CN105085696A - Method for extracting Anoectochilus roxburghii polysaccharides - Google Patents
Method for extracting Anoectochilus roxburghii polysaccharides Download PDFInfo
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Abstract
The invention discloses a method for extracting Anoectochilus roxburghii polysaccharides. The method comprises the steps: (1) cleanly washing Anoectochilus roxburghii herbs, then, baking the Anoectochilus roxburghii herbs at the temperature of 40-60 DEG C, then, crushing and sieving the baked Anoectochilus roxburghii herbs, soaking the sieved Anoectochilus roxburghii herbs with 80-95% ethanol according to the ratio (v: w) of (8-12): 1 for 10-15 hours, carrying out filtration, carrying out repeated treatment for 2-3 times, concentrating the filtrate for later use, and baking filtration residue so as to obtain pretreated powder; and (2) weighing the pretreated powder, adding distilled water into the weighed pretreated powder according to the liquid/material ratio of 1: (25-35), carrying out water bath for 60-90 minutes at the temperature of 60-70 DEG C, carrying out suction filtration, carrying out repeated extraction for 3-5 times, and merging the filtrates, thereby obtaining a polysaccharide extract. Under the condition of the extraction process disclosed by the invention, the extraction yield of the Anoectochilus roxburghii polysaccharides is 26.68%. By applying the process disclosed by the invention, the polysaccharides are high in yield and simple and easy in regulation and control and have production and application values.
Description
Technical field
The present invention relates to a kind of extracting method of anoectochilus roxburghii polyose.
Background technology
Herba Anoectochili roxburghii (Anoectochilusroxburghii (Wall.) Lindl.), also known as Shorthairy Antenoron, gold grass, Herba Anoectochili roxburghii, Herba Colei Pumili, Longhairy Antenoron Herb etc.For the orchid family (Orchidaceae) Anoectochilus Blume (Anoectochilus) per nnial herb
[1].The traditional valuable ingredient of traditional Chinese medicine of China, the good reputation have " king of medicine ", " gold grass " among the people.Its sweet and slightly bitter taste, property is flat to be slightly cold, there is clearing heat and cooling blood, expelling wind and removing dampness, cardiac stimulant, reinforce the kidney, diuresis, flat liver, the effect such as hypotensive, be mainly used in the diseases such as treatment hypertension, diabetes, heart trouble, pneumonia, acute, chronic hepatitis, ephritis
[2].Polysaccharide is one of important activity composition in Herba Anoectochili roxburghii, and research display anoectochilus roxburghii polyose tool is anti-oxidant
[3-4], immunomodulatory
[5-8], protect the liver
[9], anti-glycosuria sugar
[10-13]and antitumor action
[14-15]etc. multiple efficacies.In view of the multiple biological activity of anoectochilus roxburghii polyose tool, it serves to show the important Development volue of its tool.Therefore, the High-efficient Production of anoectochilus roxburghii polyose seems particularly important.
Extraction method of polysaccharides has multiple, and Hot water extraction is wherein the traditional technology of Polyose extraction, this method mild condition, survivable polysaccharide original structure and biological activity, and this technique is easy to control, and is suitable for large-scale industrialized production.At present, extract technology optimization report less about anoectochilus roxburghii polyose, the Polyose extraction yield of the research display Herba Anoectochili roxburghii of forefathers is 1.73%-8.23%
[16-18].
[1] Chinese Academy of Sciences's Chinese Plants will editorial board. Chinese Plants will (the 17th volume) [M]. Beijing: Science Press, 2006.220.
[2] ZhangYH, CaiJY, RuanHL, etal.Antihyperglycemicactivityofkinsenoside, ahighyieldingconstituentfromAnoectochilusroxburghiiinstr eptozotocindiabeticrats [J] .JournalofEthnopharmacology, 2007,114:141-145. // Fujian Chinese medicine study compiled. Fujian medicine will. and Foochow: Fujian Science Press .1982:215-217.
