WO2022111331A1 - 一种灵芝孢子提取物及其应用 - Google Patents
一种灵芝孢子提取物及其应用 Download PDFInfo
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- WO2022111331A1 WO2022111331A1 PCT/CN2021/130818 CN2021130818W WO2022111331A1 WO 2022111331 A1 WO2022111331 A1 WO 2022111331A1 CN 2021130818 W CN2021130818 W CN 2021130818W WO 2022111331 A1 WO2022111331 A1 WO 2022111331A1
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- Prior art keywords
- ganoderma lucidum
- lucidum spore
- extract
- extraction
- ethanol
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Links
- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 53
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 53
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 44
- 239000000843 powder Substances 0.000 claims abstract description 38
- 239000000706 filtrate Substances 0.000 claims abstract description 15
- 239000006228 supernatant Substances 0.000 claims abstract description 15
- 238000001556 precipitation Methods 0.000 claims abstract description 9
- 239000000341 volatile oil Substances 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims abstract description 8
- 238000003809 water extraction Methods 0.000 claims abstract description 5
- 238000000605 extraction Methods 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000012141 concentrate Substances 0.000 claims description 9
- 238000005238 degreasing Methods 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 6
- 230000029087 digestion Effects 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims 1
- 238000010298 pulverizing process Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 150000004676 glycans Chemical class 0.000 abstract description 6
- 239000005017 polysaccharide Substances 0.000 abstract description 6
- 229920001282 polysaccharide Polymers 0.000 abstract description 6
- 238000001914 filtration Methods 0.000 abstract description 5
- 239000003921 oil Substances 0.000 abstract description 4
- 239000000047 product Substances 0.000 abstract description 3
- 230000037396 body weight Effects 0.000 description 7
- 241000699670 Mus sp. Species 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 235000013305 food Nutrition 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000222336 Ganoderma Species 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 235000012631 food intake Nutrition 0.000 description 4
- 230000037406 food intake Effects 0.000 description 4
- 210000000813 small intestine Anatomy 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- -1 compound diphenoxylate Chemical class 0.000 description 3
- 229960004192 diphenoxylate Drugs 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000003304 gavage Methods 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 210000001187 pylorus Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 235000019786 weight gain Nutrition 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000222341 Polyporaceae Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007661 gastrointestinal function Effects 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Definitions
- the invention belongs to the field of biotechnology, in particular to a Ganoderma lucidum spore extract and its application.
- Ganoderma lucidum is the fruiting body of Polyporaceae fungi Ganoderma lucidum and purple zhi. Sweet in taste and flat in nature, it functions to nourish qi and blood, reassure the mind, and invigorate the spleen and stomach. It was first recorded in "Shen Nong's Materia Medica" and listed as top grade. It is said to be beneficial to the heart, nourish the middle, increase wisdom and not forget, protect the spirit, benefit the essence, benefit the joints, and strengthen the muscles and bones. Jiufu has many functions such as lightening the body and prolonging life. The main diseases recorded in the ancient Chinese pharmacopoeia are all diseases of physical exhaustion and fatigue, indicating that it is their specialty to strengthen the body.
- Ganoderma lucidum is included in the 2000 edition of the Pharmacopoeia of the People's Republic of China and subsequent editions of the Pharmacopoeia. Modern pharmacology also recognizes its role in improving the body's non-specific immunity and enhancing the body's ability to resist disease.
- Ganoderma lucidum spore powder is the spores ejected from the cap when Ganoderma lucidum matures. With the advancement of modern technology, there is a new understanding of Ganoderma lucidum spores. It concentrates the essence of Ganoderma lucidum, the righteousness of heaven and earth, and its medicinal efficacy and clinical application effect have been significantly improved.
- Ganoderma lucidum spore powder including broken wall Ganoderma lucidum spore powder, has been included in the processing specifications of traditional Chinese medicine in Zhejiangzhou, Anhui province and other places. It is also widely used in the field of health food. It has been included in the list of raw materials for health food filing, and its safety and immunity-enhancing effects are also widely recognized.
