CN103923221B - A kind of preparation method of Japanese Ardisia Herb polysaccharide and application - Google Patents
A kind of preparation method of Japanese Ardisia Herb polysaccharide and application Download PDFInfo
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Abstract
The invention discloses a kind of preparation method and application of Japanese Ardisia Herb polysaccharide, effectively can solve the preparation of Japanese Ardisia Herb polysaccharide, and the problem being effective to antifatigue and improving in immunizing power medicine, its technical scheme solved is, the Japanese Ardisia Herb adding distil water of drying is soaked, ultrasonic, microwave intermittent-heating, filtration, obtain filtrate, again filtrate concentrated and add ethanol, centrifugal, washing precipitation, obtains Japanese Ardisia Herb polysaccharide crude; Again crude product is carried out purification and obtain Japanese Ardisia Herb polysaccharide.Japanese Ardisia Herb polysaccharide prepared by the present invention take natural traditional Chinese medicine as raw material, can guarantee that produced Japanese Ardisia Herb polysaccharide has no side effect, be applied in medicine, can effective antifatigue and improve the immunizing power of body, having good Social benefit and economic benefit, is the innovation on Chinese medicine.
Description
Technical field
The present invention relates to field of medicaments, particularly relate to a kind of preparation method and application of Japanese Ardisia Herb polysaccharide.
Background technology
Japanese Ardisia Herb belongs to the dry herb of the Myrsinacea plant Japanese Ardisia Herb (Ardisiajaponica (Thunb.) Blume), has another name called HERBA ARDISIAE JAPONICAE, always greatly, does not go out woods, Suffrutescent Securinega Twig.There is effect of relieving cough and reducing sputum, the stasis of blood of dispelling removing toxic substances, diuresis, pain relieving; Cure mainly swelling and pain in the throat, arthralgia, amenorrhoea, pulmonary tuberculosis, chronic bronchitis, jaundice type hepatitis and de-power internal lesion caused by overexertion.Coumarins is related generally to the research of its effective constituent, flavones and Polyphenols, triterpenes, volatile oil etc. now, and there is not been reported for the research of Japanese Ardisia Herb polysaccharide.Polysaccharide is present in the polymkeric substance that natural aldose and (or) ketose linked together by glycosidic link, and it is the requisite compositions of all lived organisms, and it and all physiological functions sustained life have close contacting.Along with people's going deep into polysaccharide researches, have been found that polysaccharide has many biological actions.From anticancer, improve the aspects such as immune, antiviral, anti-inflammatory, hypoglycemic, reducing blood-fat, antioxygenation, anti-ageing and union of fracture and all present gratifying effect.Therefore the research of Japanese Ardisia Herb polysaccharide is significant.
Summary of the invention
For above-mentioned situation, for overcoming the defect of prior art, the object of the present invention is just to provide a kind of preparation method and application of Japanese Ardisia Herb polysaccharide, effectively can solve the preparation of Japanese Ardisia Herb polysaccharide, and the problem being effective to antifatigue and improving in immunizing power medicine.
The technical scheme that the present invention solves is that a kind of preparation method of Japanese Ardisia Herb polysaccharide, comprises the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:10-20, soak 25-35min, ultrasonic 30-90min under the power of 300-600W, then microwave intermittent-heating 5-50min under identical power, filter, filtrate is concentrated into the 1.5-2 of Japanese Ardisia Herb weight doubly, adding ethanol, to make ethanol content reach volumetric concentration be 60-80%, >=12h is left standstill under 4-6 DEG C of condition, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 0.5-1%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into the 1.5-2 of Japanese Ardisia Herb polysaccharide crude weight doubly, adding ethanol, to make ethanol content reach volumetric concentration be 60-80%, >=12h is left standstill under 4-6 DEG C of condition, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide.
In step (1), microwave intermittent-heating is that 70-80 DEG C of heating 55-65s, interval 110-130s heat 55-65s in 70-80 DEG C again, then interval 110-130s, so repeats.
Japanese Ardisia Herb is the dregs of a decoction after Japanese Ardisia Herb alcohol extracting.
In step (1), ultrasonic and order that is microwave intermittent-heating can be exchanged.
Technical program of the present invention also lies in providing the application of a kind of Japanese Ardisia Herb polysaccharide adopting preparation method of the present invention to prepare in preparation antifatigue and raising immunizing power medicine.
The present invention studies for the preparation method of Japanese Ardisia Herb polysaccharide first, and method is simple, and abundant raw material, product application is wide, and be that Japanese Ardisia Herb polysaccharide prepared by raw material with Japanese Ardisia Herb, the step without the need to complexity can obtain product.Meanwhile, Japanese Ardisia Herb polysaccharide prepared by the present invention take natural traditional Chinese medicine as raw material, can guarantee that produced Japanese Ardisia Herb polysaccharide has no side effect, be applied in medicine, can effective antifatigue and improve the immunizing power of body, having good Social benefit and economic benefit, is the innovation on Chinese medicine.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is elaborated.
