CN101288762B - Content determination method of medicine composition for treating natal lochioschesis - Google Patents
Content determination method of medicine composition for treating natal lochioschesis Download PDFInfo
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Abstract
The invention discloses a drug combination for treating lochia after post partum, a preparation method and a quality control method thereof. The drug combination of the invention comprises the following raw materials: angelica, Szechwan Lovage Rhizome, semen persicae, dried ginger, dry carbon, leonurus and dipsacus root. The preparation method is that the raw materials are grinded into course powder; 8-12 times amount of water is added for extracting volatile oil for 5-8 hours; water solution after the distillation is collected by another vessel; dregs of a decoction is boiled for 2-3 times after being added with water, wherein, 6-8 times amount of water is added to be boiled for 1-2h for every time; the boiled liquids are mixed and filtered; the filtered liquid is mixed with the solution to be concentrated into thick paste which is added with brown sugar in optimal dose to be mixed uniformly and prepared into particles; the particles are dried at 50-70 DEG C, are grinded and prepared into particles with ethanol; the volatile oil is sprayed and then the drug combination is obtained. High performance liquid chromatography is adopted by the invention for the content determination of ferulic acid. The drug combination of the invention is provided with good efficacy in treating lochia after post partum and abdomen pain.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition and preparation method thereof and method of quality control, particularly relate to a kind of Chinese medicine composition for the treatment of disease that postpartum lochia can't go and preparation method thereof and method of quality control.
Background technology
Lochia, be meant that fetus is given birth to after, the blood sample secretion that vagina flows out.Lochia can be divided into sanguinous lochia, lochia serosa and lochia alba.
Normal lochia has blood smell, but not smelly, begins to be cerise or peony, contains a large amount of blood, little clot and downright bad demoulding tissue, and this lochia is called as sanguinous lochia, and sanguinous lochia generally continues 3-7 days.After 1 week, lochia will become faint yellow, contains oligemia, and more vagina and cervical mucus are arranged, and contains bacterium, and this lochia is called as lochia serosa.After 2 weeks, lochia includes a large amount of leucocytes, decidual cell and epidermal cell by the faint yellow white that becomes, and this kind lochia is called as lochia alba.Healthy lochia is clean about general 3 weeks.Come off with uterine decidua postpartum, contains blood, downright bad decidua and organize transvaginal to discharge, and is called lochia.Continue above more than person of 3 weeks as lochia, be called " post partum lochiorrhea ", claim again " disease that postpartum lochia can't go ".Be equivalent to the late puerperal hemorrhage of doctor trained in Western medicine.Hemorrhage reason has that involution of uterus is bad, infection or placental remnants (placenta retension) etc., and the latter needs the dilatation and curettage treatment sometimes.Lochia is dripping may to bring out infection incessantly, causes endometritis, also may be to nourish the leaf cell disease.
The traditional Chinese medical science thinks that the not all right disharmony of Chong and Conception Channels that belong to of lochia due to QI-blood circulation is not normal more.Its cause of disease is many by collapse from qi deficiency, and the debility of chong and ren channels can not be taken the photograph blood; Or blood system has heat, and heat is disturbed to dash and appointed, and it is descending to compel blood; Or blood-stasis internal-depression, the escape of blood from vessels and channels causes.Can be divided into three types of the deficiency of vital energy, blood stasis, blood-head etc. according to clinical symptom performance.1. consumption gas when the deficiency of vital energy is owing to product, the deficiency of vital energy can not be received and take the photograph, and makes the not good and dripping haemophilia of involution of uterus.Week in postpartum 3, above lochia was more than, and look light is not smelly, and complexion Anhui is white, and the burnout weak breath is received few loose stool.Tongue is thin, and tongue is light, and arteries and veins is thin and delicate.2. catch cold when blood stasis produces, cold and the blood knot of fighting mutually forms stasis of blood resistance, and newly blood is not living and dripping without cease.Or part afterbirth tissue residue also can cause lochiorrhea in uterine cavity.Postpartum 3, all lochia were more than, and amount less or has been held clot under the arm more, and the look purple is dim, lower abdominal distention pain.Tongue is purple dim or purple plague purpura arranged, small and wiry pulse and puckery.3. the plain body deficiency of Yin of blood-head puerpera, because of the childbirth thin fluid injury, cloudy liquid more loses and causes yin asthenia generating intrinsic heat.Or have some setbacks when producing, depressed liver and heat transmission, heat is compeled the absurd row of blood, and lochia is exceeded the time limit incessantly.Or because of postpartum weakness, damp and hot disease and evil is taken advantage of a weak point, and is detained intrauterine and to cause lochia dripping more than.Postpartum 3, all lochia were more than, more or less, as dark reddish brown, accompany dirty smelly, the lower abdominal distention pain tenderness, low-heat is persistent, dry is drunk less, it is red to urinate.Yellow and greasy fur or thin Huangshi are red, thready rapid pulse.
Invent a kind of disease that postpartum lochia can't go that is used for, few abdomen pain also can be tried out the colporrhagia that causes, menorrhalgia after being fitted with a contraceptive ring.
Summary of the invention
The object of the invention is to provide a kind of Chinese medicine composition for the treatment of disease that postpartum lochia can't go;
The object of the invention also is to provide a kind of Chinese medicine composition preparation method who treats disease that postpartum lochia can't go;
The object of the invention also is to provide a kind of method of quality control for the treatment of the Chinese medicine composition of disease that postpartum lochia can't go.
The present invention seeks to be achieved through the following technical solutions:
The Chinese medicine composition of treatment disease that postpartum lochia can't go of the present invention is to be made by the bulk drug of following weight ratio:
Radix Angelicae Sinensis 100-300 weight portion Ligusticum wallichii 40-180 weight portion
Peach kernel 10-90 weight portion rhizoma zingiberis (charcoal) 10-50 weight portion
The 10-90 weight portion breaks in motherwort 200-400 weight portion river
The Chinese medicine composition of treatment disease that postpartum lochia can't go of the present invention can be made by the bulk drug of following weight ratio:
Radix Angelicae Sinensis 100-300 weight portion Ligusticum wallichii 40-180 weight portion
Peach kernel 10-90 weight portion rhizoma zingiberis (charcoal) 10-50 weight portion
Motherwort 200-400 weight portion safflower 10-50 weight portion
Radix Glycyrrhizae (processing) 10-50 weight portion;
The above-mentioned raw materials optimum ratio is:
Radix Angelicae Sinensis 100-200 weight portion Ligusticum wallichii 120-170 weight portion
Peach kernel 40-80 weight portion rhizoma zingiberis (charcoal) 20-40 weight portion
Motherwort 320-380 weight portion safflower 30-50 weight portion
Radix Glycyrrhizae (processing) 30-50 weight portion;
The above-mentioned raw materials optimum ratio is:
Radix Angelicae Sinensis 140 weight portion Ligusticum wallichiis 130 weight portions
Peach kernel 60 weight portion rhizoma zingiberis (charcoal) 30 weight portions
Motherwort 350 weight portion safflowers 40 weight portions
Radix Glycyrrhizae (processing) 40 weight portions;
Composition of the present invention technology adding auxiliary material is routinely made clinical acceptable forms such as tablet, capsule, oral liquid, dripping pill, spray, granule; Described auxiliary material comprises solvent, disintegrant, flavouring, antiseptic, colorant, bonding agent, lubricant, matrix etc.
