CN105726623B - Astragalus membranaceus ultramicro decoction pieces as well as preparation method and quality testing method thereof - Google Patents

Astragalus membranaceus ultramicro decoction pieces as well as preparation method and quality testing method thereof Download PDF

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CN105726623B
CN105726623B CN201610106958.8A CN201610106958A CN105726623B CN 105726623 B CN105726623 B CN 105726623B CN 201610106958 A CN201610106958 A CN 201610106958A CN 105726623 B CN105726623 B CN 105726623B
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radix astragali
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吕承洋
吕随民
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GANSU FUXINGHOU BIOLOGICAL MEDICAL TECHNOLOGY Co Ltd
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Abstract

The invention belongs to the field of traditional Chinese medicines, and in particular relates to astragalus membranaceus ultramicro decoction pieces as well as a preparation method and a quality testing method thereof. The astragalus membranaceus ultramicro decoction pieces are prepared from the following raw materials in parts by weight: water extract of 10-20% of an astragalus membranaceus decoction piece and superfine crushed powder of 80-90% of the astragalus membranaceus decoction piece. The preparation method comprises the following steps: (1) adding water with the amount being 5-15 times of the weight of the astragalus membranaceus ultramicro decoction pieces into 10-20% of the astragalus membranaceus ultramicro decoction pieces, boiling for 1-4 times for 0.5-2.5 hours each time, mixing the decoctions, and concentrating to an extract, so as to obtain the water extract of the astragalus membranaceus ultramicro decoction pieces; (2) performing superfine crushing on 80-90% of the astragalus membranaceus ultramicro decoction pieces, so as to obtain the superfine crushed powder of the astragalus membranaceus ultramicro decoction pieces; (3) mixing the water extract of the astragalus membranaceus ultramicro decoction pieces with the superfine crushed powder of the astragalus membranaceus ultramicro decoction pieces, pelletizing, and drying, so as to obtain the astragalus membranaceus ultramicro decoction pieces.

Description

A kind of Radix Astragali ultramicro decoction piece and preparation method thereof and quality determining method
Technical field
The present invention relates to the field of Chinese medicines is and in particular to a kind of Radix Astragali ultramicro decoction piece and preparation method thereof and quality testing side Method.
Background technology
Chinese medicine ultramicro decoction piece is to process and extract, through science, the Chinese medicine drink that its active ingredient is made by single medicinal material medicine materical crude slice Piece novel form, has the advantages that taking convenience, accurate measurement, safety and sanitation, is easy to allocate.
Ultramicro decoction piece of the present invention is with Radix Astragali medicine materical crude slice as raw material, through technique systems such as water extraction, concentration, drying, ultramicro grinding Standby Chinese medicine ultramicro decoction piece, under the specific preparation technology of here, the ultramicro decoction piece of the present invention of preparation is different to treatment endometrium Potential energy enough plays unexpected prominent treatment advantage.
Endometriosis is one of gynaecology's common disease, the incidence of disease in the women of child-bearing age about 10, and falls ill Rate is in rising trend.Because the multiple, recurrent of endometriosis and the difficulty for the treatment of are big, and canceration is had to be inclined to, For one of difficult and complicated cases in gynecological disease, mainly adopt operative treatment and hormone therapy at present.Endometriosis causes Pelvic pain, dysmenorrhoea and the infertile quality of life having had a strong impact on patient and health, caused the attention of numerous doctors.Traditional Chinese medicine One of main method for the treatment of endometriosis, have compared with Western medicine that toxic action is little, can take for a long time, curative effect stable The features such as.
There is no the report of content assaying method at present with regard to ultramicro decoction piece of the present invention.Lupenone is the activity one-tenth in the Radix Astragali / mono-, assay is carried out to it is ensured that clinical application safely, effectively.The invention provides to ultra micro of the present invention The method of lupenone assay in medicine materical crude slice.
The base of the Radix Astragali of the present invention is the root of pulse family Astragalus astragalus mongolicus and Astragalus membranacus.
Content of the invention
It is an object of the invention to provide a kind of Radix Astragali ultramicro decoction piece.
It is a further object of the present invention to provide the preparation method of this Radix Astragali ultramicro decoction piece.
Present invention also offers the quality determining method of this Radix Astragali ultramicro decoction piece.
Present invention also offers the pharmaceutical applications of this Radix Astragali ultramicro decoction piece.
The purpose of the present invention is realized by the following manner：
A kind of Radix Astragali ultramicro decoction piece is to be made up of the raw material of following weight portion：The water of Radix Astragali medicine materical crude slice 10%～20% weight portion Extract, the Ultramicro-powder of Radix Astragali medicine materical crude slice 80%～90% weight portion mince.
The preparation method of this Radix Astragali ultramicro decoction piece is as follows：
（1）Take Radix Astragali medicine materical crude slice 10%～20% weight portion, plus 5～15 times amount water, decoct 1～4 time, 0.5～2.5 is little every time When, merge decocting liquid, be condensed into medicinal extract, obtain the water extract of Radix Astragali medicine materical crude slice；
（2）Take Radix Astragali medicine materical crude slice 80%～90% weight portion, carry out ultramicro grinding, the Ultramicro-powder obtaining Radix Astragali medicine materical crude slice minces；
（3）The water extract of above-mentioned Radix Astragali medicine materical crude slice is minced with the Ultramicro-powder of Radix Astragali medicine materical crude slice and merges, pelletize, be dried, obtain yellow Stilbene ultramicro decoction piece.
The preferred preparation method of the water extract of Radix Astragali medicine materical crude slice in this Radix Astragali ultramicro decoction piece is as follows：Take Radix Astragali medicine materical crude slice 10% ～20% weight portion, plus 8～12 times amount water, decocts 2～3 times, 1～2 hour every time, merges decocting liquid, be condensed into medicinal extract, obtain The water extract of Radix Astragali medicine materical crude slice.
The preparation method being more highly preferred to of the water extract of Radix Astragali medicine materical crude slice in this Radix Astragali ultramicro decoction piece is as follows：The Radix Astragali is taken to drink Piece, plus 10 times amount water, decoct 2 times, 1 hour every time, merge decocting liquid, be condensed into medicinal extract, obtain the water extract of Radix Astragali medicine materical crude slice.
The preparation method that the Ultramicro-powder of the Radix Astragali medicine materical crude slice in this Radix Astragali ultramicro decoction piece minces is as follows：Take Radix Astragali medicine materical crude slice 80%～ 90% weight portion, carries out ultramicro grinding using pulverizer to Radix Astragali medicine materical crude slice, and after ultramicro grinding, the Ultramicro-powder of Radix Astragali medicine materical crude slice minces Particle diameter is 10～75 μm.
