CN103920012A - Dendrobe pure micro-powder buccal tablet and preparation method thereof - Google Patents

Abstract

The invention relates to a dendrobe micro-powder buccal tablet and a preparation method thereof, belongs to the field of medicines and health-care products, and aims to provide a tablet which does not contain any auxiliary material except the dendrobe medicinal micro-powder and has appearance property and hardness meeting the tablet quality standard. The dendrobe micro-powder tablet is a tablet preparation which is prepared from dendrobe micro-powder with water content of 10-18 percent and average volume grain diameter D[4,3] in a range of 10-40mu m, no auxiliary material is added in the preparation process, and the dendrobe micro-powder is the unique component in the tablet preparation. According to the preparation method, the tablet has good appearance, low rate of pitted faces and cracked tablets, can meet the tablet quality requirement for tablet hardness, disintegration and friability, can be subjected to coating and laminating treatment in later phase due to the hardness of the tablet, and cannot be cracked in the preparation process. The bioavailability of the buccal tablet is improved by one time as compared with that of common dendrobe powder.

Description

Dendrobium pure powder buccal tablet and preparation method thereof

Technical field

The present invention relates to Chinese herbal medicine manufacture field, be specifically related to dendrobium pure powder buccal tablet and preparation method thereof, belong to health product and field of medicaments.

Background technology

Herba Dendrobii is that < < Pharmacopoeia of People's Republic of China > > records kind.Medicinal part is fresh or dry stem.Main containing bioactive substances such as dendrobium polysaccharide, dendrobine, dendramine, Herba Dendrobii time alkali, dendroxine, dendrine, 6-hydroxydendroxines.Japan waits country in Southeast Asia, Hong Kong, Macao and Taiwan is regional and coastal developed regions, and useful Herba Dendrobii Herba Dendrobii extract is for tea or as Baoshang material, and as high-class healthy product, having makes light of one's life by commiting suicide prolongs life, the effect of improving eyesight, the sore-throat relieving that hawks.Take Herba Dendrobii in recent years as the increasing high-class healthy product of raw material production and Chinese patent medicine.

Existing Herba Dendrobii is processed to Herba Dendrobii extract or Herba Dendrobii extractum conventionally, then directly soaked, and when the Herba Dendrobii dry product of this Herba Dendrobii extract form is soaked, dissolution rate is low, and dissolution rate is slow, and bioavailability is low, is unfavorable for the utilization of resources of rare medical material.

At present Herba Dendrobii is made in the technique of tablet and generally all needed to mix with other medical materials use or add a large amount of adjuvants in order to figuration, never occurred making with pure Herba Dendrobii micropowder form the form of buccal tablet.In pharmaceutics, during tabletting, generally all can add adjuvant, to reach filling, bonding, disintegrate, lubricated, painted, taste masking and beauty function etc., not use pure Chinese drugs powder tabletting.Such as, the patent No. is: 201210431691.1 patent discloses a kind of pure Herba Dendrobii buccal tablet, but in its composition, has added multiple auxiliary materials to carry out excipient, not that pure Herba Dendrobii powder is suppressed and obtained, add the Herba Dendrobii tablet of adjuvant, there will be mixing inhomogeneous, the defect that tablet medicine content is inhomogeneous; In addition, do not contain disintegrating agent in the adjuvant of this patent, the probability that tablet exists not disintegrate to disperse in gastrointestinal tract, affects the absorption of effective ingredient, again, after a large amount of adjuvant of long-term taking, unfavorable to health.Application number is that 200610050206.0 and 201210213474.5 the disclosed Herba Dendrobii buccal tablet of patent is all compound preparation, except Herba Dendrobii, also has other medical materials and adjuvant, with the present invention relates to containing the dendrobium pure powder buccal tablet of any adjuvant, do not have essential difference.

Inventor in research process, find the stripping of effective ingredient of Herba Dendrobii and the size of particle diameter closely related; particle diameter is less; effective ingredient dissolution rate is faster, and the amount of stripping is more, is more conducive to the absorbing of effective ingredient of Herba Dendrobii; but at volume average particle size D[4; 3] within the scope of=10～40 μ m, the surface tension of Herba Dendrobii micropowder is very large, and general fluidized bed granulation or wet granule compression tablet technique are difficult to direct compression; be prone to the technical problem of sticking, need to add adjuvant to regulate.

