CN101313938B - Magnolia flower volatile oil spray for nose - Google Patents
Magnolia flower volatile oil spray for nose Download PDFInfo
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- CN101313938B CN101313938B CN2008100509586A CN200810050958A CN101313938B CN 101313938 B CN101313938 B CN 101313938B CN 2008100509586 A CN2008100509586 A CN 2008100509586A CN 200810050958 A CN200810050958 A CN 200810050958A CN 101313938 B CN101313938 B CN 101313938B
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Abstract
The invention relates to a Flos Magnoliae volatile oil nasal spray, belonging to the Chinese traditional medicine preparation field. The nasal spray is prepared by the following raw material medicines and auxiliary materials in volume portion: 40 to 60ml of Flos Magnoliae volatile oil, 40 to 60ml of polysorbate 80 and 20 to 32ml of glycerin. The preparation method comprises the following steps that: the polysorbate 80 and the glycerin in the formula are taken and added with 100 to 500ml of absolute ethyl alcohol and stirred so as to dissolve the polysorbate 80 and the glycerin in the absoluteethyl alcohol; under the stirring at a rotary speed of between 900 and 1,100r/min, the mixed solution is slowly added with the Flos Magnoliae volatile oil and stirred for 10 minutes at a rotary speedof between 900 and 1,100r/min; and the mixed solution is added with distilled water until the constant volume is 1,000ml, and is filtered through a micro-hole filter membrane with a hole diameter of 0.22mu m. The nasal spray of the invention has the effects of dispelling chill, removing nasal obstruction and resisting allergy; moreover, the nasal spray is used to treat acute and chronic rhinitis,allergic rhinitis, sinusitis, nasal obstruction, nasosinusitis and nasal discharge. The nasal spray is used to spray four times once, twice a day.
Description
Technical field
The invention belongs to field of traditional Chinese, is the nasal spray that is used for the treatment of the Flos Magnoliae single preparations of ephedrine of acute, chronic rhinitis.
Background technology
Rhinitis is a kind of common multiple respiratory tract disease.Its morbidity no obvious region, seasonality, age, sex difference, therefore throughout the year, all parts of the country, men and women, old and young all can suffer from rhinitis.Its sickness rate accounts for the 5-6% of population.And allergic rhinitis are the most common, account for 30% of various cacorhinias.Its predisposing factors is mainly factors such as antibacterial, virus attack, pollen sample material allergy, cold air allergy, environmental pollution, physics and chemistry.Clinical manifestation is symptoms such as nasal obstruction, watery nasal discharge, hypopnea, the acid of nose eye are itched, anoxia.If untimely treatment, outbreak repeatedly, because of anoxia can make concha nasalis, deviation of nasal septum hypertrophy, plump degeneration, seriously then must operative treatment.
Along with atmospheric pollution increases the weight of day by day, cause this sick sickness rate to be ascendant trend year by year, especially obvious in recent years.People's orthobiosis and work have been had a strong impact on during its outbreak.
Rhinitis particularly allergic rhinitis is a kind of commonly encountered diseases frequently-occurring disease, and along with the industrial expansion that reaches of society, its sickness rate is more and more higher, and does not still have safe, quick-acting medicine at present.For this reason, a large amount of literature survey and the project screening work of my company's process determines that Magnoliacea plant Flos Magnoliae (Flos Magnoliae) is the research crude drug, it is developed to treats new drug acute, chronic, allergic rhinitis.
The theoretical foundation of Flos Magnoliae treatment rhinitis
1, the antiinflammatory action of Flos Magnoliae volatile oil composition
The rat foot that the Flos Magnoliae volatile oil lumbar injection can obviously resist due to dimethylbenzene induced mice auricle edema and the Ovum Gallus domesticus album is opened up swelling; The isolated ileum segments in guinea pigs contraction that slow reacting substance, histamine, acetylcholine are caused has antagonism.
2, the antiinflammatory mechanism of Flos Magnoliae volatile oil composition
With the Flos Magnoliae total volatile oil is raw material, select different inflammation-causing substances and animal, bring out various acute inflammatory model (acetic acid causes that the mouse peritoneal capillary permeability increases, mice caused by dimethylbenzene xylene skin heart permeability increases, mice caused by dimethylbenzene xylene auricle edema, carrageenin cause the mice foot swelling, Ovum Gallus domesticus album causes rat paw edema, carrageenin causes the rat pleuritis), mice granuloma induced by implantation of cotton pellets chronic inflammation model, the rat assist agent arthritis model has been observed the antiinflammatory action of Flos Magnoliae total volatile oil.The result shows, the Flos Magnoliae total volatile oil to the hyperemia in the inflammatory process, ooze out, swelling, leukocyte is swum out of and pathological change such as cell infiltration, tissue necrosis, granulation tissue hyperplasia, inhibitory action is all arranged, illustrate that the Flos Magnoliae total volatile oil has good antiinflammatory action.Its antiinflammatory action has following characteristics: (1) suppresses arachidonic generation and metabolism by the activity that reduces phospholipase A2, suppresses the generation or the release of acid lipid inflammatory mediator (as PGE2).(2) can not only suppress the synthetic release of vasoactive amines-histamine, can also cause the edema effect antihistaminic.(3) the unusual rising of IL-1 and TNF in the prevention inflammatory reaction process, the damage that the reaction pair body that reduces inflammation causes.(4) secretion of glucocorticoid is not had obvious influence, its antiinflammatory action does not rely on adrenal existence yet.
Domestic and international present Research
1, the characteristics of the clinical usage of Flos Magnoliae
(1) it is for oral administration that soup is fried in shallow oil in traditional Flos Magnoliae clinical practice more, and the magnolia flower rhinitis-treating pill-type that companies such as Guangzhou Zhongyi Medicine Industry Co., Ltd produce is pill (concentrated watered pill), and the endo-medicine onset is slow, and it is long to reach the disease sites time, and increases the weight of the burden to body excretes.
(2) useful clinically now Flos Magnoliae distillate collunarium, but the distillate sorptive power is poor, and be subjected to powder long-pending inhomogeneous, and also having the monomeric usage of Flos Magnoliae volatile oil is injection, the patient can not self-administration.
(3) patent of invention of Liu Ge Yin of Zhengzhou City, Henan Province " a kind of medicine spray for the treatment of rhinitis " is a raw material with Chinese medicine and western medicine such as Flos Magnoliae volatile oil, chlorobutanol, Mentholum, sodium benzoate, wherein contains chemical drugs, and is bigger to the side effect of human body.
(4) now the Chinese medicine spray is compound preparation mostly, and medical material is co-extracted volatile oil, and Flos Magnoliae volatile oil can not extract fully, and is accurate inadequately to the research of pharmacodynamics.