Embodiment 1
The preparation method of the Japanese Ardisia Herb polysaccharide of the present embodiment, comprises the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:10, soak 30min, ultrasonic 50min under the power of 450W, then under identical power, 60s are heated at 70 DEG C, interval 120s is again in 70 DEG C of heating 60s, interval 120s again, so repeat common 5min, filter, filtrate is concentrated into 1.5 times of Japanese Ardisia Herb weight, adding ethanol, to make ethanol content reach volumetric concentration be 80%, 12h is left standstill under 4 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 0.5%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into 1.5 times of Japanese Ardisia Herb polysaccharide crude weight, adding ethanol, to make ethanol content reach volumetric concentration be 80%, 12h is left standstill under 4 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide.
Embodiment 2
The preparation method of the Japanese Ardisia Herb polysaccharide of the present embodiment, comprises the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:15, soak 35min, ultrasonic 30min under the power of 300W, then under identical power, 55s are heated at 80 DEG C, interval 130s is again in 80 DEG C of heating 55s, interval 130s again, so repeat common 50min, filter, filtrate is concentrated into 2 times of Japanese Ardisia Herb weight, adding ethanol, to make ethanol content reach volumetric concentration be 60%, 16h is left standstill under 5 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 1%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into 2 times of Japanese Ardisia Herb polysaccharide crude weight, adding ethanol, to make ethanol content reach volumetric concentration be 60%, 16h is left standstill under 5 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide.
Embodiment 3
The preparation method of the Japanese Ardisia Herb polysaccharide of the present embodiment, comprises the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:20, soak 25min, ultrasonic 90min under the power of 600W, then under identical power, 65s are heated at 75 DEG C, interval 110s is again in 75 DEG C of heating 65s, interval 110s again, so repeat common 30min, filter, filtrate is concentrated into 1.8 times of Japanese Ardisia Herb weight, adding ethanol, to make ethanol content reach volumetric concentration be 70%, 14h is left standstill under 6 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 0.8%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into 1.8 times of Japanese Ardisia Herb polysaccharide crude weight, adding ethanol, to make ethanol content reach volumetric concentration be 70%, 14h is left standstill under 6 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide.
Japanese Ardisia Herb polysaccharide prepared by aforesaid method can be used for the preparation of medicine, and especially for preparation antifatigue and the immunizing power medicine improving body, and obtain sufficient proof through test, related tests data is as follows:
Assay
Adopt phend-sulphuric acid to identify the Japanese Ardisia Herb polysaccharide component that embodiment 1 prepares, result is positive, and polysaccharide content is 89.76%.
The experiment of Japanese Ardisia Herb polysaccharide antifatigue effect
Get the Kunming mouse 40 that body weight is 20 ± 2g, be divided into 4 groups at random by body weight: Normal group, low (200mg/kg), in (500mg/kg), high dosage (800mg/kg) administration group; Every day gastric infusion once, Normal group is with the physiological saline gavage of equivalent, successive administration 30 days, 30min after last administration, be put in by mouse in swimming trunk, water temperature is 25 ± 5 DEG C, and bear a heavy burden the sheet lead of 5% body weight in mouse root of the tail portion, record mouse 8s in swimming starts to submerge water to head does not go out the time of the water surface, i.e. the walking weight load of mouse.Mouse is after swimming terminates to recover 24h, often organize mouse respectively eye socket get blood, separation of serum, with superoxide-dismutase (SOD) measure test kit, mda (MDA) measures test kit and measures SOD in mice serum respectively, MDA value, in table 1.
The impact of SOD and MDA in table 1 Japanese Ardisia Herb Polysaccharides on Mice walking weight load, serum
* P<0.05 compared with blank group is represented
As can be seen from Table 1, Japanese Ardisia Herb polysaccharide is low, in, high dosage all can significantly improve body SOD vigor, reduces MDA content, thus shows Japanese Ardisia Herb polysaccharide and have good antifatigue effect.
The immunization experiment of Japanese Ardisia Herb polysaccharide:
Get the Kunming mouse 40 that body weight is 20 ± 2g, be divided into 4 groups at random by body weight: Normal group, low (40mg/kg), in (60mg/kg, high dosage (80mg/kg) administration group, every day gastric infusion once, Normal group is with the physiological saline gavage of equivalent, successive administration 7 days, after last administration 24h, mouse is pulled cervical vertebra to put to death, take out liver, spleen, lymphoglandula is weighed, and calculates the ratio that each immune organ weight accounts for Mouse Weight, observe the change of immune organ weight, the results are shown in Table 2.
The impact of each immune organ weight of table 2 Japanese Ardisia Herb Polysaccharides on Mice
* P<0.05 compared with blank group is represented
As can be seen from Table 2, Japanese Ardisia Herb polysaccharide is low, in, high dosage all can change the quality of each immune organ of mouse, thus shows Japanese Ardisia Herb polysaccharide and have and strengthen immunization preferably.