Inventor finds in research process, contains a large amount of polysaccharide in Herba Dendrobii medical material, and Dendrobium is met the water capacity and easily formed thick-pasty, if Herba Dendrobii is developed to conventional tablet, water is swallowed, and tablet is easily attached on throat and gastrointestinal tract surface, easy nausea and vomiting while taking, is unfavorable for absorbing.For overcoming the tablet problem of easy disintegrating not in gastrointestinal tract that does not add any disintegrating agent adjuvant, inventor is developed to buccal tablet dosage form by Herba Dendrobii powder, can make Herba Dendrobii micropowder first in oral cavity, melt, effective ingredient is easier stripping under the effect of saliva, avoided conventional tablet drawback of easy disintegrating not in gastrointestinal tract, can improve the utilization rate of human body to the active component of Herba Dendrobii, there is the feature of accurate measurement, steady quality, taking convenience simultaneously.

The dendrobium pure micropowder buccal tablet that does not add at present any adjuvant has no report, and the technical staff of our company is through years of researches, develops the dendrobium pure micropowder that does not add any adjuvant and makes buccal tablet.

Summary of the invention

For current Herba Dendrobii, exist directly with medical material form, to take utilization rate of active components not high, and be developed to the problem that conventional tablet contains a large amount of adjuvants, the object of the present invention is to provide a kind of dendrobium pure powder buccal tablet, do not add any adjuvant, its accurate measurement, steady quality, taking convenience, and appearance character and hardness all can meet the quality standard containing tablet, the invention discloses the preparation technology of this dendrobium pure powder buccal tablet.

Another object of the present invention is that the dendrobium pure powder buccal tablet providing can effectively improve the bioavailability of Herba Dendrobii active component.

The technical solution used in the present invention is for achieving the above object:

Dendrobium pure powder buccal tablet, is characterized in that: it is to be formed by pure Herba Dendrobii micropowder compacting, does not contain any other adjuvant, and the water content of described pure Herba Dendrobii micropowder is 10-18%, volume average particle size D[4,3]=5～40 μ m.

The volume average particle size D[4 of described dendrobium pure micropowder, 3] be 5～20 μ m.

The volume average particle size D[4 of described dendrobium pure micropowder, 3] be 15～30 μ m.

The volume average particle size D[4 of described dendrobium pure micropowder, 3] be 25～40 μ m.

Described dendrobium pure micropowder water content is 10-18%.

Described tablet is buccal tablet.

The preparation method of described dendrobium pure powder buccal tablet, comprises the following steps:

1) Herba Dendrobii fresh goods stem section is cleaned, drained away the water, then Herba Dendrobii is placed in to freeze drying equipment, at-30 ± 2 ℃ of pre-freeze 1-2h, when temperature drops to-30 ℃, constant temperature 3-4h, is reduced between 10-15% to Herba Dendrobii moisture;

2) in the situation that of low temperature, the Herba Dendrobii being dried is pulverized with micron mechanical crusher, require particle size volume average particle size D[4,3] be distributed between 10-40 μ m, after sieving, obtain Herba Dendrobii micropowder;

3) first the Herba Dendrobii micropowder making is carried out to tabletting one time, precompressed 1KN-30KN during tabletting, main pressure 10KN-40KN; After the slice, thin piece being pressed into is pulverized, then carry out secondary tabletting aftershaping, precompressed 5KN-30KN during secondary tabletting, main pressure 20KN-50KN; Or Herba Dendrobii micropowder is carried out after dry granulation, more directly carry out tabletting aftershaping, precompressed 5KN-30KN during direct compression, main pressure 20KN-50KN.

Described Herba Dendrobii is a kind of in Herba Dendrobii, Herba Dendrobii.

Beneficial effect

1) this buccal tablet does not add any adjuvant, the quality standard that tablet appearance is good, hardness meets conventional tablet that adopts special tablet forming technique to make; Owing to not adding any adjuvant, can avoid user too much to take the adjuvant of inanimate object activity, can effectively prevent the certified products of pretending to be of fake and forged product that active constituent content is few simultaneously.

2) Herba Dendrobii micropowder is developed to buccal tablet dosage form, can make Herba Dendrobii micropowder first in oral cavity, melt, effective ingredient is easier stripping under the effect of saliva, by Sublingual and oral cavity buccal surface mucosa, tentatively absorb, have and absorb fast, onset feature rapidly, avoided conventional tablet drawback of easy disintegrating not in gastrointestinal tract, can improve the utilization rate of human body to the active component of Herba Dendrobii, fully improve bioavailability, there is the feature of accurate measurement, steady quality, taking convenience simultaneously.

3) the present invention controls the volume average particle size D[4 of Dendrobium, 3] be distributed between 10～40 μ m, small particle diameter is conducive to discharging completely rapidly of Herba Dendrobii effective ingredient on the one hand, on the other hand, when the mean diameter of Herba Dendrobii micropowder is less than 40 μ m, buccal tablet has the mouthfeel characteristic of good fine and smooth China Resources in oral cavity.

