CN100591339C - Method for preparing the total sennoside in senna - Google Patents
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Abstract
A medical application of the general sennoside extracted from senna leaf in preparing the medicines for treating osteoporosis is disclosed. Its preparing process includes such steps as immersing, seeping, alkaline macroreticular resin adsorption, acidic macroreticular resin adsorption, recovering and drying.
Description
Technical field:
The present invention relates to new medical use of carboxylic anthraquinone class and dianthrone class in the plant and preparation method thereof, exactly is the new medical use and the preparation method of (belonging to carboxylic anthraquinone class and dianthrone class on the chemical constitution) of total sennoside in the Chinese medicine Folium Sennae.
Background technology:
Folium Sennae (Folium sennae) is one of conventional Chinese medicine, one of intestinal stasis relieving medicine down that rushes down commonly used for Chinese Pharmacopoeia one one of version in 2000 is recorded is narrow leaf Folium Sennae of leguminous plant (Cassia angustifolia Vahl) or cassia acutifolia Delile leaf C.acutifolia Delile) dried leaves
[1]
Folium Sennae sweet in the mouth, hardship, cold in nature having and purging heat that row stagnates, the function of relieving constipation.Be mainly used in constipation with heat retention, stagnant abdominal distention.Found in the last few years that Folium Sennae can treat epidemic hemorrhagic fever, acute pancreatitis and toxical paralytic ileus etc.
[2] [3]This product relies on import for a long time, and main body entrance state is an India.Twentieth century is since the eighties, China in Hainan, Guangxi etc. economizes successful introduction, but the commodity price only is about 1/3～1/2 of import
[4]Total sennoside is the main active in the Chinese medicine Folium Sennae, but less to its activity research report, mainly concentrates on to rush down intestinal stasis relieving aspect down.This seminar has carried out more comprehensive pharmacological experiment study to it, and finds that first total sennoside has estrogen sample function of resisting osteoporosis.
Osteoporosis (osteoporosis) is common complaint among the elderly, frequently-occurring disease.Women after climacteric is because osteoporosis is easily suffered from the minimizing of estrogen in the body.Along with the aging of population, osteoporosis will become the significant problem of medical treatment and society
[5]Though the osteoporotic medicament categories of present alternative treatment is many, curative effect is not good enough, and some import drug price costlinesses embarrass Most patients to accept in fact.The beginning of this century, a kind of vegetable complexing agent that contains plant estrogen (phytoestrogen) becomes the situation of selling well medicine because of its side effect is little in the U.S. and Japan.Middle-aged and elderly people has senile constipation more than 80%, osteoporosis is arranged again simultaneously.Therefore, total sennoside has high using value and prospect in the Folium Sennae.
Total sennoside has medical usage widely, high using value and prospect.Therefore the expert, the scholar that comprise many countries of China have carried out extensive and deep research to the method for extracting total sennoside from Folium Sennae, obtain gratifying achievements.But the most frequently used method of conclude is alcohol extract or the water extract with Folium Sennae to suspend in water, and uses the solvent extraction of opposed polarity successively, and solvent commonly used has ethyl acetate and water saturated n-butyl alcohol.Total sennoside focuses mostly in n-butanol layer.The reclaim under reduced pressure n-butyl alcohol; Extractum concentration is 95% dissolve with ethanol, and makes it pass through the aluminium sesquioxide chromatographic column
[6]Collect effluent and decompression recycling ethanol, make finished product.The subject matter of utilizing this method to produce the total sennoside existence is: (1) product cost is higher, and per kilogram finished product cost is up to 2800 yuans; (2) impurity in the product, particularly content of harmful is higher, wherein content of beary metal surpasses 5ppm, persticide residue is greater than 8ppm, surpass prescribed limit, toxic solvent, n-butanol residual inevitably also simultaneously, the content of effective ingredient Sennoside A and B only have technology of the present invention preparation method 1/5～1/7.
Summary of the invention:
The present invention is exactly at the problems referred to above, provides in a kind of Folium Sennae total sennoside to treat osteoporotic medicine; Provide and from Folium Sennae, extract the preparation method that the total sennoside cost is low, impurity content is few and environmental pollution is little.
Total sennoside has estrogen sample function of resisting osteoporosis in the Folium Sennae, can be proved from the test of following three aspects.
Folium Sennae is the conventional Chinese medicine that China's pharmacopeia (1995～2005 years versions) is recorded, and its main component is compositions such as Sennoside A, B, C, D and rheinoside, is referred to as total sennoside (Sennosides).We study the function of resisting osteoporosis of total sennoside:
Animal: Kunming kind female mice, body weight 20.0 ± 2.0g.