[3] Lin Liqing, Huang Liying, Zheng Yanjie, etc. the extraction of anoectochilus roxburghii polyose and the research [J] of scavenging activated oxygen effect. Colleges Of Traditional Chinese Medicine Of Fujian's journal, 2006,16 (5): 37-39.
[4] Liu Qing, Liu Zhenling, Zhou Juan. the antioxidation activity in vitro [J] of anoectochilus roxburghii polyose. Huaqiao University's journal (natural science edition), 2010,31 (6): 718-720.
[5] Jiang Jianxing. the composition of Herba Anoectochili roxburghii water-soluble polysaccharide differentiation and analysis of immunogenicity [D]. the Taibei: Univ Nat Taiwan, 2003.
[6] Wu Yizhen. anoectochilus roxburghii polyose is to the research [D] of mouse immune reaction adjustment. the Taibei: Taipei Medical University, 2005.
[7] Liao Siying. tumor activities of Anoectochilus formosanus polysaccharide Body components is on the impact [D] of innate immune response. in platform: China Medical University, 2005.
[8] Xu Danni, Li Jian, Ma Yufang. anoectochilus roxburghii polyose is on the impact [J] of immunosuppressed mice spleen lymphocyte proliferation and immune organ. Chinese veterinarian's medical magazine, 2011 (3): 15-17.
[9] Zhang Xiaohui. anoectochilus roxburghii polyose structural analysis and anti-diabetic activity research [D]. Shantou: University Of Shantou, 2008.
[10] Cui Shichao. the extraction of Herba Anoectochili roxburghii active substance and Study on Physiological Activity [D]. Shantou: University Of Shantou, 2010.
[11] Yu Jie, Li Wei, Zhang Xiaohui. the research [J] of anoectochilus roxburghii polyose antidiabetic effect. Strait Pharmaceutical Journal, 2013,25 (11): 37-40
[12] Deng Yanqun, Li Wei, Zhang Xiaohui, etc. the impact [J] on alpha-glucosidase activity and blood glucose in diabetic mice of. anoectochilus roxburghii polyose. University Of Shantou's journal (natural science edition), 2014,39 (3): 41-45.
[13]JGZhang,QLiu,ZLLiu,LLi,LTYi.AntihyperglycemicactivityofAnoectochilusroxburghiipolysaccharoseindiabeticmiceinducedbyhigh-fatdietandstreptozotocin[J].JournalofEthnopharmacology,2015,164:180-185.
[14] Wang Changqing, Yan Chengqi, Wang Yong, etc. the separation and purification of tumor activities of Anoectochilus formosanus polysaccharide and external antitumor activity research [J] thereof. Chinese biochemical drug magazine, 2008,29 (2): 93-95
[15] Weng Xiuhua, Wang Changlian, Yuan Xi, etc. Herba Anoectochili roxburghii is on the impact [J] of proliferation of human prostate carcinoma PC-3 cell ability. institute of traditional Chinese medicine's pharmaceutical journal, 2011,31 (13): 1083-1087
[16] Li Dehua, Huang Qiaowen. anoectochilus roxburghii polyose Study on extraction [J]. Strait Pharmaceutical Journal, 2007,19 (6): 26-27.
[17] Zhao Baofa, Zhang Zhixin. the research [J] of anoectochilus roxburghii polyose extraction process. Yunnan Normal University's journal, 2008,28 (4): 60-63.
[18] Yu Jie, Cui Shichao, Zhang Xiaohui, etc. Responds Surface Methodology optimizes the research [J] of Herba Anoectochili roxburghii Polyose extraction process. University Of Shantou's journal (natural science edition), 2009,24 (4): 55-60.
Summary of the invention
The object of the invention is to solve the too low problem of current polysaccharide extract rate, provide a kind of hot water extraction to extract the method for anoectochilus roxburghii polyose.