4) the Herba Dendrobii raw material of this buccal tablet adopts deep cooling crush when pulverizing, and can effectively preserve the biological activity of Herba Dendrobii and the loss of minimizing nutritional labeling, has effectively prevented that Herba Dendrobii from because equipment heats up, rotten phenomenon occurring.

Accompanying drawing explanation:

Fig. 1 is the grain size analysis report of Herba Dendrobii I level powder (volume average particle size D[4,3]=749 μ m)

Fig. 2 is the grain size analysis report of Herba Dendrobii II level powder (volume average particle size D[4,3]=286 μ m)

Fig. 3 is the grain size analysis report of Herba Dendrobii III level powder (volume average particle size D[4,3]=28 μ m)

Fig. 4 dendrobine and typical GC chromatogram interior mark naphthalene (peak of retention time 10.1min is interior mark naphthalene, and 17.9min place is dendrobine)

Fig. 5 is that LC-MS/MS method is measured the dendrobine typical color spectrogram (A: blank plasma in Dog Plasma; B: blank plasma adds dendrobine reference substance (blood drug level 5.025ngmL ^-1); C: mark atropine sulfate reference substance in blank plasma adds; D: tested dog A gavage is subject to the plasma sample after test preparation dendrobium pure powder buccal tablet 6h)

Fig. 6 is the average blood drug level-time graph of dendrobine after 6 Beagle dog single gavage Herba Dendrobii II level powder and Herba Dendrobii III level powder

The specific embodiment

1 one kinds of pure powder buccal tablets of Herba Dendrobii of embodiment, preparation method is:

1) get iron-sheet dendrobe fresh product stem section, cleaned, drain away the water, then Herba Dendrobii is placed in to freeze drying equipment, at-30 ± 2 ℃ of pre-freeze 1.5h, when temperature drops to-30 ℃, constant temperature 3h, to Herba Dendrobii moisture reduction 12-15%;

2) in the situation that of 10 ℃ of left and right, the Herba Dendrobii being dried is pulverized with micropowder mechanical crusher, require the volume average particle size D[4 of Dendrobium, 3] be distributed between 10～40 μ m;

3) by the Herba Dendrobii micropowder direct compression making,, wherein, tablet specification 0.25g/ sheet, 100,

First the Herba Dendrobii micropowder making is carried out to tabletting one time, precompressed 1KN～30KN during tabletting, main pressure 10KN～40KN.After the slice, thin piece being pressed into is pulverized, carry out secondary tabletting aftershaping, precompressed 5KN～30KN during tabletting, main pressure 20KN～50KN or Herba Dendrobii micropowder is carried out after dry granulation, carry out direct compression aftershaping, precompressed 5KN～30KN during tabletting, main pressure 20KN～50KN.

2 one kinds of pure powder buccal tablets of rice dry measure used in former times of embodiment, preparation method is:

1) get a meter dry measure used in former times fresh goods stem section, cleaned, drain away the water, then rice dry measure used in former times is placed in to freeze drying equipment, at-30 ± 2 ℃ of pre-freeze 2h, when temperature drops to-30 ℃, constant temperature 4h, is reduced to 15-18% to Herba Dendrobii moisture;

2) in temperature, lower than 15 ℃ of left and right in the situation that, the Herba Dendrobii being dried is pulverized with micron mechanical crusher, is required volume average particle size D[4,3] be distributed between 10～40 μ m;

3) tablet specification 0.25g/ sheet, 100.

First the Herba Dendrobii micropowder making is carried out to tabletting one time, precompressed 1KN～30KN during tabletting, main pressure 10KN～40KN.After the slice, thin piece being pressed into is pulverized, carry out secondary tabletting aftershaping.Precompressed 5KN～30KN during tabletting, main pressure 20KN～50KN.

Or Herba Dendrobii micropowder is carried out, after dry granulation, carrying out direct compression aftershaping.Precompressed 5KN～30KN during tabletting.Main pressure 20KN～50KN.

The process results that the present invention produces two kinds of tablettings contrasts in Table 1(in 100 tablets of pure powder buccal tablets).

The process results contrast test data of two kinds of tablettings of table 1

Bioavailability test example in the Herba Dendrobii micropowder In Vitro Dissolution test of different-grain diameter and body

For beneficial effect of the present invention is described, below in conjunction with accompanying drawing, bioavailability situation in the stripping situation of Herba Dendrobii micropowder in the present invention and body is further described to explanation.