The preferred preparation method that the Ultramicro-powder of the Radix Astragali medicine materical crude slice in this Radix Astragali ultramicro decoction piece minces is as follows：Using air-flow crushing Machine carries out ultramicro grinding to Radix Astragali medicine materical crude slice, and the stream pressure of airslide disintegrating mill is 1000～1200kPa, and charging rate is 160～ 200r·min-1, grade frequency is 30～40Hz, and grinding time is 40～50min, and the Ultramicro-powder obtaining Radix Astragali medicine materical crude slice minces.
The preparation method being more highly preferred to that the Ultramicro-powder of the Radix Astragali medicine materical crude slice in this Radix Astragali ultramicro decoction piece minces is as follows：Using gas Stream pulverizer carries out ultramicro grinding to Radix Astragali medicine materical crude slice, and the stream pressure of airslide disintegrating mill is 1100kPa, and charging rate is 180r min-1, grade frequency is 35Hz, and grinding time is 45min, and the Ultramicro-powder obtaining Radix Astragali medicine materical crude slice minces.
Described Radix Astragali ultramicro decoction piece can be using high performance liquid chromatography to the lupenone in this Radix Astragali ultramicro decoction piece Carry out assay, assay method is：
（1）Chromatographic condition：Chromatographic column：C18Post；Mobile phase：Ratio is 83：17 acetonitrile -0.5% phosphoric acid solution；Flow velocity： 1.0mL/min；Detection wavelength：210nm；Column temperature：35℃；Sample size：20μL；Number of theoretical plate is calculated and should be not less than by lupenone 5000；
（2）The preparation of reference substance solution：Take lupenone reference substance appropriate, accurately weighed, plus methyl alcohol makes every 1mL and contains The solution of 0.0773mg, as reference substance storing solution；Again in accurate absorption reference substance storing solution 2mL to 25mL measuring bottle, plus methyl alcohol It is diluted to scale, shakes up, obtain final product；
（3）The preparation of need testing solution：Take ultramicro decoction piece of the present invention, finely ground, accurately weighed, put in conical flask with cover, essence Close addition methyl alcohol 25mL, close plug, weighed weight, ultrasonically treated 30min, let cool, more weighed weight, the weight of less loss is supplied with methyl alcohol Amount, shakes up, filtration, takes subsequent filtrate, obtains final product；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph.
Described Radix Astragali ultramicro decoction piece can also be using high performance liquid chromatography to the different dimpling in this Radix Astragali ultramicro decoction piece Sword-like leave Sha's alcohol carries out assay, and assay method is：
（1）Chromatographic condition：Chromatographic column is C18Post；Mobile phase：Ratio is 10：90 acetonitrile -0.1% phosphoric acid；Column temperature：30℃； Flow velocity：1.0 ml/min；Detection wavelength：207 nm；Sample size：20μL；Number of theoretical plate presses different dimpling sword-like leave Sha's alcohol calculating should not Less than 5000；
（2）The preparation of reference substance solution：Precision weighs different dimpling sword-like leave Sha's alcohol reference substance 10.13 mg, puts 100 ml measuring bottles In, dissolved with methyl alcohol and be diluted to scale, shake up；Precision measures 2 ml, puts in 25 ml measuring bottles, is diluted to scale with mobile phase, Shake up, then every 1 ml contains different dimpling sword-like leave Sha's alcohol 8.1 μ g；
（3）The preparation of need testing solution：Precision weighs in Radix Astragali ultramicro decoction piece about 0.2 g to 100 ml measuring bottle, plus methyl alcohol The ultrasonic 15min of 80 ml, with methanol constant volume to scale after cooling, shakes up, and pipettes in 1 ml to 10 ml measuring bottle, with the phase dilution that flows To scale, shake up, 0.45 μm of filter membrane filtration, take subsequent filtrate as need testing solution；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph.
Described Radix Astragali ultramicro decoction piece can be used for preparing the medicine for the treatment of endometriosis.
Described Radix Astragali ultramicro decoction piece can be additionally used in preparing the medicine for the treatment of gonitis.
Verify the technique effect of the present invention by following experimental study：
Experiment one：Ultramicro decoction piece of the present invention treats the experimental study of endometriosis
1 experiment material
1.1 animal
Health does not mate female Wistar rat, and body weight 200～250 g, purchased from Gansu university of TCM animal used as test Center.
1.2 medicine
1.2.1 Radix Astragali ultramicro decoction piece of the present invention：Prescription：The water extract of Radix Astragali medicine materical crude slice 200g, Radix Astragali medicine materical crude slice 800g super Micro mist minces.Preparation method：（1）Take Radix Astragali medicine materical crude slice 200g, add water 2000mL, decoct 2 times, 1 hour every time, merge decocting liquid, concentrate Become medicinal extract, obtain the water extract of Radix Astragali medicine materical crude slice；（2）Take Radix Astragali medicine materical crude slice 800g, using airslide disintegrating mill, Radix Astragali medicine materical crude slice is carried out Ultramicro grinding, the stream pressure of airslide disintegrating mill is 1100kPa, and charging rate is 180r min-1, grade frequency is 35Hz, powder The broken time is 45min, measures the particle diameter distribution of powder, the D of fine powder using laser particle instrument wet method50, D90It is respectively 29.758 μ M, 58.376 μm, the Ultramicro-powder obtaining Radix Astragali medicine materical crude slice minces；（3）Radix Astragali water extract is minced with Radix Astragali Ultramicro-powder and mixes, obtain To Radix Astragali ultramicro decoction piece of the present invention.It is configured to 0.5g/mL drug suspension, standby.
1.2.2 contrast Radix Astragali ultramicro decoction piece A：Prescription：The water extract of Radix Astragali medicine materical crude slice 200g, the ultra micro of Radix Astragali medicine materical crude slice 800g Crushed material A.Preparation method：（1）Take Radix Astragali medicine materical crude slice 200g, add water 2000mL, decoct 2 times, 1 hour every time, merge decocting liquid, be condensed into Medicinal extract, obtains the water extract of Radix Astragali medicine materical crude slice；（2）Take Radix Astragali medicine materical crude slice 800g, using airslide disintegrating mill, Radix Astragali medicine materical crude slice is surpassed Crushing of Ultrafine, the stream pressure adopting airslide disintegrating mill is 1300kPa, and charging rate is 150r min-1, grade frequency is 50Hz, powder The broken time is 60min, measures the particle diameter distribution of powder, the D of fine powder using laser particle instrument wet method50, D90It is respectively 4.658 μm, 8.951 μm, the Ultramicro-powder obtaining Radix Astragali medicine materical crude slice minces A；（3）Radix Astragali water extract and the Radix Astragali Ultramicro-powder A that minces is mixed, obtains Contrast Radix Astragali ultramicro decoction piece A.It is configured to 0.5g/mL drug suspension, standby.
