CN108272998A - A kind of Mongolian compound preparation and preparation method thereof for treating insomnia - Google Patents

Abstract

The invention discloses a kind of preparation method of the Mongolian compound preparation for the treatment of of insomnia patients, Mongolian compound preparation is solid particle agent, and preparation method specifically includes following steps：Active compound crushing, the extraction of active compound coarse powder, inclusion essential oil, the drying of active compound extraction filtrate, granule molding；The preparation method takes full advantage of the medical value of medicinal material volatile ingredient and soluble component, strengthens drug effect；Mongolian compound preparation prepared by the present invention meets following quality requirement：When thin-layered chromatography differentiates the Amomum cardamomum in Mongolian compound preparation, Mongolian compound preparation shows identical blue spot in eucalyptol reference substance corresponding position, negative noiseless；When high performance liquid chromatography differentiates stir-baked SEMEN ZIZIPHI SPINOSAE in Mongolian compound preparation, Mongolian compound preparation has absorption peak in Spine Date Seed jujubosideB standard items corresponding position, negative noiseless.Mongolian compound preparation dissolves in 2min；Eucalyptol content, which is set to its content, must not be less than 1.5mg/g.The requirement of the above index is to ensure the important indicator of Mongolian compound preparation drug effect.

Description

A kind of Mongolian compound preparation and preparation method thereof for treating insomnia

Technical field

The present invention relates to anaesthetic technical fields, and in particular to a kind of Mongolian compound preparation that treating insomnia and its preparation side Method.

Background technology

With the development of society, the accelerating rhythm of life, prodigious change, external world's pressure also has occurred in people’s lives mode Power is big, mental labour is more, bad life habits etc. make the illness rate of insomnia rise violently year by year, the continuous rejuvenation of insomnia group. Insomnia is divided into primary insomnia and insomnia secondary, and insomniac will appear difficulty falling asleep, sleep quality declines, sleeping time subtracts Less, the clinical manifestations such as memory function decline cause be easy daytime sleepy since patient is bad in sleeping at night quality, and work energy Power declines, or even has certain harmful effect to the quality of life of patient, emotion.At present not only using western medicine insomnia With a variety of adverse reactions, it is also easy to that patient is made to generate dependence, therapeutic effect is undesirable, and Prescription of Mongolian Medicine has in its treatment There is some superiority.

Invention content

First of the present invention is designed to provide a kind of preparation method of Mongolian compound preparation that treating insomnia, fully profit With the medical value of medicinal material volatile ingredient and soluble component, drug effect is strengthened；Second object of the present invention is to provide A kind of Mongolian compound preparation for treating insomnia, moisture, granularity, dissolubility meet《Chinese Pharmacopoeia》（Version four in 2015） Required standard；Third purpose of the present invention provides a kind of method of evaluation Mongolian compound preparation qualification,

The present invention is realized by following technical solution：

A kind of preparation method of Mongolian compound preparation that treating insomnia, it is characterised in that：Mongolian compound preparation is solid particle agent, Preparation method specifically includes following steps：

Active compound crushes：After taking 6-12 parts 1-3 parts of fructus piperis longi, 2-4 parts of white lettuce victory, 3-6 parts of Amomum cardamomum, stir-fry of jujube to crush 24 mesh sieve Active compound coarse powder is made；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 9-11 parts of water is added, after mixing, impregnates 1-3 hours, then uses Volatile oil is collected after extraction by steam distillation 5-7h and active compound extracts filtrate；

Inclusion essential oil：Using polishing：Take volatile oil and β-CD by 1:9-1:10 ratio be mixed it is even after, add 3-4 times to measure It is filtered after water grinding 70-90min, filter residue 60-90 DEG C of petroleum ether 2-3 times, drying 3-5h under the conditions of 40-45 DEG C, system At volatile oil medicinal powder；

Active compound extracts filtrate drying：After active compound extracts the 1/9-1/11 that concentrating filter liquor extracts filtrate volume to active compound, then at 55- Extracting solution medicinal powder is made in dry 5-7h in 65 DEG C of drying box；The volatile oil medicinal powder is made after mixing with extracting solution medicinal powder At anaesthetic composition medicinal powder；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:0.8-1:1.2 ratio after mixing, uses 65-75% Ethanol wet, the dosage of the ethyl alcohol of 65-75% is the 15-25% of anaesthetic composition medicinal powder and excipient quality sum, after wetting Mongolian compound preparation is made under the conditions of 55-65 DEG C, after dry 3-5h.

Moreover, the Mongolian compound preparation is solid particle agent, preparation method specifically includes following steps：

Active compound crushes：Take 6 parts 1 part of fructus piperis longi, 2 parts of white lettuce victory, 3 parts of Amomum cardamomum, stir-fry of jujube that active compound was made after crushing 24 mesh sieve Coarse powder；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 10 parts of water is added, after mixing, impregnates 2 hours, then water is used to steam Steam distillation method collects volatile oil after extracting 6h and active compound extracts filtrate；

Inclusion essential oil：Using polishing：Take volatile oil and β-CD by 1:10 ratio be mixed it is even after, add 3 times amount water grinding It is filtered after 80min, filter residue 80 DEG C of petroleum ethers 2 times, dry 4h, is made volatile oil medicinal powder under the conditions of 40 DEG C；

Active compound extracts filtrate drying：After active compound extracts concentrating filter liquor to the 1/10 of active compound extraction filtrate volume, then at 60 DEG C Extracting solution medicinal powder is made in dry 6h in drying box；Anaesthetic group is made in the volatile oil medicinal powder and extracting solution medicinal powder after mixing Close object medicinal powder；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:1 ratio after mixing, with 70% ethanol wet, The dosage of 70% ethyl alcohol is the 20% of anaesthetic composition medicinal powder and excipient quality sum, after wetting under the conditions of 60 DEG C, drying Mongolian compound preparation is made after 4h.

Moreover, the excipient is soluble starch.

The present invention also provides a kind of Mongolian compound preparations for treating insomnia, multiple by a kind of above-mentioned anaesthetic for treating insomnia The preparation method of square preparation is prepared, and the content for treating eucalyptol in the Mongolian compound preparation of insomnia is more than 1.5mg/g.

The present invention also provides a kind of methods of evaluation Mongolian compound preparation qualification, it is characterised in that：It is prepared by the above method Mongolian compound preparation, following index request need to be met：

（1）When thin-layered chromatography differentiates the Amomum cardamomum in Mongolian compound preparation, Mongolian compound preparation is corresponded in eucalyptol reference substance Place shows identical blue spot, negative noiseless；

（2）When high performance liquid chromatography differentiates stir-baked SEMEN ZIZIPHI SPINOSAE in Mongolian compound preparation, Mongolian compound preparation is marked in jujuboside-B There is absorption peak in quasi- product corresponding position, negative noiseless；

（3）The moisture of Mongolian compound preparation, granularity, dissolubility meet《Chinese Pharmacopoeia》（Version four in 2015）Required standard；

（4）Eucalyptol content in gas chromatography measurement Mongolian compound preparation, which is set to its content, must not be less than 1.5mg/g；

The Mongolian compound preparation for meeting requirements above is qualified products；The Mongolian compound preparation for being unsatisfactory for requirements above is unqualified Product.The above method can be used for that identification is assisted to be expected the true and false of the Mongolian compound preparation listed in the future.

The application method of Mongolian compound preparation：Granule boiling water impulsive motion, stirring can take after mixing well.

The composition drug and chemical composition of Mongolian compound preparation：

Amomum cardamomum is the fruit of zingiberaceous plant Amomum cardamomum and amomum compactum Soland ex Maton.Main product is in the ground such as Vietnam, Thailand, Guangdong, Guangxi, cloud Also there is cultivation on the ground such as south.7-8 months pick.Acrid flavour, warm-natured, promoting the circulation of qi, middle benefit gas, preventing or arresting vomiting.Volatile oil is its principle active component, K containing organic/inorganic substance, Ca, Mg, Zn, Mn and fat, starch, protein etc..