In the present invention, first inventor is that the breaking method of Herba Dendrobii micropowder and the medicinal powder size of acquisition have been carried out sufficient research to preparing unique constitutive material of dendrobium pure micropowder tablet.Result of study shows, all comminution process all can be smashed cell on a certain amount of, but Ordinary pulverization method and cellular level pulverizing method have significant difference in Herba Dendrobii plant cell wall breaking rate: the powder diameter that Ordinary pulverization obtains is larger than cell dia, cell wall breaking rate is just lower, in Dendrobium, most cells are complete, its powder particle is comprised of several, the even more intact cells of dozens of, the stripping of the effective ingredient in cell need just can enter solvent through several even dozens of cells, can not can only waste through the part of cell wall; Cellular level pulverizing Dendrobium situation is completely different, and Herba Dendrobii cell wall breaking rate is high, and the effective ingredient in cell can directly contact solvent, is conducive to stripping and absorption of human body.Therefore inventor has selected cellular level pulverizing method, utilize the brute force vibration of vibration source, make Herba Dendrobii medical material in crushing chamber under fluidized state, be subject to the high-intensity effect of hitting, cut, grind, rubbing with the hands resultant force of frotton, make Herba Dendrobii medical material reach at short notice micron order and pulverize or cell wall breaking effect.The intensity of vibrating by adjustment, can conveniently control by the fineness of comminuting matter.Adopt cryogenics simultaneously, can reduce the temperature in crushing chamber, be beneficial to the pulverizing of heat sensitive material.

The volume average particle size D[4 when Herba Dendrobii micropowder, 3 are found in inventor's research] during=28 μ m rank, the dissolution time of effective ingredient is fast, and dissolution rate is high, can realize the object that the effective ingredient of Herba Dendrobii buccal tablet discharges fast in oral cavity.If volume average particle size less (<10 μ m), the specification requirement to disintegrating apparatus in actual production is higher, and cost straight line rises.In addition, inventor finds under study for action, and when the mean diameter of Herba Dendrobii micropowder surpasses 40 μ m, buccal tablet just can obviously be felt harsh feeling in oral cavity; When particle mean size is during in 25 μ m left and right, buccal tablet has the mouthfeel characteristic of good fine and smooth China Resources.Therefore, the present invention controls volume average particle size D[4,3]=10～40 μ m, can take into account the mouthfeel of the absorption of active component, the control of cost and buccal tablet.

By different other active substance stripping experimental data powder of level of pulverizing of Herba Dendrobii, test to illustrate the volume average particle size D[4 controlling in the present invention, 3 below] in the selection foundation of 10～40 μ m.Experiment is provided with 4 and pulverizes rank, is respectively 1. Herba Dendrobii herb; 2. Herba Dendrobii I level powder volume average particle size D[4,3]=749 μ m; 3. Herba Dendrobii II level powder volume average particle size D[4,3]=286 μ m; 4. Herba Dendrobii III level powder volume average particle size D[4,3]=28 μ m; Fig. 1 to Fig. 3 is shown in grain size analysis report.

1, the mensuration of extractum

Press " hot dipping " operation under " water-soluble extractives " item in < < Chinese Pharmacopoeia > 2010 editions one appendix XA of > " Extract mensuration ".Get respectively Herba Dendrobii herb, Herba Dendrobii I level powder, Herba Dendrobii II level powder and each about 2g of Herba Dendrobii III level powder test sample, accurately weighed, to put in the conical flask of 100ml, precision adds water 50ml, close plug, weighed weight, after standing 1 hour, connect reflux condensing tube, be heated to boiling, and keep micro-and boil, respectively micro-5min that boils, 15min, 30min and 60min.After letting cool, take off conical flask, close plug, weighed weight again, water is supplied the weight of less loss, shakes up, with dry filter, filter, precision measures filtrate 25ml, puts in the evaporating dish that is dried to constant weight, in water-bath after evaporate to dryness, in 105 ℃ dry 3 hours, put in exsiccator rapid accurately weighed weight cooling 30 minutes, the content (%) that calculates water-soluble extractives in test sample with dry product, experimental result is in Table 2.

Different other Herba Dendrobii of the level leaching content (%) in time of pulverizing of table 2

Experimental result shows: each pulverizes other extract content of level progressively increases in time, and 30min respectively organizes extractum later and reaches balance.Each pulverizes other extract content of level reducing and increase with granularity, granularity is thinner, more be conducive to the leaching of effective ingredient, when 5min, the leaching content of Herba Dendrobii III level powder is about 4 times of Herba Dendrobii herb, be about 2 times of Herba Dendrobii I level powder, the results show Herba Dendrobii granule size has important impact to the stripping of effective ingredient.