Experimental technique: under etherization take out the mice bilateral ovaries, postoperative began to do vaginal smear in 5 days, every day 1 time, continuous 6 days.With the mice random packet of no oestrous cycle, 20 every group.Pseudo-castration mice in control group is under etherization done two stringer otch in the twelfth rib bone lower edge and the outside, spinal column sacrococcygeal joint place, is about 1 centimetre, does not remove ovary.To irritate the administration of stomach mode.Make the mouse vagina smear and observe the situation that occurs rutting period every day in the administration process.Successive administration 49 days is got blood in 24 hours posterior orbits of last administration, and separation of serum is surveyed the content of blood calcium, serium inorganic phosphorus and calcitonin, and record mice body weight, the gonophore uterus is heavy and the left side femur is heavy, calculates organ coefficient.The results are shown in Table 1～3.
(1) total sennoside (Sennosides) the results are shown in Table 1 to the influence of castration female mice oestrous cycle
Table 1. total sennoside is to the influence of castration female mice oestrous cycle
Annotate: every treated animal number average is 20.
Experimental result shows that total sennoside can make castration mouse reappear the oestrous cycle, shows that total sennoside has the effect of estrogen sample.
(2) influence that total sennoside is heavy to castration female mice uterus and fl is heavy the results are shown in Table 2
The influence that table 2. total sennoside is heavy to castration female mice uterus and fl is heavy
Compare with the castration group #p＜0.05 ##p＜0.01;
*P＜0.05
*Compare with model group p＜0.01
Experimental result shows between castration model group uterus coefficient and fl coefficient and pseudo-castration group significant difference is arranged all; Between the high low dose group of diethylstilbestrol group and total sennoside uterus coefficient and fl coefficient and the castration model group significant difference is arranged all.Illustrate that total sennoside has the estrogen sample to promote uterus hypertrophy and the effect that the sclerotin calcification is increased the weight of.
(3) total sennoside the results are shown in Table 3 to the influence of castration mouse blood calcium, serium inorganic phosphorus and calcitonin content
Table 3. total sennoside is to the influence of castration mouse blood calcium, serium inorganic phosphorus and calcitonin content
Compare with the castration group #p＜0.05;
*Compare with model group p＜0.05
With the blood calcium is index, between castration model group and pseudo-castration group significant difference is arranged; Between high low dose group of total sennoside and castration model group significant difference is arranged, illustrate that total sennoside can reduce blood calcium, makes the sclerotin calcification.
With the serium inorganic phosphorus is index, does not have significant difference between pseudo-castration group and administration group and the castration model group.
With the calcitonin is index, compares with pseudo-castration group, and castration model group mice serum calcitonin descends, and total sennoside group and castration model group relatively serum calcitonin are in rising trend, but do not have significant difference.
More than the experimental result of three aspects show that fully the total sennoside for preparing has estrogen sample function of resisting osteoporosis from the Chinese medicine Folium Sennae.
The preparation method of total sennoside is as follows in the Folium Sennae of the present invention:
The method for preparing total sennoside provided by the invention comprises the immersion of Folium Sennae and percolation, recovery, exsiccant step (above be common process), its main feature is after immersion and percolation step, before recovery, exsiccant step, increase macroporous adsorptive resins absorption impurity and noxious substance under the alkalescence successively, wherein total sennoside is not adsorbed and flows out, macroporous adsorptive resins absorption total sennoside and desalination under acidity; The liquid of producing behind described immersion and the percolation respectively by the macroporous adsorptive resins after the activated processing or be mixed the back by the macroporous adsorptive resins after the activated processing, the collection effluent, be adjusted to acidity; The macroporous adsorptive resins of acid medicinal liquid after by activated processing, total sennoside in the liquid is adsorbed by macroporous adsorptive resins, discard effluent, press 0.1～4 times distilled water flushing macroporous adsorptive resins of macroporous adsorptive resins retention volume again, water lotion discards, continuing with concentration is 10～50% ethanol or washed with methanol macroporous adsorptive resins, makes absorption total sennoside thereon by eluting fully, and collects eluent.
Compared with the prior art the present invention has the following advantages: (1) product cost is low, and the per kilogram finished product is about 600 yuans; (2) impurity content in the finished product is low: content of beary metal is less than 5ppm, and persticide residue is less than 10ppb; (3) eliminate the residual of toxic solvents n-butyl alcohol; (4) production process is almost pollution-free to environment.