1.2.3 medicine materical crude slice B pulverized by the contrast Radix Astragali：Prescription：The water extract of Radix Astragali medicine materical crude slice 200g, the pulverizing of Radix Astragali medicine materical crude slice 800g Thing B.Preparation method：（1）Take Radix Astragali medicine materical crude slice 200g, add water 2000mL, decoct 2 times, 1 hour every time, merge decocting liquid, be condensed into leaching Cream, obtains the water extract of Radix Astragali medicine materical crude slice；（2）Take Radix Astragali medicine materical crude slice 800g, pulverized using conventional tooth-like medicinal herb grinder, mistake 120 mesh sieves, measure the particle diameter distribution of powder, the D of fine powder using laser particle instrument wet method50, D90It is respectively 95.658 μm, 118.951 μm, obtain the crushed material B of Radix Astragali medicine materical crude slice；（3）The crushed material B of the water extract of Radix Astragali medicine materical crude slice and Radix Astragali medicine materical crude slice is mixed Close, obtain contrasting Radix Astragali pulverizing medicine materical crude slice B.It is configured to 0.5g/mL drug suspension, standby.
1.2.4 danazol capsule：200 mg/ grains, Jiangsu Lianhuan Pharmaceutical Co., Ltd. produces.
1.3 reagent and instrument
Estradiol（E2）Put and exempt from medicine box, progesterone（P）Put and exempt from medicine box, carried by Tianjin Jiuding Medical Biological Engineering Co., Ltd For.ER, PR, VEGF, NK cellular immunity group kit, is provided by Wuhan Boster Biological Technology Co., Ltd..2008 G/r Automatic counter for counting.Microscope：OLYMPUS, model IX70.
2 experimental techniques
2.1 modeling
Animal Model takes the adult female rats vaginal smear examination oestrous cycle.Take estrus rat bibliography Method（Chang Nuan. the foundation of Endometriosis Model in Rats and its pathological observation [J]. clinical and experimental pathology magazine, 1998,14（1）: 67-69.）, yellow Jackets 40 mg/kg intraperitoneal injection of anesthesia, does one long above lower abdomen center keeps left About 1.5 cm otch, choose left uterine angle, ligation mesometrium blood vessel, then ligature the two ends of the uterus section that need to excise, cut The uterus of one section of 1.5 cm length.In the aseptic inner membrance worn separating uterus inner membrance in Crohn liquid, then cut 5 mm × 5 mm Tissue fragment, by epithelial layer against stomach wall, is sutured on stomach wall.3 weeks after modeling, select estrus rat, open abdomen and see transplanting uterus Interior membrane volume increases, and has liquid to gather and has angiopoietic transparent vesicles, with volume 20 mm in formation3, interior membrane vesicle It can be seen that transparent hydrops is modeling Success criteria.
2.2 packet and administration
Packet and administration take and model successfully 60 rats, are randomly divided into 6 groups by Ectopic Endometrium volume size, that is, model group, Medicine materical crude slice B pulverized by danazol positive drug group, Radix Astragali ultramicro decoction piece group of the present invention, contrast Radix Astragali ultramicro decoction piece A group, the contrast Radix Astragali Group, separately takes with batch normal 10 rats as normal control, the daily gastric infusion of each group once, gavage volume 5mL/kg, red that Azoles group daily 100mg/kg danazol gavage；Radix Astragali ultramicro decoction piece group of the present invention, contrast Radix Astragali ultramicro decoction piece A group, the contrast Radix Astragali Pulverize medicine materical crude slice B group, be 5g/kg gavage daily.Normal group and model group such as give at the physiological saline of capacity, all in modeling art 35d starts to be administered afterwards, and one time a day, continuous 30 d.
2.3 index determining
1 h after last dose, each group animal is anaesthetized with yellow Jackets, and arteria carotis takes blood, centrifugation（2000 r/min） 30 min, separate serum, to be checked.Followed by execution animal, the endometriotic tissues taking normal group endometrium and respectively making film group, Make the section of tissue paraffin standard（Thick 3 μm）.5 groups of section point, for detecting ER, PR, VEGF, NK cytoactive and HE Dyeing, measurement rat Ectopic Endometrium volume of cutting open the belly, calculate inhibiting rate.
（1）Serum measured by radioimmunoassay E2、P：Operated by reagent kit determination step.
（2）Immunohistochemical Method measures ER, PR, VEGF, NK cell：According to the operation of SABC immunohistochemical staining kit Routine is carried out.Microscope inspection.Graphical analysis：Every sample after immunohistochemical staining, color using U.S. CMS800 Color image analysis system, arbitrarily takes the optical density in 3 visuals field on glandular epithelium surface（Optical density/area）Mean value represent The content of ER, PR, VEGF, NK cell in this section glandular epithelium.
3 results
3.1 antithetical phrase endometriosis in rats serum E2, the impact of P（It is shown in Table 1-1）
Table 1-1 antithetical phrase endometriosis in rats E2, the impact of P（± s, n=10）
Note：Compare with model control group, #P<0.05, ##P<0.01, ###P<0.001；
Compare with Normal group, * P<0.05, * * P<0.01, * * * P<0.001
Test result indicate that, the action effect of Radix Astragali ultramicro decoction piece group of the present invention is suitable with positive drug danazol group, bright Show and pulverize medicine materical crude slice B group better than contrast Radix Astragali ultramicro decoction piece A group and the contrast Radix Astragali.
3.2 pairs of experimental ectopic endometriums organize the impact of ER, PR content（It is shown in Table 1-2）
Table 1-2 organizes the impact of ER, PR content to experimental ectopic endometrium（± s, n=10）
Note：Compare with model control group, #P<0.05, ##P<0.01, ###P<0.001；
Compare with Normal group, * P<0.05, * * P<0.01, * * * P<0.001
Test result indicate that, the action effect of Radix Astragali ultramicro decoction piece group of the present invention is suitable with positive drug danazol group, bright Show and pulverize medicine materical crude slice B group better than contrast Radix Astragali ultramicro decoction piece A group and the contrast Radix Astragali.
3.3 Radix Astragali ultramicro decoction pieces of the present invention organize the impact of VEGF, NK cytoactive to experimental ectopic endometrium （It is shown in Table 1-3）
Table 1-3 organizes the impact of VEGF, NK cell to experimental ectopic endometrium（± s, n=10）
Note：Compare with model control group, #P<0.05, ##P<0.01, ###P<0.001；
Compare with Normal group, * P<0.05, * * P<0.01, * * * P<0.001
Test result indicate that, the action effect of Radix Astragali ultramicro decoction piece group of the present invention is suitable with positive drug danazol group, bright Show and pulverize medicine materical crude slice B group better than contrast Radix Astragali ultramicro decoction piece A group and the contrast Radix Astragali.