The preparation method of Chinese medicine composition granular preparation of the present invention is:
Bulk drug is ground into meal, adds 8-12 times of water gaging and extracts volatile oil 5-8 hour, and the aqueous solution after distillation device is in addition collected, dregs of a decoction boiling 2-3 time again adds 6-8 times of water gaging at every turn and fried in shallow oil collecting decoction 1-2 hour, filter, filtrate and above-mentioned solution merge, and are condensed into thick paste, it is an amount of that other goes into brown sugar, and mixing is made particle, in 50-70 ℃ of oven dry, pulverize, make particle with ethanol, spray into volatile oil, promptly.
Pharmaceutical composition method of quality control of the present invention comprises one or more in following discrimination method and/or the assay:
Differentiate:
(1) gets preparation and be equivalent to described composition material medicine 3g, the ammonification water number drip make moistening, add chloroform 15-25ml again, in water-bath backflow 10-25 minute, cooling, filter, filtrate evaporate to dryness, residue add 1% hydrochloric acid 1ml dissolving, get 3～4 of hydrochloric acid solutions respectively on two surface plates, one adds 1 of bismuth potassium iodide test solution, salmon precipitation promptly takes place; Another adds 1 of silico-tungstic acid test solution, and white precipitate promptly takes place;
(2) get preparation and be equivalent to described composition material medicine 9g,, add an amount of zeyssatite and grind evenly, put in the apparatus,Soxhlet's, an amount of with methyl alcohol, reflux is closely colourless to extract, reclaim methyl alcohol, residue adds water 50ml, and heating makes dissolving, put coldly, be transferred in the separating funnel, with extracted by ether 4-6 time, merge extract, extract 4-6 time, merge alkali lye with 2% sodium carbonate liquor, wash 2-4 time with ethyl acetate, each 10-20ml discards ethyl acetate liquid, alkali lye add hcl acidifying to PH be 2～3, wash 2-4 time each 10-20ml again with benzene, discard benzene liquid, continue and use extracted by ether 4-6 time, merge ether solution, volatilize, residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the forulic acid reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, compares product solution; According to the thin-layered chromatography test, draw above-mentioned need testing solution 20 μ l, reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with 85-95:20-30:3-5 benzene-dioxane-acetic acid is developping agent, launches, and takes out, dry, put under the ultraviolet lamp and inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show identical mazarine fluorescence spot;
(3) get preparation and be equivalent to described composition material medicine 9g,, add the 40ml absolute ethyl alcohol, divide 2-4 jolting to extract, extract evaporate to dryness, residue add the 1.5ml dissolve with ethanol, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds an amount of ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Drawing above-mentioned two kinds of solution each 15 μ l, 5 μ l puts respectively on same silica gel H plate, with the 0.5-1.3:0.2-1.2 chloroform-methanol is developping agent, put a small beaker of containing ammoniacal liquor in the developing tank and carry out the presaturation of ammonia steam, launch, after drying, secrete potassium with the improvement iodate, the electric heating blowing is to dry, spray 10% ethanol solution of sulfuric acid again, clear to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
Assay:
According to high performance liquid chromatography: chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; 20-30:70-80:1-2.5 methanol-water-acetic acid is moving phase; The detection wavelength is 330nm; Number of theoretical plate calculates by forulic acid should be not less than 2000; The preparation of reference substance solution: it is an amount of that precision takes by weighing the forulic acid reference substance, adds the mixed solution of 90-100:4-7 methyl alcohol-formic acid, makes the solution that contains 5ug among every 1ml, filters with miillpore filter (0.45um), promptly; The preparation of need testing solution: get preparation and be equivalent to described composition material medicine 4.5g,, put in the tool plug Erlenmeyer flask the accurate mixed solution 25ml that adds methyl alcohol-formic acid (90-100:4-7), claim to decide weight, sonicated 25-35 minute, put coldly, claim to decide weight again, after supplying the weight that subtracts mistake with the mixed solution of methyl alcohol-formic acid (90-100:4-6), shake up, filter with miillpore filter, promptly; Assay method: accurate respectively reference substance solution, each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.