Embodiment 1
Step 1: Folium Sennae is soaked and percolation:
(A) Folium Sennae and aqueous alkali (5%NaHCO
3, or pH is the Na of 8-10
2CO
3(or NaOH, K
2CO
3, KOH aqueous solution)) be 1: 6～24 by ratio of weight and the number of copies, in Folium Sennae, add aqueous alkali, soak at room temperature 0.5～2 hour, centrifugal then, residue obtained liquid, or percolation is got liquid;
(B) be 1: 6～12 in the slag that step (A) leaches, to add aqueous alkali by the ratio of weight and number of described Folium Sennae and aqueous alkali, centrifugal, residue obtained liquid, or percolation is got liquid;
(C) be 1: 6～12 in the slag that step (B) leaches, to add aqueous alkali by the ratio of weight and number of described Folium Sennae and aqueous alkali, centrifugal, residue obtained liquid, or percolation is got liquid;
Step 2: alkalescence is absorption impurity down:
The liquid that step 1 is produced respectively by the macroporous adsorptive resins after the activated processing or the back that is mixed by the macroporous adsorptive resins after the activated processing, the collection effluent is adjusted to pH1～4 with acid;
Step 3: absorption total sennoside and desalination:
With the acid medicinal liquid of step 2 macroporous adsorptive resins after by activated processing, total sennoside in the liquid is adsorbed by macroporous adsorptive resins, discard effluent, press 0.1～4 times distilled water flushing macroporous adsorptive resins of macroporous adsorptive resins retention volume again, water lotion discards, continuing with concentration is 10～50% ethanol or washed with methanol macroporous adsorptive resins, makes absorption total sennoside thereon by eluting fully, and collects eluent;
Step 4: recovery, drying:
Ethanol in the eluent that recycling step 3 makes after the remaining material drying, can obtain total sennoside finished product in the Folium Sennae.
Embodiment 2
Step 1: Folium Sennae is soaked:
(A) get narrow leaf Folium Sennae (Folium sennae) 1Kg and put into container, in container, add 5%NaHCO again
38 kilograms of aqueous solutions, soak at room temperature is 1 hour under stirring at any time, then, after the centrifuge centrifugalize, collects filtrate;
(B) in the slag that step (A) leaches, add 5%NaHCO
36 kilograms of aqueous solutions, soak at room temperature is 40 minutes under stirring at any time, then, after the centrifuge centrifugalize, collects filtrate;
(C) in the slag that step (B) leaches, add 5%NaHCO
36 kilograms of aqueous solutions, soak at room temperature is 30 minutes under stirring at any time, afterwards, centrifugalize, residue obtained liquid;
Step 2: alkalescence is absorption impurity down:
The akaline liquid that above steps (A), (B), (C) are produced by 2 kilograms after the activated processing D101 type macroporous adsorptive resins, is collected about 15.5 kilograms of effluent respectively, is adjusted to pH2.5 with 10% hydrochloric acid;
Step 3: acid absorption total sennoside down desalination:
D101 type macroporous adsorptive resins with the acid medicinal liquid of step 2 2Kg after by activated processing, total sennoside in the liquid is adsorbed by macroporous adsorptive resins, discard effluent, press 1 times distilled water 3000ml flushing macroporous adsorptive resins of macroporous adsorptive resins retention volume again, water lotion discards, continuing with concentration is that 30% ethanol cleans macroporous adsorptive resins, makes absorption total sennoside thereon by eluting fully, and collects eluent;
Step 4: recovery, drying:
Ethanol in the eluent that recycling step 3 makes, remaining material promptly obtain total sennoside finished product 33.6g in the Folium Sennae after drying below 80 ℃.