Fructus piperis longi is the nearly mature fruit cluster of drying of Piperaceae fructus piperis longi.Originate in the Yunnan southeast to the west and south, Guangxi, Guangdong and Fujian There is cultivation.The 9-8 months pick.Acrid flavour, warm-natured, warming spleen and stomach for dispelling cold, lower gas analgesic.Containing pipering, Piperyline and a small amount of（1%）Wave Hair oil etc..

White lettuce victory is the fruit of composite family lettuce.Taste is slightly sweet, cold in nature, reduces phlegm and internal heat, diuresis, lactagogue.Containing volatile oil, amino acid, phenol Class and carbohydrate etc..

Stir-baked SEMEN ZIZIPHI SPINOSAE is to remove pulp, nucleocapsid by Rhamnaceae wild jujube ripening fruits, collects seed, dries.It is born in Yangshan The ground such as Henan, Hebei, Shaanxi, Liaoning, Shanxi, Shandong, Yunnan are mainly originated in slope, roadside.It harvests, goes when the fruit matures in the fall Pulp and stone take seed, raw use or micro- stir-fry to use.Taste is dry, sour, mild-natured, controls restlessness of asrhenia type and insomnia, palpitation of palpitating with fear, polydipsia, abnormal sweating.It is main Contain Saponin A, Spine Date Seed jujubosideB and a small amount of volatile oil.

The positive effect of the present invention is：

1, Mongolian compound preparation prepared by the method for the present invention has town " He Yi " effect, cures mainly insomnia.It is imitated by Clinical practice for many years Fruit is evaluated, and the therapeutic effect of insomnia can be good.And the therapeutic effect of granule is better than decoction pulvis.

2, Mongolian compound preparation prepared by the method for the present invention, is a kind of granule, extracted through active compound crushing, active compound coarse powder, Inclusion essential oil, active compound extraction filtrate is dry, granule molding and etc. ensure that the content of Mongolian compound preparation active ingredient Reach higher content, ensure that drug effect plays the dose requirement of effect, and Mongolian compound preparation meets《Chinese Pharmacopoeia》 （Version four in 2015）Requirement to granule.

3, the present invention using gas phase survey spectrometer measure eucalyptol in preparation content specificity it is strong, precision, stability, again Renaturation and the data of sample recovery rate are reliable, and linear relationship is good.The content of eucalyptol is in gas phase survey spectrometer measurement preparation Use internal standard method.Internal standard method can reduce sample pre-treatments and the error of sample size.Internal standard compound is added when extraction to can avoid by extractor, Error caused by temperature, measurement result is accurate, and the content of eucalyptol is big in Mongolian compound preparation prepared by the method for the present invention In 1.5mg/g.

4, Mongolian compound preparation prepared by the present invention is granule, except satisfaction《Chinese Pharmacopoeia》（Version four in 2015）It is right Outside the requirement of granule, following quality requirement is also needed to meet：

（1）When thin-layered chromatography differentiates the Amomum cardamomum in Mongolian compound preparation, Mongolian compound preparation is corresponded in eucalyptol reference substance Place shows identical blue spot, negative noiseless.

（2）When high performance liquid chromatography differentiates stir-baked SEMEN ZIZIPHI SPINOSAE in Mongolian compound preparation, Mongolian compound preparation is in jujuboside- There is absorption peak in B standard items corresponding position, negative noiseless.

（3）Mongolian compound preparation can dissolve in 2min.It is better than《Chinese Pharmacopoeia》（Version four in 2015）Granule is wanted The requirement dissolved in 5min is asked, the time that patient waits for drug of taking after mixing it with water is greatly reduced.

（4）Eucalyptol content in gas chromatography measurement Mongolian compound preparation, which is set to its content, must not be less than 1.5mg/g.

The requirement of the above index is to ensure the important indicator of Mongolian compound preparation drug effect.Meet the anaesthetic compound of requirements above Preparation is qualified products；The Mongolian compound preparation for being unsatisfactory for requirements above is substandard product.The above method can be used for assisting Identification is expected the true and false of the Mongolian compound preparation of listing in the future.

5, Mongolian compound preparation of the present invention is the granule type preparation developed in conjunction with modern pharmacy technology, can be to avoid The disadvantage of decoction has the advantages such as curative effect is fast, easily absorbs, is easy to be accepted by patients.

Description of the drawings

Fig. 1 is to test 1 specificity test result, in figure：A standard solutions；B test solutions；C negative controls are molten Liquid.

Fig. 2 eucalyptol thin-layer chromatograms, in figure：1-eucalyptol reference substance solution, 2-Amomum cardamomum list medicines,

3-negative controls solution, 4-Mongolian compound preparations.

Fig. 3 is HPLC discriminating Spine Date Seed jujubosideBs as a result, in figure：A reference substance solutions；B test solutions；C feminine genders are right According to product solution.

Specific implementation mode

With reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto, Protection scope of the present invention cannot be limited with following embodiments.

Experimental method used in following embodiments is conventional method unless otherwise specified；Used material, Reagent etc., is commercially available unless otherwise specified.

Chemical reagent and standard items are in specific implementation mode：

Chemical reagent：Eucalyptol, Spine Date Seed jujubosideB, toluene, n-hexane, hexamethylene, dichloromethane, ethyl acetate, ethyl alcohol, first Alcohol, petroleum ether, sodium chloride, cyclohexanone, acetonitrile, natrium carbonicum calcinatum, dextrin, lactose, soluble starch, beta-cyclodextrin etc., it is above Reagent is that analysis is pure.

Standard items：Eucalyptol（Lot number：110788-201105）, Spine Date Seed jujubosideB（110814-200607）Equal standard items Bought from Chinese food and medicine calibrating institute.

The assay method of specific implementation mode Biochemical Indexes is as follows：

1, in extraction process Testing index measurement

It presses《Chinese Pharmacopoeia》（Version four in 2015）Volatile oil extracting method（2204 first method of general rule）Volatile oil is extracted, calculating is waved Hair oil recovery rate, yield of extract.

Volatile oil extracting rate（%）=volatilization oil mass/active compound quality × 100%；

Yield of extract（%）=（The volume of medicinal extract quality × totality taking liquid volume/actual amount taking liquid）/ active compound quality × 100%

Volatile oil extracting method（First method）Shown in being as follows：

The test sample of measurement is taken, unless otherwise specified, must crush and enable through No. two to No. three sieves, and be uniformly mixed.

Determination of volatile oil：This law is suitable for measuring relative density in 1.0 volatile oil below.Take test sample appropriate（About phase When in containing 0.5~1.0mL of volatile oil）, weighed weight（Accurately to 0.01g）, set in flask, add 300 ~ 500mL of water（Or it is appropriate） With bead number volatile oil determination apparatus and reflux condensing tube are connected after shaking mixing.Autocondensation pipe upper end Jia Shui, which makes to be full of, to be waved The scale part of hair oil analyzer, and when overflow enters flask until.It sets in electric jacket or other proper methods is slowly heated to boiling, And kept for slightly boiling certain time, until oil mass is not further added by analyzer, stop heating, place a moment, opens analyzer lower end Piston slowly releases water, until above oil reservoir upper end arrival 0 line of scale at 5mm.Place 1 hour or more, it is then turned on piston So that oil reservoir is dropped to the upper end just concordant with 0 line of scale, reads volatilization oil mass, and calculate the content of volatile oil in test sample（%）.

2, in inclusion essential oil technique Testing index measurement

（1）The calculating of the volatile oil blank rate of recovery：

According to《Chinese Pharmacopoeia》（Version four in 2015）Volatile oil extracting method（2204 first method of general rule）Carry out determination of volatile oil. Precision pipettes volatile oil 1mL, is placed in round-bottomed flask, adds distilled water 200mL, and heating keeps slightly boiling, until when oil mass is not further added by Stop heating, places room temperature, the blank rate of recovery is calculated according to the following formula.As a result the blank rate of recovery 91%, RSD 1.02%, this Method is feasible.

The blank rate of recovery=recycling volatile oil oil mass/throwing oil mass × 100%.

（2）Volatile oil clathrate compound inspection target

The inclusion effect expedition of volatile oil clathrate compound can be analyzed by inclusion essential oil rate, inclusion compound recovery rate.Volatile oil The leading indicator that inclusion rate is screened as clathrate process, inclusion rate is higher, and inclusion effect is better；Inclusion compound yield contains with inclusion compound Oil cut rate has certain influence to the inclusion effect of volatile oil, inclusion compound yield and inclusion compound oil content in the case where input amount is certain Higher, the inclusion effect of volatile oil is also better.