Pharmaceutical composition method of quality control of the present invention is preferably as follows one or more in discrimination method and/or the assay:
Differentiate:
(1) gets preparation and be equivalent to described composition material medicine 3g,, the ammonification water number drips and makes moisteningly, adds chloroform 20ml again, in water-bath, refluxed 15 minutes, cooling filters the filtrate evaporate to dryness, residue adds 1% hydrochloric acid 1ml dissolving, get 3～4 of hydrochloric acid solutions respectively on two surface plates, one adds 1 of bismuth potassium iodide test solution, salmon precipitation promptly takes place; Another adds 1 of silico-tungstic acid test solution, and white precipitate promptly takes place;
(2) get preparation and be equivalent to described composition material medicine 9g,, add an amount of zeyssatite and grind evenly, put in the apparatus,Soxhlet's, an amount of with methyl alcohol, reflux is closely colourless to extract, reclaim methyl alcohol, residue adds water 50ml, and heating makes dissolving, puts cold, be transferred in the separating funnel, with extracted by ether 5 times, each ether amount is respectively 20,20,15,15,10ml merges extract, extract 5 times with 2% sodium carbonate liquor, each 2% sodium carbonate liquor amount is respectively 20,20,15,15,10ml merges alkali lye, washes 3 times with ethyl acetate, each 15ml, discard ethyl acetate liquid, alkali lye add hcl acidifying to PH be 2～3, wash 3 times with benzene again, each 15ml, discard benzene liquid, continue and use extracted by ether 5 times, each ether amount is respectively 20,20,15,15,10ml, merge ether solution, volatilize, residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the forulic acid reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, compares product solution; According to the thin-layered chromatography test, draw above-mentioned need testing solution 20 μ l, reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with 90:25:4 benzene-dioxane-acetic acid is developping agent, launches, and takes out, dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show identical mazarine fluorescence spot;
(3) get preparation and be equivalent to described composition material medicine 9g,, add the 40ml absolute ethyl alcohol, divide 3 joltings to extract, extract evaporate to dryness, residue add the 1.5ml dissolve with ethanol, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds an amount of ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Drawing above-mentioned two kinds of solution each 15 μ l, 5 μ l puts respectively on same silica gel H plate, with the 1:1 chloroform-methanol is developping agent, put a small beaker of containing ammoniacal liquor in the developing tank and carry out the presaturation of ammonia steam, launch, after drying, secrete potassium with the improvement iodate, the electric heating blowing is to dry, spray 10% ethanol solution of sulfuric acid again, clear to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
Assay:
According to high performance liquid chromatography: chromatographic condition and system suitability test are filling agent (4.6 * 150mm, 5 μ m) with octadecylsilane chemically bonded silica; 25:75:1.8 methanol-water-acetic acid is moving phase; The detection wavelength is 330nm; Number of theoretical plate calculates by forulic acid should be not less than 2000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing the forulic acid reference substance, adds the mixed solution of 95:5 methyl alcohol-formic acid, makes the solution that contains 5ug among every 1ml, filters with the 0.45um miillpore filter, promptly; The preparation of need testing solution: get preparation and be equivalent to described composition material medicine 4.5g,, put in the tool plug Erlenmeyer flask, the accurate mixed solution 25ml that adds methyl alcohol-formic acid (95:5), claim to decide weight, sonicated (power 250W, frequency 40KHZ) 30 minutes, put cold, claim again to decide weight, supply the weight that subtracts mistake with the mixed solution of 95:5 methyl alcohol-formic acid after, shake up, filter with the 0.45um miillpore filter, promptly; Assay method: accurate respectively reference substance solution, each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.
The present composition has good drug effect, have well invigorate blood circulation, the dispel stasis of blood, analgesic effect, the method of quality control of Chinese medicine composition provided by the present invention, be by obtaining behind the creative experiment sieving of big measuring, by the screening to sample treatment, the selection of developping agent makes and differentiates that specificity is fine in the discrimination method, and method is economic and practical, the result is quick, and can both use different thin layer plates.Pass through screening in the content assaying method to sample, test sample disposal route, the selection of developping agent, make content assaying method effectivelyly to carry out quality control, and will compare more stable that product that additive method measures shows on pharmacological effect with the product that this method is measured to product.
Following experimental example and embodiment are used to further specify but are not limited to the present invention.
Experimental example 1 pharmacological testing
Choose the present invention and test the medicine group:
Medicine group I of the present invention (embodiment 6 preparations)
Medicine group II of the present invention (pressing embodiment 5 preparations)
Medicine group III of the present invention (pressing embodiment 1 preparation)
Medicine group IV of the present invention (pressing embodiment 7 preparations)
1 analgesia
1.1 influence to hot plate method induced mice pain
Selecting body weight for use is 1822g and first with 55 ℃ of female mices of measuring threshold of pains health in 30s, and all mouse that does not lick rear solid end or escape in 30s are given it up.60 of the mouse that the threshold of pain is qualified are divided into 6 groups at random, first group hypodermic injection antondin 0.1ml/10g; Second group hypodermic injection physiological saline 0.1ml/10g; The third, fourth, penta, oneself group gavage the threshold of pain of measuring different time after medicine group I of the present invention, II, III, the IV0.10ml/10g. administration and change, and see Table 1
Table 1 dysmenorrhoea is relaxed to the influence of hot plate method induced mice pain
Annotate: 1. respectively organize P before the medicine〉0.05,3. with 4. compare P＜0.05,6. with 7. compare P＜0.05,8. with 9. compare P＜0.05,2. with 4. P〉0.05,5. with 7. compare P＜0.05.
The results are shown in Table, each organizes the preceding threshold of pain of medicine does not have significant difference (P〉0.05), medicine group 0.10ml/10g of the present invention, when 30min, can significantly improve the mouse threshold of pain (P＜0.05), in 1h, significant difference is more all arranged with the physiological saline group, 15min and antondin group be the indifference opposite sex relatively, and relatively there were significant differences (P＜0.05) for 30min and antondin group, illustrates that medicine group of the present invention has remarkable analgesic activity.
1.2 glacial acetic acid induced mice writhing response
Get 70 of the healthy mices of body weight 18～22g, divide 7 groups at random, the first group gavages antondin 0.1ml/10g; The second group gavages physiological saline 0.1ml/10g; The third, fourth, penta, oneself group gavage medicine group I of the present invention, II, III, IV0.10ml/10g; Gavage the 20min pneumoretroperitoneum and inject 0.7% glacial acetic acid 0.1ml/10g.Turn round the body number of times in the observed and recorded mouse 20min, the results are shown in following table 2
The inhibiting effect of table 2 pair mouse writhing reaction
| Group | The writhing response number |
| Antondin | 7.9±2.8 |
| Physiological saline | 46.5±12.3 |
| Medicine group I of the present invention | 7.4±2.1 |
| Medicine group II of the present invention | 7.2±3.0 |
| Medicine group III of the present invention | 7.0±2.6 |
| Medicine group IV of the present invention | 6.7±2.4 |
Annotate: blank is compared P＜0.01 with antondin and medicine group of the present invention, and antondin and medicine group of the present invention are compared P〉0.05.
The result shows: medicine group of the present invention has its effect of the caused writhing response of obvious suppression glacial acetic acid similar to the antondin group.
2PGF
2aAnalog or oxytocin effect are to the influence in uterus
2.1 to the influence of rat in the body uterus
Get 20 of healthy unpregnancy female rats, be divided into two groups, press 0.2mg/100g hypodermic injection diethylstilbestrol before the experiment, every day 1 time, inject after 3 days, with the yellow Jackets intraperitoneal anesthesia, lie on the back and on experiment table, cut open the belly, find out a side cornua uteri, the vagina end and the ovary end at angle, palace are sutured in respectively on the fulcrum of two ends, special fixed cover bottom,, draught line are connected on the sensor by pulley at angle, the palace centre joint one fine rule, then stomach wall is sewed up around the fixed cover bottom, closed abdominal cavity is by MS-302 multimedization bio signal record analysis system record experimentation.
