Group dosage (g/kg) left and right sides ear weight difference (mg)

Control group---24.4 ± 1.5

The clear sheet group 6 18.0 ± 2.3 of throat

Dripping pill group 6 17.1 ± 1.2 of the present invention

Dispersing tablet group 6 17.4 ± 3.0 of the present invention

Soft capsule group 6 17.2 ± 0.8 of the present invention

5.2 influence to ankle swelling in rat

Get 50 of rats, male and female dual-purpose, body weight 180～220g are divided at random: control group, the clear sheet group of throat, preparation group of the present invention, 10 every group.Clear sheet group of throat and formulation components of the present invention are not pressed the dosage gastric infusion in the table 5, every day 1 time, 3d continuously.1h after the last administration, right back sufficient plantar subcutaneous injection 1% carrageenan solution 0.1ml/ only causes inflammation rat, and before the Yu Zhiyan and cause scorching back 1,2,5,8h, measure the girth of the right back ankle-joint of rat respectively with inelastic tape, and the difference of the girth of consequently scorching front and back ankle-joint the results are shown in following table as the swelling degree.

Ankle swelling in rat degree (mm)

Group dosage (g/kg) 1h 2h 5h

Control group---3.56 ± 1.87 9.75 ± 4.21 12.98 ± 1.46

The clear sheet group 6 1.82 of throat ± 0.54 6.48 ± 0.51 7.86 ± 2.05

Dripping pill group 6 1.70 of the present invention ± 0.18 6.20 ± 1.22 7.77 ± 2.29

Dispersing tablet group 6 1.71 of the present invention ± 0.54 6.19 ± 2.03 7.75 ± 0.98

Soft capsule group 6 1.69 of the present invention ± 0.66 6.23 ± 1.88 7.79 ± 3.04

More than two experimental results show that preparation of the present invention can obviously alleviate mice ear due to the dimethylbenzene and the ankle swelling in rat due to the carrageenan, action intensity is not less than the clear sheet of throat of same dose.

The thin-layer chromatography discrimination method of japanese avens root research in experimental example 6 pills

Feature for outstanding japanese avens root, selected the characteristic component spot in the japanese avens root medicinal material to contrast as it, but owing to there is the composition that the characteristic component structure is close or polarity is similar in more and the japanese avens root medicinal material in the preparation, the liposoluble constituent in peristrophe, the bicolor for example.Have only the interference of getting rid of these compositions, could obtain desirable chromatogram effect.The key factor of thin-layered chromatography effect quality is the composition of unfolding condition, particularly developping agent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:

The thin-layer chromatography discrimination method of japanese avens root research in the pill

Conditional outcome

Sherwood oil (30～60 ℃)-ethanol=15: 4

The silica gel H thin layer plate separates unintelligible

Methenyl choloride-acetone-glacial acetic acid=silica GF254 thin layer plate feminine gender had interference in 10: 1: 1

Normal hexane-ethyl acetate=silica gel g thin-layer plate Rf value was on the low side in 9: 1

Methenyl choloride-acetone=silica GF254 thin layer plate feminine gender had interference in 5: 0.5

Toluene-methyl alcohol=the silica GF254 thin layer plate separated unintelligible in 5: 1

Methenyl choloride-acetone=silica gel g thin-layer plate separation in 6: 0.3 is clear, and the Rf value is low slightly, and is negative noiseless

Methenyl choloride-acetone=silica gel g thin-layer plate separation in 4: 0.7 is clear, and the Rf value is high slightly, and is negative noiseless

Methenyl choloride-acetone=silica gel g thin-layer plate separation in 5: 0.5 is the most clear, and the Rf value is moderate, and is negative noiseless

Through screening, determined top condition: be stationary phase with the silica gel g thin-layer plate, methenyl choloride-acetone=5: 0.5 be developping agent, and with this understanding, the Rf value of japanese avens root medicinal material feature spot is moderate, and it is the most clear to separate with other spot, and feminine gender is noiseless.

The thin-layer chromatography discrimination method of bicolor medicinal material, Bergenin research in experimental example 7 Diskets

For the feature of outstanding bicolor, selected Bergenin as its characteristic component, but owing to had more or composition that polarity similar close, for example saponin component in balloonflower root, the tinosporae in the preparation to the Bergenin structure.Have only the interference of getting rid of these compositions, could obtain desirable chromatogram effect.The key factor of thin-layered chromatography effect quality is the composition of unfolding condition, particularly developping agent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:

The thin-layer chromatography discrimination method of bicolor medicinal material, Bergenin research in the Disket

Conditional outcome

Sherwood oil (30～60 ℃)-ethanol=silica gel g thin-layer plate separated unintelligible in 8: 1

Methenyl choloride-acetone-methyl alcohol=silica GF254 thin layer plate feminine gender had interference in 10: 1: 1

Normal hexane-ethyl acetate=silica GF254 thin layer plate Rf value was on the low side in 9: 1

Methenyl choloride-acetone=the silica gel g thin-layer plate feminine gender had interference in 5: 1

Methenyl choloride-ethyl acetate-ethanol=silica gel g thin-layer plate separated unintelligible in 8: 3: 4

Methenyl choloride-ethyl acetate-ethanol=silica gel g thin-layer plate separation in 6: 3: 4 is clear, and the Rf value is low slightly, and is negative noiseless

Methenyl choloride-ethyl acetate-ethanol=silica gel g thin-layer plate separation in 4: 5: 2 is clear, and the Rf value is high slightly, and is negative noiseless

Methenyl choloride-ethyl acetate-ethanol=silica gel g thin-layer plate separation in 5: 4: 3 is the most clear, and the Rf value is moderate, and is negative noiseless

Through screening, determined top condition: be stationary phase with the silica gel g thin-layer plate, methenyl choloride-ethyl acetate-ethanol=5: 4: 3 be a developping agent, and with this understanding, the Rf value of Bergenin feature spot is moderate, and it is the most clear to separate with other spot, and feminine gender is noiseless.