Under 3.4 light microscopes, each group endometrial morphology is observed
（1）Normal group：Normal rats endometrium is simple columnar epithelium it is seen that body of gland, blood vessel, interstitial etc. are normally tied Structure.
（2）Model control group：There is an endometrial tissue by striated muscle, Glandular Dilatation, galandular epithelium is simple columnar, short cube Shape or flat；Interstitial rich blood vessel, dilatation and congestion is it is seen that a small amount of hemosiderin and neutrophil infiltration.
（3）Danazol group：Compare with model group, lumen of gland reduces；There is quantity granulocyte infiltration not etc. in interstitial and lumen of gland And hemosiderin.
（4）Radix Astragali ultramicro decoction piece group of the present invention：Part lumen of gland reduces, and part galandular epithelium comes off, in part lumen of gland visible relatively Many neutrophil leucocytes and foam cells, interstitial is relatively reduced, wherein blood vessel number and caliber relatively reduced it is seen that more iron content blood Flavine.
（5）Contrast Radix Astragali ultramicro decoction piece A group：Part lumen of gland relative decrease, is shown in neutrophil leucocyte, interstitial blood in part lumen of gland Enlargement of pipe hyperemia, wherein visible hemosiderin and a small amount of lymphocyte, neutrophil leucocyte.
（6）Medicine materical crude slice B group pulverized by the contrast Radix Astragali：Part galandular epithelium comes off, visible neutrophil leucocyte and histocyte in lumen of gland, Interstitial blood vessel number and caliber are relatively reduced, and in interstitial, inflammatory cell reduces it is seen that hemosiderin.
Test result indicate that, the action effect of Radix Astragali ultramicro decoction piece group of the present invention is suitable with positive drug danazol group, bright Show and pulverize medicine materical crude slice B group better than contrast Radix Astragali ultramicro decoction piece A group and the contrast Radix Astragali.
The impact to Endometriosis Model in Rats for 3.5 Radix Astragali ultramicro decoction pieces of the present invention（It is shown in Table 1-4）
The impact to Endometriosis Model in Rats for the table 1-4（± s, n=10）
Note:Compare with before medication, * P<0.05, * * P<0.01;Compare with model group, #P<0.05, ##P<0.01.
Test result indicate that, the action effect of Radix Astragali ultramicro decoction piece group of the present invention is suitable with positive drug danazol group, bright Show and pulverize medicine materical crude slice B group better than contrast Radix Astragali ultramicro decoction piece A group and the contrast Radix Astragali.
4 discussion
Endometriosis is a kind of sex hormone-dependent disease, performs the operation and the postoperative application women such as GnRH, nemestran Gonad axis inhibitor, to causing low Estrogen and progestin environment in vivo, makes residual dystopy stove atresia be current primary treatment side Method.Traditional Chinese medicine is studied to it and is still in the starting stage.
4.1 regulation endocrines
4.1.1 reducing Estrogen and progestin level
Increasing experiment proves that endometriosis is estrogen-dependent diseases.In recent years it was found that pregnant swash Element equally has considerable effect during in uterus endometriosis develop：P can promote endometriotic tissues Hyperplasia and differentiation.Estrogen and progestin all can promote the expression of VEGF mRNA, in the high expression of Ectopic Endometrium VEGF contributes to Film obtains necessary nutriment using the vascularization that VEGF mediates and survives.In this experiment, model control group serum is female, Apparently higher than normal group, danazol and Radix Astragali ultramicro decoction piece group of the present invention can effectively reduce mullerianosis to progestogen content Disease rat blood serum E2, P level, illustrate Radix Astragali ultramicro decoction piece of the present invention can by suppress rat Estrogen and progestin abnormal secretion Reach the purpose for the treatment of endometriosis, exercising result is similar to danazol.
4.1.2 reducing Estrogen and progesterone receptors content
Ectopic Endometrium growth and sex hormone necessary to maintenance need to play a role by sex hormone receptor.Ectopic Endometrium group ER, PR is had to express in knitting, its content changed with the menstrual cycle.In this experiment, endometriosis rat estrus ER, The expression of PR is slightly below same period normal endometrium.Radix Astragali ultramicro decoction piece of the present invention can be by reducing ER, PR in Ectopic Endometrium Expression, thus blocking Ectopic Endometrium to E2, the reaction of P, suppression Ectopic Endometrium growth.
4.2 improve immunologic function
4.2.1 improving NK cytoactive
NK cell be a class in uterus play in endometriosis generating process important immunosurveillance immunity thin Born of the same parents.NK cell is Endometriosis incidence mechanism to the defect of endometriotic tissues specific recognition and lethal effect In one of key link.In an experiment, Radix Astragali ultramicro decoction piece of the present invention has notable liter and proposes effect to the activity of NK cell, Effect is similar to danazol.The raising of NK Cell-mediated Immunity contributes to the identification to Ectopic Endometrium and removing, and this is also this Bright Radix Astragali ultramicro decoction piece treats one of mechanism of endometriosis.
4.2.2 the Angiogenesiss of ectopic focus are suppressed
The foundation of new blood vessel network is that the endometrial implantation of adverse current survives and develops into the pass of endometriosis Key.VEGF is the most key vascularization stimulating factor having now been found that, stimulation vascular endothelial cell that can be direct and special, Induce its propagation, migration, and increase vascular endothelial cell permeability, induction new vessels is formed.Research finds, endometrium is different Position disease ectopic endometrium inner membrance VEGF mRNA is in high expression, and Estrogen and progestin can promote the expression of VEGF.For this reason, in animal In experiment, VEGF is studied.Result shows：In experimental rat endometriotic tissues, vegf expression strengthens；See under light microscopic Examine endometrial morphology：Model control group interstitial medium vessels enriches, and dilatation and congestion is it was demonstrated that surviving of Ectopic Endometrium needs rete vasculosum The support of network.And Radix Astragali ultramicro decoction piece group of the present invention and danazol group all can reduce the expression of VEGF, light microscopic Xia Ge treatment group blood The minimizing of pipe quantity is inconspicuous, but caliber reduces, and points out the dilatation and congestion of Ectopic Endometrium blood vessel to be inhibited, treats with through GnRH-a Afterwards blood vessel surface amass minimizing result consistent.Consider to be probably that medicine passes through to suppress the secretion of Estrogen and progestin, reduce VEGF Generation so that the formation of Ectopic Endometrium blood vessel is inhibited.
Experiment two：HPLC method measures the content of lupenone in ultramicro decoction piece of the present invention
1 instrument and reagent
LC-10A type high performance liquid chromatograph；SPD-10A UV-detector；Anastar chromatographic work station.Acetonitrile is chromatogram Pure, water is distilled water, and it is pure that other reagent are analysis.Lupenone reference substance（Tianjin Danxi Guoyao Institute, content ＞ 98%）.Ultramicro decoction piece of the present invention is prepared according to the embodiment 1 in description of the invention specific embodiment.