Above-mentioned each experimental result clearly shows, Japanese Ardisia Herb polysaccharide of the present invention can effective antifatigue and improve the immunizing power of body.
Claims (6)
1. a preparation method for Japanese Ardisia Herb polysaccharide, is characterized in that, comprises the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:10-20, soak 25-35min, ultrasonic 30-90min under the power of 300-600W, then microwave intermittent-heating 5-50min under identical power, filter, filtrate is concentrated into the 1.5-2 of Japanese Ardisia Herb weight doubly, adding ethanol, to make ethanol content reach volumetric concentration be 60-80%, >=12h is left standstill under 4-6 DEG C of condition, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 0.5-1%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into the 1.5-2 of Japanese Ardisia Herb polysaccharide crude weight doubly, adding ethanol, to make ethanol content reach volumetric concentration be 60-80%, >=12h is left standstill under 4-6 DEG C of condition, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide; In described step (1), microwave intermittent-heating is that 70-80 DEG C of heating 55-65s, interval 110-130s heat 55-65s in 70-80 DEG C again, then interval 110-130s, so repeats.
2. the preparation method of Japanese Ardisia Herb polysaccharide according to claim 1, is characterized in that, Japanese Ardisia Herb is the dregs of a decoction after Japanese Ardisia Herb alcohol extracting.
3. the preparation method of Japanese Ardisia Herb polysaccharide according to claim 1, is characterized in that, comprise the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:10, soak 30min, ultrasonic 50min under the power of 450W, then under identical power, 60s are heated at 70 DEG C, interval 120s is again in 70 DEG C of heating 60s, interval 120s again, so repeat common 5min, filter, filtrate is concentrated into 1.5 times of Japanese Ardisia Herb weight, adding ethanol, to make ethanol content reach volumetric concentration be 80%, 12h is left standstill under 4 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 0.5%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into 1.5 times of Japanese Ardisia Herb polysaccharide crude weight, adding ethanol, to make ethanol content reach volumetric concentration be 80%, 12h is left standstill under 4 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide.
4. the preparation method of Japanese Ardisia Herb polysaccharide according to claim 1, is characterized in that, comprise the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:15, soak 35min, ultrasonic 30min under the power of 300W, then under identical power, 55s are heated at 80 DEG C, interval 130s is again in 80 DEG C of heating 55s, interval 130s again, so repeat common 50min, filter, filtrate is concentrated into 2 times of Japanese Ardisia Herb weight, adding ethanol, to make ethanol content reach volumetric concentration be 60%, 16h is left standstill under 5 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 1%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into 2 times of Japanese Ardisia Herb polysaccharide crude weight, adding ethanol, to make ethanol content reach volumetric concentration be 60%, 16h is left standstill under 5 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide.
5. the preparation method of Japanese Ardisia Herb polysaccharide according to claim 1, is characterized in that, comprise the following steps:
(1) get Japanese Ardisia Herb and dry, pulverize powdered, take Japanese Ardisia Herb, distilled water is added in Japanese Ardisia Herb, the weight ratio of Japanese Ardisia Herb and distilled water is 1:20, soak 25min, ultrasonic 90min under the power of 600W, then under identical power, 65s are heated at 75 DEG C, interval 110s is again in 75 DEG C of heating 65s, interval 110s again, so repeat common 30min, filter, filtrate is concentrated into 1.8 times of Japanese Ardisia Herb weight, adding ethanol, to make ethanol content reach volumetric concentration be 70%, 14h is left standstill under 6 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, the moisture in precipitation is washed away with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, the acetone in precipitation is finally washed away with anhydrous diethyl ether, drying under reduced pressure, obtain Japanese Ardisia Herb polysaccharide crude,
(2) the Japanese Ardisia Herb polysaccharide crude of step (1) is added in distilled water dissolve, be mixed with the polysaccharide soln A that Japanese Ardisia Herb polysaccharide crude mass concentration is 0.8%, with the albumen in Sevag method removing polysaccharide soln A, obtain polysaccharide soln B; Polysaccharide soln B is concentrated into 1.8 times of Japanese Ardisia Herb polysaccharide crude weight, adding ethanol, to make ethanol content reach volumetric concentration be 70%, 14h is left standstill under 6 DEG C of conditions, centrifugal segregation supernatant liquor, collecting precipitation, washes away the moisture in precipitation with dehydrated alcohol, the dehydrated alcohol in precipitation is washed away again with acetone, finally wash away the acetone in precipitation with anhydrous diethyl ether, drying under reduced pressure, obtains Japanese Ardisia Herb polysaccharide.
6. the application of the Japanese Ardisia Herb polysaccharide that as described in any one of claim 1-5 prepared by preparation method in preparation antifatigue and raising immunizing power medicine.
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