The thin-layer chromatography discrimination method of balloonflower root research in experimental example 8 micropill preparations

For the feature of outstanding balloonflower root, selected the characteristic component spot of balloonflower root medicinal material to be contrast, but owing to had more or composition that polarity similar close, for example saponin component in tinosporae, the peristrophe in the preparation to the Bergenin structure.Have only the interference of getting rid of these compositions, could obtain desirable chromatogram effect.The key factor of thin-layered chromatography effect quality is the composition of unfolding condition, particularly developping agent.Therefore, experiment sieving multiple unfolding condition, part unfolding condition and result are as follows:

The thin-layer chromatography discrimination method of balloonflower root research in the micropill preparation

Conditional outcome

Methenyl choloride-ethanol=silica gel H thin layer plate Rf value was higher in 9: 1

Methenyl choloride-acetone-methyl alcohol=silica GF254 thin layer plate feminine gender had interference in 15: 3: 1

Normal hexane-ethyl acetate=the silica GF254 thin layer plate separated unintelligible in 7: 1

Methenyl choloride-acetone=the silica gel g thin-layer plate feminine gender had interference in 5: 1

Methenyl choloride-ethyl acetate-ethanol=silica gel g thin-layer plate separated unintelligible in 8: 3: 4

Methenyl choloride-ether=silica gel g thin-layer plate separated unintelligible in 3: 7

Methenyl choloride-ether=silica gel g thin-layer plate separation in 5: 5 is clear, and the Rf value is low slightly, and is negative noiseless

Methenyl choloride-ether=silica gel g thin-layer plate separation in 4: 6 is the most clear, and the Rf value is moderate, and is negative noiseless

Through screening, determined top condition: be stationary phase with the silica gel g thin-layer plate, methenyl choloride-ether=4: 6 be a developping agent, and with this understanding, the Rf value of balloonflower root medicinal material characteristic component spot is moderate, and it is the most clear to separate with other spot, and feminine gender is noiseless.

Peppermint medicinal material, the research of menthol thin-layer chromatography discrimination method in experimental example 9 microcapsuless

Feature for outstanding peppermint, selected menthol as its characteristic component, but owing to there are more or composition that polarity similar close, for example the volatile oil composition in the medicinal materials such as peristrophe, bicolor, japanese avens root, tinosporae in the preparation to the menthol structure.Have only the interference of getting rid of these compositions, could obtain desirable chromatogram effect.The key factor of thin-layered chromatography effect quality is the composition of unfolding condition, particularly developping agent.Therefore, the test silica gel g thin-layer plate is a stationary phase, has screened multiple developping agent, and part developping agent and result are as follows:

Peppermint medicinal material, the research of menthol thin-layer chromatography discrimination method in the microcapsules

Conditional outcome

Benzene-methenyl choloride=silica gel g thin-layer plate separated unintelligible in 9: 1

Methenyl choloride-acetone-methyl alcohol=feminine gender had interference in 10: 1: 1

Normal hexane-ethyl acetate=the Rf value was on the low side in 9: 1

Methenyl choloride-acetone=feminine gender had interference in 5: 1

Methenyl choloride-ethyl acetate-ethanol=separate unintelligible at 8: 3: 4

Cyclohexane-ethyl acetate-methenyl choloride=separation in 8: 1.5: 1.2 is clear, and the Rf value is low slightly, and is negative noiseless

Cyclohexane-ethyl acetate-methenyl choloride=separation in 10: 2.5: 0.8 is clear, and the Rf value is high slightly, and is negative noiseless

Cyclohexane-ethyl acetate-methenyl choloride=separation in 9: 2: 1 is the most clear, and the Rf value is moderate, and is negative noiseless

Through screening, determined top condition: be stationary phase with the silica gel g thin-layer plate, cyclohexane-ethyl acetate-methenyl choloride=9: 2: 1 be developping agent, and with this understanding, the Rf value of menthol feature spot is moderate, and it is the most clear to separate with other spot, and feminine gender is noiseless.

The high performance liquid chromatography assay of Bergenin research in experimental example 10 pills

1 instrument and reagent

1.1 key instrument

High performance liquid chromatograph 1100Series Agilent

The general all purpose instrument company limited of analysing in ultraviolet spectrophotometer TU-1800SPC Beijing

Electronic analytical balance BP211D SARTORIUS

Supersonic wave cleaning machine KQ250B Kunshan Ultrasonic Instruments Co., Ltd.

1.2 reagent

Methyl alcohol is analyzed pure Beijing Chemical Plant

Acetonitrile chromatographically pure CALEDON

The pure Beijing of phosphoric acid top grade chemical reagents corporation

The pure water Wahaha

It is an amount of that the 2 selection precisions that detect wavelength take by weighing the Bergenin reference substance, adds methyl alcohol and make the solution that every 1ml contains 50 μ g, in the interscan of 200～400nm wavelength coverage.The result shows that Bergenin has absorption maximum at the 275nm place, therefore selects 275nm as the detection wavelength of measuring Bergenin content in the featherleaf rodgersflower and pyrola herb contained tablet for treating mammary disease.