2, the mensuration of dendrobium polysaccharide

Experimental principle: adopting phenol sulfuric acid process is to measure polyoses content, utilizes polysaccharide under the effect of sulphuric acid, be hydrolyzed into monosaccharide molecule, and dehydration generates alditol derivant rapidly, then forms colored compound with phenol, then detects with spectrophotometer.

Experimental technique: the preparation of reference substance solution: precision takes 105 ℃ of glucose reference substance 10mg that are dried to constant weight, is placed in the measuring bottle of 100ml, with water, is settled to scale, shakes up and obtain reference substance solution.

Detect determining of wavelength: precision pipettes reference substance solution 0.4ml, be placed in 10ml tool plug test tube, add water to 2ml and shake up, obtain reference substance diluent, add phenol reagent 1mL, shake up, then add rapidly concentrated sulphuric acid 5mL, fully shake up, place 5min, put in boiling water bath and heat 15min, put into the cooling 10min of frozen water.With method, take distilled water as blank solution.In 300-800nm wave-length coverage, take blank reactant liquor as reference, example reaction liquid is scanned, from scintigram, λ max=489.5nm, will check wavelength set is 489nm.

The preparation of standard curve: precision takes 105 ℃ of glucose reference substance 0.01g that are dried to constant weight, is dissolved in water, and 100mL volumetric flask standardize solution obtains the glucose reference substance liquid of 0.1mgmL-1.Precision measures reference substance liquid 0.3,0.4,0.5,0.6,0.7,0.8mL, is placed in tool plug test tube, adds water to respectively 2.0mL, with detecting the middle method operation of determining of wavelength, under 489nm, measures absorbance.Take concentration as abscissa, and absorbance is vertical coordinate, drawing standard curve.Result shows, glucose content is good in 0.015mg/ml～0.045mg/ml scope internal linear relation.The results are shown in Table 3:

Table 3 glucose reference substance absorbance

The mensuration of conversion factor: take appropriate Herba Dendrobii, use petroleum ether 100ml, reflux, extract, 1h, petroleum ether is flung in water-bath, residue adds 80% ethanol 100ml, at ultrasonic power 300w, pretreatment 6min under 45HZ condition, filtered while hot after lixiviate 1h under 80 ℃ of water bath condition again, washing with alcohol 3 times of defecator and residue, residue adds water 100ml, supersound extraction 30min, filter to get filtrate 1., residue adds water 100ml, ultrasonic 30min, filter to get filtrate 2., residue continues to add water 100ml, ultrasonic 30min, filter to get filtrate 3., merging filtrate 1. 2. 3., be concentrated into 50ml, add long-pending 95% ethanol of triploid, placement is spent the night, separating precipitate is dissolved in distilled water, with Sevay method deproteinization: chloroform adds by polysaccharide solution 1/5 volume, then the n-butyl alcohol that adds chloroform volume 1/5, violent jolting 20min, centrifugal 30min, the Denatured protein of branch vibration layer and solution layer intersection, collect supernatant, in supernatant, add 3 times of volumes, 95% ethanol precipitation, precipitate is used dehydrated alcohol successively, acetone, ether washing, be dried to constant weight, obtain dendrobium polysaccharide.Precision takes 60 ℃ of dendrobium polysaccharide 1mg that are dried to constant weight, is placed in 10mL volumetric flask, to scale, shakes up, as polysaccharide stock solution with distilled water diluting.Precision measures this polysaccharide stock solution 0.3mL, measures absorbance.According to standard curve, obtain the concentration of glucose in dendrobium polysaccharide solution, be calculated as follows conversion factor (F).

The concentration of F=polysaccharide solution (μ gmL-1)/(concentration of glucose in polysaccharide solution (μ gmL-1) * dilution factor), calculates: F=1.284

The preparation of need testing solution: precision takes Herba Dendrobii herb, Herba Dendrobii I level powder, Herba Dendrobii II level powder and each 0.25g of Herba Dendrobii III level powder, add petroleum ether 100mL, 40 ℃ of reflux 1h, discard petroleum ether liquid, residue is put water-bath Back stroke ether to the greatest extent, add 80% ethanol 100mL, 80 ℃ of heating and refluxing extraction 1h, while hot sucking filtration, 80% ethanol 20mL washing 3 times for filtering residue and filter, filtering residue is put in flask, and residue adds water 100ml, supersound extraction dendrobium polysaccharide, time is respectively 5min, 15min, 30min and 60min, in 1000mL volumetric flask standardize solution, mix.Each accurate 2.0mL that draws, in tool plug test tube, by " detecting determining of a wavelength " lower operation, records and respectively manages absorbance, and the regression equation of substitution gained respectively calculates the dendrobium polysaccharide content of each pipe.