Embodiment 3
Step 1: Folium Sennae is soaked:
(A) the dried leaves 1Kg that gets cassia acutifolia Delile leaf (C.acutifolia Delile) puts into the percolation container, progressively adds pH and be 8 kilograms of 8.5 alkaline aqueous solutions in container, press the percolation method, regulates and control flow velocity for per hour about 1 kilogram, collects filtrate;
(B) adding pH in step (A) slag that for the first time percolation leaches is 6 kilograms of 8.5 alkaline aqueous solutions, press the percolation method, regulates and control flow velocity for per hour about 0.7 kilogram, collects filtrate;
(C) adding pH in step (B) slag that for the second time percolation leaches is 6 kilograms of 8.5 alkaline aqueous solutions, press the percolation method, regulates and control flow velocity for per hour about 0.5 kilogram, collects filtrate;
Step 2: alkalescence is absorption impurity down:
The akaline liquid that step 1 is produced by the AB-8 type macroporous adsorptive resins of the 2Kg after the activated processing, is collected about 15 kilograms of effluent respectively, is adjusted to pH3.0 with 10% hydrochloric acid;
Step 3: acid absorption total sennoside down desalination:
AB-8 type macroporous adsorptive resins with the acid medicinal liquid of step 2 2Kg after by activated processing, total sennoside in the liquid is adsorbed by macroporous adsorptive resins, discard effluent, press 2 times distilled water 6000ml flushing macroporous adsorptive resins of macroporous adsorptive resins retention volume again, water lotion discards, continuing with concentration is 40% washed with methanol macroporous adsorptive resins, makes absorption total sennoside thereon by eluting fully, and collects eluent;
Step 4: recovery, drying:
Ethanol in the eluent that recycling step 3 makes, remaining material promptly obtain total sennoside finished product 43.6g in the Folium Sennae after drying below 80 ℃.
Total sennoside product in the Folium Sennae of producing by the present embodiment method:
1. yield (percentage by weight): be 4.1～4.5%.
2. detect by thin layer chromatography scanning: this product total sennoside (in Sennoside A, B) 〉=20%.
3. press spectrophotography: this product general anthraquinone class is 56-60% in chrysophanic acid content.
4. pressing the HPLC method detects: contained Sennoside A of this product and sennoside B content sum 〉=20%
5. composition: Sennoside A, sennoside B, sennoside C, sennoside D, chrysophanic acid, chrysophanic acid-8-O-glucoside, kaempferol etc.
6. impurity content: heavy metal 5ppm, persticide residue, both the residual quantity of " 666 ", " DDT ", " 5 chloronitrobenzene " all≤10ppb.
List of references:
[1] 2000 editions .285. of Pharmacopoeia of People's Republic of China
[2] Xiao Peigen chief editor. newly organized Chinese medicinal herbal (the 3rd volume) .509.
[3] Yao KaiBing etc. Journal of Traditional Chinese Medicine, 1988,29:535.
[4] commodity price catalogue (and decoction pieces). June calendar year 2001 is compiled .248. by Chinese company
[5] fourth Ramulus Cinnamomi etc. prevention and treatment of osteoporosis and research direction thereof. clinical department of internal medicine magazine, 1991,8 (2): 44-45.
[6]Atzom?R，et?al.Planta?Med.1981，41：1-8.
Claims (3)
1, the preparation method of total sennoside in a kind of Folium Sennae is characterized in that
Step 1: Folium Sennae is soaked and percolation:
(A) Folium Sennae and aqueous alkali are 1: 6～24 by ratio of weight and the number of copies, add aqueous alkali in Folium Sennae, and soak at room temperature 0.5～2 hour is centrifugal then, residue obtained liquid, or percolation is got liquid;
(B) be 1: 6～12 in the slag that step (A) leaches, to add aqueous alkali by the ratio of weight and number of described Folium Sennae and aqueous alkali, centrifugal, residue obtained liquid, or percolation is got liquid;
(C) be 1: 6～12 in the slag that step (B) leaches, to add aqueous alkali by the ratio of weight and number of described Folium Sennae and aqueous alkali, centrifugal, residue obtained liquid, or percolation is got liquid;
Step 2: alkalescence is absorption impurity down:
The liquid that step 1 is produced respectively by the macroporous adsorptive resins after the activated processing or the back that is mixed by the macroporous adsorptive resins after the activated processing, the collection effluent is adjusted to pH1～4 with acid;
Step 3: absorption total sennoside and desalination:
With the acid medicinal liquid of step 2 macroporous adsorptive resins after by activated processing, total sennoside in the liquid is adsorbed by macroporous adsorptive resins, discard effluent, press 0.1～4 times distilled water flushing macroporous adsorptive resins of macroporous adsorptive resins retention volume again, water lotion discards, continuing with concentration is 10～50% ethanol or washed with methanol macroporous adsorptive resins, makes absorption total sennoside thereon by eluting fully, and collects eluent;
Step 4: recovery, drying:
Ethanol in the eluent that recycling step 3 makes after the remaining material drying, can obtain total sennoside finished product in the Folium Sennae.