2, the chemical constituent of Flos Magnoliae
The Flos Magnoliae main component has volatile oil, alkaloid, and Lignanoids compounds etc., wherein the index composition is a volatile oil.Adopt GC-MS computer multiple techniques, isolation identification goes out 91 compositions from Flos Magnoliae volatile oil, accounts for 97.37% of total oil; In the YULAN volatile oil isolation identification 71 compositions, account for 97.66% of total oil; In the Flos Magnoliae volatile oil isolation identification 88 compositions, account for 95.68% of total oil.Flos Magnoliae volatile oil not only content tourney is worked as the YULAN height, and some active component are worked as the YULAN height as the content tourney of 1,8 cineole, linalool, Camphora, alpha-terpineol, farnesol etc.
Summary of the invention
The invention provides a kind of magnolia flower volatile oil spray for nose, slow to solve present endo-medicine onset, it is long to reach the disease sites time, and increases the weight of the problem to the burden of body excretes.
The present invention takes technical scheme to be: it is by making by the crude drug and the adjuvant of following volume parts ratio:
Flos Magnoliae volatile oil 40～60ml, polyoxyethylene sorbitan monoleate 40～60ml,
Glycerol 20～32ml.
Be preferably:
Flos Magnoliae volatile oil 50ml, polyoxyethylene sorbitan monoleate 50ml, glycerol 26ml.
Preparation method of the present invention is:
Get polyoxyethylene sorbitan monoleate and glycerol in the prescription, add 100～500ml dehydrated alcohol, stirring makes it dissolving, under stirring with the rotating speed of 900～1100r/min, slowly add Flos Magnoliae volatile oil after, with the rotating speed stirring of 900～1100r/min 10 minutes, add distilled water again to being settled to 1000ml, through 0.22 μ m microporous filter membrane, filter packing.
Drug quality control method of the present invention comprise differentiate and content assaying method as follows:
Discrimination method:
(1) gets this product 2ml, put in the 5ml measuring bottle, add dehydrated alcohol and be diluted to scale, shake up, as need testing solution; Other gets eucalyptole reference substance 1ml, puts in the 100ml measuring bottle, adds dehydrated alcohol and is diluted to scale, shakes up, in contrast product solution.Test according to thin layer chromatography (Chinese Pharmacopoeia version appendix in 2005 IV B), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-cyclohexane extraction (1: 9) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the same color speckle;
(2) in the chromatogram that writes down under eucalyptole assay item, the main peak retention time of eucalyptole should be consistent with the main peak retention time of eucalyptole reference substance in the test sample.
Assay:
Total volatile oil is measured according to determination of volatile oil method (Chinese Pharmacopoeia version appendix in 2005 X D first method);
Algoscopy is got this product 20ml, puts in the flask, adds water 300ml and bead number, after jolting mixes, connects volatile oil determination apparatus and reflux condensing tube.Autocondensation pipe upper end adds water makes the scale part that is full of volatile oil determination apparatus, and overflow when going into flask till.Put slowly to be heated in the electric jacket and boil, and keep little and boiled about 5 hours, oil mass no longer increases to the determinator, stops heating, places a moment, opens the piston of determinator lower end, and water is slowly emitted, and arrives to the oil reservoir upper end above scale 0 line till the 5mm place.Place more than 1 hour, opening piston again, to make oil reservoir drop to its upper end just concordant with scale 0 line, reads the volatilization oil mass, and volatile oil contents in the calculating test sample.
This product contain volatile oil should be labelled amount 90.0%～110.0% between.
Eucalyptole is measured according to gas chromatography (appendix VI E).
Chromatographic condition and system suitability test GC-7900 gas chromatograph; Chromatographic column is HP-519091J-413 (30m*0.32mm) quartz capillary column.Fid detector (280 ℃); Temperature of vaporization chamber is: 260 ℃; Injector temperature is 220 ℃; Heating schedule: 80 ℃ of column temperatures, kept 25 minutes, the speed with 20 ℃/min rises to 220 ℃ (keeping 3 minutes) again; Split ratio is 10: 1.
The about 0.25g of eucalyptole reference substance is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 50ml measuring bottle, adds absolute methanol and is diluted to scale, shakes up, in contrast product solution;
This product 2ml is got in the preparation of need testing solution, puts in the 5ml measuring bottle, adds absolute methanol and is diluted to scale, shakes up, as need testing solution.
Algoscopy is got reference substance solution and each 1 μ l of need testing solution, and inject gas chromatograph writes down chromatogram respectively, by external standard method, with calculated by peak area, promptly.
This product contains eucalyptole (C
18H
18O) must not be less than 1.0% (g/ml).
The inventor mainly aspect chemical constituent, clinical practice and the compound preparation, still rests on basic research stage to the exploitation of Flos Magnoliae single preparation and the research of anti-allergic effects at the research of Flos Magnoliae.On the basis of using for reference ancient books and domestic and international similar research, explore the Flos Magnoliae volatile oil preparation technology and the nasal spray dosage form that are fit to suitability for industrialized production, and prove that it has definite antiinflammatory, anti-allergic effects through pharmacological experiment, develop the magnolia flower volatile oil spray for nose new drug thus.
This product is flaxen clear liquid, fragrant odour, slightly foreign odor.
The present invention has dispersing wind and cold, clearing the nasal passage, antiallergic.Be used for acute, chronic rhinitis, allergic rhinitis, sinusitis, nasal obstruction, nasal sinusitis, turbid nasal discharge.Once spray 2 times on the one 4 times.
Specification, 10ml: contain the 0.5ml Flos Magnoliae volatile oil.
Pharmacodynamic test of active extract data of the present invention:
One, experiment material
1.1 medicine
Be subjected to the reagent thing
Title: Flos Magnoliae volatile oil, hereinafter to be referred as: VOMP.
Product batch number: 040312,
The unit of providing: Jilin Huinan Tiantai Pharmaceutical Co., Ltd..
Test sample preparation: the time spent is 10% the medicinal liquid that 0.5%Tween-80, glycerol, second alcohol and water are mixed with normal saline preparation desired concn per os experimental drug thing.
Preservation condition: room temperature preservation
Title: magnolia flower volatile oil spray for nose
Product batch number: 040401
The unit of providing: Jilin Huinan Tiantai Pharmaceutical Co., Ltd.
Specification: 6ml/ bottle, 10% volatile oil
Preservation condition: room temperature preservation
Positive control drug
Title: triamcinolone acetonide nasal spray (BINUO)
Product batch number: 20050203
Specification: the 6ml/ bottle, every ml contains triamcinolone acetonide 1.1mg, presses for every bottle 120, whenever presses 55 μ l.
Manufacturer: the auspicious pharmaceutical Co. Ltd in source, Kunming produces
Title: aspirin (ASP)
Product batch number: 050327
Specification: 0.3g/ sheet
Growth producer: Shaanxi white deer pharmaceutical factory
Title: prednisone acetate tablets (Pns)
Product batch number: 20050620
Specification: 5mg/ sheet
Growth producer: Shandong XinHua Pharmacy stock Co., Ltd
Title: dexamethasone acetate tablets (Dex)
Product batch number: 20050413
Specification: 0.75mg/ sheet
Growth producer: Shandong XinHua Pharmacy stock Co., Ltd
The negative control medicine
Title: sodium chloride injection (NS)
Product batch number: 050914
Manufacturer: Shangdong Hualu Pharmaceutical Co., Ltd.