2 methods and result
The determination of 2.1 Detection wavelengths
Take appropriate lupenone reference substance, add methyl alcohol to make the reference substance solution of certain mass concentration, with methyl alcohol as sky In vain, carry out length scanning in 200～400nm wave-length coverage with ultraviolet-visible spectrophotometer.Scanning result shows, feather fan Ketenes has absorption maximum near 210nm, therefore selecting 210nm is Detection wavelength.
2.2 chromatographic condition
Chromatographic column：Di Ma company Diamonsil C18（Specification：250mm × 4.6mm, 5 μm）；Mobile phase：Acetonitrile -0.5% phosphorus Acid solution（83：17）；Flow velocity：1.0mL/min；Detection wavelength：210nm；Column temperature：35℃；Sample size：20μL.Number of theoretical plate is pressed Lupenone calculates and should be not less than 5000.
2.3 solution preparations
2.3.1 the preparation of reference substance solution
Take lupenone reference substance appropriate, accurately weighed, plus methyl alcohol makes the solution that every 1mL contains 0.0773mg, as right According to product storing solution.Again in accurate absorption reference substance storing solution 2mL to 25mL measuring bottle, plus methanol dilution, to scale, shakes up, and obtains final product.
2.3.2 the preparation of need testing solution
Take ultramicro decoction piece of the present invention, finely ground, take appropriate, accurately weighed, put in conical flask with cover, accurate addition methyl alcohol 25mL, close plug, weighed weight, ultrasonically treated 30min, let cool, more weighed weight, supply the weight of less loss with methyl alcohol, shake up, filter Cross, take subsequent filtrate, obtain final product.Precision measures reference substance solution, each 20 μ L of need testing solution, by chromatographic condition sample introduction under " 2.1 " item Measure, record chromatogram, see Fig. 1.
2.4 linear relationship examinations
Take 0.0773mg/mL lupenone reference substance storing solution, accurate absorption 0.5,1.0,2.0,3.0,4.0,6.0,8.0 Put in 25mL measuring bottle with 10.0mL, with methanol dilution to scale, shake up.By above-mentioned chromatographic condition sample introduction respectively, measure peak area Value.With concentration（μg/mL）For abscissa, corresponding peak area value is ordinate, carries out linear regression, obtains regression equation Y= 570352.3X-47.58,（r=0.9998）.Result shows that lupenone is in good in the range of 1.546～30.92 μ g/mL Linear relationship.
2.5 reappearance test
Take 6 parts of the sample of same lot number, by the preparation method preparation of need testing solution, measure, result records sample in accordance with the law The average mass fraction of middle lupenone is 0.02741%, RSD is 1.35%（n=6）.
2.6 stability test
Take ultramicro decoction piece need testing solution of the present invention, measure peak face by above-mentioned chromatographic condition respectively at 0,2,4,6,8 and 24h Product value, its RSD value is 0.74%, and it is stable that result shows that need testing solution measures in 24h.
2.7 precision test
The concentration that line taking sexual intercourse tests under item is the reference substance solution of 0.00618mg/mL, continuous by above-mentioned chromatographic condition Sample introduction 6 times, measures peak area value, its RSD value is 0.73%, and result shows that instrument precision is good.
2.8 average recovery tests
Take the ultramicro decoction piece of the present invention of above-mentioned same lot number, precision weighs 250mg, parallel 6 parts, accurate addition feather fan respectively Ketenes reference substance solution（Concentration is 0.077mg/mL）1.0mL, adds methyl alcohol to 25mL, by the preparation side of above-mentioned need testing solution Method operates.Measure peak area value by chromatographic condition under " 2.2 " item, calculate the rate of recovery, average recovery rate for 100.06%, RSD is 1.42%, the results are shown in Table 2-1.
Table 2-1 lupenone average recovery result of the test（n=6）
2.9 sample determination
Take each batch sample of ultramicro decoction piece of the present invention, all prepare by the preparation method of need testing solution.By above-mentioned chromatostrip Part sample introduction respectively, measures peak area value, calculates the amount being often equivalent to the ultramicro decoction piece of the present invention of 1g medicine materical crude slice containing lupenone（μg/ g）, the results are shown in Table 2-2.
Table 2-2 sample size measurement result（n=6）
3 discuss and conclusion
This result of the test shows, the content of ultramicro decoction piece lupenone of the present invention should be greater than 20 μ g/g.
The content of the high effective liquid chromatography for measuring lupenone that this test is set up, examines multiple chromatographic conditions, makes plumage Fan ketenes peak reaches baseline separation with other impurities peak.This method is simple to operate, specificity strong, favorable reproducibility, can be used as this The method with controlling for the quality testing of bright ultramicro decoction piece.
Experiment three：HPLC method measures the content of different dimpling sword-like leave Sha's alcohol in ultramicro decoction piece of the present invention
1 instrument and reagent
SP8800 efficient liquid phase pump, SepctraFocus UV-detector, Weil-McLain dragon Data Processing in Chromatography Workstation, different dimpling sword Leaf Sha's alcohol reference substance（By the calibrating of Chinese pharmaceutical biological product, research institute provides）；Acetonitrile, phosphoric acid are that analysis is pure；Ultra micro drink of the present invention Piece is prepared according to the embodiment 1 in description of the invention specific embodiment.
2 assays
2.1 chromatographic condition：Chromatographic column is Diamonsil TM C18Post（Specification：4.6 mm × 200 mm, 5 μm）；Flowing Phase：Acetonitrile -0.1% phosphoric acid（10：90）；Column temperature：30℃；Flow velocity：1.0 ml/min；Detection wavelength：207 nm, sample size：20μ L；Number of theoretical plate is calculated by different dimpling sword-like leave Sha's alcohol and should be not less than 5000.
The preparation of 2.2 reference substance solution：Precision weighs different dimpling sword-like leave Sha's alcohol reference substance 10.13 mg, puts 100 ml amounts In bottle, dissolved with methyl alcohol and be diluted to scale, shake up.Precision measures 2 ml, puts in 25 ml measuring bottles, is diluted to quarter with mobile phase Degree, shakes up, then every 1 ml contains different dimpling sword-like leave Sha's alcohol 8.1 μ g.
The preparation of 2.3 need testing solutions：Precision weighs in Radix Astragali ultramicro decoction piece about 0.2 g to 100 ml measuring bottle, plus methyl alcohol The ultrasonic 15min of 80 ml, with methanol constant volume to scale after cooling, shakes up, and pipettes in 1 ml to 10 ml measuring bottle, with the phase dilution that flows To scale, shake up, 0.45 μm of filter membrane filtration, take subsequent filtrate as need testing solution.