Get above-mentioned rat and inject Rockwell nutrient solution 10ml to the abdominal cavity, trace the normal contraction curve, first group abdominal cavity injects medicine group I0.5ml of the present invention, and the 2min pneumoretroperitoneum injects PGF
2aAnalog 0.05mg/0.5ml traces 10min; The second group is given PGF earlier
2aAnalog 0.05mg/0.5ml, make the uterus be contraction after, the medicine group I0.5ml of the present invention that reinjects, the record result, see the following form 3:
Table 3 pair rat is in the influence in body uterus
| Group | Uterus (propping up) | Antagonism not | Antagonism | Not blocking-up | Blocking-up |
| First | 10? | 2? | 8? | ? | ? |
| Second | 10 | ? | ? | 0 | 10 |
The result shows: medicine group I of the present invention is to PGF
2aHysterotrismus contraction due to the analog has tangible antagonism and blocking effect.
2.2 at PGF
2aMedicine is to the influence of isolated uterine smooth muscle under analog or the oxytocin effect
Select 70 of healthy unpregnancy female rats, be divided into 7 groups, 0.2mg/100g hypodermic injection diethylstilbestrol before the experiment, every day 1 time, inject after 3 days, cut open rapidly with ether inhalation anesthesia and get the uterus, clip uterus 2cm, be placed on and fill 30ml, in the sulculus of Rockwell liquid, be connected on the sensor, water-bath keeps (36 ± 0.5) ℃, by ventilation hook aerating oxygen in groove,, add Cloprostenol (PGF then with MS-302 multimedization bio signal record analysis system record normal contraction curve
2aAnalog) 5ug/0.5ml or oxytocin inj 0.5ug/5ml, uterine smooth muscle is shunk strongly, stable back adds medicine group 0.6ml of the present invention or dysmenorrhoea conditioning liquid 0.4ml, and 10min uterine smooth muscle shrink tension, frequency the results are shown in following table 4 and table 5 after the observed and recorded administration
Table 4 is at PGF
2aMedicine is to the influence of isolated uterine smooth muscle tension force under analog or the oxytocin effect
Annotate: 1. with 2. 3. 4. compare P＜0.05,2. with 3. 5. with 4. 6. with 7. compare P＜0.01,8. 9. 10. P between group 0.05
Table 5 is at PGF
2aMedicine is to the influence of isolated uterine smooth muscle frequency under analog or the oxytocin effect
Annotate: 1. with 4., 2. with 5., 3. with 6. compare P＜0.05,7. 8. 9. P between group 0.05
The result shows: at PGF
2aAnalog or oxytocin rise not obvious to the rat uterus smooth muscle contraction, but tension force is strengthened, frequency is accelerated, the uterine smooth muscle shrinkage amplitude is descended for medicine group of the present invention, tension force descends, and frequency reduces, behind the processing solution, its tension force can recover gradually, and medicine group I of the present invention, II, III, IV have obvious inhibiting effect to isolated rat uterine smooth muscle due to Cloprostenol and the oxytocin, and significant difference is arranged.Medicine group I of the present invention, II, III, IV can obviously increase shrink tension, contraction frequency and the contractive amplitude in isolated rat uterus.
3 tests that activate blood circulation and disperse blood clots
Animal model is set up and grouping is blood stasis animal model [1] with the male rat.Select 48 of 6-8 monthly age Wistar male rats for use, body weight 550 ± 56g is divided into the positive drug group at random, medicine group I of the present invention, II, III, IV and model group.More than be divided into 6 groups, 8 every group, gavage every day respectively and give medicine group I of the present invention, II, III, IV1.24gkg
-1, positive drug 5.5mgkg
-1, 22d takes a blood sample behind the last administration 1h continuously.
3.1 platelet aggregation (PAG) is measured
Through arteria carotis blood sampling type 3mL, add 3.8 sodium citrate solution anti-freezings (1: 9) in plastic tube, mixing, 2h is interior with 1000rmin
-1Centrifugal 10min, blood platelet blood plasma (RRP) and sucking-off are rich in preparation, and remainder is with 3000rmin
-1Centrifugal 15min gets platelet-poor plasma (PPP), and ADP is a derivant, measures 1min platelet aggregation rate [PAG (1)], 5min platelet aggregation rate [PAG (5)], MA [PAG (Max)] with TYXN291 intelligence blood agglutometer.The results are shown in Table 6
Table 6 couple platelet aggregation influence (x ± s) (n=8)
| Grouping | PAG(1) | PAG(5) | PAG(Max) |
| Positive controls | 31.95±3.94① | 35.29±6.54① | 38.68±7.46① |
| Medicine group I of the present invention | 35.41±5.13? | 43.76±3.75①? | 50.59±3.66①? |
| Medicine group II of the present invention | 28.56±9.66①? | 13.96±1.66①③ | 34.36±9.14①? |
| Medicine group III of the present invention | 7.96±2.17①③? | 6.38±3.02①③? | 14.33±2.27①③ |
| Medicine group IV of the present invention | 7.96±2.17①③? | 6.38±3.02①③? | 14.33±2.27①③ |
| Model group | 52.89±5.81 | 66.20±11.39 | 70.86±8.09 |
Annotate: compare with model group: 1. P＜0.01, compare with the positive drug group: 3. P＜0.01
As seen from the above table, medicine group I of the present invention, II, III, IV all can obviously suppress platelet aggregation, and wherein the high dose group effect is better than positive drug (P＜0.01).
3.2 plasma fibrinogen (FIB) is measured
Arteria carotis blood sampling 3mL adds 3.8 liquor sodii citratis anti-freezings (1:9), mixing, 1000rmin
-1Centrifugal 10min, blood platelet blood plasma is rich in preparation, and that draws preparation is rich in blood platelet blood plasma, measures with semi-automatic biochemical analyzer by immune turbidimetry, and experimental result sees Table 7,8,9.
The influence of table 7 pair whole blood viscosity (x ± s) (n=8)
Annotate: compare: 1. P＜0.05,2. P＜0.01 with model group; Compare with the positive drug group: 3. P＜0.01.
3.3 hemorheology index is measured.