- the recognized active ingredients of Ganoderma lucidum spore powder are polysaccharides and triterpenes.
- the utilization of its volatile oils and polysaccharides has been relatively mature, and other substances are usually discarded as waste residues.
- the invention fully utilizes the remaining products of the production of Ganoderma lucidum spore oil and Ganoderma lucidum spore polysaccharide, and comprehensively develops the Ganoderma lucidum spore powder, which conforms to the concept of green environmental protection, and at the same time significantly reduces the production cost of the enterprise.
- the present invention provides a Ganoderma lucidum spore extract that fully utilizes Ganoderma lucidum spore powder in all directions.
- Another object of the present invention is to provide the application of the Ganoderma lucidum spore extract in promoting digestion.
- the Ganoderma lucidum spore extract is obtained by the following method: the Ganoderma lucidum spore powder is broken and crushed, defatted to obtain the defatted spore powder and volatile oil, the defatted spore powder is extracted with water, filtered, and combined After the filtrate is concentrated, ethanol is added to stand for precipitation, and the supernatant is obtained by filtration. The supernatant is recovered with ethanol and then concentrated to a density of 1.1-1.2 to obtain an extract.
- the above-mentioned degreasing is extracted by supercritical CO 2 , and the extraction conditions are: pressure 30MPa, flow rate 350-400L/h, temperature 40-45°C, and time 4-5h.
- the above water extraction is performed for 1 to 2 times, each time adding 8 to 10 times the amount of water, the extraction time for each time is 6 to 10 hours, and the extraction temperature is 70 to 80°C.
- the above-mentioned adding ethanol to stand for precipitation is to concentrate the filtrate to a density of 1.10 ⁇ 1.12, control the temperature not to exceed 80 ° C, slowly add 2 times of the concentrated solution by weight concentration of 95% ethanol while stirring, and leave standstill at 4 ° C overnight after adding.
- the preparation method of the above-mentioned Ganoderma lucidum spore extract comprises the following steps: breaking the wall of Ganoderma lucidum spore powder, degreasing to obtain degreasing spore powder and volatile oil, extracting the degreasing spore powder with water, filtering, combining filtrates, and concentrating the filtrate After that, ethanol is added to stand for precipitation, and the supernatant is obtained by filtration. The supernatant is recovered with ethanol and then concentrated to a density of 1.1-12 to obtain an extract.
- the degreasing adopts supercritical CO2 extraction, and the extraction conditions are: pressure 30MPa, flow rate 350-400L/h, temperature 40-45°C, and time 4-5h.
- the water extraction is performed for 1 to 2 times, 8 to 10 times the amount of water is added each time, the extraction time is 6 to 10 hours each time, and the extraction temperature is 70 to 80°C.
- the described adding ethanol to stand for precipitation is to concentrate the filtrate to a density of 1.10-1.12, control the temperature not to exceed 80 ° C, slowly add 2 times the ethanol of the concentrated solution with a weight concentration of 95% while stirring, and leave it to stand at 4 ° C after adding overnight.
- the Ganoderma lucidum spore extract prepared by the method of the present invention has remarkable effect in the preparation of medicines for promoting digestion.
- the present invention re-enrichs the substances after extracting Ganoderma lucidum spore oil and polysaccharide, and finds that the Ganoderma lucidum spore extract has obvious effects on regulating gastrointestinal function.
- the process used is to make full use of the remaining products from the production of Ganoderma lucidum spore oil and Ganoderma lucidum spore polysaccharide, which is in line with the concept of green environmental protection, and at the same time significantly reduces the production cost of the enterprise.
- the Ganoderma lucidum spore powder is broken and pulverized, it is granulated and extracted with supercritical CO 2 .