2.4 linear relationships are investigated：Draw 1,5,10,15,17 μ L reference substance solution injection high performance liquid chromatographs successively, With sample size（C）For abscissa, chromatographic peak area（A）For ordinate, try to achieve regression equation：A=4 763.3 C-5826.2, r= 0.9999.Show that sample size linear relationship in 0.0081～0.1377 μ g range is good.
2.5 Precision Experiment：The above-mentioned need testing solution 10 μ L of accurate absorption, repeats sample introduction 5 times, by above-mentioned chromatographic condition Measure, record peak area.Calculate RSD=0.47%（n=6）.
2.6 stability experiment：The above-mentioned need testing solution 10 μ L of accurate absorption, enters in placement 0,2,4,6,12,24 h respectively Sample, is measured the peak area of different dimpling sword-like leave Sha's alcohol, calculates RSD=0.50% by above-mentioned chromatographic condition（n=6）, result shows, supplies Test sample solution is stable in 24 h.
2.7 reappearance experiments：Precision weighs 5 parts of same batch sample, prepares need testing solution by 2.3 methods, by upper State the content that chromatographic condition measures different dimpling sword-like leave Sha's alcohol, calculate RSD=0.98%（n=6）.
2.8 average recovery experiments：Weigh Radix Astragali ultramicro decoction piece powder about 0.03 g, accurately weighed, accurate addition is certain Different dimpling sword-like leave Sha's alcohol reference substance of amount, measures different dimpling sword-like leave Sha's alcohol content by 2.3 with method operation, result averagely reclaims Rate is 100.65%, RSD=2.24%, n=6.
The assay of 2.9 finished products：Weigh Radix Astragali ultramicro decoction piece of the present invention about 0.1 g of different lot numbers respectively, accurate title Fixed, prepare need testing solution by 2.3 methods, measure by above-mentioned chromatographic condition, the results are shown in Table 3-1.
Table 3-1 six batch sample assay
3 discuss and conclusion
This result of the test shows, the content of ultramicro decoction piece of the present invention different dimpling sword-like leave Sha's alcohol should be greater than 20 μ g/g.
Different dimpling sword-like leave Sha's alcohol is one of active component in Milkvetch Root, using different dimpling sword-like leave Sha's alcohol as quality control Index is reasonable it is ensured that the quality of ultramicro decoction piece.
Brief description：
Fig. 1：Reference substance（A）Test sample（B）HPLC chromatogram, wherein No. 1 peak is lupenone
Specific embodiment：
Embodiment 1：
Prescription：The water extract of Radix Astragali medicine materical crude slice 200g, the Ultramicro-powder of Radix Astragali medicine materical crude slice 800g mince.
Preparation method：
（1）Take Radix Astragali medicine materical crude slice 200g, add water 2000mL, decoct 2 times, 1 hour every time, merge decocting liquid, be condensed into medicinal extract, Obtain the water extract of Radix Astragali medicine materical crude slice；
（2）Take Radix Astragali medicine materical crude slice 800g, ultramicro grinding, the gas of airslide disintegrating mill are carried out using airslide disintegrating mill to Radix Astragali medicine materical crude slice Flowing pressure is 1100kPa, and charging rate is 180r min-1, grade frequency is 35Hz, and grinding time is 45min, using laser Particle instrument wet method measures the particle diameter distribution of powder, the D of fine powder50, D90It is respectively 29.758 μm, 58.376 μm, obtain Radix Astragali medicine materical crude slice Ultramicro-powder mince；
（3）The water extract of above-mentioned Radix Astragali medicine materical crude slice is minced with the Ultramicro-powder of Radix Astragali medicine materical crude slice and merges, pelletize, be dried, obtain yellow Stilbene ultramicro decoction piece.
Detection method：
Assay, assay method are carried out to the lupenone in this Radix Astragali ultramicro decoction piece using high performance liquid chromatography For：
（1）Chromatographic condition：Chromatographic column：C18Post；Mobile phase：Ratio is 83：17 acetonitrile -0.5% phosphoric acid solution；Flow velocity： 1.0mL/min；Detection wavelength：210nm；Column temperature：35℃；Sample size：20μL；Number of theoretical plate is calculated and should be not less than by lupenone 5000；
（2）The preparation of reference substance solution：Take lupenone reference substance appropriate, accurately weighed, plus methyl alcohol makes every 1mL and contains The solution of 0.0773mg, as reference substance storing solution；Again in accurate absorption reference substance storing solution 2mL to 25mL measuring bottle, plus methyl alcohol It is diluted to scale, shakes up, obtain final product；
（3）The preparation of need testing solution：Take ultramicro decoction piece of the present invention, finely ground, accurately weighed, put in conical flask with cover, essence Close addition methyl alcohol 25mL, close plug, weighed weight, ultrasonically treated 30min, let cool, more weighed weight, the weight of less loss is supplied with methyl alcohol Amount, shakes up, filtration, takes subsequent filtrate, obtains final product；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph；
（5）Measurement result：The content of ultramicro decoction piece lupenone of the present invention is 22.89 μ g/g.
Assay is carried out to the different dimpling sword-like leave Sha's alcohol in this Radix Astragali ultramicro decoction piece using high performance liquid chromatography, measures Method is：
（1）Chromatographic condition：Chromatographic column is C18Post；Mobile phase：Ratio is 10：90 acetonitrile -0.1% phosphoric acid；Column temperature：30℃； Flow velocity：1.0 ml/min；Detection wavelength：207 nm；Sample size：20μL；Number of theoretical plate presses different dimpling sword-like leave Sha's alcohol calculating should not Less than 5000；
（2）The preparation of reference substance solution：Precision weighs different dimpling sword-like leave Sha's alcohol reference substance 10.13 mg, puts 100 ml measuring bottles In, dissolved with methyl alcohol and be diluted to scale, shake up；Precision measures 2 ml, puts in 25 ml measuring bottles, is diluted to scale with mobile phase, Shake up, then every 1 ml contains different dimpling sword-like leave Sha's alcohol 8.1 μ g；
（3）The preparation of need testing solution：Precision weighs in Radix Astragali ultramicro decoction piece about 0.2 g to 100 ml measuring bottle, plus methyl alcohol The ultrasonic 15min of 80 ml, with methanol constant volume to scale after cooling, shakes up, and pipettes in 1 ml to 10 ml measuring bottle, with the phase dilution that flows To scale, shake up, 0.45 μm of filter membrane filtration, take subsequent filtrate as need testing solution；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph.
（5）Measurement result：The content of ultramicro decoction piece lupenone of the present invention is 29.86 μ g/g.
Embodiment 2：
Prescription：The water extract of Radix Astragali medicine materical crude slice 100g, the Ultramicro-powder of Radix Astragali medicine materical crude slice 900g mince.