1.2 reagent
Glacial acetic acid: Laiyang City is the chemical industry company limited in pairs, lot number: 20041115
Azovan blue: Shanghai chemical reagent purchasing and supply station provides lot number: 20030306.
Dimethylbenzene: Laiyang City is the chemical industry company limited in pairs, lot number: 20011023
Histamine phosphate: the beautiful pearl east wind in Shanghai Bioisystech Co., Ltd produces, lot number: 20050816
2,4-toluene-2,4-diisocyanate (TDI): Shanghai reagent one factory, lot number 20040816.
Benzylpenicillin sodium for injection: promise pharmaceutcal corporation, Ltd in the stone medicine group, lot number 20051020.
Carrageenin: Sigma company product, lot number 45F-0427.
Total protein quantitative test box: bio-engineering research institute is built up in Nanjing.
1.3 instrument
The LD5-2A centrifuge, Beijing Medical Centrifugal Machine Factory.
The FA1004 electronic balance, Shanghai balance equipment factory.
721 spectrophotometers, Shanghai the 3rd analytical tool factory.
YLS-7A toes volumetric measurement instrument, Shandong Academy of Medical Sciences equipment station.
Micro sample adding appliance, German Eppendorf company
Inverted phase contrast microscope (Japanese OLYMPUS),
BX50 type optical microscope (Japanese OLYMPUS)
FJ-2008P type automatic register (Xi'an 262 factories),
1.4 laboratory animal
The Wistar rat, the SPF level, male and female have concurrently, body weight 180～200g; Available from Shandong University's Experimental Animal Center, production licence number: SCXK (Shandong) 20030004.
Kunming mouse, the SPF level, male, body weight 20 ± 2g is provided by Shandong University's Experimental Animal Center, laboratory animal production licence number: SCXK (Shandong) 2003004.
Cavia porcellus, male and female half and half, body weight 300～350g is available from plant of academy of agricultural sciences, Shandong Province herding institute [credit number: SCXK (Shandong) 20030013].
Two, behind the Flos Magnoliae volatile oil oral administration to the influence of acute inflammation animal model
1. experimental technique
1.1 Flos Magnoliae volatile oil is to the influence of mouse peritoneal capillary permeability
Get 50 of healthy male mouse of kunming, be divided into 5 groups, be i.e. VOMP high, medium and low (30,15,7.5ml/kg) dosage group, aspirin (200mg/kg) group, NS group by the body weight stratified random.Every day, gastric infusion was 1 time, successive administration 3 days.Half an hour after the last administration, the normal saline solution of each mouse tail vein injection 2% azovan blue (0.10ml/10g body weight), the while is lumbar injection 0.6% acetum 0.20ml immediately.Take off cervical vertebra after 20 minutes and put to death animal, with 6ml normal saline flushing abdominal cavity.With centrifugal 15 minutes of the flushing liquor 3000rpm that collects, get supernatant and measure absorbance (OD value representation) in 590nm place, comparison between organizing.
1.2 Flos Magnoliae volatile oil is to the influence of mouse skin capillary permeability
Get 50 of healthy kunming mices, male, be divided into 5 groups at random by body weight, i.e. VOMP high, medium and low (30,15,7.5ml/kg) dosage group, Pns (10mg/kg) group, NS group.Every day, gastric infusion was 1 time, successive administration 3 days.After the last administration 1 hour, the normal saline of each mouse tail vein injection 1% azovan blue (0.10ml/10g body weight), and evenly smear 4ml dimethylbenzene (about 1.5 * 1.5cm in abdominal part depilation district immediately
2) cause inflammation.Put to death animal after 20 minutes, peel off abdominal part locus coeruleus skin, (diameter=1.2cm) takes off skin with card punch to select blue bosom.After it is shredded, be soaked in and the 4ml aqueous acetone solution be housed (acetone: water=7: 3 contains 0.3% anhydrous Na
2SO
4) teat glass in, capping test tube mouth.Soaked jolting every day 3～4 times two days.3000rpm is centrifugal 15 minutes two days later, gets supernatant in 721 spectrophotometer 590nm place colorimetrics, does not soak the aqueous acetone solution zeroing of skin, measures its absorbance, compares between organizing.
1.3 the influence of Flos Magnoliae volatile oil xylol induced mice auricle edema
Get 50 of healthy male mouse of kunming, be divided into 5 groups at random, be i.e. VOMP high, medium and low (30,15,7.5ml/kg) dosage group, Dex (5mg/kg) group, NS group by body weight.Every day gastric infusion once, successive administration 3 days (Dex group lumbar injection dexamethasone).After the art time administration 1 hour, every wide two sides of mouse right ear was evenly smeared dimethylbenzene 30 μ l and is caused inflammation, and left ear is not coated with and compares.Cause scorching back 45min and take off cervical vertebra execution animal.Card punch with diameter 8mm takes off left and right sides auricle, weighs respectively.Represent the ear swelling degree with left and right sides auricle weight difference, compare between organizing.
1.4 the Flos Magnoliae total volatile oil is to the influence of carrageenin induced mice foot swelling
Get 50 of healthy male mouse of kunming, be divided into 5 groups at random, be i.e. VOMP high, medium and low (30,15,7.5ml/kg) dosage group, Pns (10mg/kg) group, NS group by body weight.Every day gastric infusion once, successive administration 5 days.Half an hour after the art time administration, every sufficient plantar subcutaneous injection 1% carrageenin 0.03ml in a mice left side causes inflammation.Put to death animal after causing scorching 4 hours, the neat ankle of each Mus is cut two metapedes, weighs respectively, represents the swelling degree with two sufficient weight differences, compares between organizing.
1.5 Flos Magnoliae volatile oil causes the influence of rat paw edema to Ovum Gallus domesticus album
Get 50 of healthy male Wistar rats, be divided into 5 groups, be i.e. VOMP high, medium and low (20,10,5ml/kg) dosage group, Pns (10mg/kg) group, NS group by the body weight stratified random.Mark in the right back ankle of each Mus joint before the experiment.Measure the right back sufficient volume twice of rat with volumetric method, average and make the preceding normal volume of administration.Be administered once in first day, half an hour behind second day gastric infusion, every subcutaneous inserting needle of the right back sufficient sole of the foot portion of rat is injected 10% Ovum Gallus domesticus album normal saline solution 0.1ml and is caused inflammation to ankle.Respectively at cause scorching back 30,60,120,180,240,300min measures right back sufficient volume, calculate each rat cause scorching before and after right back sufficient volume-variation value, judge the medicine anti-inflammatory effect with the swelling degree, between organizing relatively.