Inclusion compound recovery rate=inclusion amount of substance/（β-CD quality+throwing oil mass）× 100%

The practical receipts oil mass of inclusion essential oil rate=inclusion compound/（Throw oil mass+blank rate of recovery）×100%

3, the evaluation index of granulate quality

Hygroscopicity, ratio of briquetting, melting, brittleness, water content, the granularity of granule are to evaluate the important ginseng of granule quality Number, specifically measures with the following method：

（1）Hydroscopicity：The glass desicator for filling sodium chloride supersaturated solution is put into 25 DEG C of constant incubator and is cultivated For 24 hours, the wetting balance for making it internal to relative humidity is 75%.The herb in granule form of about 2g is put into the measuring cup of constant-quality Bottom, and it is made uniformly to spread out, the bottom that constant humidity is put into after precise fills the glass of sodium chloride supersaturated solution In drier, and weigh the cap is opened, weighed after placing 48h, calculates medicinal material hydroscopicity.

Hydroscopicity（%）Medicinal powder quality-moisture absorption prodrug silty amount/moisture absorption prodrug silty amount × 100% after=moisture absorption

（2）Ratio of briquetting：Granule is crossed to the medicine sieve of 10 mesh and 60 mesh successively, conforming particle is that can pass through 10

Mesh sieves, but the particle that cannot be sieved by 60 mesh.

Ratio of briquetting（%）=conforming particle quality/sample particle quality × 100%.

（3）Melting：According to《Chinese Pharmacopoeia》（Version four in 2015）Granule（General rule 0104）Solvable under Grain inspection Check methods, weigh granule 10g and add 5 min of 200mL hot water stirs, observe immediately, soluble particles should all dissolve or slightly It is muddy；And record the time all dissolved.

（4）Brittleness：It weighs after granule 10g is placed in 100 turns of brittleness instrument turn and crosses 80 mesh sieve, the medicinal powder meter of 80 mesh of weighed mistake sieve Calculate brittleness.

（5）Moisture：According to aquametry（《Chinese Pharmacopoeia》Four general rules 0832 of version in 2015）4th method（Toluene method） It measures.

Accurately weighed test sample 25g, sets in round-bottomed flask, adds toluene 200mL, and dry, clean zeolite is added, and shines《In State's pharmacopeia》（Version four in 2015）Aquametry（The 4th method of general rule 0832）Apparatus figure connects instrument portion, from Toluene is added in condenser pipe top, until the narrow thin portion full of determination of moisture pipe point.Round-bottomed flask is set in electric jacket, waits for that toluene starts When boiling, temperature is adjusted, makes per minute to distillate 2 drops.Wait for that moisture distillates completely, that is, the water for measuring pipe scale part is not further added by When, toluene rinse will be first used inside condenser pipe, then pushed the toluene adhered on tube wall with the full long brush for dipping in toluene, continue to distill It 5 minutes, lets cool to room temperature, provision for disengagement, is attached on if any water on the tube wall of determination of moisture pipe, can be pushed away with the copper wire for dipping in toluene Under, it places, moisture is made to be kept completely separate with toluene（Methylenum careuleum powder can be added a small amount of, water is made to dye blue, to observe）.Review water Amount, and calculate the water content in test sample（%）,

（6）Granularity：According to granularity and determination of particle size distribution（《Chinese Pharmacopoeia》Four general rules of version in 2015,0,982 second method is double Sieve method）It measures.

Take test sample 30g, weighed weight, set as defined in medicine sieve, keep horizontality sieving, left and right is round-trip, while sieving Side tapping 3 minutes.The particle and powder that cannot be crossed No.1 sieve and can be sieved by No. five, weighed weight is taken to calculate percentage. The summation that cannot be sieved and can be sieved by No. five by No.1, must not cross 15%.

《Chinese Pharmacopoeia》（Version one in 2015）Regulation：（1）Most coarse powder：Finger can be sieved all by No.1, but be mixed with and can be led to Cross No. three powder of the sieve no more than 20%.（2）Coarse powder：Referring to all to be sieved by No. two, but is mixed with and can be no more than by No. four sieves 40% powder.（3）Middle powder：Referring to all to be sieved by No. four, but being mixed with can be by No. five powder of the sieve no more than 60%.（4） Fine powder：Finger can be sieved all by No. five, and containing the powder that can pass through No. six sieves no less than 95%.（5）Most fine powder：Finger can all lead to Cross No. six sieves, and containing the powder that can pass through No. seven sieves no less than 95%.（6）Impalpable powder：Referring to all to be sieved by No. eight, and contains energy Pass through the powder of No. nine sieves no less than 95%.

4, the assay of eucalyptol

Using the content of the eucalyptol measured with reference to gas chromatography in Mongolian compound preparation.Eucalyptol is Amomum cardamomum volatile oil master Ingredient is wanted, is had and is removed free radical, calms the nerves the effect of calm；Another Amomum cardamomum is the monarch drug in a prescription of this granule, therefore combines this granule The pharmacological effect of the monarch rule and monarch drug in a prescription main active of prescription, screening eucalyptol refer to as our quality control Mark, has carried out assay.

Chromatographic condition：Stationary phase：Polyethylene glycol（2000）；Chromatographic column： OV-1701 ( 30m ×0.32mm )；Column temperature： 80 ℃；Detect temperature：From 80 DEG C of beginning（Continue 3min）It is warming up to 100 DEG C（16min）Split ratio：10:1；It carries Gas：Nitrogen；Flow rate of carrier gas：2mL/min；Detector：Hydrogen flame detector；Detector temperature：230℃；Sample size：0.5ul；Into Sample temperature：230℃；Internal standard method.

The preparation of solution：

（1）The preparation of inner mark solution：Precision weighs appropriate cyclohexanone, adds n-hexane that the internal standard of every 1mL n-hexanes containing 50mg is made Solution.

（2）The preparation of reference substance solution：Take eucalyptol reference substance appropriate, it is accurately weighed, add n-hexane that every 1mL is made and contains The solution of 25mg n-hexanes adds 1mL inner mark solutions to obtain the final product.

（3）The preparation of test solution：Mongolian compound preparation 5g is taken, it is accurately weighed, it sets in round-bottomed flask plus water 200mL, Volatile oil determination apparatus is connected, scale 3mL is added water to from analyzer upper end, then add 2~3mL of n-hexane, connects reflux condensing tube, add Heat is kept for 2 hours to slightly boiling, is let cool, and is divided and is taken n-hexane liquid, and the funnel by being covered with lg anhydrous sodium sulfates filters, and filtrate is set In 5mL measuring bottles, volatile oil determination apparatus inner wall is washed on a small quantity with n-hexane, in washing lotion and the same measuring bottle of people, adds 1mL inner mark solutions, Be diluted to scale with n-hexane, shake up, filter, take subsequent filtrate to get.

（4）The preparation of negative controls solution：Take the Mongolian compound preparation for not adding Amomum cardamomum（Only in active compound pulverising step In without plus Amomum cardamomum medicinal material, the preparation method is the same as that of Example 1 for other）5g, it is accurately weighed, by the preparation method system of test solution It is standby.

Mongolian compound preparation prepared by the present invention is granule, except satisfaction《China》（Version four in 2015）To granule It is required that it is outer, also need to meet following quality requirement：

（1）When thin-layered chromatography differentiates the Amomum cardamomum in Mongolian compound preparation, Mongolian compound preparation is corresponded in eucalyptol reference substance Place shows identical blue spot, negative noiseless.

（2）When high performance liquid chromatography differentiates stir-baked SEMEN ZIZIPHI SPINOSAE in Mongolian compound preparation, Mongolian compound preparation is in jujuboside- There is absorption peak in B standard items corresponding position, negative noiseless.

（3）The moisture of Mongolian compound preparation, granularity, dissolubility meet《Chinese Pharmacopoeia》（Version four in 2015）Regulation Standard, the Mongolian compound preparation can melt in 2min.