3 chromatographic conditions

Chromatographic column: Diamonsil (diamond) C ₁₈(250 * 4.6mm, 5 μ m);

Moving phase: methanol-water (18: 82);

Detect wavelength: 275nm;

Flow velocity: 1ml/min;

Test and Selection Bergenin as its index components, but owing to have for example saponin component in balloonflower root, the tinosporae of more or composition that polarity similar close in the preparation to the Bergenin structure.Have only the interference of getting rid of these compositions, could obtain desirable chromatogram effect.The key factor of high performance liquid chromatography effect quality is the composition of elution requirement, particularly moving phase.Therefore, experiment sieving multiple moving phase, part moving phase and result are as follows:

The investigation of chromatographic condition

The moving phase conditional outcome

Methanol-water=separate not exclusively at 50: 50

Acetonitrile-water=separate not exclusively at 60: 20

Acetonitrile-0.05mol/L sodium dihydrogen phosphate=appearance time was longer in 10: 90

Methanol-water=separate not exclusively at 30: 70

Methanol-water=separation in 21: 79 is clear, negative noiseless

Methanol-water=retention time was long slightly in 15: 85, and it is clear to separate, and is negative noiseless

Methanol-water=retention time was moderate in 18: 82, and it is the most clear to separate, and is negative noiseless

Through screening, determined with the octadecylsilane chemically bonded silica to be stationary phase, methanol-water=18: 82 be a moving phase, and with this understanding, the Bergenin retention time is moderate, and the peak is capable sharp-pointed, symmetry, it is the most clear to separate with adjacent peak, and feminine gender is noiseless.

4 determination methods

Get this product, porphyrize is got about 0.2g, the accurate title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight, sonicated (power 250W, frequency 33KHz) 60 minutes, take out, put to room temperature, claim to decide weight, supply the weight that subtracts mistake, shake up with methyl alcohol, filter with miillpore filter (0.45 μ m), get subsequent filtrate, as need testing solution; It is an amount of to get the Bergenin reference substance, and accurate the title decides, and adds moving phase and makes the solution that every 1ml contains 0.05mg, in contrast product solution; Accurate respectively each the 10 μ l of above-mentioned two kinds of solution that draw inject liquid chromatograph, measure, promptly.

The investigation precision of 5 linear relationships is measured Bergenin reference substance solution (0.5136mg/ml) 0.4ml, 0.8ml, 1.2ml, 1.6ml, 2.0ml, split in the 10ml measuring bottle, add methyl alcohol and be diluted to scale, shake up, be mixed with the reference substance solution of 0.020544mg/ml, 0.041088mg/ml, 0.061632mg/ml, 0.082176mg/ml, 0.10272mg/ml, the therefrom accurate respectively 10 μ l that draw inject liquid chromatograph, according to high effective liquid chromatography for measuring.With the peak area is horizontal ordinate, and Bergenin sample size (μ g) is figure for ordinate, the drawing standard curve.The result is as follows:

The Bergenin linear relationship

Numbering peak area Bergenin (μ g)

1 262.23 0.20544

2 530.14 0.41088

3 801.55 0.61632

4 1073.15 0.82176

5 1320.08 1.02720

Regression equation: Y=0.000772X+0.000325

Related coefficient: γ=0.9999

The result shows that Bergenin linear relationship between 0.20544 μ g～1.0270 μ g is good.

Through calculating, the Bergenin typical curve is one to cross the straight line of initial point, therefore selects one point external standard method to measure the content of Bergenin in the throat clearing dropping pill.

The test of 6 precision is accurate draws with a Bergenin reference substance solution 10 μ l, injects liquid chromatograph, and replication 5 times is investigated reference substance solution precision, and measurement result is as follows:

The precision test

Test number (TN) 12345 mean value RSD (%)

Peak area 680.70 685.23 681.16 688.94 675.38 682.28 0.74

The result shows that reference substance solution precision is good.

7 need testing solution stability tests are accurate draws with a need testing solution 10 μ l, injects liquid chromatograph, measures at 0,2,4,8,24 hour sample introduction respectively, and measurement result is as follows:

Need testing solution stability test result

Test durations (h) 0248 24 mean value RSD (%)

Content (mg/ grain) 0.3812 0.3774 0.3805 0.3826 0.3769 0.3797 0.65

The result shows that need testing solution is good at 24 hours internal stabilities.

8 replica tests are got same lot number this product, and porphyrize is got about 0.2g (totally 5 parts), and accurate the title decides, and press operation under chromatographic condition and the determination method item.The result is as follows:

Replica test

Number 12345 mean value RSD (%)

Content (mg/ grain) 0.3783 0.3821 0.3856 0.3746 0.3805 0.3802 1.08

The result shows that repeatability is good.

The application of sample absorption method is adopted in the test of 9 average recoveries, gets this product under the weight differential item, and porphyrize is got about 0.1g, and accurate the title decides, and puts in the tool plug conical flask; Precision is measured Bergenin (0.6092mg/ml) 0.8mg, 1.0ml, 1.2ml (each 2 parts) split in the above-mentioned tool plug conical flask, and precision adds methyl alcohol to 25ml, close plug, claim to decide weight, sonicated (power 250W, frequency 33KHz) 60 minutes, take out, put, claim to decide weight to room temperature, supply the weight that subtracts mistake with methyl alcohol, shake up, filter with miillpore filter (0.45 μ m), get subsequent filtrate, press operation under chromatographic condition and the determination method item, measure, promptly.Measurement result is as follows:

The test of Bergenin average recovery

The white Bergenin measured value of the rock recovery in the weighing test sample

Numbering

(g) plain (mg) addition (mg) (mg) (%) of measuring of dish

1 0.10525 0.6415 0.48736 1.1205 98.28

2 0.11073 0.6749 0.48736 1.1497 97.42

3 0.10594 0.6457 0.60920 1.2480 98.87

4 0.11215 0.6836 0.60920 1.2803 97.95

5 0.10958 0.6679 0.73104 1.3853 98.13

6 0.10737 0.6544 0.73104 1.3783 99.02

Average recovery rate=98.27%, RSD=0.60%.