1.6 Flos Magnoliae volatile oil is to the influence of rat pleuritis model
Get 60 healthy male Wistar rats, be divided into 6 groups, be i.e. VOMP high, medium and low (20,10,5ml/kg) dosage group, Pns (10mg/kg) group, NS group, normal control group by the body weight stratified random.Gastric infusion every day once (NS group and normal control group filling stomach normal saline), successive administration 7 days.After the last administration 1 hour, the physiological saline solution solution of all the other each treated animal intrapleural injection 1% carrageenin (0.20ml/100g body weight) caused inflammation except that the normal control group, normal control group thoracic cavity injection equivalent physiological saline solution.
Cause scorching back 8 hours, with the animal sacrificed by exsanguination.Measure the amount of thoracic cavity transudate, use the protein content in the Coomassie brilliant blue method mensuration transudate, and it is carried out differential blood count, compare between organizing.
1.7 Flos Magnoliae volatile oil is to the influence of normal mouse inflammatory reaction
Get 50 of healthy male mouse of kunming, be divided into 5 groups at random, be i.e. VOMP high, medium and low (30,15,7.5ml/kg) dosage group, Pns (10mg/kg) group, NS group by body weight.Perfusion every day once, successive administration 5 days.Before the last administration, survey its left back sufficient volume (getting twice meansigma methods) with mice foot volume determination device.Half an hour after the administration, every left back sufficient plantar subcutaneous injection 0.2% normal saline solution 0.03ml of histamine phosphate of mice causes inflammation.Measure mice inflammation foot volume respectively causing scorching back 30,60,90 and 120min, calculate the swelling degree, between organizing relatively.
1.8 Flos Magnoliae volatile oil is to the influence of adrenalectomy mice inflammatory reaction
Get 50 of male mouse of kunming, be divided into 5 groups at random, be i.e. VOMP high, medium and low (30,15,7.5ml/kg) dosage group, Pns (10mg/kg) group, NS group by body weight.Under aseptic condition, surgical removal mice both sides adrenal gland.3 days infection of postoperative intramuscular injection penicillin, and with the raising of sugar-salt-water replacement tap water.The beginning administration of one week of operation back, once a day, continuous 5 days.Cause inflammation by 1.7 methods, mensuration causes scorching front and back mice foot volume, calculates paw swelling, compares between organizing.
2. experimental result
2.1 Flos Magnoliae volatile oil is to the influence of mouse peritoneal capillary permeability
Result of the test shows that the middle and high dosage of VOMP can significantly reduce the azovan blue absorbance, compares with the NS group, significant difference (P＜0.01 or P＜0.05) is arranged, and be certain dose-effect relationship.The aspirin matched group also can significantly reduce the azovan blue absorbance, the results are shown in Table 1.
Table 1.VOMP is to the influence of mouse peritoneal capillary permeability (n=10, X ± SD)
Annotate: compare with the NS matched group;
*P＜0.05,
*P＜0.01
2.2 Flos Magnoliae volatile oil is to the influence of mouse skin capillary permeability
Result of the test shows that the remarkable reduction absorbance of the middle and high dosage energy of VOMP dose dependent is compared with the NS group, and significant difference (P＜0.01 or P＜0.05) is arranged.The Pns matched group also can significantly reduce the azovan blue absorbance, the results are shown in Table 2.
Table 2.VOMP is to the influence of mouse skin capillary permeability (n=10, X ± SD)
Annotate: compare with the NS matched group;
*P＜0.05,
*P＜0.01
2.3 the influence of Flos Magnoliae volatile oil xylol induced mice auricle edema
Result of the test shows that the middle and high dosage of Flos Magnoliae volatile oil all can make auricle swelling degree descend, and with the NS group significant difference (p＜0.05 or p＜0.01) is arranged relatively, and is the dose-effect dependency.The Dex matched group also can significantly reduce auricle swelling degree, the results are shown in Table 3.
Table 3.VOMP is to the influence of mouse peritoneal capillary permeability (n=10, X ± SD)
Annotate: compare with the NS matched group;
*P＜0.05,
*P＜0.01
2.4 the Flos Magnoliae total volatile oil is to the influence of carrageenin induced mice foot swelling
Result of the test shows that each dosage of VOMP all can significantly reduce swelling degree of the paw, compares with the NS group, and significant difference (P＜0.01 or P＜0.05) is arranged, and it is remarkable wherein to reduce the swelling degree of the paw effect with the VOMP high dose especially.
The Pns matched group also can significantly reduce swelling degree of the paw, the results are shown in Table 4.
Table 4.VOMP is to the influence of carrageenin induced mice pedal swelling (n=10, X ± SD)
Annotate: compare with the NS matched group;
*P＜0.05,
*P＜0.01
2.5 Flos Magnoliae volatile oil causes the influence of rat paw edema to Ovum Gallus domesticus album
Result of the test shows that VOMP high dose 30～300min after causing inflammation can significantly reduce swelling degree of the paw, compares with the NS group, and significant difference (P＜0.01) is arranged, and dosage 180～300min after causing inflammation can significantly reduce swelling degree of the paw (P＜0.01) among the VOMP.The Pns matched group also can significantly reduce swelling degree of the paw after causing inflammation, the results are shown in Table 5.
Table 5.VOMP causes influence (n=10, the X ± SD) of pedal swelling to rat Ovum Gallus domesticus album
Annotate: compare with the NS matched group;
*P＜0.05,
*P＜0.01
2.6 influence to rat pleuritis model
Result of the test shows, protein content and total white blood cells in model control group rat thoracic cavity seepage discharge, the transudate, all be significantly higher than the normal control group, and the neutrophilic granulocyte ratio obviously raises, be typical acute inflammation performance, illustrate that thoracic cavity injection carrageenin causes the rat pleuritis, the modeling success.
Result of the test shows, the VOMP high dose can significantly reduce total white blood cells in the transudate after causing inflammation, reduce the amount of thoracic cavity transudate, compare with the NS group, significant difference (P＜0.01) is arranged, and wherein dosage can obviously reduce protein content and total white blood cells (P＜0.01) in the transudate among the VOMP after causing inflammation.The Pns matched group also reduces protein content and the total white blood cells in the transudate after causing inflammation, suppress to ooze out, and the results are shown in Table 6.
Table 6 VOMP is to the influence of rat pleuritis transudate total amount, protein content and quantity of leucocyte (n=10, X ± SD)
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
2.7 Flos Magnoliae volatile oil is to the influence of normal mouse inflammatory reaction
Result of the test shows, VOMP high dose 60～150min after causing inflammation can significantly reduce the normal mouse paw swelling, compare with the NS group, significant difference (P＜0.05) is arranged, dosage 30min after causing inflammation can significantly reduce normal mouse swelling degree of the paw (P＜0.05) among the VOMP.The Pns matched group also can significantly reduce paw swelling after causing inflammation, the results are shown in Table 7.