（4）Eucalyptol content in gas chromatography measurement Mongolian compound preparation, which is set to its content, must not be less than 1.5mg/g.

The requirement of the above index is to ensure the important indicator of Mongolian compound preparation drug effect.The measurement side of These parameters simultaneously Method, index value and attached drawing 2 and 3, the true and false for the Mongolian compound preparation that can be used for that identification is assisted to be expected listing in the future.

Implement 1

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Active compound crushes：Take 6 parts 1 part of fructus piperis longi, 2 parts of white lettuce victory, 3 parts of Amomum cardamomum, stir-fry of jujube that active compound was made after crushing 24 mesh sieve Coarse powder；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 10 parts of water is added, after mixing, impregnates 2 hours, then water is used to steam Steam distillation method collects volatile oil after extracting 4-8h and active compound extracts filtrate；

Inclusion essential oil：Using polishing：Take volatile oil and β-CD by 1:10 ratio be mixed it is even after, add 3 times amount water grinding 80min is filtered, and filter residue 80 DEG C of petroleum ethers 2 times dry 4h under the conditions of 40 DEG C, volatile oil medicinal powder is made；

Active compound extracts filtrate drying：After active compound extracts concentrating filter liquor to the 1/10 of active compound extraction filtrate volume, then at 60 DEG C Extracting solution medicinal powder is made in dry 6h in drying box；Anaesthetic group is made in the volatile oil medicinal powder and extracting solution medicinal powder after mixing Close object medicinal powder；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:1 ratio after mixing, with 70% ethanol wet, The dosage of 70% ethyl alcohol is the 20% of anaesthetic composition medicinal powder and excipient quality sum, after wetting under the conditions of 60 DEG C, drying Mongolian compound preparation is made after 4h.

The steam distillation refers to shining《Chinese Pharmacopoeia》（Version four in 2015）Volatile oil extracting method（General rule 2204 First method）Volatile oil is extracted, volatile oil extracting rate, yield of extract are calculated.

The active compound extraction filtrate refers to the filtrate that the extracting solution after the extraction of active compound coarse powder is collected after filtered through gauze.

Embodiment 2

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Active compound crushes：Take 12 parts 3 parts of fructus piperis longi, 4 parts of white lettuce victory, 6 parts of Amomum cardamomum, stir-fry of jujube that active compound was made after crushing 24 mesh sieve Coarse powder；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 9 parts of water is added, after mixing, impregnates 3 hours, then water is used to steam Steam distillation method collects volatile oil after extracting 5h and active compound extracts filtrate；

Inclusion essential oil：Using polishing：Take volatile oil and β-CD by 1:9 ratio be mixed it is even after, add 3.5 times amount water grind 90min is ground, is filtered, filter residue 60 DEG C of petroleum ethers 3 times dry 3h under the conditions of 45 DEG C, volatile oil medicinal powder is made；

Active compound extracts filtrate drying：After active compound extracts concentrating filter liquor to the 1/9 of active compound extraction filtrate volume, then at 55 DEG C of baking Extracting solution medicinal powder is made in dry 5h in dry case；Anaesthetic is made with extracting solution medicinal powder in the volatile oil medicinal powder after mixing to combine Object medicinal powder；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:1.2 ratio after mixing, is moistened with 75% ethyl alcohol Wet, the dosage of 75% ethyl alcohol is the 25% of anaesthetic composition medicinal powder and excipient quality sum, after wetting, under the conditions of 65 DEG C, is done Mongolian compound preparation is made after dry 3h.

Embodiment 3

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Active compound crushes：Take 10 parts 3 parts of Piper longum, 2 parts of white lettuce victory, 4 parts of Amomum cardamomum, stir-fry of jujube that active compound was made after crushing 24 mesh sieve Coarse powder；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 11 parts of water is added, after mixing, impregnates 1 hour, then water is used to steam Steam distillation method collects volatile oil after extracting 7h and active compound extracts filtrate；

Inclusion essential oil：Using polishing：Take volatile oil and β-CD by 1:10 ratio be mixed it is even after, add 4 times amount water grinding 70min is filtered, and filter residue 90 DEG C of petroleum ethers 2 times dry 5h under the conditions of 42 DEG C, volatile oil medicinal powder is made；

Active compound extracts filtrate drying：After active compound extracts concentrating filter liquor to the 1/11 of active compound extraction filtrate volume, then at 65 DEG C Extracting solution medicinal powder is made in dry 7h in drying box；Anaesthetic group is made in the volatile oil medicinal powder and extracting solution medicinal powder after mixing Close object medicinal powder；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:0.8 ratio after mixing, is moistened with 65% ethyl alcohol Wet, the dosage of 65% ethyl alcohol is the 15% of anaesthetic composition medicinal powder and excipient quality sum, after wetting under the conditions of 55 DEG C, it is dry Mongolian compound preparation is made after dry 5h.

Comparative example 1

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Active compound crushes：With embodiment 1；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 8 parts of water is added, after mixing, impregnates 10 hours, then water is used to steam Steam distillation method collects volatile oil after extracting 4h and active compound extracts filtrate；

Inclusion essential oil：Using ultrasonic method：Precision weighs the β-CD of 8g, sets in 150mL conical flask with cover, and 100mL is added and steams Distilled water, boiling water bath make dissolving, are down to 40 DEG C；Precision measures volatile oil：Ethyl alcohol (1:1) mixed solution 2mL slowly injects β-CD In saturated solution, ultrasonic 1h, set refrigerated in refrigerator for 24 hours, filter, filter residue with petroleum ether (60-90 DEG C) wash 2 times (15mL, 10mL), dry 4h under the conditions of 40 DEG C, is made volatile oil medicinal powder；

Active compound extracts filtrate drying：With embodiment 1；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:1.5 ratio after mixing, is moistened with 80% ethyl alcohol Wet, the dosage of 80% ethyl alcohol is the 20% of anaesthetic composition medicinal powder and excipient quality sum,

Mongolian compound preparation is made under the conditions of 40 DEG C, after dry 6h again after wetting.

Comparative example 2

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Active compound crushes：With embodiment 1；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 6 parts of water is added, after mixing, impregnates 2 hours, then water is used to steam Steam distillation method collects volatile oil after extracting 2h and active compound extracts filtrate；

Inclusion essential oil：Saturated water solution method：Precision weighs β-CD 8g, adds distilled water 100mL, boiling water bath to make dissolving, is down to It 40 DEG C, sets in constant temperature blender with magnetic force, 2mL volatile oil is slowly added dropwise：Ethyl alcohol（1:1）Mixed solution, 40 DEG C of stirring 1h of constant temperature Afterwards, taking-up is cooled to room temperature, and is put into refrigerator cold-storage for 24 hours, filter, filter residue with petroleum ether (60-90 DEG C) wash 2 times (15mL, 10mL), volatile oil medicinal powder is made in dry 4h under the conditions of 40 DEG C；

Active compound extracts filtrate drying：With embodiment 1；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:1 ratio after mixing, with 80% ethanol wet, The dosage of 80% ethyl alcohol is the 20% of anaesthetic composition medicinal powder and excipient quality sum, after wetting under the conditions of 60 DEG C, drying Mongolian compound preparation is made after 4h.

Comparative example 3

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Active compound crushes：With embodiment 1；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 10 parts of water is added, after mixing, impregnates 6 hours, then water is used to steam Steam distillation method collects volatile oil after extracting 6h and active compound extracts filtrate；

Inclusion essential oil：With embodiment 1

Active compound extracts filtrate drying：With embodiment 1；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:05 ratio after mixing, is moistened with 90% ethyl alcohol Wet, the dosage of 90% ethyl alcohol is the 20% of anaesthetic composition medicinal powder and excipient quality sum, is made under the conditions of 60 DEG C, after dry 6h Obtain Mongolian compound preparation.

Comparative example 4

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Polishing in inclusion essential oil step is replaced with into supercritical ultrasonics technology（Supercritical ultrasonics technology step is the same as comparative example 1）, other steps With embodiment 1.

Comparative example 5

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Polishing in inclusion essential oil step is replaced with into saturated water solution method（Saturated water solution method step is the same as comparative example 2）, Other steps are the same as embodiment 1.

Comparative example 6

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Soluble starch in granule forming step is replaced with into lactose, other steps are the same as embodiment 1.