10 sample sizes are measured and are pressed chromatographic condition and the operation down of determination method item, measure ten batch samples, and the result is as follows:

The content of Bergenin in the throat clearing dropping pill

Lot number Bergenin average content (mg/ grain)

1 0.3774

2 0.3851

3 0.3695

4 0.3901

5 0.3887

6 0.3795

7 0.3824

8 0.3885

9 0.3793

10 0.3815

The vapor-phase chromatography assay of menthol research in experimental example 11 Diskets

1 instrument and reagent

1.1 key instrument

Gas chromatograph day island proper Tianjin company

Electronic analytical balance BP211D SARTORIUS

Supersonic wave cleaning machine KQ250B Kunshan Ultrasonic Instruments Co., Ltd.

1.2 reagent

Ethanol is analyzed pure Beijing Chemical Plant

The pure water Wahaha

2 chromatographic conditions

Glass column (3.2mm * 2.1m); With PEG-20M is stationary phase; Coating concentration is 10%; Column temperature is 140 ℃; Fid detector; Nitrogen (50ml/min), hydrogen (50ml/min), air (500ml/min).

Test and Selection menthol as its index components, but owing to have more or composition that polarity similar close, for example the volatile oil composition in the medicinal materials such as peristrophe, bicolor, japanese avens root, tinosporae in the preparation to the menthol structure.Have only the interference of getting rid of these compositions, could obtain desirable chromatogram effect.The key factor of gas phase color method effect quality is factors such as the kind, coating concentration, post of elution requirement, particularly stationary phase.Therefore, test is screened above factor related levels, and partial condition and result are as follows:

The investigation of chromatographic condition

The moving phase conditional outcome

PEG-20M is a stationary phase; Coating concentration is 5%; Separate not exclusively

Column temperature is 200 ℃

PEG-20M is a stationary phase; Coating concentration is 5%;

Separate not exclusively

Column temperature is 110 ℃

SE-54 is a stationary phase; Coating concentration is 20%;

Separate not exclusively

Column temperature is 130 ℃

SE-54 is a stationary phase; Coating concentration is 10%;

Separate not exclusively

Column temperature is 140 ℃

PEG-20M is a stationary phase; Coating concentration is

It is more clear to separate, negative noiseless

12%; Column temperature is 150 ℃

PEG-20M is a stationary phase; Coating concentration is 8%;

It is more clear to separate, negative noiseless

Column temperature is 110 ℃

PEG-20M is a stationary phase; Coating concentration is that retention time is moderate, and it is the most clear to separate, and negative nothing is done

10%; Column temperature is 140 ℃ and disturbs

Through screening, determined top condition: be stationary phase with PEG-20M, coating concentration is 10%, and column temperature is 140 ℃; With this understanding, the menthol retention time is moderate, and the peak is capable sharp-pointed, symmetry, and it is the most clear to separate with adjacent peak, negative noiseless.

3 determination methods

It is an amount of to get this product, and porphyrize is got 1g, and accurate the title decides, put in the tool plug conical flask, the accurate absolute ethyl alcohol 20ml that adds claims to decide weight, ice-bath ultrasonic was handled 30 minutes, was placed to room temperature, claimed to decide weight again, supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, centrifugal, precision is measured supernatant 5ml, puts in the 10ml measuring bottle, the accurate inner mark solution 2ml that adds, add absolute ethyl alcohol to scale, shake up, as need testing solution; Get naphthalene 150mg, the accurate title, decide, and puts in the 100ml measuring bottle, adds anhydrous alcohol solution and be diluted to scale, as inner mark solution; Other gets menthol reference substance 10mg, and accurate the title decides, and puts in the 10ml measuring bottle, and the accurate inner mark solution 2ml that adds adds absolute ethyl alcohol to scale, shakes up, in contrast product solution; Draw reference substance solution 1 μ l, inject gas chromatograph calculates than positive divisor; Draw need testing solution 1 μ l, inject gas chromatograph is measured.

The investigation of 4 linear relationships

Get the reference substance solution of variable concentrations, sample introduction 1 μ l does linear regression with sample size and peak area respectively.The result shows that menthol sample size linear relationship between 0.314 μ g～3.143 μ g is good, related coefficient γ＞0.9999.

The test of 5 precision

Test shows that the accurate reference substance solution 10 μ l that draw inject liquid chromatograph, calculate than positive divisor replication 5 times.RSD=0.91% shows that instrument precision is good as a result.

6 need testing solution stability tests are accurate draws with a need testing solution 10 μ l, injects liquid chromatograph, measures at 0,2,4,8,24 hour sample introduction respectively, calculates than positive divisor.RSD=0.97% shows that need testing solution is good at 24 hours internal stabilities as a result.

7 replica tests are got same lot number this product, and porphyrize is got about 1g, accurate claim surely, press under chromatographic condition and the determination method item and operate, and prepare and measure 5 duplicate samples, menthol content in the calculation sample.RSD=2.15% shows that repeatability is good as a result.