Arteria carotis blood sampling 5mL, add test tube, (every test tube adds 0.5 heparin-saline solution 0.2mL to anticoagulant heparin before the experiment, 100 ℃ of following dry for standby), mixing accelerates with full-automatic blood flow and to survey instrument and measure whole blood viscosity, blood plasma viscosity, whole blood reduced viscosity, packed cell volume (RBC hematocrit), erythrocyte aggregation index (RBC aggregate index), erythrocyte electrophoretic time (RBC electrophoresis time).
The influence of table 8 pair blood plasma viscosity, whole blood reduced viscosity, FIB (x ± s) (n=8)
| Group | Blood plasma viscosity (mPaS) | Whole blood reduced viscosity (mPaS) | FIB(dl/mg)? |
| Positive controls | 1.350±0.186 | 11.791±1.324 | 404.85±22.38 |
| Medicine group I of the present invention | 1.325±0.074? | 11.052±0.745①? | 350.46±36.06② |
| Medicine group II of the present invention | 1.342±0.155? | 10.992±0.850①? | 377.00±32.31① |
| Medicine group III of the present invention | 1.397±0.045? | 10.957±1.262①? | 399.48±26.52? |
[0102]?
| Medicine group IV of the present invention | 1.401±0.049? | 10.917±1.285①? | 404.44±25.72? |
| Model group | 1.478±0.191 | 12.780±1.278 | 426.40±21.60 |
Annotate: compare: 1. P＜0.05,2. P＜0.01 with model group.
Table 9 pair RBC hematocrit, RBC aggregate index, RBC electrophoresis time influence (x ± s) (n=8)
| Group | RBC hematocrit (L/L) | The RBC aggregate index | RBC electrophoresis time (S) |
| Positive controls | 0.423±0.025? | 11.770±1.360? | 22.361±1.622① |
| Medicine group I of the present invention | 0.341±0.046① | 9.395±1.312? | 18.232±1.749②③ |
| Medicine group II of the present invention | 0.361±0.039① | 9.695±1.212? | 18.732±1.949②③ |
| Medicine group III of the present invention | 0.434±0.038? | 10.825±0.370? | 21.380±0.704② |
| Medicine group IV of the present invention | 0.406±0.070? | 10.793±1.984? | 20.438±2.771② |
| Model group | 0.431±0.024 | 11.953±1.696 | 24.341±1.460 |
Annotate: compare: 1. P＜0.05,2. P＜0.01 with model group; Compare with positive controls: 3. P＜0.01.
By table 7,8 results as seen, medicine group I of the present invention, II, III, IV all can significantly reduce whole blood viscosity; Each group all can reduce the whole blood reduced viscosity.By table 8,9 results as seen, medicine group I of the present invention, II, III, IV all can obviously shorten rat RBC electrophoresis time, and each group can reduce plasma F IB content.Effect relatively has positive effect with positive controls.
4 antithrombotics form
4.1 thromboxane B2 (TXB2) and 62 ketone, 2 PGF1s (62keto2PGF1 α) are measured
Through arteria carotis blood sampling 3mL, medicine group 2ED2TANa of the present invention
2The about 0.2mL anti-freezing of solution, mixing, 4 ℃ of centrifugal 3500rmin
-1, 15min, separated plasma, TXB2,62keto2PGF1 α adopt measured by radioimmunoassay.All operation is undertaken by the requirement of radioimmunology analysis mensuration kit instructions.Experimental result sees Table 10
The influence of table 10 couple blood plasma TXB2,62keto2PGF1 alpha content (x ± s)
| Grouping | TXB2(pg/mL)? | 62keto2PGF1α(pg/mL) | TXB2/62keto2PGF1α |
| Positive controls | 120.38±13.14② | 153.38±35.68? | 0.83±0.22? |
| Medicine group I of the present invention | 100.13±8.90②③ | 172.50±35.56? | 0.62±0.17②? |
| Medicine group II of the present invention | 108.50±14.65② | 160.00±25.63? | 0.69±0.14①? |
| Medicine group III of the present invention | 113.63±13.68② | 161.25±28.25? | 0.75±0.17①? |
| Medicine group IV of the present invention | 127.00±20.99① | 143.25±38.97? | 0.94±0.25①? |
| Model group | 143.75±12.75 | 131.38±23.35? | 1.11±0.12? |
Annotate: compare with model group: 1. P＜0.05,2. P＜0.01 is compared with low dose group: 3. P＜0.01.
By table 10 as seen, each group of medication all can reduce the content of TXB2 in the mouse body, and medicine group I wherein of the present invention, II, III, IV blood plasma TXB2 content all reduce; Each organizes blood plasma TXB2 and 62keto2PGF1 α ratio obviously reduces (P＜0.05, P＜0.01) than model group.
4.2 external thrombus is measured
Arteria carotis blood sampling 1.8mL, injecting diameter rapidly is in the 4mm thrombus ring, puts on the extracorporeal thrombosis forming device rotating disk and takes off behind the rotation 15min, thrombus is poured on the filter paper in will encircling, and inhales and removes surplus blood, measures its length, Wen Chong, dry weight.Experimental result sees Table 11
Influence (x ± s) (n=8) that table 11 pair external thrombus forms
| Grouping | Length (cm) | Weight in wet base (mg) | Dry weight (mg) |
| Positive controls | 3.43±0.65① | 36.75±6.47① | 15.13±2.74① |
| Medicine group I of the present invention | 3.78±1.03①? | 40.00±11.33① | 17.43±12.40① |
| Medicine group II of the present invention | 2.81±0.91①? | 24.25±7.46①? | 11.50±2.45①? |
[0119]?
| Medicine group III of the present invention | 1.02±0.89①②? | 9.02±7.05①②? | 4.05±2.94②? |
| Medicine group IV of the present invention | 1.10±0.92①②? | 8.94±7.65①②? | 4.25±3.64②? |
| Model group | 9.77±2.20 | 128.38±26.45 | 55.38±11.77 |
Annotate: compare with model group: 1. P＜0.01, compare with positive controls: 2. P＜0.01.
Find out that by the result each group of medication all has anti thrombotic action, with Chinese medicine high dose best results.Thrombus do not occur, and be better than positive control.
Experimental example 2 is differentiated screening experiment
We get the different preparations of medicine of the present invention, carry out microscopical identification respectively, prove that this discrimination method is stable, are applicable to that the thin layer of all preparations under this its preparation process is differentiated.
1, the thin layer discrimination method of motherwort:
1) preparation of need testing solution
The preparation method of need testing solution determines that according to the concrete effective ingredient of discriminating medicine the discriminating to motherwort in the medicine of the present invention mainly is the wherein discriminating of stachydrine hydrochloride.Adopt orthogonal test to determine effective separating and extracting process.