Comparative example 7

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Soluble starch in granule forming step is replaced with into dextrin, other steps are the same as embodiment 1.

Comparative example 8

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

Soluble starch in granule forming step is replaced with into 0.5%CMC-Na, other steps are the same as embodiment 1.

Comparative example 9

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

70% ethyl alcohol in granule forming step is replaced with into 0.5%CMC-Na suspensions, other steps are the same as embodiment 1.

Comparative example 10

A kind of Mongolian compound preparation for treating insomnia, Mongolian compound preparation are solid particle agent made of anaesthetic compound, Preparation method specifically includes following steps：

70% ethyl alcohol in granule forming step is replaced with into water, other steps are the same as embodiment 1.

The experimental result of embodiment 1-3, comparative example 6-9 are as shown in table 1, table 2, table 3：

As can be known from Table 1：

With volatile oil extracting rate（%）And yield of extract（%）It, can by embodiment 1-3 and comparative example 1-3 comparative analysis for measurement index Know：Extraction process influences than more significant, only under suitable Extraction technique, to obtain on the dissolution of anaesthetic volatile oil Higher volatile oil extracting rate（%）And yield of extract（%）；

With inclusion compound recovery rate（%）With inclusion essential oil rate（%）For measurement index, embodiment 1-3 is divided compared with comparative example 4-5 Known to analysis：Including technique influences than more significant the inclusion of volatile oil, only under suitably inclusion parameter, could obtain higher Inclusion compound recovery rate（%）With inclusion essential oil rate（%）, polishing is better than supercritical ultrasonics technology and saturated solution method.

The volatile oil extracting rate of embodiment 1-3（%）, yield of extract（%）, inclusion compound recovery rate（%）, inclusion essential oil rate （%）Obtain preferable extraction and inclusion effect, and eucalyptol content

It is all higher than 1.5mg/mL, granule can dissolve within 2min, and ratio of briquetting has reached 98% or more.

Comparative example 1 carries out inclusion essential oil with comparative example 4 using supercritical ultrasonics technology, and solution time is all higher than 2min, ratio of briquetting Less than 98%.

Comparative example 1, comparative example 5 carry out inclusion essential oil using saturated water solution method, and solution time is all higher than 2min, are molded Rate is less than 98%.

Comparative example 3 uses polishing, solution time to be more than 2min.

As can be known from Table 2：In entire pelletization, it is easiest to by the pelletization of auxiliary material of soluble starch, grain shape Preferably, melting is good, and hydroscopicity is higher, so selecting soluble starch for most suitable gift shape agent.

As known from Table 3, soft in pelletization just to have if selecting 0.5%CMC-Na suspensions and water as wetting agent respectively Viscosity is not easy sieving granulation, and character of pelletizing is uneven, selects the ethyl alcohol of proper ratio to do soft just suitable in humectant pelletization In, it is easy sieving granulation, and character of pelletizing is uneven, accordingly, it is determined that most suitable wetting agent is ethyl alcohol.

In summary：Only under conditions of suitable extraction process, clathrate process and moulding process, the anaesthetic of preparation is multiple Square preparation could meet the relevant criterion requirement of granule, have chain effect between each step, arbitrarily change a technique Parameter will lead to the decline of granule quality.

In order to prove the quality and drug effect of Mongolian compound preparation of the present invention, once tested：

Test the methodology experiment of the assay of 1 eucalyptol

The eucalyptol content of Mongolian compound preparation is measured using gas chromatography.

1 chromatographic condition

Stationary phase：Polyethylene glycol（2000）；Chromatographic column： OV-1701 ( 30m ×0.32mm )；Column temperature： 80 ℃；Detection temperature Degree：From 80 DEG C of beginning（Continue 3min）It is warming up to 100 DEG C（16min）Split ratio：10:1；Carrier gas：Nitrogen；Carrier gas stream Speed：2mL/min；Detector：Hydrogen flame detector；Detector temperature：230℃；Sample size：0.5ul；Injector temperature：230℃； Internal standard method.

The preparation of 2 solution

（1）The preparation of inner mark solution：Precision weighs appropriate cyclohexanone, adds n-hexane that inner mark solutions of every 1mL containing 50mg is made.

（2）The preparation of standard solution：Take eucalyptol reference substance appropriate, it is accurately weighed, add n-hexane that every 1mL is made and contains The solution of 25mg adds 1mL inner mark solutions to obtain the final product.

（3）The preparation of test solution：Mongolian compound preparation about 5g is taken, it is accurately weighed, it sets in round-bottomed flask plus water 200mL connects volatile oil determination apparatus, adds water to scale 3mL from analyzer upper end, then add 2~3mL of n-hexane, connects returned cold Solidifying pipe is heated to slightly boiling, and is kept for 2 hours, lets cool, and divides and takes n-hexane liquid, and the funnel by being covered with anhydrous sodium sulfate about lg is filtered It crosses, filtrate is set in 5mL measuring bottles, and volatile oil determination apparatus inner wall is washed on a small quantity with n-hexane, in washing lotion and the same measuring bottle of people, adds 1mL Inner mark solution is diluted to scale with n-hexane, shakes up, filtration, take subsequent filtrate to get.

（4）The preparation of negative controls solution：Take the Mongolian compound preparation for not adding Amomum cardamomum（Only in active compound pulverising step In without plus Amomum cardamomum medicinal material, the preparation method is the same as that of Example 1 for other）5g, it is accurately weighed, by the preparation method system of test solution It is standby.

3, specificity is tested

Take standard solution, the preparation of test solution, each 0.5mL injections chromatographic of negative controls solution.Knot Fruit is the corresponding period that test solution goes out characteristic peak in standard solution（14min-15min）Aobvious absorption peak, and it is negative Contrast solution is without peak.Illustrating inner mark solution not influences experimental result and specificity is good, as shown in Figure 1.

4, linear relationship is investigated

Precision measures eucalyptol 250mg reference substance stock solutions, draws 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL respectively, 1mL sets 5mL volumetric flasks, respectively adds the inner mark solution of 1mL, is diluted to scale with n-hexane, shakes up, and 0.5ul is taken to inject gas-chromatography Instrument is measured by above-mentioned chromatographic condition, records peak area.With a concentration of abscissa of reference substance solution, reference substance peak area and internal standard The ratio of peak area is that ordinate carries out linear regression, obtains regression equation Y=174.85X-165.62, r=0.998 (n=6).As a result Show that eucalyptol mass concentration is good with the ratio linear relationship of internal standard peak area within the scope of 2.5mg/mL-25mg/mL（A: Inner mark solution； B:Test solution； C:The ratio of A and B）, the concentration of eucalyptol reference substance solution is as shown in table 4.

5, precision test

Precision draws same reference substance solution, is measured by above-mentioned chromatographic condition, repeatedly sample introduction 6 times, with reference substance peak area with it is interior Mark the ratio calculation of peak area.Average peak area ratio is 1.679, RSD=0.15%, as a result such as table 5.

6, stability test

The test solution prepared is taken, by above-mentioned chromatographic condition respectively in 0h, 1h, 2h, 4h, 6h, 8h sample introductions measurement, calculating confession The ratio of test product peak area and internal standard peak area.Average peak area ratio is 0.642, RSD=0.21%.It the results are shown in Table 6.

7, repetitive test

It takes with batch 6 parts of sample, handles by test solution preparation method, measured under above-mentioned chromatographic condition, calculate test sample peak The ratio of area and internal standard peak area.Average peak area ratio is 1.442, RSD=1.82%, as a result such as table 7.

8, it is loaded recovery test

5 parts of the sample of known content is taken, accurate addition eucalyptol is appropriate respectively, is prepared by the preparation method of test solution, It is measured under above-mentioned chromatographic condition, calculates the rate of recovery.Average recovery rate is 97.686%, RSD=1.55%.It the results are shown in Table 8.

Methodology the results show：Surveying the content that spectrometer measures eucalyptol in preparation using gas phase has specificity strong, The advantages that precision, stability, the RSD values of repeatability and sample recovery rate are respectively less than 5%, and data are reliable, linear relationship is good Good, the experimental result measured is accurate and reliable；

Test the quality evaluation experiment of 2 Mongolian compound preparations

Take 3 batches of experiment medicinal materials（Since medicinal material batch is different, the medicinal material eucalyptol content of different batches can be variant）, using this hair Bright method is prepared into Mongolian compound preparation, carries out following quality evaluation.