The application of sample absorption method is adopted in 8 average recoveries tests; get this product; porphyrize, get about 0.5g (totally 6 parts), accurate claim fixed; split in the tool plug conical flask; the accurate menthol reference substance solution that adds, make in the sample menthol actual content suitable, press respectively again under chromatographic condition and the determination method item and operate; mensuration, the calculating menthol recovery with addition.Average recovery rate=98.7% as a result, RSD=1.26%.The result shows that the method recovery is good.

9 sample sizes are measured and are pressed chromatographic condition and the operation down of determination method item, measure ten batch samples, and the result is as follows:

The content of menthol in the throat Yiqing dispersing tablets

Lot number menthol average content (mg/ sheet)

1 5.5

2 5.6

3 5.6

4 5.5

5 5.4

6 5.5

7 5.6

8 5.4

9 5.3

10 5.5

Concrete embodiment

Embodiments of the invention 1: peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g

Get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, and dregs of a decoction boiling 1.5 hours filters, merge with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mixes and is ground into fine powder, press extract powder: matrix=1: 1.5 adding matrix PEG4000, spray into above-mentioned volatile oil behind the heating and melting, mixing is transferred to the dripping pill machine, drip 85 ℃ of system temperature, cooling medium is 15 ℃ a dimethyl silicon oil, and dripping speed is 30d/min, drips distance at 6cm, collect dripping pill and remove the dimethyl silicon oil on surface, promptly get pill, oral, one time 2～4, one hour 1 time, 5～6 times on the one.

Embodiments of the invention 2: peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g

Get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, and dregs of a decoction boiling 1.5 hours filters, merge with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mixes and is ground into fine powder, press extract powder: matrix=1: 1.5 adding matrix PEG4000, spray into above-mentioned volatile oil behind the heating and melting, mixing is transferred to the dripping pill machine, drip 85 ℃ of system temperature, cooling medium is 10 ℃ ℃ a dimethyl silicon oil, and dripping speed is 30d/min, drips distance at 4cm, collect dripping pill and remove the dimethyl silicon oil on surface, promptly get pill.

Embodiments of the invention 3: peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g

Get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, and dregs of a decoction boiling 1.5 hours filters, merge with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mixes and is ground into fine powder, press extract powder: matrix=1: 1.5 adding matrix PEG4000, spray into above-mentioned volatile oil behind the heating and melting, mixing is transferred to the dripping pill machine, drip 85 ℃ of system temperature, cooling medium is 20 ℃ a dimethyl silicon oil, and dripping speed is 30d/min, drips distance at 8cm, collect dripping pill and remove the dimethyl silicon oil on surface, promptly get pill.

Embodiments of the invention 4: peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g

Get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, and dregs of a decoction boiling 1.5 hours filters, merge with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mixes and is ground into fine powder, press extract powder: matrix=1: 1 adding matrix PEG4000, spray into above-mentioned volatile oil behind the heating and melting, mixing is transferred to the dripping pill machine, drip 70 ℃ of system temperature, cooling medium is 5 ℃ a dimethyl silicon oil, and dripping speed is 20d/min, drips distance at 2cm, collect dripping pill and remove the dimethyl silicon oil on surface, promptly get dripping pill.

Embodiments of the invention 5: peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g

Get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, and dregs of a decoction boiling 1.5 hours filters, merge with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mixes and is ground into fine powder, press extract powder: matrix=1: 3 adding matrix PEG4000, spray into above-mentioned volatile oil behind the heating and melting, mixing is transferred to the dripping pill machine, drip 90 ℃ of system temperature, cooling medium is 30 ℃ a dimethyl silicon oil, and dripping speed is 40d/min, drips distance at 12cm, collect dripping pill and remove the dimethyl silicon oil on surface, promptly

Embodiments of the invention 6: peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g

Get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours, filtering, merge with above-mentioned aqueous solution, is 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, adding ethanol makes and contains alcohol amount and reach 60%, left standstill 24 hours, and filtered, filtrate concentrates at decompression recycling ethanol below 80 ℃, dry, add menthol, mix and be ground into fine powder, spray into above-mentioned volatile oil again, mixing, airtight 2 hours, press extract powder again: matrix=1: 1.5 adds soybean oil, the mixed-matrix of soybean lecithin, heating and melting, mixing gets soft capsule content; The preparation of glue: with gelatin: glycerine: water=1: 0.7: 1.0, getting gelatin adds an amount of distilled water and makes its imbibition, in addition the water of glycerine and remainder is put and be heated to 70～80 ℃ in the glue pot, mix, add the gelatin that expands and stir, make it to dissolve into uniform glue, in 60 ℃ of insulations 2～3 hours, leave standstill, remove the come-up foam, filter standby with cloth bag.Soft capsule content and glue are pressed into soft capsule in encapsulating machine, put in the drum dryer and finalize the design, whole ball, drying promptly gets soft capsule.

Embodiments of the invention 7: peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g

Get peristrophe, bicolor, japanese avens root, root of Herba Gonostegiae hirtae, tinosporae, balloonflower root, peppermint, steam distillation is extracted volatile oil, and the aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 1.5 hours filters, and merges with above-mentioned aqueous solution, be 1.10～1.12 clear cream when being evaporated to 60 ℃ of relative densities, add ethanol and make and contain the alcohol amount and reach 60%, left standstill 24 hours, filter, filtrate concentrates at decompression recycling ethanol below 80 ℃, drying adds menthol, mix and be ground into fine powder, add 10% microcrystalline cellulose, 7% crospolyvinylpyrrolidone, mixing adds ethanol, the system softwood, granulate drying, the whole grain of 40 mesh sieves, compressing tablet behind the mixing, other gets menthol, adds ethanol and makes dissolving in right amount, sprays into, spray into above-mentioned volatile oil again, mixing airtight 2 hours, promptly gets Disket.