One of need testing solution preparation method: get this product, porphyrize adds ethanol, and sonicated filters, and filtrate volatilizes, and residue adds absolute ethyl alcohol makes dissolving, as need testing solution.The negative need testing solution that is equipped with the motherwort medicinal material with legal system.
Need testing solution preparation method's two: get this product, porphyrize adds the 40ml absolute ethyl alcohol, divides 3 joltings to extract, and extract evaporate to dryness, residue add the 1.5ml absolute ethyl alcohol makes dissolving, as need testing solution.The negative need testing solution that is equipped with the motherwort medicinal material with legal system.
2) the middle developping agent proportioning of above-mentioned discrimination method (2) is preferred: draw the need testing solution 15 μ l of preparation as stated above, the reference substance solution 5 μ l of stachydrine hydrochloride, put respectively on same silica gel g thin-layer plate, with benzene-dioxane-acetic acid proportioning be (90:25:4) (75:25:4), ethyl acetate-absolute ethyl alcohol-hydrochloric acid proportioning is (3:7:1), normal butyl alcohol-hydrochloric acid-water proportioning is (1-10:0.2-3:0.1-2), normal butyl alcohol-hydrochloric acid-ethyl acetate proportioning is that (8:3:1) is developping agent, put a small beaker of containing ammoniacal liquor in the developing tank and carry out the presaturation of ammonia steam, launch, after drying, potassium is secreted in spray improvement iodate, the electric heating blowing is sprayed 10% sulfuric acid ethanol again to dry, and is clear to the spot colour developing.Observe the effect that test sample launches on each thin layer plate, the results are shown in following table:
Developping agent is benzene-dioxane-acetic acid as can be seen from the above table, and when proportioning was (90:25:4), it is best that need testing solution launches effect, and appearance hangover, spot separate phenomenons such as bad.
3) mensuration of stachydrine hydrochloride detectability in the above-mentioned discrimination method (2) contains 0.05mg, 0.1mg, 0.2mg, 0.3mg in every 1ml reference substance solution, gets 15 μ l.Reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with benzene-dioxane-acetic acid proportioning is that (90:25:4) is developping agent, put a small beaker of containing ammoniacal liquor in the developing tank and carry out the presaturation of ammonia steam, launch, after drying, potassium is secreted in spray improvement iodate, the electric heating blowing is sprayed 10% sulfuric acid ethanol again to dry, and is clear to the spot colour developing.Observe the effect of test sample principal spot colour developing on the thin layer plate, the results are shown in following table:
| Contrast concentration/ml | 0.05mg? | 0.1mg? | 0.2mg? | 0.3mg? |
| Color developing effect | Test sample is at corresponding reference substance position immaculate | Test sample is shallow in corresponding reference substance position spot colors, but invisible sometimes | Test sample is at corresponding reference substance position clear spot | Test sample is very clear in the colour developing of corresponding reference substance position spot |
Test sample concentration is when 0.2mg/ml as can be seen from the above table, and it is clear to develop the color on thin layer plate, is fit to testing requirements.Gu is with 10
-6G is decided to be the detectability of stachydrine hydrochloride.Therefore, test sample concentration is decided to be more than the 2mg/ml point sample amount 15 μ l.
4) the middle sample solution concentration of above-mentioned discrimination method (2) is preferred, contains this product 1g, 3g, 0.5g, 0.7g in every 1ml need testing solution.Reference substance solution 1 μ l need testing solution 3 μ l, put respectively on same silica gel g thin-layer plate, with benzene-dioxane-acetic acid proportioning is that (90:25:4) is developping agent, put a small beaker of containing ammoniacal liquor in the developing tank and carry out the presaturation of ammonia steam, launch, after drying, potassium is secreted in spray improvement iodate, the electric heating blowing is sprayed 10% sulfuric acid ethanol again to dry, and is clear to the spot colour developing.Observe the effect of test sample principal spot colour developing on the thin layer plate, the results are shown in following table:
| Sample concentration/ml | 0.1g? | 0.3g? | 0.5g? | 0.7g? |
| Color developing effect | Test sample is at corresponding reference substance position immaculate | Test sample is shallow in corresponding reference substance position spot colors, but invisible sometimes | Test sample is at corresponding reference substance position clear spot | Test sample is very clear in the colour developing of corresponding reference substance position spot |
Test sample concentration is when 0.5g/ml as can be seen from the above table, and it is clear to develop the color on thin layer plate, is fit to testing requirements.Gu is decided to be 5g/ml the test sample concentration of stachydrine hydrochloride.
4) negative control test
Get the negative sample that lacks motherwort, prepare negative control solution, launch the back and corresponding spot on the reference substance solution correspondence position, do not occur, illustrate that selected identification experiment specificity is strong according to need testing solution preparation method in the above-mentioned discrimination method (2).
The experiment of experimental example 3 assays
Detecting instrument (room temperature detection):
Agilent1100 type high performance liquid chromatograph;
Octadecylsilane chemically bonded silica (4.6 * 150mm, 5 μ m)
Producer: Agilent Technologies Anjelen Sci. ﹠ Tech. Inc (China)
Moving phase: methanol-water-glacial acetic acid (25:75:1.8)
Detect wavelength: 330nm
Flow velocity: 1.0ml/min
Column temperature: room temperature
The reference substance source: forulic acid is purchased lot number: the 0773-9910 in Nat'l Pharmaceutical ﹠ Biological Products Control Institute
Assay method: the preparation method who gets by need testing solution under [assay] item prepares sample liquid; And by preparing the blank sample that lacks Ligusticum wallichii and Radix Angelicae Sinensis under [method for making] item, the preparation negative controls.Filter with miillpore filter (0.45 μ m).Accurate respectively each the 10 μ l of negative controls, reference substance liquid and need testing solution that draw inject liquid chromatograph, measure, promptly.