Technical scheme of the present invention is as follows:
An extracting method for anoectochilus roxburghii polyose, comprises the steps:
1) get Herba Anoectochili roxburghii herb, after cleaning up, 40-60 DEG C of oven dry, pulverizes and sieves, 8-12 times of volume (v:w) 80-95% alcohol immersion 10-15h, filters, and repeatedly process 2-3 time, filtrate concentrates for subsequent use, and filter residue is dried, and is pre-treatment powder;
2) get pre-treatment powder, add distilled water by the liquid ratio of 1:20-40,60-80 DEG C of water-bath 60-90min, suction filtration, repeatedly extract 3-5 time, merging filtrate obtains polysaccharide extraction liquid.
In the preferred embodiment, step 1) in, pulverize and sieve as crossing 20-60 object sieve.
The extracting method of anoectochilus roxburghii polyose of the present invention, its optimum comprises the steps:
(1) get Herba Anoectochili roxburghii herb, cleaning, dry to constant weight for 50 DEG C, pulverize, cross 40 mesh sieves, obtain Herba Anoectochili roxburghii dried powder;
(2) accurately take a certain amount of Herba Anoectochili roxburghii powder, filter, repetitive operation twice with 10 times of volume (v:w) 95% alcohol immersion 12h.Getting filter residue to dry to constant weight in 50 DEG C, is Herba Anoectochili roxburghii pre-treatment powder;
(3) accurately take a certain amount of Herba Anoectochili roxburghii pre-treatment powder, add distilled water by the liquid ratio of 1:30 (g:mL), 70 DEG C of water-bath 90min, suction filtration, repeatedly extract 4 times, merging filtrate obtains polysaccharide extraction liquid.
(4) polysaccharide extraction liquid constant volume, use phend-sulphuric acid and 3 respectively, 5-dinitrosalicylic acid system measures total reducing sugar and reducing sugar content in extracting solution, and polysaccharide content is that total sugar content deducts reducing sugar content, and Polyose extraction yield is the ratio that polysaccharide content accounts for pre-treatment powder weight.
In flooding step of the present invention, 1) extraction time: for 3-5 time.Extraction time is less than 3 times, and Polyose extraction yield is low; But when extraction time is greater than 5 times, polysaccharide extract rate does not increase, on the contrary technical process elongated, increase after the workload to concentrate etc. of thread filtrate and dependent loss, be also unfavorable for producing energy-conservation simultaneously.
2) Extracting temperature: if Extracting temperature is lower than 60 DEG C, extraction efficiency sharply declines, when Extracting temperature is higher than 80 DEG C, polysaccharide may be caused to degrade affects its activity simultaneously; And high temperature also can make impurity stripping quantity increase, for subsequent disposal is made troubles.
3) solid-liquid ratio: if solid-liquid ratio is lower than 1:20, extraction yield declines; If when being greater than 1:40, polysaccharide yield amplification is little, but can increase subsequent concentration workload and dependent loss, is unfavorable for the saving of production water resources simultaneously.
4) extraction time: if extraction time is lower than 60min, polysaccharide can not fully be leached, but the time is more than 90min, and polysaccharide yield amplification is little, but extraction time longly may affect polysaccharide structures and activity under the high temperature conditions, also improves extraction efficiency and energy-conservation consideration in unfavorable production.
Beneficial effect of the present invention is: 1) in the prior art, the Polyose extraction yield of Herba Anoectochili roxburghii is only 1.73%-8.23%, and the present invention is through 40-60 DEG C of oven dry, pulverize and sieve, 8-12 times of volume (v:w) 80-95% alcohol immersion 10-15h, after such pre-treatment, again through specific hot water extraction's condition, comprehensive action, polysaccharide yield is high, extract yield and can reach 26.68%, far exceed the yield of prior art.And hot water extraction's polysaccharide condition of the present invention is comparatively gentle, the structure and activity of survivable polysaccharide.
2) under top condition of the present invention, result difference is less, reproducible, and extraction yield is high, process stabilizing
3) present invention process requires device simple, is easy to control, and is suitable for large-scale industrialized production.