Table 7.VOMP is to the influence of normal mouse foot swelling due to the histamine (n=10, X ± SD)
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
2.8 Flos Magnoliae volatile oil is to the influence of adrenalectomy mice inflammatory reaction
Result of the test shows, VOMP high dose 30～150min after causing inflammation can significantly reduce adrenalectomy mice paw swelling, compare with the NS group, significant difference (P＜0.05 or P＜0.01) is arranged, and dosage 90～150min after causing inflammation also can significantly reduce adrenalectomy mice paw swelling (P＜0.05 or P＜0.01) among the VOMP.The Pns matched group can significantly reduce paw swelling after causing inflammation, the results are shown in Table 8.
Table 8.VOMP is to the influence of adrenalectomy mice foot swelling due to the histamine (n=10, X ± SD)
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
3. conclusion
3.1 Flos Magnoliae volatile oil Dichlorodiphenyl Acetate induced mice abdominal cavity capillary permeability increases obvious inhibitory action is arranged.
3.2 the increase of Flos Magnoliae volatile oil xylol induced mice skin capillary permeability has obvious inhibitory action.
3.3 Flos Magnoliae volatile oil can obviously suppress dimethylbenzene induced mice auricle edema.
3.4 Flos Magnoliae volatile oil has remarkable inhibitory action to the mice pedal swelling that carrageenin causes.
3.5 the rat paw edema that Flos Magnoliae volatile oil causes Ovum Gallus domesticus album presents the obvious suppression effect.
3.6 Flos Magnoliae volatile oil can reduce the amount of the pleuritic transudate of rat due to the carrageenin, reduces protein content and total white blood cells in the transudate.
Cause normal mouse and the foot swelling of adrenalectomy mice 3.7 Flos Magnoliae volatile oil can obviously alleviate histamine, the antiinflammatory action of prompting Flos Magnoliae volatile oil does not rely on adrenal existence.
Three, to the influence of chronic inflammatory disease animal model (granuloma induced by implantation of cotton pellets)
1. experimental technique
Get 50 of healthy male mouse of kunming, be divided into 5 groups at random, be i.e. VOMP high, medium and low (30,15,7.5ml/kg) dosage group, Dex (5mg/kg) group, NS group by body weight.The administration same day (Dex group lumbar injection dexamethasone), aseptic condition operation down is at a 10mg sterilization of the subcutaneous implantation of mouse armpit portion cotton balls.2 infection of postoperative intramuscular injection penicillin.Be administered once every day, and successive administration was put to death animal after 7 days, peels off the cotton balls of band granulation tissue, and baking was weighed after 12 hours in 60 ℃ of baking ovens, was calculated as follows granulation weight, compared between organizing.
Cotton balls weight-cotton balls the original weight of granulation weight=band granulation tissue
2. experimental result
Result of the test shows that the middle and high dosage of Flos Magnoliae volatile oil all can make granulation tissue weight descend, and with the NS group significant difference (p＜0.05 or p＜0.01) is arranged relatively, and is wherein the strongest with the effect of dosage minimizing granulation tissue weight among the VOMP especially.The Dex matched group also can significantly reduce granulation tissue weight, the results are shown in Table 9.
Table 9.VOMP is to the influence of mice cott on pellet-induced granuloma formation (n=10, X ± SD)
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
3. conclusion
Flos Magnoliae volatile oil has remarkable inhibitory action to the mice cott on pellet-induced granuloma formation
Four, Flos Magnoliae volatile oil is to the influence of rat assist agent arthritis
1. experimental technique
Make complete Freund ' s adjuvant by oneself by literature method.Get 120 of healthy male Wistar rats, i.e. VOMP high, medium and low (20,10,5ml/kg) dosage group, Dex (5mg/kg) group, NS group.Be administered once every day, continuous 3 times, measures the right back sufficient normal volume of rat (getting meansigma methods twice), every the right back sufficient sole of the foot intradermal injection Freund ' s of portion adjuvant 0.1ml sensitization of rat after the administration before the administration for the third time.Be administered once again in 8 hours after the sensitization, and measured right back sufficient volume, swelling degree (swelling degree=sensitization metapedes volume-sensitization front foot volume) after the calculating sensitization at 18 hours
[6], compare between organizing.
2. experimental result
Result of the test shows, the VOMP high dose can significantly reduce paw swelling in 18 hours after sensitization, compare with the NS group, and significant difference (P＜0.05) is arranged.The Dex matched group also can obviously reduce paw swelling in 18 hours after sensitization, the results are shown in Table 10.
Table 10.VOMP is to the influence of rat assist agent arthritis foot swelling (n=10, X ± SD)
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
3. conclusion: heavy dose of Flos Magnoliae volatile oil has obvious inhibitory action to the constitutional pedal swelling due to the Freund adjuvant.
Five, the VOMP nasal spray is to the therapeutical effect of zoopery allergic rhinitis
1. experimental technique
1.1 therapeutical effect to the rat allergic rhinitis
Get 70 of healthy rats, adaptability was fed 3 days, was divided into two groups according to body weight, sex stratified random.First group is modeling group (60), and the TDI olive oil solution 10 μ l with 10% splash into the two nostrils, front side (every side 5 μ l) of rat with sample injector, once a day, and continuous 7 days sensitization.Give TDI (keeping the phase) next day of changing into after 7 days, continuous 14 days.Second group is normal control group (10), uses the olive oil collunarium merely, and method is the same.From sensitization the 3rd day, the next day, once regularly observed and gives the sniffle in 30 minutes behind TDI or the olive oil, observes continuously 3 times, and according to table 1 scoring, average mark surpasses 5 and is divided into the modeling success.
Select the animal of modeling success,, be divided into four groups at random, be respectively the high, medium and low dosage group of model control group, BINUO matched group and VOMP nasal spray, every group of 10 animals according to symptom score value, body weight, sex layering.Keep the phase nose spray (promptly sensitization after 7 days) first every day and give relative medicine, NS matched group and model control group nose spray normal saline, except that VOMP nasal spray high and low dose administration every day 4 times and 1 time, all the other respectively organize administration every day 2 times, whenever press 50 μ l, successive administration 14 days.From sensitization the 9th day (being administration the 2nd day), observe sniffle in 30 minutes after administration, the next day, observe once, and mark according to table 11, observe and finish after the interior mastocyte of 5 high power field nasal mucosa lamina proprias of Toluidine blue staining counting compares between organizing.
Table 11.TDI brings out animal pattern nose allergy sniffle standards of grading
1.2 therapeutical effect to the Cavia porcellus allergic rhinitis
Get 48 of healthy guinea pigs, adaptability was fed 3 days, was divided into two groups according to body weight, sex stratified random.First group is modeling group (40), and the TDI olive oil solution 10 μ l with 10% splash into the two nostrils, front side (every side 5 μ l) of rat with sample injector, once a day, and continuous 7 days sensitization.Give TDI (keeping the phase) next day of changing into after 7 days, continuous 14 days.Second group is normal control group (10), uses the olive oil collunarium merely, and method is the same.From sensitization the 6th day, observe every day to the sniffle in behind TDI or the olive oil 30 minutes, observed continuously 2 days, according to table 11 scoring, average mark surpasses 5 and is divided into the modeling success.