1. content assaying method is investigated:
(1) stability test reference substance solution respectively at preparing the back 0,2,4,6,12,24 hour, is measured in accordance with the law, and the result shows that it is basicly stable in 24 hours, the results are shown in following table
(2) linear relationship is investigated and to be got reference substance solution (0.00526mg/ml) and shake up, accurate respectively 1,3,5,7,9, the 11 μ l of absorption inject high performance liquid chromatograph, measure peak area, the results are shown in following table, and drawing standard curve, show that forulic acid is linear between 0.00526 μ g-0.05786 μ g, its regression equation is:
Area＝4752.682781×Amt+6.03859(r＝0.999403)
(3) the accurate need testing solution of drawing of precision test, (lot number: 03110401) 10 μ l, repeat sample introduction 5 times, try to achieve relative standard deviation＜2%, the results are shown in following table
(4) the text method is pressed in the reappearance test, and (lot number: 03110502) sample is 5 parts, measures respectively, tries to achieve relative standard deviation＜2%, the results are shown in following table to get same lot number
(5) the recovery test precision takes by weighing the same lot number (lot number: accurate respectively again forulic acid reference substance solution (5.26ug/ml) 10ml that adds of sample 1.0g 03110502) of known content, press preparation method's operation of text need testing solution, measure its content, and calculate its recovery, measurement result sees the following form:
2. measurement result: (seeing figure)
According to above data, content limit is decided to be: every bag of this product contains Ligusticum wallichii and Radix Angelicae Sinensis by forulic acid (C
10H
10O
4) meter, must not be less than 0.1mg.
Following embodiment all can realize the described effect of above-mentioned experimental example
Embodiment
Embodiment 1: granule
Radix Angelicae Sinensis 140g Ligusticum wallichii 130g
Peach kernel 60g rhizoma zingiberis (charcoal) 30g
40g breaks in motherwort 350g river
Above Six-element is ground into meal, adds 10 times of water gagings and extracts volatile oil 6 hours, and the aqueous solution after distillation device is in addition collected, the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, adds 6 times of water gagings 1.5 hours, collecting decoction for the second time, filter, filtrate and above-mentioned solution merge, and are condensed into thick paste, it is an amount of that other goes into brown sugar, and mixing is made particle, in 60 ℃ of oven dry, pulverize, make particle with ethanol, spray into volatile oil, make 466g, promptly.
Embodiment 2: soft capsule
Radix Angelicae Sinensis 140g Ligusticum wallichii 130g peach kernel 60g
40g breaks in rhizoma zingiberis (charcoal) 30g motherwort 350g river
Radix Glycyrrhizae (processing) 40g;
More than seven flavors, be ground into meal, add 10 times of water gagings and extracted volatile oil 6 hours, aqueous solution after distillation device is in addition collected, and the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, for the second time add 6 times of water gagings 1.5 hours, collecting decoction filters, filtrate and above-mentioned solution merge, and are condensed into thick paste, in 60 ℃ of oven dry, pulverize, add vegetable oil and above-mentioned volatile oil, mixing, make 77, promptly.
Embodiment 3: effervescent agent
Radix Angelicae Sinensis 140g Ligusticum wallichii 130g peach kernel 60g
Rhizoma zingiberis (charcoal) 30g motherwort 350g safflower 30-50g
More than seven flavors, be ground into meal, add 10 times of water gagings and extracted volatile oil 6 hours, the aqueous solution after distillation device is in addition collected, the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, adds 6 times of water gagings 1.5 hours, collecting decoction for the second time, filter, filtrate and above-mentioned solution merge, and are condensed into thick paste, other goes into brown sugar and effervescent agent is an amount of, and mixing is made particle, in 60 ℃ of oven dry, pulverize, make particle with ethanol, spray into volatile oil, make 466g, promptly.
Embodiment 4: oral liquid
Radix Angelicae Sinensis 180g Ligusticum wallichii 150g peach kernel 50g
40g breaks in rhizoma zingiberis (charcoal) 30g motherwort 350g river
Radix Glycyrrhizae (processing) 40g;
More than seven flavors, be ground into meal, add 10 times of water gagings and extracted volatile oil 6 hours, the aqueous solution after distillation device is in addition collected, the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, adds 6 times of water gagings 1.5 hours, collecting decoction for the second time, filter, filtrate and above-mentioned solution merge, and are condensed into thick paste, and it is an amount of that other goes into brown sugar, mixing sprays into volatile oil, and adding essence and antiseptic are an amount of, filter, adjust total amount, stir evenly to 770ml, sterilization, can, promptly.
Embodiment 5: capsule
Radix Angelicae Sinensis 160g Ligusticum wallichii 120g peach kernel 70g
Rhizoma zingiberis (charcoal) 40g motherwort 360g safflower 30g
Radix Glycyrrhizae (processing) 35g;
More than seven flavors, be ground into meal, add 10 times of water gagings and extracted volatile oil 6 hours, the aqueous solution after distillation device is in addition collected, the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, adds 6 times of water gagings 1.5 hours, collecting decoction for the second time, filter, filtrate and above-mentioned solution merge, and are condensed into thick paste, and it is an amount of that other goes into brown sugar, mixing is made particle, in 60 ℃ of oven dry, pulverize, make particle, spray into volatile oil with ethanol, encapsulated, make 77, promptly.
Embodiment 6: tablet
Radix Angelicae Sinensis 140g Ligusticum wallichii 130g peach kernel 60g
Rhizoma zingiberis (charcoal) 30g motherwort 350g safflower 40g
Radix Glycyrrhizae (processing) 40g;
More than seven flavors, be ground into meal, add 10 times of water gagings and extracted volatile oil 6 hours, aqueous solution after distillation device is in addition collected, and the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, for the second time add 6 times of water gagings 1.5 hours, collecting decoction filters, filtrate and above-mentioned solution merge, and are condensed into thick paste, and it is an amount of that other goes into brown sugar, mixing is made particle with ethanol, sprays into volatile oil, in 60 ℃ of oven dry, compressing tablet, promptly.
Embodiment 7: particle
Radix Angelicae Sinensis 240g Ligusticum wallichii 90g peach kernel 24g
Radix Glycyrrhizae (processing) 15g rhizoma zingiberis (charcoal) 15g motherwort 300g
Safflower 15g
More than seven flavors, be ground into meal, add 10 times of water gagings and extracted volatile oil 6 hours, the aqueous solution after distillation device is in addition collected, the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, adds 6 times of water gagings 1.5 hours, collecting decoction for the second time, filter, filtrate and above-mentioned solution merge, and are condensed into thick paste, it is an amount of that other goes into brown sugar, and mixing is made particle, in 60 ℃ of oven dry, pulverize, make particle with ethanol, spray into volatile oil, make 466g, promptly.