The thin-layered chromatography of japanese avens root is differentiated in embodiment 8 pills

Get this product powder 3.0g, add methyl alcohol 50ml, sonicated 30 minutes filters, filtrate evaporate to dryness, residue add water 20ml makes dissolving, adds the ethyl acetate jolting and extracts 2 times, each 20ml, combined ethyl acetate liquid, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets negative sample (lack of water Chinese waxmyrtle root), makes negative sample solution with method.The Chinese waxmyrtle root control medicinal material 2g that fetches water again adds ethyl acetate 50ml, and sonicated 60 minutes filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, in contrast medicinal material solution.According to the thin-layered chromatography test, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-acetone (5: 0.5) is developping agent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to spot colour developing clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

The thin-layered chromatography of japanese avens root is differentiated in embodiment 9 Diskets

Get this product powder 2.0g, add methyl alcohol 50ml, sonicated 30 minutes filters, filtrate evaporate to dryness, residue add water 20ml makes dissolving, adds the ethyl acetate jolting and extracts 2 times, each 20ml, combined ethyl acetate liquid, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.The Chinese waxmyrtle root control medicinal material 2g that fetches water again adds ethyl acetate 50ml, and sonicated 60 minutes filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, in contrast medicinal material solution.According to thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with methenyl choloride-acetone (4: 0.7), launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to spot develop the color clear.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

The thin-layered chromatography of japanese avens root is differentiated in embodiment 10 micropill preparations

Get this product powder 2.0g, add methyl alcohol 30ml, sonicated 15 minutes filters, and filtrate evaporate to dryness, residue add water 15ml makes dissolving, adds ethyl acetate 20ml jolting and extracts, and acetic acid ethyl fluid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.The Chinese waxmyrtle root control medicinal material 2g that fetches water again adds ethyl acetate 50ml, and sonicated 60 minutes filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, in contrast medicinal material solution.According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with methenyl choloride-acetone (6: 0.3), launch, take out, dry, put under the uviol lamp and inspect; Spray with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing again.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color.

The thin-layered chromatography of bicolor medicinal material, Bergenin is differentiated in embodiment 11 Diskets

Get this product powder 2g, add methyl alcohol 20ml ultrasonic Extraction, extract is concentrated into about 1ml, as need testing solution; Get bicolor control medicinal material 0.5g, shine medicinal material solution in pairs with legal system; Other gets the Bergenin reference substance, adds methyl alcohol and makes the reference substance solution that every ml contains 0.5mg; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-ethyl acetate-ethanol=5: 4: 3 was developping agent, launched, and took out, dry, spray is with the mixed solution of 1% liquor ferri trichloridi-1% potassium ferricyanide solution (1: 1); In the test sample chromatogram, with reference substance and the corresponding position of control medicinal material chromatogram on, show the spot of same color.

The thin-layered chromatography of Bergenin is differentiated in embodiment 12 micropill preparations

Get this product powder 2g, add methyl alcohol 30ml ultrasonic Extraction, extract is concentrated into about 1ml, as need testing solution; Other gets the Bergenin reference substance, adds methyl alcohol and makes the reference substance solution that every ml contains 1mg; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-ethyl acetate-ethanol=4: 5: 2 was developping agent, launched, and took out, dry, spray is with the mixed solution of 1% liquor ferri trichloridi-1% potassium ferricyanide solution (1: 2); In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.

The thin-layered chromatography of bicolor medicinal material, Bergenin is differentiated in embodiment 13 microcapsuless

Get this product powder 1g, add methyl alcohol 25ml ultrasonic Extraction, extract is concentrated into about 1ml, as need testing solution; Get bicolor control medicinal material 1g, shine medicinal material solution in pairs with legal system; Other gets the Bergenin reference substance, adds methyl alcohol and makes the reference substance solution that every ml contains 1mg; According to the thin-layered chromatography test, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-ethyl acetate-ethanol=6: 3: 4 was developping agent, launched, and took out, dry, spray is with the mixed solution of 1% liquor ferri trichloridi-1% potassium ferricyanide solution (2: 1); In the test sample chromatogram, with reference substance and the corresponding position of control medicinal material chromatogram on, show the spot of same color.

The thin-layered chromatography of balloonflower root is differentiated in embodiment 14 Diskets

Get this product powder 3g, add the mixed solution 30ml of 7% ethanol solution of sulfuric acid-water=1: 3, reflux 3 hours, put coldly, add the methenyl choloride jolting and extract 2 times, each 20ml, merge methenyl choloride liquid, add water 30ml washing, discard water liquid, methenyl choloride liquid anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the balloonflower root control medicinal material, shines medicinal material solution in pairs with legal system; According to thin-layered chromatography test, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with methenyl choloride-ether=4: 6, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to spot develop the color clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color.

The thin-layered chromatography of balloonflower root is differentiated in embodiment 15 pills

Get this product powder 2g, add the mixed solution 25ml of 10% ethanol solution of sulfuric acid-water=1: 4, reflux 2 hours, put coldly, add the methenyl choloride jolting and extract 2 times, each 20ml, merge methenyl choloride liquid, add water 30ml washing, discard water liquid, methenyl choloride liquid anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the balloonflower root control medicinal material, shines medicinal material solution in pairs with legal system; According to thin-layered chromatography test, draw each 3 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with methenyl choloride-ether=5: 5, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to spot develop the color clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the principal spot of same color.