- the supercritical CO 2 extraction conditions are: pressure 30MPa, flow rate 400L/h, temperature 40°C, time 4h, to obtain degreasing spore powder and volatile oil,
- the defatted spore powder was extracted twice with water, the first time was with 10 times the amount of water, stirred at 80 °C for 10 hours, the second time was with 8 times the amount of water, stirred at 80 °C for 6 hours, filtered, and the filtrate was combined and concentrated to density in vacuo 1.10, control the temperature not to exceed 80°C, slowly add 2 times of ethanol with a concentration of 95% by weight of the concentrate, stir while adding, let stand at 4°C overnight after adding, separate the precipitate and supernatant, and recover the supernatant After ethanol concentration to a relative density of 1.2, an extract was obtained.
- the total sugar content of the extract was 120
- the extraction conditions of supercritical CO 2 are: pressure 30MPa, flow rate 350L/h, temperature 45°C, time 5h, to obtain degreased spore powder and volatile oil , the defatted spore powder was extracted twice with water, the first time was 10 times the amount of water, and the 80 °C stirring was used for 10 hours, and the second time was 8 times the amount of water, which was stirred and extracted at 80 °C for 6 hours, filtered, and the filtrate was combined, Concentrate in vacuo to a density of 1.10, control the temperature not to exceed 80 °C, slowly add 2 times of the concentrated solution with 95% ethanol by weight, stir while adding, and leave it to stand at 4 °C overnight after adding, to separate the precipitate and the supernatant. The supernatant was recovered with ethanol and then concentrated to a relative density of 1.1 to obtain an extract.
- mice were randomly divided into blank control group, model control group, sample group (prepared according to the method of Example 1), wall-broken Ganoderma lucidum spore powder group and defatted Ganoderma lucidum spore powder group (after the Ganoderma lucidum spore powder was broken and pulverized, Granulation, using supercritical CO2 extraction, supercritical CO2 extraction conditions: pressure 30MPa, flow rate 400L/h, temperature 40°C, time 4h, discard the volatile oil, and take the defatted spore powder).
- the sample group was given 10 times the recommended dose for adults, that is, 0.5g/kg ⁇ bw, by gavage.
- the blank control group and the model control group were given distilled water, and at the same time, the dose of 2.0g/kg ⁇ bw was given by gavage to break the wall.
- Ganoderma lucidum spore powder and defatted Ganoderma lucidum spore powder once a day for 30 days.
- the sample group, model, blank control group, broken ganoderma spore powder group and defatted ganoderma spore powder group were given the test sample or distilled water again on the day of measurement. After 30 minutes, each sample group and model The control group was given compound diphenoxylate, the blank control group was given distilled water, and each group was given ink indicator after 30 min. After 25 min, the animals were sacrificed by cervical dislocation. The intestinal tube, placed on a flat plate, was gently drawn into a straight line, and the distances from the pylorus to the ileocecal and from the pylorus to the front of the ink were measured respectively, and the ink propulsion rate was calculated:
- ** means comparison with blank control group (P ⁇ 0.01)
- # means comparison with model control group (P ⁇ 0.01).
- Table 1 shows that the small intestine advancement rate of the model control group was significantly lower than that of the blank control (P ⁇ 0.01), indicating that the model was successfully established.
- the small intestine ink propulsion rate of the mice in the sample group was significantly higher than that in the compound diphenoxylate model control group (P ⁇ 0.01). It indicated that Ganoderma lucidum spore extract could improve the reduction of intestinal propulsion caused by compound diphenoxylate.
- the sample group was given 0.5g/kg ⁇ bw
- the ganoderma spore powder group and the defatted ganoderma spore powder group were administered by gavage at a dose of 2.0g/kg ⁇ bw for 30 consecutive days.
- the body weight and food intake of mice were measured twice a week, and the sample amount was adjusted according to changes in body weight. Animal weight gain, food intake and food utilization were calculated at the end of the experiment.
- the experimental results are shown in Table 2.
- Table 2 shows that the total food intake of the rats in the sample group was significantly higher than that in the control group (P ⁇ 0.05), and there was no significant difference in other indicators compared with the control group (P>0.05).