Preparation method：
（1）Take Radix Astragali medicine materical crude slice 100g, add water 500mL, decoct 3 times, 0.5 hour every time, merge decocting liquid, be condensed into leaching Cream, obtains the water extract of Radix Astragali medicine materical crude slice；
（2）Take Radix Astragali medicine materical crude slice 900g, ultramicro grinding, the gas of airslide disintegrating mill are carried out using airslide disintegrating mill to Radix Astragali medicine materical crude slice Flowing pressure is 1100kPa, and charging rate is 200r min-1, grade frequency is 30Hz, and grinding time is 50min, using laser Particle instrument wet method measures the particle diameter distribution of powder, the D of fine powder50, D90It is respectively 30.216 μm, 61.624 μm, obtain Radix Astragali medicine materical crude slice Ultramicro-powder mince；
（3）The water extract of above-mentioned Radix Astragali medicine materical crude slice is minced with the Ultramicro-powder of Radix Astragali medicine materical crude slice and merges, pelletize, be dried, obtain yellow Stilbene ultramicro decoction piece.
Detection method：
Assay, assay method are carried out to the lupenone in this Radix Astragali ultramicro decoction piece using high performance liquid chromatography For：
（1）Chromatographic condition：Chromatographic column：C18Post；Mobile phase：Ratio is 83：17 acetonitrile -0.5% phosphoric acid solution；Flow velocity： 1.0mL/min；Detection wavelength：210nm；Column temperature：35℃；Sample size：20μL；Number of theoretical plate is calculated and should be not less than by lupenone 5000；
（2）The preparation of reference substance solution：Take lupenone reference substance appropriate, accurately weighed, plus methyl alcohol makes every 1mL and contains The solution of 0.0773mg, as reference substance storing solution；Again in accurate absorption reference substance storing solution 2mL to 25mL measuring bottle, plus methyl alcohol It is diluted to scale, shakes up, obtain final product；
（3）The preparation of need testing solution：Take ultramicro decoction piece of the present invention, finely ground, accurately weighed, put in conical flask with cover, essence Close addition methyl alcohol 25mL, close plug, weighed weight, ultrasonically treated 30min, let cool, more weighed weight, the weight of less loss is supplied with methyl alcohol Amount, shakes up, filtration, takes subsequent filtrate, obtains final product；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph；
（5）Measurement result：The content of ultramicro decoction piece lupenone of the present invention is 23.08 μ g/g.
Assay is carried out to the different dimpling sword-like leave Sha's alcohol in this Radix Astragali ultramicro decoction piece using high performance liquid chromatography, measures Method is：
（1）Chromatographic condition：Chromatographic column is C18Post；Mobile phase：Ratio is 10：90 acetonitrile -0.1% phosphoric acid；Column temperature：30℃； Flow velocity：1.0 ml/min；Detection wavelength：207 nm；Sample size：20μL；Number of theoretical plate presses different dimpling sword-like leave Sha's alcohol calculating should not Less than 5000；
（2）The preparation of reference substance solution：Precision weighs different dimpling sword-like leave Sha's alcohol reference substance 10.13 mg, puts 100 ml measuring bottles In, dissolved with methyl alcohol and be diluted to scale, shake up；Precision measures 2 ml, puts in 25 ml measuring bottles, is diluted to scale with mobile phase, Shake up, then every 1 ml contains different dimpling sword-like leave Sha's alcohol 8.1 μ g；
（3）The preparation of need testing solution：Precision weighs in Radix Astragali ultramicro decoction piece about 0.2 g to 100 ml measuring bottle, plus methyl alcohol The ultrasonic 15min of 80 ml, with methanol constant volume to scale after cooling, shakes up, and pipettes in 1 ml to 10 ml measuring bottle, with the phase dilution that flows To scale, shake up, 0.45 μm of filter membrane filtration, take subsequent filtrate as need testing solution；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph.
（5）Measurement result：The content of ultramicro decoction piece lupenone of the present invention is 28.96 μ g/g.
Embodiment 3：
Prescription：The water extract of Radix Astragali medicine materical crude slice 150g, the Ultramicro-powder of Radix Astragali medicine materical crude slice 850g mince.
Preparation method：
（1）Take Radix Astragali medicine materical crude slice 150g, add water 3000mL, decoct 1 time, 2 hours, merge decocting liquid, be condensed into medicinal extract, obtain The water extract of Radix Astragali medicine materical crude slice；
（2）Take Radix Astragali medicine materical crude slice 850g, ultramicro grinding, the gas of airslide disintegrating mill are carried out using airslide disintegrating mill to Radix Astragali medicine materical crude slice Flowing pressure is 1200kPa, and charging rate is 160r min-1, grade frequency is 40Hz, and grinding time is 40min, using laser Particle instrument wet method measures the particle diameter distribution of powder, the D of fine powder50, D90It is respectively 32.568 μm, 64.954 μm, obtain Radix Astragali medicine materical crude slice Ultramicro-powder mince；
（3）The water extract of above-mentioned Radix Astragali medicine materical crude slice is minced with the Ultramicro-powder of Radix Astragali medicine materical crude slice and merges, pelletize, be dried, obtain yellow Stilbene ultramicro decoction piece.
Detection method：
Assay, assay method are carried out to the lupenone in this Radix Astragali ultramicro decoction piece using high performance liquid chromatography For：
（1）Chromatographic condition：Chromatographic column：C18Post；Mobile phase：Ratio is 83：17 acetonitrile -0.5% phosphoric acid solution；Flow velocity： 1.0mL/min；Detection wavelength：210nm；Column temperature：35℃；Sample size：20μL；Number of theoretical plate is calculated and should be not less than by lupenone 5000；
（2）The preparation of reference substance solution：Take lupenone reference substance appropriate, accurately weighed, plus methyl alcohol makes every 1mL and contains The solution of 0.0773mg, as reference substance storing solution；Again in accurate absorption reference substance storing solution 2mL to 25mL measuring bottle, plus methyl alcohol It is diluted to scale, shakes up, obtain final product；
（3）The preparation of need testing solution：Take ultramicro decoction piece of the present invention, finely ground, accurately weighed, put in conical flask with cover, essence Close addition methyl alcohol 25mL, close plug, weighed weight, ultrasonically treated 30min, let cool, more weighed weight, the weight of less loss is supplied with methyl alcohol Amount, shakes up, filtration, takes subsequent filtrate, obtains final product；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph；
（5）Measurement result：The content of ultramicro decoction piece lupenone of the present invention is 22.85 μ g/g.