1, the measurement of sample eucalyptol content

Take above-mentioned medicine compound preparation（Embodiment 1, embodiment 2 is respectively adopted, prepared by 3 method of embodiment）It is molten to prepare 9 parts of test samples Liquid calculates the content of eucalyptol according to being measured under 1 GC conditions of experiment.Experimental result is as shown in table 9, and it is multiple to measure anaesthetic The content of eucalyptol is all higher than 1.5mg/g in square preparation；Eucalyptol contains in drug quality requirement gas phase survey spectrometer measurement preparation Amount is to use internal standard method.Because internal standard method can reduce sample pre-treatments and the error of sample size.When extraction be added internal standard compound can avoid by Extractor, the error caused by temperature.

2, in Mongolian compound preparation whether the thin-layer chromatography qualitative analysis containing Amomum cardamomum

（1）The preparation of reference substance solution：Take eucalyptol reference substance appropriate, it is accurately weighed, add n-hexane that every 1mL eucalyptus containing 25mg is made The solution of olein to get.

（2）The preparation of test solution：It randomly selects Mongolian compound preparation 5g prepared by above-mentioned 2 batches of medicinal materials and prepares 2 parts of confessions Test sample solution, it is accurately weighed, it sets in round-bottomed flask, adds water 200mL, connect volatile oil determination apparatus, added water to from analyzer upper end Scale 3mL, then add 2~3mL of n-hexane, connects reflux condensing tube, is heated to slightly boiling, and kept for 2 hours, let cool, point take just oneself Alkane liquid, the funnel by being covered with lg anhydrous sodium sulfates filter, and filtrate is set in 5mL measuring bottles, volatile oil determination apparatus inner wall n-hexane A small amount of washing, washing lotion are incorporated in same measuring bottle, are diluted to scale with n-hexane, shake up, and filter, take subsequent filtrate to get.

（3）The preparation of Amomum cardamomum list medicine test solution：It randomly selects Amomum cardamomum list medicine 5g and prepares Amomum cardamomum list medicine for examination Product solution, it is accurately weighed, it sets in round-bottomed flask, adds water 200mL, connect volatile oil determination apparatus, quarter is added water to from analyzer upper end 3mL is spent, then adds 2~3mL of n-hexane, connects reflux condensing tube, slightly boiling is heated to, and kept for 2 hours, lets cool, divide and take n-hexane Liquid, the funnel by being covered with lg anhydrous sodium sulfates filter, and filtrate is set in 5mL measuring bottles, and volatile oil determination apparatus inner wall n-hexane is few Amount washing, washing lotion is incorporated in same measuring bottle, is diluted to scale with n-hexane, shakes up, and filters, take subsequent filtrate to get.

（4）Negative control solution：Take the Mongolian compound preparation for not adding Amomum cardamomum（Only without adding in active compound pulverising step Amomum cardamomum medicinal material, the preparation method is the same as that of Example 1 for other）5g, it is accurately weighed, it is prepared by the preparation method of test solution.

（5）Differentiate：According to《Chinese Pharmacopoeia》（Version four in 2015）Thin-layered chromatography (general rule 0502) is tested, in absorption Each l0 μ l of solution are stated, are put respectively on same silica gel g thin-layer plate, with hexamethylene-dichloromethane-ethyl acetate (15：5：0.5) it is Solvent is unfolded, and takes out, dries, and with 5% vanillin-sulfuric acid solution, it is clear to be heated to spot development at 105 DEG C, examines immediately for spray Depending on.

Experimental result is as shown in Figure 2：In test sample chromatography, on position corresponding with reference substance chromatography, same color is shown Spot, and negative controls solution do not show spot.This proves there is Amomum cardamomum in Mongolian compound preparation agent.This method can also be used Whether contain Amomum cardamomum progress qualitative analysis in the drug of identification in the market.

3, the high performance liquid chromatography qualitative analysis of stir-baked SEMEN ZIZIPHI SPINOSAE

Chromatographic condition：Chromatographic column：Agilent Eclipse XDB-C18（4.6 × 250mm.5μm ）；Mobile phase：Acetonitrile （A）, water（B）；Condition of gradient elution：A when 0-7min:B=28:A when 72,13min:B=35:A when 65,13-22min:B =35:A when 65,25min:B=45:Terminate when 55,35min, balances 20min；Flow velocity：1.0mL/min；Detection wavelength： 201nm；Column temperature：25℃.

It is prepared by solution：

（1）The preparation of reference substance solution：Spine Date Seed jujubosideB reference substance is taken, it is accurately weighed, it sets 10mL volumetric flasks and adds methanol to quarter Degree, is made the solution of every 1mL Spine Date Seed jujubosideBs containing 0.15mg, shakes up, and the filtration of 0.45um miillpore filters, product are molten as a contrast Liquid.

（2）The preparation of test solution：7.5 g of Mongolian compound preparation prepared by above-mentioned 3 batches of medicinal materials is taken to prepare 9 parts respectively Test solution is set in beaker, and 5% 50 mL of sodium hydroxide solution is added to be allowed to be completely dissolved, and is fully transferred to point In liquid funnel, ether defatting 3 times, 30 mL, discards ether layer, after subnatant waves most ether, use is water saturated every time Extracting n-butyl alcohol 3 times, every time 30 mL merge the water washing 2 times that n-butanol liquid n-butanol is saturated, every time 10 mL, N-butanol liquid water bath method, residue methanol are dissolved and are settled in 10mL measuring bottles, shakes up, the filter of 0.45um miillpore filters It crosses, discards primary filtrate, subsequent filtrate is as test solution.

（3）Negative control solution：Take 7.5g not plus the Mongolian compound preparation of stir-baked SEMEN ZIZIPHI SPINOSAE（Do not have in active compound pulverising step only Have and add stir-baked SEMEN ZIZIPHI SPINOSAE medicinal material, the preparation method is the same as that of Example 1 for other）Negative controls solution is prepared with test solution preparation method.

Experimental result：Above-mentioned 9 parts of test solutions differentiate stir-baked SEMEN ZIZIPHI SPINOSAE in Mongolian compound preparation using high performance liquid chromatography When, there is absorption peak in jujuboside-B standard items corresponding position（Appearance time is between 21-22min）, negative noiseless. Enclose the high-efficient liquid phase chromatogram of a sample（See Fig. 3）.

4, the quality evaluation of Mongolian compound preparation moisture, melting, granularity

For ensure granule quality according to《Chinese Pharmacopoeia》（Four general rules 0104 of version in 2015）Granule check item to Grain carries out the detection of moisture, melting, granularity.

（1）Moisture

According to aquametry（《Chinese Pharmacopoeia》The 4th general rule 0832 of version in 2015）4th method（Toluene method）It measures.Three Criticize the water content in test sample（%）As a result such as table 10.As known from Table 17, three batches of sample moisture are less than 5%, meet《Middle traditional Chinese medicines Allusion quotation》（Version in 2015）Regulation（Moisture must not exceed 8 .0%）.

（2）Melting

It takes test sample 1g, heats water 200ml, stir 5 minutes, observe immediately, should all dissolve or be in suspension shape, as a result such as table 11.As known from Table 11, three batches of sample meltings all dissolve in 2min, meet《Chinese Pharmacopoeia》（Version in 2015）Regulation （Granule dissolves in 5 minutes）.

（3）Granularity

According to granulometry（0,982 second method of general rule, double sieve methods）It measures.As known from Table 12, three batches of sample granularities are without super 15% is crossed, is met《Chinese Pharmacopoeia》（Version portion four editions in 2015）Regulation（The summation that cannot be sieved and can be sieved by No. five by No.1 It must not exceed 15%）.

It summarizes：Mongolian compound preparation prepared by the present invention is granule, except satisfaction《Chinese Pharmacopoeia》（Version portion four editions in 2015）It is right Outside the requirement of granule, following quality requirement is also needed to meet：

（1）When thin-layered chromatography differentiates the Amomum cardamomum in Mongolian compound preparation, Mongolian compound preparation is corresponded in eucalyptol reference substance Place shows identical blue spot, negative noiseless.