Differentiate:
(1) get this product 2g, the ammonification water number drips and makes moisteningly, adds chloroform 20ml again, refluxes 15 minutes in water-bath, cooling filters, and filtrate evaporate to dryness, residue add 1% hydrochloric acid 1ml dissolving, get 3～4 of hydrochloric acid solutions respectively on two surface plates, one adds 1 of bismuth potassium iodide test solution, salmon precipitation promptly takes place; Another adds 1 of silico-tungstic acid test solution, and white precipitate promptly takes place;
(2) get this product 6g, add an amount of zeyssatite and grind evenly, put in the apparatus,Soxhlet's, an amount of with methyl alcohol, reflux is closely colourless to extract, reclaims methyl alcohol, residue adds water 50ml, heating makes dissolving, puts coldly, is transferred in the separating funnel, with extracted by ether 5 times (20,20,15,15,10ml), merge extract, extract 5 times (20 with 2% sodium carbonate liquor, 20,15,15,10ml), merge alkali lye, wash 3 times with ethyl acetate, each 15ml discards ethyl acetate liquid, alkali lye add hcl acidifying to PH be 2～3, wash 3 times with benzene again, each 15ml discards benzene liquid, continues and uses extracted by ether 5 times (20,20,15,15,10ml), merge ether solution, volatilize, residue adds methyl alcohol 1ml makes dissolving, as need testing solution; Other gets the forulic acid reference substance, adds methyl alcohol and makes the solution that every 1ml contains 1mg, compares product solution; According to the thin-layered chromatography test, draw above-mentioned need testing solution 20 μ l, reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, with benzene-dioxane-acetic acid (90:25:4) is developping agent, launches, and takes out, dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show identical mazarine fluorescence spot;
(3) get this product 6g, add the 40ml absolute ethyl alcohol, divide 3 joltings to extract, extract evaporate to dryness, residue add the 1.5ml dissolve with ethanol, as need testing solution; Other gets the stachydrine hydrochloride reference substance, adds an amount of ethanol and makes the solution that every 1ml contains 2mg, in contrast product solution; Drawing above-mentioned two kinds of solution each 15 μ l, 5 μ l puts respectively on same silica gel H plate, with chloroform-methanol (1:1) is developping agent, put a small beaker of containing ammoniacal liquor in the developing tank and carry out the presaturation of ammonia steam, launch, after drying, secrete potassium with the improvement iodate, the electric heating blowing is to dry, spray 10% ethanol solution of sulfuric acid again, clear to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
Assay:
According to high performance liquid chromatography: chromatographic condition and system suitability test are filling agent (4.6 * 150mm, 5 μ m) with octadecylsilane chemically bonded silica; Methanol-water-acetic acid (25:75:1.8) is moving phase; The detection wavelength is 330nm; Number of theoretical plate calculates by forulic acid should be not less than 2000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing the forulic acid reference substance, adds the mixed solution of methyl alcohol-formic acid (95:5), makes the solution that contains 5ug among every 1ml, filters with miillpore filter (0.45um), promptly; The preparation of need testing solution: it is an amount of to get under this product content uniformity item particle, porphyrize, and precision takes by weighing about 3g, put in the tool plug Erlenmeyer flask, the accurate mixed solution 25ml that adds methyl alcohol-formic acid (95:5) claims to decide weight, sonicated (power 250W, frequency 40KHZ) 30 minutes, put coldly, claim to decide weight again, after supplying the weight that subtracts mistake with the mixed solution of methyl alcohol-formic acid (95:5), shake up, filter with miillpore filter (0.45um), promptly; Assay method: accurate respectively reference substance solution, each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.Every bag of this product contains Ligusticum wallichii and Radix Angelicae Sinensis with forulic acid (C
10H
10O
4) meter, must not be less than 0.1mg.
Function with cure mainly: invigorate blood circulation, the dispel stasis of blood, pain relieving.Be used for disease that postpartum lochia can't go, few abdomen pain also can be tried out the colporrhagia that causes, menorrhalgia after being fitted with a contraceptive ring.
Specification: every packed 6g is equivalent to crude drug 9g.
Claims (1)
1. content assaying method for the treatment of the pharmaceutical composition of disease that postpartum lochia can't go is characterized in that this method is:
Choose following material medicine and make granule:
Radix Angelicae Sinensis 240 weight portions, Ligusticum wallichii 90 weight portions, peach kernel 24 weight portions, honey-fried licorice root 15 weight portions, carbonized Rhizoma Zingiberis 15 weight portions, motherwort 300 weight portions, safflower 15 weight portions; More than seven flavors, be ground into meal, add 10 times of water gagings and extracted volatile oil 6 hours, the aqueous solution after distillation device is in addition collected, the dregs of a decoction are the boiling secondary again, adds 8 times of water gagings 2 hours for the first time, adds 6 times of water gagings 1.5 hours, collecting decoction for the second time, filter, filtrate and above-mentioned solution merge, and are condensed into thick paste, it is an amount of that other goes into brown sugar, and mixing is made particle, in 60 ℃ of oven dry, pulverize, make particle with ethanol, spray into volatile oil, make 466 weight portions, promptly;
According to high performance liquid chromatography: chromatographic condition and system suitability test are filling agent with the octadecylsilane chemically bonded silica of 4.6 * 150mm, 5 μ m; 25: 75: 1.8 methanol-water-acetic acid is moving phase; The detection wavelength is 330nm; Number of theoretical plate calculates by forulic acid should be not less than 2000; The preparation of reference substance solution: it is an amount of that precision takes by weighing the forulic acid reference substance, adds the mixed solution of methyl alcohol-formic acid of 95: 5, makes the solution that contains 5 μ g among every 1ml, filters with 0.45 μ m miillpore filter, promptly; The preparation of need testing solution: it is an amount of to get under this product content uniformity item particle, porphyrize, and precision takes by weighing 3g, put in the tool plug Erlenmeyer flask, the accurate mixed solution 25ml that adds methyl alcohol-formic acid of 95: 5 claims to decide weight, sonicated, power 250W, frequency 40KHZ, 30 minutes, put coldly, claim to decide weight again, after supplying the weight that subtracts mistake with the mixed solution of methyl alcohol-formic acid of 95: 5, shake up, filter with 0.45 μ m miillpore filter, promptly; Assay method: accurate respectively reference substance solution, each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.
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Effective date of registration: 20160226 Address after: 102600, Daxing District Zhongguancun science and Technology Park, Daxing pharmaceutical industry base, Tianhe West Road, No. 19, Beijing Patentee after: Beijing rich church Pharmaceutical Technology Co., Ltd. Address before: 102200, 8, Zhenxing Road, Zhongguancun science and Technology Park, Beijing, Changping District Patentee before: Beijing Asia-East Bio-Pharmaceutical Co., Ltd. |