Assay is carried out to the different dimpling sword-like leave Sha's alcohol in this Radix Astragali ultramicro decoction piece using high performance liquid chromatography, measures Method is：
（1）Chromatographic condition：Chromatographic column is C18Post；Mobile phase：Ratio is 10：90 acetonitrile -0.1% phosphoric acid；Column temperature：30℃； Flow velocity：1.0 ml/min；Detection wavelength：207 nm；Sample size：20μL；Number of theoretical plate presses different dimpling sword-like leave Sha's alcohol calculating should not Less than 5000；
（2）The preparation of reference substance solution：Precision weighs different dimpling sword-like leave Sha's alcohol reference substance 10.13 mg, puts 100 ml measuring bottles In, dissolved with methyl alcohol and be diluted to scale, shake up；Precision measures 2 ml, puts in 25 ml measuring bottles, is diluted to scale with mobile phase, Shake up, then every 1 ml contains different dimpling sword-like leave Sha's alcohol 8.1 μ g；
（3）The preparation of need testing solution：Precision weighs in Radix Astragali ultramicro decoction piece about 0.2 g to 100 ml measuring bottle, plus methyl alcohol The ultrasonic 15min of 80 ml, with methanol constant volume to scale after cooling, shakes up, and pipettes in 1 ml to 10 ml measuring bottle, with the phase dilution that flows To scale, shake up, 0.45 μm of filter membrane filtration, take subsequent filtrate as need testing solution；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph.
（5）Measurement result：The content of ultramicro decoction piece lupenone of the present invention is 29.67 μ g/g.
Moreover, it will be appreciated that although this specification is been described by according to embodiment, not each embodiment only wraps Containing an independent technical scheme, only for clarity, those skilled in the art should for this narrating mode of specification Using specification as an entirety, the technical scheme in each embodiment can also form those skilled in the art through appropriately combined Understandable other embodiment.

Claims (5)

1. a kind of Radix Astragali ultramicro decoction piece, this Radix Astragali ultramicro decoction piece is to be made up of the raw material of following weight portion：Radix Astragali medicine materical crude slice 10%～ The water extract of 20% weight portion, the Ultramicro-powder of Radix Astragali medicine materical crude slice 80%～90% weight portion mince it is characterised in that this Radix Astragali ultra micro The preparation method of medicine materical crude slice is as follows：
（1）Take Radix Astragali medicine materical crude slice 10%～20% weight portion, plus 10 times amount water, decoct 2 times, 1 hour every time, merge decocting liquid, concentrate Become medicinal extract, obtain the water extract of Radix Astragali medicine materical crude slice；
（2）Take Radix Astragali medicine materical crude slice 80%～90% weight portion, carry out ultramicro grinding, ultra micro is carried out to Radix Astragali medicine materical crude slice using airslide disintegrating mill Pulverize, the stream pressure of airslide disintegrating mill is 1000～1200kPa, and charging rate is 160～200r min-1, grade frequency is 30～40Hz, grinding time is 40～50min, and the Ultramicro-powder obtaining Radix Astragali medicine materical crude slice minces, after ultramicro grinding, Radix Astragali medicine materical crude slice super The particle diameter that micro mist minces is 10～75 μm；
（3）The water extract of above-mentioned Radix Astragali medicine materical crude slice is minced with the Ultramicro-powder of Radix Astragali medicine materical crude slice and merges, pelletize, be dried, obtain the Radix Astragali and surpass Micro- medicine materical crude slice.
2. Radix Astragali ultramicro decoction piece as claimed in claim 1 it is characterised in that Radix Astragali medicine materical crude slice in this Radix Astragali ultramicro decoction piece super The preparation method that micro mist minces is as follows：Ultramicro grinding, the air-flow of airslide disintegrating mill are carried out using airslide disintegrating mill to Radix Astragali medicine materical crude slice Pressure is 1100kPa, and charging rate is 180r min-1, grade frequency is 35Hz, and grinding time is 45min, obtains Radix Astragali drink The Ultramicro-powder of piece minces.
3. Radix Astragali ultramicro decoction piece as claimed in claim 1 is it is characterised in that adopt high performance liquid chromatography to this Radix Astragali ultra micro Lupenone in medicine materical crude slice carries out assay, and assay method is：
（1）Chromatographic condition：Chromatographic column：C18Post；Mobile phase：Ratio is 83：17 acetonitrile -0.5% phosphoric acid solution；Flow velocity：1.0mL/ min；Detection wavelength：210nm；Column temperature：35℃；Sample size：20μL；Number of theoretical plate is calculated by lupenone and should be not less than 5000；
（2）The preparation of reference substance solution：Take lupenone reference substance appropriate, accurately weighed, plus methyl alcohol makes every 1mL and contains The solution of 0.0773mg, as reference substance storing solution；Again in accurate absorption reference substance storing solution 2mL to 25mL measuring bottle, plus methyl alcohol It is diluted to scale, shakes up, obtain final product；
（3）The preparation of need testing solution：Take ultramicro decoction piece, finely ground, accurately weighed, put in conical flask with cover, accurate addition methyl alcohol 25mL, close plug, weighed weight, ultrasonically treated 30min, let cool, more weighed weight, supply the weight of less loss with methyl alcohol, shake up, filter Cross, take subsequent filtrate, obtain final product；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph.
4. Radix Astragali ultramicro decoction piece as claimed in claim 1 is it is characterised in that adopt high performance liquid chromatography to this Radix Astragali ultra micro Different dimpling sword-like leave Sha's alcohol in medicine materical crude slice carries out assay, and assay method is：
（1）Chromatographic condition：Chromatographic column is C18Post；Mobile phase：Ratio is 10：90 acetonitrile -0.1% phosphoric acid；Column temperature：30℃；Stream Speed：1.0 ml/min；Detection wavelength：207 nm；Sample size：20μL；Number of theoretical plate presses different dimpling sword-like leave Sha's alcohol calculating should not be low In 5000；
（2）The preparation of reference substance solution：Precision weighs different dimpling sword-like leave Sha's alcohol reference substance 10.13 mg, puts in 100 ml measuring bottles, Dissolved with methyl alcohol and be diluted to scale, shake up；Precision measures 2 ml, puts in 25 ml measuring bottles, is diluted to scale with mobile phase, shakes Even, then every 1 ml contains different dimpling sword-like leave Sha's alcohol 8.1 μ g；
（3）The preparation of need testing solution：Precision weighs in Radix Astragali ultramicro decoction piece about 0.2 g to 100 ml measuring bottle, plus methyl alcohol 80 ml Ultrasonic 15min, with methanol constant volume to scale after cooling, shakes up, and pipettes in 1 ml to 10 ml measuring bottle, is diluted to quarter with mobile phase Degree, shakes up, 0.45 μm of filter membrane filtration, takes subsequent filtrate as need testing solution；
（4）Measure：Precision measures reference substance solution, each 20 μ L of need testing solution, measures in injection high performance liquid chromatograph.
5. application in preparation treatment endometriosis medicine for the Radix Astragali ultramicro decoction piece as claimed in claim 1.
CN201610106958.8A 2016-02-28 2016-02-28 Astragalus membranaceus ultramicro decoction pieces as well as preparation method and quality testing method thereof Active CN105726623B (en)

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