（2）When high performance liquid chromatography differentiates stir-baked SEMEN ZIZIPHI SPINOSAE in Mongolian compound preparation, Mongolian compound preparation is in jujuboside- There is absorption peak in B standard items corresponding position, negative noiseless.

（3）Mongolian compound preparation can dissolve in 2min.

（4）Eucalyptol content in gas chromatography measurement Mongolian compound preparation, which is set to its content, must not be less than 1.5mg/g.

The requirement of the above index is to ensure the important indicator of Mongolian compound preparation drug effect.The measurement side of These parameters simultaneously Method, index value and attached drawing 2 and 3 can be used for that identification is assisted to be expected the true and false of the Mongolian compound preparation listed in the future.Experiment knot Fruit shows that Mongolian compound preparation prepared by this 3 batches of medicinal materials is up-to-standard.

Test the pharmacodynamic experiment and acute toxicity testing of 3 Mongolian compound preparations

1 experiment material

1. reagent：Medicinal material yellow Jackets (Guangzhou Chemical Reagent Factory, product batch number:950601);CMC-Na (concentration 0.5%);It is raw Manage brine (packet header Kang Li medicine companies limited liability company, product batch number:060222103).

1.2 animal：Kunming mouse is purchased from University of the Inner Mongol's animal experimental center, cleans two level, quality certification number:《Capital is dynamic Word》No. 8806M035;20 ± 2g of weight, half male and half female divide cage to feed, and laboratory is tested after adapting to 3 days.

1.3 instrument and equipment:Upper ware electronic balance (Shanghai balance equipment factory, JA2003), mouse scale (weigh and consider in order to uphold justice instrument and meter by Shanghai Factory, ATZ-01), syringe, irrigation stomach device, mortar, stopwatch.

1.4 statistical method：Data are indicated with x ± s, are calculated with SPSS17.0 softwares.

2 experimental methods

2.1 hypnosis are tested

2.1.1 the preparation method of laboratory sample

Mongolian compound preparation group suspension：It is prepared according to embodiment 1, when use takes 5g, adds 25mL boiling water to rush molten, crude drug is dense Degree is 2g/mL.

Stable suspension:Take diazepam tablet appropriate, with 0.5%CMC-Na solution allocations at the suspension of a concentration of 0.75mg/mL Liquid；

- 3 taste soup of Amomum cardamomum dissipates：It takes 1 part of Piper longum, white lettuce to win 2 parts, 3 parts of Amomum cardamomum, was mixed i.e. after crushing 24 mesh sieve respectively .It takes -3 taste soup of Amomum cardamomum to dissipate 5g when use, adds 200mL water, decoct to volume 25mL filterings and filtrate progress gavage, filtrate is made A concentration of 2g/mL of crude drug.

- 4 taste of Amomum cardamomum dissipates：6 parts 1 part of Piper longum, 2 parts of white lettuce victory, 3 parts of Amomum cardamomum, stir-fry of jujube is taken to crush 24 mesh respectively It is mixed after sieve to obtain the final product.Take -3 taste soup of Amomum cardamomum to dissipate 5g when use, add 200mL water, decoct to volume 25mL filterings be made filtrate into Row gavage, a concentration of 2g/mL of filtrate crude drug.

Animal packet

Healthy Kun Ming mice 60 is taken, is randomly divided into 5 groups, every group 12, half male and half female.

Respectively blank group, positive controls, -3 taste soup of Amomum cardamomum dissipate group, -4 taste of Amomum cardamomum dissipates group, Mongolian compound preparation Group.Each group medicine feed situation is as follows：

Blank control group gavages 0.5%CMC-Na solution；

Positive controls gavage stable suspension；

The scattered group of -3 taste soup of Amomum cardamomum gavages -3 taste soup of Amomum cardamomum and dissipates；

The scattered group of -4 taste of Amomum cardamomum gavages -4 taste soup of Amomum cardamomum and dissipates；

Mongolian compound preparation group gavages Mongolian compound preparation suspension.

The dosage that gavages of above-mentioned each experimental group is 0.2mg/10g（Crude drug quality/mice weights）.

2.1.2 experimental result

Each group first according to dosage gives drug, is administered 1 day.After 15 min of perfusion, give each group animal intraperitoneal injection 50mg/kg penta bar of ratio Appropriate sodium, injection volume 0.1mL/10g, using righting reflex loss as sleep criterion, whether observation tested material extends amobarbital Sodium Sleep latency and sleeping time, as a result such as table 13-14.

From table 13 and table 14 it is found that for Sleep latency, compared with blank group, each dosage group has very significant poor It is different（P ＜ 0.01）；Compared with -3 taste soup of Amomum cardamomum dissipates group, -4 taste soup of Amomum cardamomum dissipates the significant difference of group（P ＞ 0.05）, and cover The significant difference of medicine compound preparation group（P ＜ 0.05）；With -4 taste soup dephasing ratio of Amomum cardamomum, Mongolian compound preparation group is significant Difference（P ＜ 0.05）.For sleeping time, compared with blank group, each dosage group has significant differences（P ＜ 0.01）； Compared with -3 taste soup of Amomum cardamomum dissipates group, -4 taste soup of Amomum cardamomum dissipates the significant difference of group（P ＜ 0.01）, Mongolian compound preparation group has Significant differences（P ＜ 0.01）；With -4 taste soup dephasing ratio of Amomum cardamomum, the significant difference of Mongolian compound preparation group（P ＜ 0.05）.Show -3 taste soup of Amomum cardamomum dissipate, -4 taste soup of Amomum cardamomum dissipate and Mongolian compound preparation can extend yellow Jackets induction it is small The effect of mouse Sleep latency and sleeping time, wherein Mongolian compound preparation is most apparent, to be significantly higher than and be dissipated with -3 taste soup of Amomum cardamomum It is dissipated with -4 taste soup of Amomum cardamomum.

The experimental results showed that the dosage form of drug and composition have significant impact, only guaranteed suitable drug to drug effect Composition and suitable pharmaceutical formulation, could preferably play drug effect.

2.2 acute toxicity test

2.2.1 median lethal dose (LD₅₀) measurement

Kunming mouse 10 is taken, male and female have concurrently, and before testing after fasting 12h, anaesthetic is given every 4 h after on-test Compound preparation suspension (0.2 g/mL of crude drug concentration) is gavaged by 0.2mL/10g meterings, is continued after successive administration 3 times conventional Raising, is observed continuously 7 d.As a result experimental animal does not occur abnormal response and the phenomena of mortality in the observation period.

2.2.2 maximum dosage experiment

Kunming mouse 40 is taken, male and female have concurrently, are randomly divided into control group and observation group, every group 20.Fasting before experiment 12 h, observation group's gavage gives 0.2 mL/10g of Mongolian compound preparation suspension (0.2 g/ of crude drug concentration after on-test ML), every 4 h gavages 1 time, continuous 3 times, the sodium carboxymethylcellulose that control group gavage gives isometric 0.5% is molten Liquid.Administration terminates in 6 h, is observed 1 time every 30 min, in 6~12 h, is observed 1 time every 1 h, from the Start within 2 days, observation 1 time, is observed continuously 14 d daily.Main detection mouse appearance sign, behavioral activity, breathing, body of gland The indexs such as secretion, feed, drinking-water, excrement and existing state, the 7th day and the 14th day weight before record administration and after administration, As a result such as table 3.Normal diet during observation, if finding, dead mouse carries out dissect in time.To the 14th day, dislocation was put to death small Mouse carries out dissect, visually observes main organs whether there is or not abnormal change, and coring, liver and kidney are fixed with neutral formalin solution, conventional Materials dehydration, paraffin embedding, slice row HE are dyed, and pathological examination is carried out under light microscopic.