Dendrobium polysaccharide content W=(C*S*V*F/M) * 100%

(W: dendrobium polysaccharide content/%; C: dendrobium polysaccharide concentration mg/mL in the sample being drawn by regression equation; S: extension rate; V: sample extracting solution volume mL; M: test sample sample weighting amount mg)

Experimental result: the results are shown in Table 4.

Different other dendrobium polysaccharide of the level changes of contents (%) in time of pulverizing of table 4

Experimental result shows: the stripping that ultrasonic time is pulverized other dendrobium polysaccharide of level to difference has certain influence, and 30min left and right can reach extraction balance; Different other Herba Dendrobiis of level of pulverizing have significant impact to the stripping of total polysaccharides, and Dendrobium granularity is thinner, is more conducive to the stripping of total polysaccharides, and wherein Herba Dendrobii III level powder dissolution rate is the fastest, and 5min left and right can reach the contents level that Herba Dendrobii herb extracts 1 hour.

3, the mensuration of dendrobine

Press " assay " lower operation in < < Chinese Pharmacopoeia > > 2010 editions " Herba Dendrobii ".

Instrument and condition: Agilent6890N gas chromatograph, chromatographic column: Agilent DB-1(column length 15m, internal diameter is 0.53mm, film thickness is 0.5 μ m), temperature programming: initial temperature is 80 ℃, is warming up to 250 ℃ with the speed of 10 ℃ per minute, keeps 5 minutes.Injector temperature: 250 ℃, detector temperature: 250 ℃ of split ratio: 10.0:1, nitrogen flow: 1.0ml/min, detector air-flow: hydrogen: 45.0ml/min; Air: 400ml/min, make-up gas (nitrogen): 40ml/min.

Correction factor is measured: get the about 25mg of naphthalene reference substance, and accurately weighed, put in 25ml measuring bottle, add dissolve with methanol and be diluted to scale and make every 1ml containing the solution of 1mg, as inner mark solution.Get the about 5mg of dendrobine reference substance, accurately weighed, put in 10ml measuring bottle, add dissolve with methanol and be diluted to scale, the accurate 2ml that draws, is transferred to 20ml measuring bottle, adds methanol and is diluted to scale, makes every 1ml containing the solution of 50 μ g, in contrast product solution.Precision measures reference substance solution 2ml, puts in 5ml measuring bottle, and precision adds inner mark solution 0.5ml, adds methanol to scale, shakes up, and draws 1 μ l, inject gas chromatograph, the calculation correction factor.

The preparation of need testing solution: get Herba Dendrobii herb, Herba Dendrobii I level powder, Herba Dendrobii II level powder and each about 0.25g of Herba Dendrobii III level powder, accurately weighed, to put in round-bottomed flask, precision adds the methanol solution 25ml of 0.05% formic acid, weighed weight, 70 ℃ of heating in water bath backflow 10min, 30min, 1 hour and 3 hours, let cool, more weighed weight, with the methanol solution of 0.05% formic acid, supply the weight of less loss, shake up, filter.Precision is got subsequent filtrate 2ml, puts in 5ml measuring bottle, and precision adds inner mark solution 0.5ml, adds methanol to scale, shakes up, and draws 1 μ l, and inject gas chromatograph is measured, and obtains.The results are shown in Table 4, the typical gas chromatogram of dendrobine is shown in Fig. 4.

The different changes of contents (%) of other dendrobine of level with extraction time of pulverizing of table 5

Experimental result shows: the stripping that extraction time is pulverized other dendrobine of level to difference has certain influence, about 3 hours, can reach extraction balance; Different other Herba Dendrobiis of level of pulverizing have significant impact to the stripping of dendrobine, and Dendrobium granularity is thinner, is more conducive to the stripping of dendrobine, and Herba Dendrobii III level powder dissolution rate is the fastest, also the most easily reaches stripping balance.

4, Herba Dendrobii II level powder and the bioavailability study of Herba Dendrobii III level powder in beasle dog body

Through above-mentioned basic research, there is notable difference in effective ingredient dissolution rate and the dissolution rate that can find varigrained Dendrobium, but above-mentioned experiment is experiment in vitro, still the situation after taking in the proof that lacks persuasion body, therefore select Herba Dendrobii II level powder and Herba Dendrobii III level powder, entrust China Medicine University to carry out bioavailability study in beasle dog body.