Select the animal of modeling success,, be divided into four groups at random, be respectively the high, medium and low dosage group of model control group, BINUO matched group and VOMP nasal spray, every group of 8 animals according to symptom score value, body weight, sex layering.Keep the phase nose spray (promptly sensitization after 7 days) first every day and give relative medicine, NS matched group and model control group nose spray normal saline, except that VOMP nasal spray high and low dose administration every day 4 times and 1 time, all the other respectively organize administration every day 2 times, whenever press 50 μ l, successive administration 14 days.From sensitization the 9th day (being administration the 2nd day), observe sniffle in 30 minutes after administration, the next day, observe once, and mark according to table 1, observe and finish after the interior eosinophilic granulocyte of 5 high power field nasal mucosa lamina proprias of Toluidine blue staining counting compares between organizing.
2. result of the test
2.1VOMP nasal spray is to the therapeutical effect of rat experiment allergic rhinitis
2.1.1 rat experiment allergic rhinitis symptom score
Model control group has all occurred the symptom of rhinocnesmus and sneeze in various degree, but has not had obvious thin nasal discharge after giving TDI the 1st time in the 10min, along with time lengthening, above-mentioned symptom increases the weight of gradually and thin nasal discharge occurs.Typical rhinocnesmus, sneeze and thin nasal discharge come across to TDI after 5 days.
After the Drug therapy, each dosage group of VOMP nasal spray is the 1st～4 day and model control group no significant difference after administration, administration after 6 days VOMP nasal spray height, middle dosage group rhinitis symptom begin obviously to alleviate, its symptom score has been compared significant difference (P＜0.01 or P＜0.05) (table 13) with model control group.The BINUO matched group also can obviously alleviate the rhinitis symptom in administration after 4 days, the results are shown in Table 12.
Table 12.VOMP nasal spray is to rat experiment allergic rhinitis symptom score (n=10, X ± SD)
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
2.1.2 nasal mucosa mast cell counts
Nasal mucosa shows that through Toluidine blue staining mastocyte is high-visible in the model control group rat nasal mucosa lamina propria, and cell number is compared showed increased than normal control group, and significant difference (P＜0.01) is arranged; VOMP nasal spray height, middle dosage group mastocyte number are compared obvious minimizing with model control group, its difference has significance statistical significance (P＜0.01); BINUO matched group mastocyte number also obviously is less than model control group (P＜0.01), the results are shown in Table 13.
Table 13.VOMP nasal spray soaks into influence (the n=10 X ± SD) of cell number to the rat experiment allergic rhinitis
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
2.2 the VOMP nasal spray is to the therapeutical effect of Cavia porcellus allergic rhinitis
2.21 Cavia porcellus allergic rhinitis symptom score
Result of the test shows, model control group does not have tangible rhinocnesmus, sneeze and thin nasal discharge occurring typical rhinocnesmus, sneeze and thin nasal discharge blank group after 5 days for TDI.After the Drug therapy, each dosage group of VOMP nasal spray is the 1st～3 day and model control group no significant difference after administration, administration after 3 days VOMP nasal spray height, middle dosage group Cavia porcellus rhinitis symptom begin obviously to alleviate, its symptom score has been compared significant difference (P＜0.01 or P＜0.05) with model control group.The BINUO matched group begins to alleviate rhinitis symptom (P＜0.05) in administration after 2 days, to administration after the 6th day symptom obviously alleviate (P＜0.01), the results are shown in Table 14.
Table 14.VOMP nasal spray is to Cavia porcellus experimental allergic rhinitis symptom score (n=8, X ± SD)
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
2.2.2 nasal mucosa eosinophil count
Nasal mucosa shows through Toluidine blue staining, the mucosa place of the false multiple layer cilium columnar epithelium of model control group Cavia porcellus lining, visible obviously eosinophilic granulocyte is soaked in the lamina propria, and the normal matched group of cell number is compared showed increased, and significant difference (P＜0.01) is arranged; VOMP nasal spray height, middle dosage group cell number are compared obvious minimizing with model control group, its difference has significance statistical significance (P＜0.01); BINUO cellular control unit number also obviously is less than model control group (P＜0.01), the results are shown in Table 15.
Table 15.VOMP nasal spray soaks into the influence of cell number to Cavia porcellus experimental allergic rhinitis
Annotate: compare according to group with model;
*P＜0.05,
*P＜0.01
3. conclusion
3.1 magnolia flower volatile oil spray for nose can obviously reduce experimental allergic rhinitis rat sniffle score value, can alleviate symptoms such as nose rhinocnesmus, sneeze, thin nasal discharge, also can reduce mastocyte in the partial infiltration of inflammation.
3.2 magnolia flower volatile oil spray for nose can obviously reduce experimental allergic rhinitis Cavia porcellus sniffle score value, can alleviate symptoms such as nose rhinocnesmus, sneeze, thin nasal discharge, also can reduce the eosinophilic granulocyte in the partial infiltration of inflammation.
Six, evaluation and conclusion
Allergic rhinitis is a kind of commonly encountered diseases frequently-occurring disease, along with the change of society, and industrial expansion, its sickness rate is more and more higher, and does not still have safe, quick-acting medicine at present.This paper has studied the therapeutical effect of magnolia flower volatile oil spray for nose to allergic rhinitis, at first, observed the peroral administration antiinflammatory action of VOMP at various acute inflammatory animal model (acetic acid causes that the mouse peritoneal capillary permeability increases, mice caused by dimethylbenzene xylene skin heart permeability increases, mice caused by dimethylbenzene xylene ear swelling, carrageenin cause the mice foot swelling, Ovum Gallus domesticus album causes rat paw edema, carrageenin causes the rat pleuritis), mice granuloma induced by implantation of cotton pellets chronic inflammation model, rat assist agent arthritis (immune inflammation) model.On rat allergic rhinitis model, Cavia porcellus allergic rhinitis model, observed the therapeutical effect of magnolia flower volatile oil spray for nose to allergic rhinitis.Result of study shows, Flos Magnoliae volatile oil can be by reducing the vascular permeability of inflammation tissue, the volumetric expansion of inflammation-inhibiting tissue and weight increase, suppress granulation tissue hyperplasia, alleviate the local anaphylaxis symptom, reduce eosinophilic granulocyte and mastocyte and bring into play its antiinflammatory anti-allergic effects, can use in the acute stage and the later stage of allergic rhinitis in approach such as the partial infiltrations of inflammation.
Acute toxicity testing
1, chmice acute toxicity test
Behind the magnolia flower volatile oil spray for nose gastric infusion animal toxicity reaction and the time of being poisoned to death many after administration the same day.The LD of mouse stomach single-dose
50Be 13.6ml/kg, 95% credibility interval is 10.38～15.30ml/kg.