Embodiment
Embodiment 1
(1) get Herba Anoectochili roxburghii herb, after cleaning up, the first drain well in shady and cool ventilation place, 50 DEG C of oven dry, cross 40 mesh sieves, obtain Herba Anoectochili roxburghii powder.
(2) accurately take a certain amount of Herba Anoectochili roxburghii herb powder 50.00g, 10 times of volume (v:w) 95% alcohol immersion 12h, filter, repeatedly process 2 times, merge twice filtrate and concentrate for subsequent use; Filter residue is positioned over shady and cool ventilation place and volatilizes ethanol, then dries in 50 DEG C of baking ovens, is Herba Anoectochili roxburghii pre-treatment powder.
(3) accurately take Herba Anoectochili roxburghii pre-treatment powder 15.00g, add distilled water by the liquid ratio of 1:30 (g:mL), 70 DEG C of water-bath 90min, suction filtration, repeatedly extract 4 times.Merge 4 filtrates and obtain polysaccharide extraction liquid.
(4) by polysaccharide extraction liquid constant volume, use phend-sulphuric acid and 3 respectively, 5-dinitrosalicylic acid (DNS) method measures total reducing sugar and reducing sugar content in extracting solution, and polysaccharide content is that total sugar content deducts reducing sugar content, and polysaccharide extract rate is 26.60%.
Embodiment 2
(1) get Herba Anoectochili roxburghii herb, after cleaning up, the first drain well in shady and cool ventilation place, 50 DEG C of oven dry, cross 40 mesh sieves, obtain Herba Anoectochili roxburghii powder.
(2) accurately take a certain amount of Herba Anoectochili roxburghii herb powder 50.00g, 10 times of volume (v:w) 95% alcohol immersion 12h, filter, repeatedly process 2 times, merge twice filtrate and concentrate for subsequent use; Filter residue is positioned over shady and cool ventilation place and volatilizes ethanol, then dries in 50 DEG C of baking ovens, is Herba Anoectochili roxburghii pre-treatment powder.
(3) accurately take Herba Anoectochili roxburghii pre-treatment powder 20.00g, add distilled water by the liquid ratio of 1:30 (g:mL), 70 DEG C of water-bath 90min, suction filtration, repeatedly extract 4 times.Merge 4 filtrates and obtain polysaccharide extraction liquid.
(4) by polysaccharide extraction liquid constant volume, use phend-sulphuric acid and 3 respectively, 5-dinitrosalicylic acid (DNS) method measures total reducing sugar and reducing sugar content in extracting solution, and polysaccharide content is that total sugar content deducts reducing sugar content, and polysaccharide extract rate is 26.58%.
Embodiment 3
(1) get Herba Anoectochili roxburghii herb, after cleaning up, the first drain well in shady and cool ventilation place, 50 DEG C of oven dry, cross 40 mesh sieves, obtain Herba Anoectochili roxburghii powder.
(2) accurately take a certain amount of Herba Anoectochili roxburghii herb powder 50.00g, 10 times of volume (v:w) 95% alcohol immersion 12h, filter, repeatedly process 2 times, merge twice filtrate and concentrate for subsequent use; Filter residue is positioned over shady and cool ventilation place and volatilizes ethanol, then dries in 50 DEG C of baking ovens, is Herba Anoectochili roxburghii pre-treatment powder.
(3) accurately take Herba Anoectochili roxburghii pre-treatment powder 25.00g, add distilled water by the liquid ratio of 1:30 (g:mL), 70 DEG C of water-bath 90min, suction filtration, repeatedly extract 4 times.Merge 4 filtrates and obtain polysaccharide extraction liquid.
(4) by polysaccharide extraction liquid constant volume, use phend-sulphuric acid and 3 respectively, 5-dinitrosalicylic acid (DNS) method measures total reducing sugar and reducing sugar content in extracting solution, and polysaccharide content is that total sugar content deducts reducing sugar content, and polysaccharide extract rate is 26.70%.