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Abstract
本发明公开了一种灵芝孢子提取物及其应用，灵芝孢子提取物是通过以下方法制备得到的：将灵芝孢子粉破壁粉碎、脱脂后得到脱脂后的孢子粉和挥发油，将脱脂后的孢子粉水提、过滤，合并滤液，滤液浓缩后加乙醇静置沉淀，过滤得到上清液，将上清液回收乙醇后浓缩至相对密度为1.1-1.2，得到提取物。本发明将生产灵芝孢子油、灵芝孢子多糖所余剩产物充分利用，对灵芝孢子粉进行全方位开发，符合绿色环保理念，同时显著降低了企业的生产成本。
Description
本发明属于生物技术领域，具体涉及一种灵芝孢子提取物及其应用。
灵芝是多孔菌科真菌赤芝、紫芝的子实体。味甘性平，功能益气血、安心神、健脾胃。始载于《神农本草经》，列为上品。称其有益心气、补中、增智慧不忘、保神、益精气、利关节、坚筋骨。久服轻身延年不老等多种作用。历代中药典籍所收载主治病症均属体虚劳损诸疾，表明以补虚强体为其特长。2000版《中华人民共和国药典》及之后历版药典均收载了灵芝。现代药理学也认知其提高机体非特异性免疫力，增强机体抗病能力的作用。
灵芝孢子粉是灵芝成熟时从菌盖弹射的孢子。随着现代科技手段的进步，对灵芝孢子有了新的认识。它浓缩了灵芝的精华、禀天地之正气，其药效及临床应用效果有显著提高。灵芝孢子粉包括破壁灵芝孢子粉均已被列入浙江省、安徽省等多地中药饮片炮制规范，性平味甘、具有补气安神健脾益肺等多种功效。在保健食品领域中也被广泛应用。已被纳入保健食品备案原料目录，其安全性和增强免疫力功效也被广泛认可。
灵芝孢子粉目前比较公认的有效成分是多糖类和三萜类，对其挥发油及多糖的利用已经比较成熟，其他的物质通常作为废渣弃去。本发明将生产灵芝孢子油、灵芝孢子多糖所余剩产物充分利用，对灵芝孢子粉进行全方位开发，符合绿色环保理念，同时显著降低了企业的生产成本。
发明内容
本发明针对上述不足之处提供一种对灵芝孢子粉进行全方位充分利用的灵芝孢子提取物。
本发明的另一目的是提供该灵芝孢子提取物在促消化方面的应用。
本发明的目的是通过以下方式实现的：
一种灵芝孢子提取物，该灵芝孢子提取物是通过以下方法得到的：将灵芝孢子粉破壁 粉碎、脱脂后得到脱脂后的孢子粉和挥发油，将脱脂后的孢子粉水提、过滤，合并滤液，滤液浓缩后加乙醇静置沉淀，过滤得到上清液，将上清液回收乙醇后浓缩至密度为1.1～1.2，得到提取物。
上述脱脂是采用超临界CO
2萃取，萃取条件为：压力30MPa、流速350～400L/h，温度40～45℃，时间4～5h。
上述水提取为提取1～2次，每次加水8～10倍量，每次提取时间为6～10小时，提取温度为70～80℃。
上述加乙醇静置沉淀为将滤液浓缩至密度1.10～1.12，控温不超过80℃，缓慢加入浓缩液2倍重量浓度为95％的乙醇边加边搅拌，加完后静置4℃过夜。
上述灵芝孢子提取物的制备方法，该方法包括以下步骤：将灵芝孢子粉破壁粉碎、脱脂后得到脱脂后的孢子粉和挥发油，将脱脂后的孢子粉水提、过滤，合并滤液，滤液浓缩后加乙醇静置沉淀，过滤得到上清液，将上清液回收乙醇后浓缩至密度为1.1～12，得到提取物。所述的脱脂是采用超临界CO2萃取，萃取条件为：压力30MPa、流速350～400L/h，温度40～45℃，时间4～5h。所述的水提取为提取1～2次，每次加水8～10倍量，每次提取时间为6～10小时，提取温度为70～80℃。所述的加乙醇静置沉淀为将滤液浓缩至密度1.10～1.12，控温不超过80℃，缓慢加入浓缩液2倍重量浓度为95％的乙醇边加边搅拌，加完后静置4℃过夜。