2, rabbit acute toxicity testing
The magnolia flower volatile oil spray for nose nasal membrane sprays into after the administration animal toxicity reaction and only shows as nasal discharge and increase, and occurs at once after the administration.With maximum dosage-feeding, promptly the 12h successive administration is 24 times, per half an hour once, each 50 μ l, animal does not have death after the administration, no serious toxicity reaction symptom.
The repeat administration toxicity test
1, rabbit nose spray repeat administration toxicity test
With the new zealand white rabbit is animal subject, and magnolia flower volatile oil spray for nose is to rabbit spray nasal administration one month, and dosage is respectively high dose group and solvent matched group every day 8 times, low dose group every day 4 times, each 50ul, continuous one month.24h, 7 days dead 3 animals in every component other places after the drug withdrawal.
Generally reaction after the administration: animal subject cough, restlessness occur, grabs the nose phenomenon after the high dose group administration, recovers normal behind about 2h.The low dose group individual animal has cough, restlessness once in a while, grabs the nose phenomenon, but very fast recovery is normal, no asthma, local excitation symptom such as suffocate.The solvent matched group does not have and to cough, suffocate, restlessness, grab phenomenon such as nose generally in order.Each is organized animal subject and ingests, drinks water normal.
Pathological anatomy and histopathological examination: administration finishes back 24h and recovers all no abnormal change of nasal mucosa, trachea, bronchus, lung tissue of each group execution animal after 7 days.
Conclusion: with the new zealand white rabbit is animal subject, and magnolia flower volatile oil spray for nose is to rabbit spray nasal administration one month, being no more than every day and spraying nose 8 times under (each 50ul) condition, rabbit nasal cavity and respiratory mucosa is not had the overt toxicity effect.
2, rat is repeated the gastric infusion toxicity test
With the rat is animal subject, and magnolia flower volatile oil spray for nose gastric infusion, dosage are respectively 6,4,2ml/kg, and successive administration is 7 days weekly, altogether 5 weeks of successive administration; After the last administration, put to death 10 animals for every group; Remaining continuation observed for 2 weeks, put to death then.Observed content book according to plan requires to carry out.
Generally reaction after the administration: rat oral gavage gave magnolia flower volatile oil spray for nose 1 month, and the performance of the toxicity of high dose group animal is mainly low volt, movement disorder, symptom such as prostrate; In the same high dose group of dosage group symptom, but the symptom number of animals appears and duration of symptoms all is lower than high dose group; Above-mentioned symptom appears in low dose group only individual animal, but very fast recovery is normal.Respectively organize rat food ration, amount of drinking water and body weight in the experimentation and do not have marked difference.
Blood cell is learned and blood biochemical is learned: the hematological indices of each administration treated animal, blood biochemistry index and matched group are not relatively all found the significantly abnormal change relevant with drug effect.
Pathological anatomy and histopathological examination: dead animal postmortem naked eyes show no obvious abnormalities in the experimentation, and estimating may be because misoperation causes rats death.Administration was dissected when finishing, and each treated animal internal organs general appearance inspection is not found all that naked eyes are visible and obviously changed, histopathologic examination's kidney generation pathological changes, and high, middle dosage group lesion degree is relevant with dosage.Above-mentioned change recovers normal substantially after 2 convalescent periods in week.After administration finished, high, middle dosage group rat liver organ coefficient was significantly higher than matched group, and convalescent period when finishing the high dose group organ coefficient still be higher than matched group.According to this experiment condition and result, infer that it may be kidney and liver that rat oral gavage gives magnolia flower volatile oil spray for nose toxicity target organ.
Under this experiment condition, it is 2ml/kg that rat oral gavage gives 1 month non-toxic reaction dosage of magnolia flower volatile oil spray for nose, and toxic amounts of reactants is 4ml/kg.Results suggest, the oral administration Flos Magnoliae volatile oil is bigger to rat toxicity.
Five, local irritation experiment
1, magnolia flower volatile oil spray for nose nose irritant experiment
For investigating magnolia flower volatile oil spray for nose, spray at the rabbit nose and tried thing 50ul, every day 2 times, successive administration 7 days the partial zest of rabbit nose.The result shows that magnolia flower volatile oil spray for nose is to rabbit nose nonirritant.
2, magnolia flower volatile oil spray for nose eye irritant experiment
For investigating magnolia flower volatile oil spray for nose to the partial zest of tame lagophthalmos, splash into the magnolia flower volatile oil spray for nose of 50ul at the rabbit left eye, every day 3 times, 4 weeks of successive administration, the result shows that magnolia flower volatile oil spray for nose has slight zest to the eye part, but can disappear no secretions in administration conjunctival congestion after 24 hours, edema symptom.
To sum up, magnolia flower volatile oil spray for nose pharmacodynamics, safety research are shown the local nose spray of magnolia flower volatile oil spray for nose drug safety, effective to the allergic rhinitis treatment.
The specific embodiment
The Flos Magnoliae volatile oil production method: can adopt prior art to be prepared, the present invention is exemplified below:
Adopt technologies such as steam distillation, oil-water separation from Flos Magnoliae, to extract volatile oil and make single preparations of ephedrine, and existing product is compound preparation.Get the Flos Magnoliae alabastrum, pulverize, cross 20 mesh, be added in the vapor distillation still, add 10% water with the medical material part by weight and carry out moisteningly, feed steam then, the control steam pressure is 0.1Mpa, continuous still 10h.Volatile oil adds the anhydrous CaC of 10% (weight ratio) after the oil-water separation
12The desiccant dry filter promptly gets volatile oil.
Embodiment 1:
It is by making by the crude drug and the adjuvant of following volume parts ratio:
Flos Magnoliae volatile oil 40ml, polyoxyethylene sorbitan monoleate 40ml,
Glycerol 20ml.
Preparation method of the present invention is:
Get polyoxyethylene sorbitan monoleate and glycerol in the prescription, add the 100ml dehydrated alcohol, stirring makes it dissolving, under stirring with the rotating speed of 900r/min, slowly add Flos Magnoliae volatile oil after, stirred 10 minutes with the rotating speed of 900r/min, add distilled water again to being settled to 1000ml, through 0.22 μ m microporous filter membrane, filter packing.
Embodiment 2:
Flos Magnoliae volatile oil 50ml, polyoxyethylene sorbitan monoleate 50ml, glycerol 26ml.
Preparation method of the present invention is:
Get polyoxyethylene sorbitan monoleate and glycerol in the prescription, add the 300ml dehydrated alcohol, stirring makes it dissolving, under stirring with the rotating speed of 1000r/min, slowly add Flos Magnoliae volatile oil after, stirred 10 minutes with the rotating speed of 1000r/min, add distilled water again to being settled to 1000ml, through 0.22 μ m microporous filter membrane, filter packing.
Embodiment 3:
Flos Magnoliae volatile oil 60ml, polyoxyethylene sorbitan monoleate 60ml,
Glycerol 32ml.