(1) research approach

Adopt dual crossing EXPERIMENTAL DESIGN, selecting 6 beagle dogs is experimental animal, male and female half and half, be divided at random two groups, every group 3, carry out 2 cycle cross-over experiments, respectively single gavage Herba Dendrobii II level powder (volume average particle size D[4,=286 μ m) and Herba Dendrobii III level powder (volume average particle size D[4,3]=28 μ m) 3]; Between cycle through 6 days cleaning phases.Adopt LC-MS (LC-MS-MS) method measure after administration in dog plasma the blood drug level of dendrobine through time process, calculate pharmacokinetic parameter, relatively the bioavailability difference of the Herba Dendrobii micropowder of two kinds of different-grain diameters.

(2) animal experiment method

It is Herba Dendrobii dry product 6～12g/ days that < < Chinese Pharmacopoeia > > records people's taking dose, get 10g/ days, by body weight for humans 70kg, calculate dosage conversion between animal species, Beagle dog (calculating by 12kg) dosage is multiplied by coefficient 0.32.Therefore calculating Beagle dog dosage is 10g * 0.32/12kg=0.267g/kg.Take respectively Herba Dendrobii II level powder and Herba Dendrobii III level powder is appropriate, put in graduated cylinder, add a certain amount of 0.5%CMC-Na solution, limit edged stirs, make Dendrobium dispersed gavage solution, administration volume is that 4mL/kg(is equivalent to dendrobine 0.56mgkg ^-1).Tested Beagle dog fasting 12h in the tested evening before yesterday, morning next day 8:00 gastric infusion.Before taking medicine (0h) and take medicine after 0.167,0.333,0.5,0.75,1,1.5,2,3,4,5,6, and 8h is respectively through the about 3.0mL of forelimb venous blood sampling; Put in heparinization centrifuge tube, the centrifugal 10min of 3000r/min divides and gets blood plasma, and blood plasma is put into the Refrigerator store of-20 ℃ immediately, and the blood plasma (in 24 hours) that gathered the same day is put household freezer and in half an hour, sent to pharmaceutical analysis teaching and research room of China Medicine University, put-20 ℃ of Refrigerator stores, for determination of plasma concentration.

(3) blood drug level method of testing

Instrument: Shimadzu2010CHT high performance liquid chromatograph (being contained in line vacuum degasser, quaternary gradient pump, constant temperature automatic sampler, column oven), Finnigan TSQ triple quadrupole bar mass spectrograph (electric spray ion source ESI), Xcalibur1.4 data workstation; Spectrum post: Lichrosphere C6H6 (4.6 * 150mm, 5 μ m), Hanbon Sci. & Tech. Co., Ltd., mobile phase: methanol-water-formic acid (35:65:0.1) flow velocity: 1.0mL/min column temperature: 35 ℃.Ionizing mode: ESI; Scan mode: mass spectrum multiple-reaction monitoring (SRM); Spray voltage 4500eV; Sheath atmospheric pressure 35psi; Ion cleaning air pressure 0.5psi; Assisted gas pressure 5psi; 350 ℃ of capillary temperatures; Source CID-10eV; Collision atmospheric pressure 1.0mTorr; Ion polarity: cation; Detect ion: dendrobine selective reaction detects ion [M+H] ⁺m/z264.1@18eV → m/z264.1; Atropine sulfate selective reaction detects ion [M+H] ⁺m/z290.1@18eV → m/z124.1;

Plasma sample processing method: the accurate plasma sample 200 μ L that draw, put in 2mL plastic centrifuge tube, precision adds methanol 20 μ L(standard curves and quality-control sample to add the dendrobine 20 μ L reference substance solution of respective concentration), precision adds inner mark solution 20 μ L, and vortex mixes for 30 seconds; Add again 120 μ L6% perchloric acid precipitation albumen, vortex 3min, 12000r/min high speed centrifugation 10min, gets supernatant 20 μ L and carries out LC-MS/MS analysis.Press the concentration of dendrobine in internal standard method calculating plasma sample.

Under the chromatographic condition adopting in this experiment, dendrobine appearance time is in 4.8min left and right, and interior mark atropine sulfate appearance time is in 4.1min left and right, and peak shape is good, and baseline is steady.6 Beagle dog blank plasmas have been investigated, 6 Beagle dog blank plasmas add dendrobine reference substance, 6 Beagle dog blank plasmas add interior mark reference substance, and the plasma sample chromatogram (see figure 5) of tested dog after taking medicine, result show determinand and in mark behind peak and have no effluent in 15min.For avoiding perchloric acid and polarity endogenous contaminating impurity mass spectrum, the chromatographic effluent of 0～3.0min does not enter mass spectrum by transfer valve.