Embodiment 4 repeated experiment
Accurately take certain 3 parts, Herba Anoectochili roxburghii herb dry powder (dry powder preparation is with embodiment above), solid-liquid ratio 1:30, bath temperature 70 DEG C, extraction time 90min, extract the optimum extraction condition of 4 times under, test, the extraction yield of anoectochilus roxburghii polyose is (26.68 ± 0.12) % (n=3) (table 4).Illustrative experiment is respond well, and 3 result differences are less, reproducible, demonstrate this process stabilizing feasible.
Table 4 proof test
Note: in table
adata are " mean value ± SD ".
Claims (3)
1. an extracting method for anoectochilus roxburghii polyose, comprises the steps:
1) get Herba Anoectochili roxburghii herb, after cleaning up, 40-60 DEG C of oven dry, pulverizes and sieves, 8-12 times of volume (v:w) 80-95% alcohol immersion 10-15h, filters, and repeatedly process 2-3 time, filtrate concentrates for subsequent use, and filter residue is dried, and is pre-treatment powder;
2) get pre-treatment powder, add distilled water by the liquid ratio of 1:20-40,60-80 DEG C of water-bath 60-90min, suction filtration, repeatedly extract 3-5 time, merging filtrate obtains polysaccharide extraction liquid.
2. the extracting method of anoectochilus roxburghii polyose as described in claim 1, is characterized in that, step 1) in, pulverize and sieve as crossing 20-60 object sieve.
3. the extracting method of anoectochilus roxburghii polyose as described in claim 1, is characterized in that, comprise the steps:
(1) get Herba Anoectochili roxburghii herb, cleaning, dry to constant weight for 50 DEG C, pulverize, cross 40 mesh sieves, obtain Herba Anoectochili roxburghii dried powder;
(2) accurately take a certain amount of Herba Anoectochili roxburghii powder, filter, repetitive operation twice with 10 times of volume (v:w) 95% alcohol immersion 12h; Getting filter residue to dry to constant weight in 50 DEG C, is Herba Anoectochili roxburghii pre-treatment powder;
(3) accurately take a certain amount of Herba Anoectochili roxburghii pre-treatment powder, add distilled water by the liquid ratio of 1:30 (g:mL), 70 DEG C of water-bath 90min, suction filtration, repeatedly extract 4 times, merging filtrate obtains polysaccharide extraction liquid.
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Cited By (5)
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| CN107337741A (en) * | 2017-06-17 | 2017-11-10 | 福建中医药大学 | A kind of preparation method and application of beautiful flower bud Shorthairy Antenoron polyoses extract |
| CN108456259A (en) * | 2018-05-02 | 2018-08-28 | 福建医科大学 | Application of the anoectochilus roxburghii polyose in acute alcohol-induced hepatic injury drug |
| CN109010609A (en) * | 2018-09-30 | 2018-12-18 | 华中科技大学 | A kind of preparation method of the Anoectochilus containing anoectochilus roxburghii glycosides and the application of extract |
| CN110037331A (en) * | 2019-03-20 | 2019-07-23 | 福建绿色黄金生物科技有限公司 | A kind of processing method and its application of roxburgh anoectochilus terminal bud |
| CN113087816A (en) * | 2021-05-14 | 2021-07-09 | 南昌大学 | Preparation method of high-purity anoectochilus roxburghii polysaccharide |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107337741A (en) * | 2017-06-17 | 2017-11-10 | 福建中医药大学 | A kind of preparation method and application of beautiful flower bud Shorthairy Antenoron polyoses extract |
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| CN109010609A (en) * | 2018-09-30 | 2018-12-18 | 华中科技大学 | A kind of preparation method of the Anoectochilus containing anoectochilus roxburghii glycosides and the application of extract |
| CN110037331A (en) * | 2019-03-20 | 2019-07-23 | 福建绿色黄金生物科技有限公司 | A kind of processing method and its application of roxburgh anoectochilus terminal bud |
| CN113087816A (en) * | 2021-05-14 | 2021-07-09 | 南昌大学 | Preparation method of high-purity anoectochilus roxburghii polysaccharide |
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