Preparation method of the present invention is:
Get polyoxyethylene sorbitan monoleate and glycerol in the prescription, add the 500ml dehydrated alcohol, stirring makes it dissolving, under stirring with the rotating speed of 1100r/min, slowly add Flos Magnoliae volatile oil after, stirred 10 minutes with the rotating speed of 1100r/min, add distilled water again to being settled to 1000ml, through 0.22 μ m microporous filter membrane, filter packing.
Drug quality control method of the present invention comprise differentiate and content assaying method as follows:
Discrimination method:
(1) gets this product 2ml, put in the 5ml measuring bottle, add dehydrated alcohol and be diluted to scale, shake up, as need testing solution; Other gets eucalyptole reference substance 1ml, puts in the 100ml measuring bottle, adds dehydrated alcohol and is diluted to scale, shakes up, in contrast product solution.Test according to thin layer chromatography (Chinese Pharmacopoeia version appendix in 2005 IV B), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-cyclohexane extraction (1: 9) is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to the speckle colour developing clear; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the same color speckle;
(2) in the chromatogram that writes down under eucalyptole assay item, the main peak retention time of eucalyptole should be consistent with the main peak retention time of eucalyptole reference substance in the test sample.
Assay:
Total volatile oil is measured according to determination of volatile oil method (Chinese Pharmacopoeia version appendix in 2005 X D first method);
Algoscopy is got this product 20ml, puts in the flask, adds water 300ml and bead number, after jolting mixes, connects volatile oil determination apparatus and reflux condensing tube.Autocondensation pipe upper end adds water makes the scale part that is full of volatile oil determination apparatus, and overflow when going into flask till.Put slowly to be heated in the electric jacket and boil, and keep little and boiled about 5 hours, oil mass no longer increases to the determinator, stops heating, places a moment, opens the piston of determinator lower end, and water is slowly emitted, and arrives to the oil reservoir upper end above scale 0 line till the 5mm place.Place more than 1 hour, opening piston again, to make oil reservoir drop to its upper end just concordant with scale 0 line, reads the volatilization oil mass, and volatile oil contents in the calculating test sample.
This product contain volatile oil should be labelled amount 90.0%～110.0% between.
Eucalyptole is measured according to gas chromatography (appendix VI E).
Chromatographic condition and system suitability test GC-7900 gas chromatograph; Chromatographic column is HP-519091J-413 (30m*0.32mm) quartz capillary column.Fid detector (280 ℃); Temperature of vaporization chamber is: 260 ℃; Injector temperature is 220 ℃; Heating schedule: 80 ℃ of column temperatures, kept 25 minutes, the speed with 20 ℃/min rises to 220 ℃ (keeping 3 minutes) again; Split ratio is 10: 1.
The about 0.25g of eucalyptole reference substance is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 50ml measuring bottle, adds absolute methanol and is diluted to scale, shakes up, in contrast product solution;
This product 2ml is got in the preparation of need testing solution, puts in the 5ml measuring bottle, adds absolute methanol and is diluted to scale, shakes up, as need testing solution.
Algoscopy is got reference substance solution and each 1 μ l of need testing solution, and inject gas chromatograph writes down chromatogram respectively, by external standard method, with calculated by peak area, promptly.
This product contains eucalyptole (C
18H
18O) must not be less than 1.0% (g/ml).
Claims (1)
1. the detection method of a magnolia flower volatile oil spray for nose, this magnolia flower volatile oil spray for nose is by making by the crude drug and the adjuvant of following volume parts ratio: Flos Magnoliae volatile oil 40～60ml, polyoxyethylene sorbitan monoleate 40～60ml, glycerol 20～32ml, it is characterized in that the discrimination method in this method is as follows:
(1) gets this product 2ml, put in the 5ml measuring bottle, add dehydrated alcohol and be diluted to scale, shake up, as need testing solution; Other gets eucalyptole reference substance 1ml, puts in the 100ml measuring bottle, adds dehydrated alcohol and is diluted to scale, shakes up, in contrast product solution; Test according to thin layer chromatography, see Chinese Pharmacopoeia version appendix in 2005 VIB, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-cyclohexane extraction at 1: 9, launches, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100 ℃ to be heated to speckle colour developing clear; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the same color speckle;
(2) in the chromatogram that writes down under eucalyptole assay item, the main peak retention time of eucalyptole should be consistent with the main peak retention time of eucalyptole reference substance in the test sample;
Content assaying method in this method is as follows:
Total volatile oil is measured according to the determination of volatile oil method, sees Chinese Pharmacopoeia version appendix in 2005 X D first method;
Algoscopy is got this product 20ml, puts in the flask, adds water 300ml and bead number, after jolting mixes, connects volatile oil determination apparatus and reflux condensing tube; Autocondensation pipe upper end adds water makes the scale part that is full of volatile oil determination apparatus, and overflow when going into flask till; Put slowly to be heated in the electric jacket and boil, and keep little and boiled 5 hours, oil mass no longer increases to the determinator, stops heating, places a moment, opens the piston of determinator lower end, and water is slowly emitted, and arrives to the oil reservoir upper end above scale 0 line till the 5mm place; Place more than 1 hour, opening piston again, to make oil reservoir drop to its upper end just concordant with scale 0 line, reads the volatilization oil mass, and volatile oil contents in the calculating test sample;
This product contain volatile oil should be labelled amount 90.0%～110.0% between;
Eucalyptole is seen Chinese Pharmacopoeia version appendix in 2005 VI E according to gas chromatography determination;
Chromatographic condition and system suitability test GC-7900 gas chromatograph; Chromatographic column is HP-519091J-413,30m*0.32mm, quartz capillary column; Fid detector, its temperature are 280 ℃; Temperature of vaporization chamber is: 260 ℃; Injector temperature is 220 ℃; Heating schedule: 80 ℃ of column temperatures, kept 25 minutes, the speed with 20 ℃/min rises to 220 ℃ again, keeps 3 minutes; Split ratio is 10: 1;
Eucalyptole reference substance 0.25g is got in the preparation of reference substance solution, and accurate the title decides, and puts in the 50ml measuring bottle, adds absolute methanol and is diluted to scale, shakes up, in contrast product solution;
This product 2ml is got in the preparation of need testing solution, puts in the 5ml measuring bottle, adds absolute methanol and is diluted to scale, shakes up, as need testing solution;
Algoscopy is got reference substance solution and each 1 μ l of need testing solution, and inject gas chromatograph writes down chromatogram respectively, by external standard method, with calculated by peak area, promptly;
This product contains eucalyptole C
18H
18O must not be less than 1.0% (g/ml).
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| Title |
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| 吉晓滨等.辛夷对实验性变应性鼻炎红细胞免疫功能的影响.《广东医学》.化学工业出版社,2006,第27卷(第8期), * |
| 王东兴等.鼻腔给药新剂型研究进展.《中国新药杂志》.2002,第11卷(第8期), * |
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