Methodological study result shows: the LC-MS/MS of foundation measures the method for the dendrobine in blood plasma, at 0.05025～201ngmL ^-1scope internal linear is good, correlation coefficient r >0.99; The preci-sion and accuracy of lower limit of quantitation all meets the requirements; The accuracy of basic, normal, high concentration is between 85%～115%.In batch and batch between sample precision good (RSD<15%); Extraction recovery and matrix effect all meet the requirements; Reference substance stock solution is placed and is had good stability for 30 days under 4 ℃ of conditions of refrigerator, and interior mark stock solution is placed and had good stability for 25 days under 4 ℃ of conditions of refrigerator; Sample introduction solution after processing is positioned on the specimen disc of 15 ℃, stable in 8 hours; Plasma sample is at room temperature placed 5 hours, stable under the multigelation condition of 3 times and-20 ℃ freezing 22 days.This method has higher specificity, can measure the concentration of dendrobine in blood plasma accurate, single-mindedly.

(4) experimental result

After 6 Beagle dog single gavage Herba Dendrobii II level powder Herba Dendrobii III level powder, dendrobine blood drug level-time graph of each tested dog is shown in Fig. 6.

Main pharmacokinetic parameters adopts professor Sun Ruiyuan to wait the DAS2.1.1 computed in software of writing.With non-chamber dependent form method, calculate.Cmax and Tmax are measured value.Pharmacokinetic parameters is in Table 6, and relative bioavailability result of calculation is in Table 7, and the relative bioavailability of Dendrobium sample IV is 196.9 ± 39.5% of sample II.

The comparison of the main pharmacokinetic parameters of table 6

Table 7 relative bioavailability table

By beasle dog bioavailability experimental result, can be found out, volume average particle size 28 μ m Herba Dendrobii III level powder are compared with the Herba Dendrobii II level powder of volume average particle size 286 μ m, bioavailability has in vivo improved nearly one times, show that absorption in vivo has important effect to micronization to the effective ingredient of this valuable medicinal of promotion Herba Dendrobii, micronization technique can effectively promote its inherent value.

Claims (8)

1. dendrobium pure powder buccal tablet, is characterized in that: it is to be formed by pure Herba Dendrobii micropowder compacting, does not contain any other adjuvant, and the water content of described pure Herba Dendrobii micropowder is 10-18%, volume average particle size D[4,3]=5～40 μ m.

2. dendrobium pure powder buccal tablet according to claim 1, is characterized in that: the volume average particle size D[4 of described dendrobium pure micropowder, 3] be 5～20 μ m.

3. dendrobium pure powder buccal tablet according to claim 1, is characterized in that: the volume average particle size D[4 of described dendrobium pure micropowder, 3] be 15～30 μ m.

4. dendrobium pure powder buccal tablet according to claim 1, is characterized in that: the volume average particle size D[4 of described dendrobium pure micropowder, 3] be 25～40 μ m.

5. dendrobium pure powder buccal tablet according to claim 1, is characterized in that: described dendrobium pure micropowder water content is 10-18%.

6. according to the dendrobium pure powder buccal tablet described in claim 1-5 any one, it is characterized in that: described tablet is buccal tablet.

7. the preparation method of dendrobium pure powder buccal tablet as claimed in claim 1, is characterized in that: comprise the following steps:

1) Herba Dendrobii fresh goods stem section is cleaned, drained away the water, then Herba Dendrobii is placed in to freeze drying equipment, at-30 ± 2 ℃ of pre-freeze 1-2h, when temperature drops to-30 ℃, constant temperature 3-4h, is reduced between 10-15% to Herba Dendrobii moisture;

2) in the situation that of low temperature, the Herba Dendrobii being dried is pulverized with micron mechanical crusher, require particle size volume average particle size D[4,3] be distributed between 10-40 μ m, after sieving, obtain Herba Dendrobii micropowder;

3) first the Herba Dendrobii micropowder making is carried out to tabletting one time, precompressed 1KN-30KN during tabletting, main pressure 10KN-40KN; After the slice, thin piece being pressed into is pulverized, then carry out secondary tabletting aftershaping, precompressed 5KN-30KN during secondary tabletting, main pressure 20KN-50KN; Or Herba Dendrobii micropowder is carried out after dry granulation, more directly carry out tabletting aftershaping, precompressed 5KN-30KN during direct compression, main pressure 20KN-50KN.

8. dendrobium pure powder buccal tablet according to claim 1, is characterized in that: described Herba Dendrobii is a kind of in Herba Dendrobii, Herba Dendrobii.