Experimental result is as shown in Table 15：As a result observation group animal spontaneous activity is reduced, and is shown to repose and be moved less, administration terminates Restore after 6 h normal.Two groups of animal appearance signs are showed no obvious abnormalities, and freely, feed, drinking-water and excrement are normal for activity, Fur is bright and clean, and mouth, nose and eye are without secretion, pudendum cleaning, without animal dead in the observation period.Before and after two groups of animal experiments weight without Notable difference, no significant difference（P ＞ 0.05）.Dissect has no that each main organs have abnormal change, Histopathology inspection The main organs such as two groups of animal hearts, liver and kidney are looked into be showed no and the relevant apparent pathologic lesion of drug.

3, it summarizes

Mongolian compound preparation is in conjunction with modern pharmacy technology and the granule type preparation developed, it is not only in terms of medication, but also It is easy to be accepted by patients.This research has carried out experimental study with regard to its main pharmacodynamics and acute toxicity.Pharmacodynamics experimental result Display -3 taste soup of Amomum cardamomum dissipates, -4 taste soup of Amomum cardamomum dissipates and Mongolian compound preparation can extend the sleep of yellow Jackets inducing mouse The effect of incubation period and sleeping time, wherein Mongolian compound preparation is the most apparent, is significantly better than and -3 taste soup of Amomum cardamomum dissipates and white peas or beans Cool -4 taste soup dissipates.Acute toxicity testing result shows that Mongolian compound preparation toxicity is few, is carried out with 15 g/kgd dosage anxious Property toxicity test have no obvious adverse reaction, this dosage is the 200 times equal of quasi- clinical dosage（The clinic of Mongolian compound preparation is often It is 3g/d with dosage）, almost without toxicity.

Test the clinical case of 4 Mongolian compound preparations

Case 1：Bar certain, man, 54 years old, go to a doctor reason:Insomnia.Condition-inference：Insomnia caused by " conspicuous according to ".Take Chinese medicine of the present invention Wakefulness state starts to improve after composition 3 days, by 15 days after basically reach ortho state, after treatment is discontinued after 1 week.

Case 2：Crow certain, man, 67 years old, go to a doctor reason:Poor sleep, dreaminess.Condition-inference:" conspicuous according to " contains partially.Clinical table It is existing:Agitation, say, not steady and sure, dreaminess of sleeping at night, morning get up dizziness more.Chinese medicine composition of the present invention is taken after 7 days, irritated, The symptoms such as more speeches start to mitigate, by 15 days after sleep quality start to improve, by 30 days after the above symptom disappear, basic recovery is just Often.Follow-up after treatment 7 days.

Case 3：Certain is breathed out, female, 38 years old.Condition-inference:Insomnia.Clinical manifestation:It has difficulty in going to sleep, dreaminess of sleeping, every night greatly Cause is difficult to fall asleep again after sleeping 3 hours.After taking 1 week of Chinese medicine composition of the present invention, sleep quality starts to improve, 2 week The above symptom equal hour after can sleeping every night afterwards 5 hours or more, 1 month, sleep are back to normal.Follow-up after treatment 7 days.

Case 4：Finish certain, man, 45 years old.Condition-inference:Insomnia caused by " conspicuous according to " contains partially.Clinical manifestation:Difficulty falling asleep, Dreaminess.After taking 2 week of Chinese medicine composition of the present invention, symptom completely disappears, and restores normal.

Experiment proves that Mongolian compound preparation clinical effectiveness of the present invention is good.

Claims (6)

1. a kind of preparation method of Mongolian compound preparation that treating insomnia, it is characterised in that：Mongolian compound preparation is solid particle Agent, preparation method specifically include following steps：

Active compound crushes：After taking 6-12 parts 1-3 parts of fructus piperis longi, 2-4 parts of white lettuce victory, 3-6 parts of Amomum cardamomum, stir-fry of jujube to crush 24 mesh sieve Active compound coarse powder is made；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 9-11 parts of water is added, after mixing, impregnates 1-3 hours, then uses Volatile oil is collected after extraction by steam distillation 5-7h and active compound extracts filtrate；

Inclusion essential oil：Using polishing：Take volatile oil and β-CD by 1:9-1:10 ratio be mixed it is even after, add 3-4 times to measure It is filtered after water grinding 70-90min, filter residue 60-90 DEG C of petroleum ether 2-3 times, drying 3-5h under the conditions of 40-45 DEG C, system At volatile oil medicinal powder；

Active compound extracts filtrate drying：After active compound extracts the 1/9-1/11 that concentrating filter liquor extracts filtrate volume to active compound, then at 55- Extracting solution medicinal powder is made in dry 5-7h in 65 DEG C of drying box；The volatile oil medicinal powder is made after mixing with extracting solution medicinal powder At anaesthetic composition medicinal powder；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:0.8-1:1.2 ratio after mixing, uses 65-75% Ethanol wet, the dosage of the ethyl alcohol of 65-75% is the 15-25% of anaesthetic composition medicinal powder and excipient quality sum, after wetting Mongolian compound preparation is made under the conditions of 55-65 DEG C, after dry 3-5h.

2. a kind of preparation method of Mongolian compound preparation that treating insomnia as described in claim 1, it is characterised in that：Anaesthetic is multiple Square preparation is solid particle agent, and preparation method specifically includes following steps：

Active compound crushes：Take 6 parts 1 part of fructus piperis longi, 2 parts of white lettuce victory, 3 parts of Amomum cardamomum, stir-fry of jujube that active compound was made after crushing 24 mesh sieve Coarse powder；

Active compound coarse powder extracts：1 part of active compound coarse powder is taken, 10 parts of water is added, after mixing, impregnates 2 hours, then water is used to steam Steam distillation method collects volatile oil after extracting 6h and active compound extracts filtrate；

Inclusion essential oil：Using polishing：Take volatile oil and β-CD by 1:10 ratio be mixed it is even after, add 3 times amount water grinding It is filtered after 80min, filter residue 80 DEG C of petroleum ethers 2 times, dry 4h, is made volatile oil medicinal powder under the conditions of 40 DEG C；

Active compound extracts filtrate drying：After active compound extracts concentrating filter liquor to the 1/10 of active compound extraction filtrate volume, then at 60 DEG C Extracting solution medicinal powder is made in dry 6h in drying box；Anaesthetic group is made in the volatile oil medicinal powder and extracting solution medicinal powder after mixing Close object medicinal powder；

Granule is molded：Take anaesthetic composition medicinal powder and excipient by 1:1 ratio after mixing, with 70% ethanol wet, The dosage of 70% ethyl alcohol is the 20% of anaesthetic composition medicinal powder and excipient quality sum, after wetting under the conditions of 60 DEG C, drying Mongolian compound preparation is made after 4h.

3. a kind of preparation method of Mongolian compound preparation that treating insomnia as claimed in claim 1 or 2, it is characterised in that：Institute It is soluble starch to state excipient.

4. a kind of Mongolian compound preparation for treating insomnia, it is characterised in that：By the anaesthetic for treating insomnia described in claim 1-2 The preparation method of compound preparation is prepared, and the content for treating eucalyptol in the Mongolian compound preparation of insomnia is more than 1.5mg/g.

5. a kind of Mongolian compound preparation for treating insomnia, it is characterised in that：It is multiple by the anaesthetic for treating insomnia described in claim 3 The preparation method of square preparation is prepared, and the content for treating eucalyptol in the Mongolian compound preparation of insomnia is more than 1.5mg/g.

6. a kind of method of evaluation Mongolian compound preparation qualification, it is characterised in that：The anaesthetic compound system as described in claim 4 or 5 Agent need to meet following index request：

（1）When thin-layered chromatography differentiates the Amomum cardamomum in Mongolian compound preparation, Mongolian compound preparation is corresponded in eucalyptol reference substance Place shows identical blue spot, negative noiseless；

（2）When high performance liquid chromatography differentiates stir-baked SEMEN ZIZIPHI SPINOSAE in Mongolian compound preparation, Mongolian compound preparation is marked in jujuboside-B There is absorption peak in quasi- product corresponding position, negative noiseless；

（3）The moisture of Mongolian compound preparation, granularity, dissolubility meet《Chinese Pharmacopoeia》（Version four in 2015）Required standard；

（4）Eucalyptol content in gas chromatography measurement Mongolian compound preparation, which is set to its content, must not be less than 1.5mg/g；

The Mongolian compound preparation for meeting requirements above is qualified products；The Mongolian compound preparation for being unsatisfactory for requirements above is unqualified Product.