本发明方法制备得到的灵芝孢子提取物应用于制备促消化的药物具有显著的效果。
本发明针对提取灵芝孢子油和多糖后的物质进行重新富集，发现该灵芝孢子提取物对调节肠胃功能有明显作用。所用工艺系将生产灵芝孢子油、灵芝孢子多糖所余剩产物充分利用，符合绿色环保理念，同时显著降低了企业的生产成本。
以下通过具体实施例对本发明进行进一步解释说明
实施例1
将灵芝孢子粉破壁粉碎后，制粒，采用超临界CO
2萃取，超临界CO
2萃取条件为：压力30MPa、流速400L/h，温度40℃，时间4h，得到脱脂孢子粉和挥发油，将脱脂孢子粉采用水提取2次，第一次采用10倍量水，80℃搅拌提取10小时，第二次采用8倍量水，80℃搅拌提取6小时，过滤，合并滤液，真空浓缩至密度1.10，控温不超过80℃，缓慢加入浓缩液2倍重量浓度为95％的乙醇，边加边搅拌，加完后静置4℃过夜，分离得到沉淀 物和上清液，上清液回收乙醇后浓缩至相对密度为1.2，得到提取物。检测该提取物总糖含量为120mg/ml，蛋白质含量为62mg/ml。
总糖含量的检测方法：GB/T 15672-2009食用菌中总糖含量的测定
蛋白质含量的检测方法：GB 5009.5-2016食品中蛋白质的测定
实施例2
将灵芝孢子粉破壁粉碎后，制粒，采用超临界CO
2萃取，超临界CO
2萃取条件为：压力30MPa、流速350L/h，温度45℃，时间5h，得到脱脂后的孢子粉和挥发油，将脱脂后的孢子粉采用水提取2次，第一次采用10倍量水，80℃搅拌提取10小时，第二次采用8倍量水，80℃搅拌提取6小时，过滤，合并滤液，真空浓缩至密度1.10，控温不超过80℃，缓慢加入浓缩液2倍重量浓度为95％的乙醇，边加边搅拌，加完后静置4℃过夜，分离得到沉淀物和上清液，上清液回收乙醇后浓缩至相对密度为1.1，得到提取物。检测该提取物总糖含量为126mg/ml，蛋白质含量为64mg/ml。
总糖含量的检测方法：GB/T 15672-2009食用菌中总糖含量的测定
蛋白质含量的检测方法：GB 5009.5-2016食品中蛋白质的测定
以下通过试验例对本发明效果进行进一步说明：
试验例1
一、小鼠小肠墨汁推进试验:
小鼠按体重大小随机分为空白对照组、模型对照组、样品组(按照实施例1方法制备得到)、破壁灵芝孢子粉组和脱脂灵芝孢子粉组(将灵芝孢子粉破壁粉碎后，制粒，采用超临界CO
2萃取，超临界CO
2萃取条件为：压力30MPa、流速400L/h，温度40℃，时间4h，弃挥发油，取脱脂孢子粉)。样品组按成人推荐量的10倍，即0.5g/kg·bw，以灌胃方式给药，空白对照组和模型对照组给蒸馏水，同时以2.0g/kg·bw剂量灌胃给药破壁灵芝孢子粉和脱脂灵芝孢子粉，每天1次，连续30d。
实验结束前禁食不禁水16h，于测定当天样品组、模型、空白对照组、破壁灵芝孢子粉组和脱脂灵芝孢子粉组再给予一次受试样品或蒸馏水，30min后各样品组和模型对照组灌复方地芬诺酯，空白对照组灌予蒸馏水，30min后各组再给予墨汁指示剂，25min后断颈处死动物，打开腹腔分离肠系膜，剪取上端自幽门、下端至回盲部的肠管，置于平板上，轻轻拉成直线，分别测量从幽门到回盲部和从幽门到墨汁前沿的距离，计算墨汁推进率:
结果见表1：
表1 对小鼠小肠推进运动的影响
注：**表示与空白对照组比较(P<0.01)，#表示与模型对照组比较(P<0.01)。
表1显示，模型对照组小肠推进率明显低于空白对照(P<0.01)，说明模型建立成功。样品组小鼠小肠墨汁推进率明显高于复方地芬诺酯模型对照组(P<0.01)。说明灵芝孢子提取物对复方地芬诺酯引起的小肠推进降低有改善作用。
二、动物体重、体重增重、摄食量和食物利用率的测定试验：
实验期间，样品组按0.5g/kg·bw、灵芝孢子粉组和脱脂灵芝孢子粉组按2.0g/kg·bw剂量以灌胃方式给药，连续30天。小鼠每周测2次体重和食物摄入量，并根据体重变化调整给样量。实验结束时计算动物增重、摄食量和食物利用率。实验结果见表2。
表2 实验前后小鼠体重的变化情况
注：*表示与空白对照组比较P<0.05，**表示P<0.01。
由表2可见，样品组小鼠初始体重与空白对照组比较，无显著性差异，给样30d后样品组小鼠体重增重均明显高于空白对照组。
表3 大鼠体重及食物利用率的变化
注：*表示与对照组比较P<0.05，**表示P<0.01。
表2显示,样品组大鼠总摄食量明显高于对照组(P<0.05)，其余指标与对照组相比无显著性差异(P>0.05)。
Claims (9)
- 一种灵芝孢子提取物，其特征在于：将灵芝孢子粉破壁粉碎、脱脂后得到脱脂后的孢子粉和挥发油，将脱脂后的孢子粉水提、过滤，合并滤液，滤液浓缩后加乙醇静置沉淀，过滤得到上清液，将上清液回收乙醇后浓缩至相对密度为1.1-1.2，得到提取物。
- 根据权利要求1所述的灵芝孢子提取物，其特征在于：所述的脱脂是采用超临界CO 2萃取，萃取条件为：压力30MPa、流速350～400L/h，温度40～45℃，时间4～5h。
- 根据权利要求1所述的灵芝孢子提取物，其特征在于：所述的水提取为提取1～2次，每次加水8～10倍量，每次提取时间为6～10小时，提取温度为70～80℃。
- 根据权利要求1所述的灵芝孢子提取物，其特征在于加乙醇静置沉淀为将滤液浓缩至密度1.10～1.12，控温不超过80℃，缓慢加入浓缩液2倍重量浓度为95％的乙醇边加边搅拌，加完后静置4℃过夜。
- 一种权利要求1所述的灵芝孢子提取物的制备方法，其特征在于该方法包括以下步骤：将灵芝孢子粉破壁粉碎、脱脂后得到脱脂后的孢子粉和挥发油，将脱脂后的孢子粉水提、过滤，合并滤液，滤液浓缩后加乙醇静置沉淀，过滤得到上清液，将上清液回收乙醇后浓缩至密度为1.1～12，得到提取物。
- 根据权利要求5所述的灵芝孢子提取物的制备方法，其特征在于所述的脱脂是采用超临界CO2萃取，萃取条件为：压力30MPa、流速350～400L/h，温度40～45℃，时间4～5h。
- 根据权利要求5所述的灵芝孢子提取物的制备方法，其特征在于所述的水提取为提取1～2次，每次加水8～10倍量，每次提取时间为6～10小时，提取温度为70～80℃。
- 根据权利要求5所述的灵芝孢子提取物的制备方法，其特征在于所述的加乙醇静置沉淀为将滤液浓缩至密度1.10～1.12，控温不超过80℃，缓慢加入浓缩液2倍重量浓度为95％的乙醇边加边搅拌，加完后静置4℃过夜。
- 权利要求1所述的灵芝孢子提取物在制备促消化的药物中的应用。
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