The high performance liquid chromatography assay of Bergenin in embodiment 16 pills

Get this product, porphyrize is got about 0.2g, accurate claims surely, puts in the tool plug conical flask, and the accurate methyl alcohol 25ml that adds claims decide weight, and sonicated 1 hour is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filtration is got subsequent filtrate, as need testing solution; Accurate title Bergenin reference substance is an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.05mg, in contrast product solution; According to high performance liquid chromatography test, be filling agent with 18 silylation bonded silica gels, be moving phase with methanol-water=18: 82; The detection wavelength is 275nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; Calculate with one point external standard method, this product contains the purple bergenia herb element with dosage and must not be less than 3.0mg every day;

The high performance liquid chromatography assay of Bergenin in embodiment 17 Diskets

Get this product powder 0.5g, accurate claim surely, put in the tool plug conical flask, the accurate methyl alcohol 30ml that adds claims decide weight, and sonicated 15 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filtration is got subsequent filtrate, as need testing solution; Accurate title Bergenin reference substance is an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.1mg, in contrast product solution; According to high performance liquid chromatography test, be filling agent with 18 silylation bonded silica gels, be moving phase with methanol-water=15: 85; The detection wavelength is 270nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; Calculate with one point external standard method, this product contains the purple bergenia herb element with dosage and must not be less than 2.0mg every day;

The high performance liquid chromatography assay of Bergenin in embodiment 18 microcapsuless

Get this product powder 0.5g, accurate claim surely, put in the tool plug conical flask, the accurate methyl alcohol 30ml that adds claims decide weight, and sonicated 15 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and filtration is got subsequent filtrate, as need testing solution; Accurate title Bergenin reference substance is an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.1mg, in contrast product solution; According to high performance liquid chromatography test, be filling agent with 18 silylation bonded silica gels, be moving phase with methanol-water=21: 79; The detection wavelength is 280nm; Accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing inject liquid chromatograph, measure; Calculate with one point external standard method, this product contains the purple bergenia herb element with dosage and must not be less than 2.0mg every day;

The vapor-phase chromatography assay of menthol in embodiment 19 micropill preparations

It is an amount of to get this product, and porphyrize is got 1g, and accurate the title decides, put in the tool plug conical flask, the accurate absolute ethyl alcohol 20ml that adds claims to decide weight, ice-bath ultrasonic was handled 30 minutes, was placed to room temperature, claimed to decide weight again, supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, centrifugal, precision is measured supernatant 5ml, puts in the 10ml measuring bottle, the accurate inner mark solution 2ml that adds, add absolute ethyl alcohol to scale, shake up, as need testing solution; Get naphthalene 150mg, the accurate title, decide, and puts in the 100ml measuring bottle, adds anhydrous alcohol solution and be diluted to scale, as inner mark solution; Other gets menthol reference substance 10mg, and accurate the title decides, and puts in the 10ml measuring bottle, and the accurate inner mark solution 2ml that adds adds absolute ethyl alcohol to scale, shakes up, in contrast product solution; According to the vapor-phase chromatography test, be stationary phase with polyglycol-20M, coating concentration is 10%; With Chromosorb W (AW-DMCS) (60-80 order) is carrier, and column temperature is 140 ℃; Draw reference substance solution 1 μ l, inject gas chromatograph calculates than positive divisor; Draw need testing solution 1 μ l, inject gas chromatograph is measured; This product contains menthol with dosage and must not be less than 20mg every day.

The vapor-phase chromatography assay of menthol in embodiment 20 Diskets

It is an amount of to get this product, and porphyrize is got 0.5g, and accurate the title decides, put in the tool plug conical flask, the accurate absolute ethyl alcohol 25ml that adds claims to decide weight, ice-bath ultrasonic was handled 30 minutes, was placed to room temperature, claimed to decide weight again, supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, centrifugal, precision is measured supernatant 5ml, puts in the 10ml measuring bottle, the accurate inner mark solution 2ml that adds, add absolute ethyl alcohol to scale, shake up, as need testing solution; Get naphthalene 100mg, the accurate title, decide, and puts in the 100ml measuring bottle, adds anhydrous alcohol solution and be diluted to scale, as inner mark solution; Other gets menthol reference substance 10mg, and accurate the title decides, and puts in the 10ml measuring bottle, and the accurate inner mark solution 2ml that adds adds absolute ethyl alcohol to scale, shakes up, in contrast product solution; According to the vapor-phase chromatography test, be stationary phase with polyglycol-20M, coating concentration is 8%; With Chromosorb W (AW-DMCS) (60-80 order) is carrier, and column temperature is 110 ℃; Draw reference substance solution 1 μ l, inject gas chromatograph calculates than positive divisor; Draw need testing solution 1 μ l, inject gas chromatograph is measured; This product contains menthol with dosage and must not be less than 10mg every day.

The vapor-phase chromatography assay of menthol in embodiment 21 micropill preparations

It is an amount of to get this product, and porphyrize is got 0.5g, and accurate the title decides, put in the tool plug conical flask, the accurate absolute ethyl alcohol 25ml that adds claims to decide weight, ice-bath ultrasonic was handled 30 minutes, was placed to room temperature, claimed to decide weight again, supply the weight that subtracts mistake with absolute ethyl alcohol, shake up, centrifugal, precision is measured supernatant 5ml, puts in the 10ml measuring bottle, the accurate inner mark solution 2ml that adds, add absolute ethyl alcohol to scale, shake up, as need testing solution; Get naphthalene 100mg, the accurate title, decide, and puts in the 100ml measuring bottle, adds anhydrous alcohol solution and be diluted to scale, as inner mark solution; Other gets menthol reference substance 10mg, and accurate the title decides, and puts in the 10ml measuring bottle, and the accurate inner mark solution 2ml that adds adds absolute ethyl alcohol to scale, shakes up, in contrast product solution; According to the vapor-phase chromatography test, be stationary phase with polyglycol-20M, coating concentration is 12%; With Chromosorb W (AW-DMCS) (60-80 order) is carrier, and column temperature is 150 ℃; Draw reference substance solution 1 μ l, inject gas chromatograph calculates than positive divisor; Draw need testing solution 1 μ l, inject gas chromatograph is measured; This product contains menthol with dosage and must not be less than 20mg every day.