2, the preparation method of total sennoside in a kind of Folium Sennae is characterized in that
Step 1: Folium Sennae is soaked:
(A) get narrow leaf Folium Sennae (Folium sennae) 1Kg and put into container, in container, add 5%NaHCO again
38 kilograms of aqueous solutions, soak at room temperature is 1 hour under stirring at any time, then, after the centrifuge centrifugalize, collects filtrate;
(B) in the slag that step (A) leaches, add 5%NaHCO
36 kilograms of aqueous solutions, soak at room temperature is 40 minutes under stirring at any time, then, after the centrifuge centrifugalize, collects filtrate;
(C) in the slag that step (B) leaches, add 5%NaHCO
36 kilograms of aqueous solutions, soak at room temperature is 30 minutes under stirring at any time, afterwards, centrifugalize, residue obtained liquid;
Step 2: alkalescence is absorption impurity down:
The akaline liquid that above steps (A), (B), (C) are produced by 2 kilograms after the activated processing D101 type macroporous adsorptive resins, is collected about 15.5 kilograms of effluent respectively, is adjusted to pH2.5 with 10% hydrochloric acid;
Step 3: acid absorption total sennoside down desalination:
D101 type macroporous adsorptive resins with the acid medicinal liquid of step 2 2Kg after by activated processing, total sennoside in the liquid is adsorbed by macroporous adsorptive resins, discard effluent, press 1 times distilled water 3000ml flushing macroporous adsorptive resins of macroporous adsorptive resins retention volume again, water lotion discards, continuing with concentration is that 30% ethanol cleans macroporous adsorptive resins, makes absorption total sennoside thereon by eluting fully, and collects eluent;
Step 4: recovery, drying:
Ethanol in the eluent that recycling step 3 makes, remaining material promptly obtain total sennoside finished product 33.6g in the Folium Sennae after drying below 80 ℃.
3, the preparation method of total sennoside in a kind of Folium Sennae is characterized in that
Step 1: Folium Sennae is soaked:
(A) the dried leaves 1Kg that gets cassia acutifolia Delile leaf (C.acutifolia Delile) puts into the percolation container, progressively adds pH and be 8 kilograms of 8.5 alkaline aqueous solutions in container, press the percolation method, regulates and control flow velocity for per hour about 1 kilogram, collects filtrate;
(B) adding pH in step (A) slag that for the first time percolation leaches is 6 kilograms of 8.5 alkaline aqueous solutions, press the percolation method, regulates and control flow velocity for per hour about 0.7 kilogram, collects filtrate;
(C) adding pH in step (B) slag that for the second time percolation leaches is 6 kilograms of 8.5 alkaline aqueous solutions, press the percolation method, regulates and control flow velocity for per hour about 0.5 kilogram, collects filtrate;
Step 2: alkalescence is absorption impurity down:
The akaline liquid that step 1 is produced by the AB-8 type macroporous adsorptive resins of the 2Kg after the activated processing, is collected about 15 kilograms of effluent respectively, is adjusted to pH3.0 with 10% hydrochloric acid;
Step 3: acid absorption total sennoside down desalination:
AB-8 type macroporous adsorptive resins with the acid medicinal liquid of step 2 2Kg after by activated processing, total sennoside in the liquid is adsorbed by macroporous adsorptive resins, discard effluent, press 2 times distilled water 6000ml flushing macroporous adsorptive resins of macroporous adsorptive resins retention volume again, water lotion discards, continuing with concentration is 40% washed with methanol macroporous adsorptive resins, makes absorption total sennoside thereon by eluting fully, and collects eluent;
Step 4: recovery, drying:
Ethanol in the eluent that recycling step 3 makes, remaining material promptly obtain total sennoside finished product 43.6g in the Folium Sennae after drying below 80 ℃.
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Non-Patent Citations (6)
| Title |
|---|
| 大黄总蒽醌提取与纯化工艺的研究. 金波，李薇，蔡伟.时珍国医国药，第16卷第8期. 2005 |
| 大黄总蒽醌提取与纯化工艺的研究. 金波，李薇，蔡伟.时珍国医国药，第16卷第8期. 2005 * |
| 美国药典：番泻苷. 国外医药，植物药分册，第19卷第1期. 2004 |
| 美国药典：番泻苷. 国外医药，植物药分册，第19卷第1期. 2004 * |
| 美国药典：番泻苷片. 国外医药，植物药分册，第19卷第1期. 2004 |
| 美国药典：番泻苷片. 国外医药，植物药分册，第19卷第1期. 2004 * |
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