The thin-layered chromatography of peppermint medicinal material, menthol is differentiated in embodiment 22 microcapsuless

Get this product powder 2g, add sherwood oil (30～60 ℃) 10ml jolting and extract, filter, filtrate is as need testing solution; Other gets peppermint medicinal material 0.5g, shines medicinal material solution in pairs with legal system; Get the menthol reference substance, add sherwood oil (30～60 ℃) and make the reference substance solution that every ml contains 2mg; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-methenyl choloride=9: 2: 1 was developping agent, launched, and took out, dry, spray is with 1% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.

The thin-layered chromatography of peppermint medicinal material, menthol is differentiated in embodiment 23 micropill preparations

Get this product powder 2g, add sherwood oil (60～90 ℃) 10ml jolting and extracted 10 minutes, centrifugal, supernatant is as need testing solution; Other gets peppermint medicinal material 0.5g, shines medicinal material solution in pairs with legal system; Get the menthol reference substance, add sherwood oil (60～90 ℃) and make the reference substance solution that every ml contains 2mg; According to the thin-layered chromatography test, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-methenyl choloride=8: 1.5: 1.2 was developping agent, launched, and took out, dry, spray is with 2% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.

The thin-layered chromatography of peppermint medicinal material, menthol is differentiated in embodiment 24 micropill preparations

Get this product powder 2g, add sherwood oil (60～90 ℃) 10ml jolting and extracted 10 minutes, centrifugal, supernatant concentration is as need testing solution; Other gets peppermint medicinal material 1g, shines medicinal material solution in pairs with legal system; Get the menthol reference substance, add sherwood oil (60～90 ℃) and make the reference substance solution that every ml contains 4mg; According to the thin-layered chromatography test, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-methenyl choloride=10: 2.5: 0.8 was developping agent, launched, and took out, dry, spray is with 2% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.

Claims (2)

1. discrimination method for the treatment of the clear preparation of throat of pharyngolaryngitis, the clear preparation of described throat is prepared from by peristrophe 105.5g, bicolor 63g, japanese avens root 79g, root of Herba Gonostegiae hirtae 79g, tinosporae 31.5g, balloonflower root 63g, peppermint 79g, menthol 2.5g, it is characterized in that: described discrimination method comprises following content: the thin-layered chromatography of japanese avens root is differentiated in a. preparation

It is an amount of to get this product powder, adds methanol extraction, and extract evaporate to dryness, residue add water makes dissolving, adds the ethyl acetate jolting and extracts, and acetic acid ethyl fluid evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Other the Chinese waxmyrtle root control medicinal material of fetching water is an amount of, adds ethyl acetate extraction, and extract evaporate to dryness, residue add methyl alcohol makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution respectively, puts on same silica gel g thin-layer plate, and with methenyl choloride-acetone=4～6: 0.3～0.7 is developping agent, launch, take out, dry, put under the uviol lamp and inspect, or spray is with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot colour developing; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

B. the thin-layered chromatography of peppermint medicinal material, menthol is differentiated in the preparation

It is an amount of to get this product powder, adds the sherwood oil jolting and extracts, and extract is as need testing solution; Other gets the peppermint medicinal material, shines medicinal material solution in pairs with legal system; Get the menthol reference substance, add sherwood oil and make reference substance solution; Test according to thin-layered chromatography, it is an amount of to draw above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-methenyl choloride=8～10: be developping agent at 1.5～2.5: 0.8～1.2, launch, take out, dry, spray is with the vanillic aldehyde sulfuric acid solution, and it is clear to be heated to the spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.

2. according to the described discrimination method of claim 1, it is characterized in that: comprise following content:

A. the thin-layered chromatography of japanese avens root is differentiated in the preparation

It is an amount of to get this product powder, adds the methyl alcohol ultrasonic Extraction, and extract evaporate to dryness, residue add water makes dissolving, adds the ethyl acetate jolting and extracts, and acetic acid ethyl fluid evaporate to dryness, residue add methyl alcohol makes dissolving, as need testing solution; Other the Chinese waxmyrtle root control medicinal material of fetching water is an amount of, adds the ethyl acetate ultrasonic Extraction, and extract evaporate to dryness, residue add methyl alcohol makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography test, it is an amount of to draw above-mentioned two kinds of solution respectively, put on same silica gel g thin-layer plate, be developping agent with methenyl choloride-acetone=5: 0.5, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to spot develop the color clear; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, show the spot of same color;

B. the thin-layered chromatography of peppermint medicinal material, menthol is differentiated in the preparation

It is an amount of to get this product powder, adds the sherwood oil jolting and extracts, and extract is as need testing solution; Other gets the peppermint medicinal material, shines medicinal material solution in pairs with legal system; Get the menthol reference substance, add sherwood oil and make reference substance solution; According to the thin-layered chromatography test, it is an amount of to draw above-mentioned three kinds of solution, puts respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate-methenyl choloride=9: 2: 1 was developping agent, launched, and took out, dry, spray is with the vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with contrast chromatogram corresponding position on, show the spot of same color.