CN103613677A - Extraction method of imperatoria polysaccharide - Google Patents
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- 238000000605 extraction Methods 0.000 title claims abstract description 50
- 150000004676 glycans Chemical class 0.000 title claims abstract description 38
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 38
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 38
- 241000510612 Imperatoria Species 0.000 title 1
- 244000025254 Cannabis sativa Species 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000000843 powder Substances 0.000 claims abstract description 18
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000012153 distilled water Substances 0.000 claims abstract description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 8
- 239000000706 filtrate Substances 0.000 claims abstract description 8
- 238000001914 filtration Methods 0.000 claims abstract description 8
- 239000008103 glucose Substances 0.000 claims abstract description 8
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000012086 standard solution Substances 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims abstract description 3
- 239000002244 precipitate Substances 0.000 claims description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 14
- 102000011759 adducin Human genes 0.000 claims description 7
- 108010076723 adducin Proteins 0.000 claims description 7
- 238000005119 centrifugation Methods 0.000 claims description 7
- 239000002994 raw material Substances 0.000 claims description 7
- 238000012360 testing method Methods 0.000 claims description 7
- 239000000047 product Substances 0.000 claims description 2
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000010998 test method Methods 0.000 abstract description 2
- 238000012869 ethanol precipitation Methods 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 3
- 240000004155 Saccharum barberi Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 1
- 241000209046 Pennisetum Species 0.000 description 1
- 244000130556 Pennisetum purpureum Species 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- 244000130503 Saccharum sinense Species 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000009970 fire resistant effect Effects 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 230000000055 hyoplipidemic effect Effects 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 239000004460 silage Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
本发明公开了一种王草多糖的提取方法，提取方法包括王草干燥、粉碎、按一定液料比加入蒸馏水和王草粉末水浴浸提、过滤后将滤液定容，以葡萄糖标准品溶液为对照，采用苯酚硫酸法测定粗多糖的含量，对提取液进行乙醇沉淀、丙酮洗涤处理，最终得到王草多糖粉末。本发明具体的有益效果为：考虑了多种因素对多糖提取的影响，采用正交试验方法对提取过程中提取剂水与王草粉末的比例、提取时间、提取次数和提取温度进行严格控制，整个提取过程操作简单、程序简化、提取准确、重复性好、提高了王草多糖的提取率。The invention discloses a method for extracting king grass polysaccharides. The extraction method includes drying and crushing king grass, adding distilled water and king grass powder water bath extraction according to a certain liquid-to-material ratio, filtering the filtrate to constant volume, and using glucose standard solution as In contrast, the content of crude polysaccharide was determined by the phenol-sulfuric acid method, and the extract was subjected to ethanol precipitation and acetone washing treatment to finally obtain Wangcao polysaccharide powder. The specific beneficial effects of the present invention are: taking into account the influence of various factors on the extraction of polysaccharides, adopting an orthogonal test method to strictly control the ratio of extractant water and Wangcao powder, extraction time, extraction times and extraction temperature during the extraction process, The whole extraction process is simple in operation, simplified in procedure, accurate in extraction and good in repeatability, and improves the extraction rate of Wangcao polysaccharide.
Description
技术领域technical field
本发明涉及以一种王草化学成分的提取方法，具体地讲是涉及一种王草多糖的提取方法。The invention relates to a method for extracting chemical components of Wangcao, in particular to a method for extracting polysaccharides from Wangcao.
背景技术Background technique
王草（拉丁学名Pennisetum purpureum K.Scbumacb、P.typhoideumRich），中文别名皇草（英文别名Chinese sugarcane、japanese cane、kinggrass、north Indian sugarcane、uva cane），为禾本目禾本科狼尾草属植物，是一种优良热带牧草，国内海南、广东、广西广泛栽培，江苏、福建、云南、湖南引种栽培。国外主要分布于中、南美洲。多年生丛生性高杆禾草。根系发达。茎杆形似甘蔗。叶呈长条形。圆锥花序密生呈穗状。颖果纺锤形，浅黄色，有光泽。喜温暖湿润的气候条件，不耐严寒，耐干旱，耐火烧，抗逆性强。饲用价值较高，一般利用方法为刈割后直接饲喂动物或调制成青贮饲料。King Grass (Latin scientific name Pennisetum purpureum K.Scbumacb, P.typhoideumRich), Chinese alias Huangcao (English aliases Chinese sugarcane, japanese cane, kinggrass, north Indian sugarcane, uva cane), is a plant of the genus Pennisetum in the Gramineae family. It is an excellent tropical forage grass, which is widely cultivated in Hainan, Guangdong, and Guangxi in China, and introduced and cultivated in Jiangsu, Fujian, Yunnan, and Hunan. Foreign countries are mainly distributed in Central and South America. Perennial bushy tall grass. The root system is well developed. The stem resembles sugar cane. The leaves are elongated. The panicles are densely spike-shaped. Caryopsis fusiform, light yellow, shiny. It likes warm and humid climate conditions, not tolerant to severe cold, drought-resistant, fire-resistant, and strong stress resistance. The feeding value is high, and the general utilization method is to directly feed animals after cutting or prepare silage.
植物多糖具有多种生物活性功能，包括免疫调节、抗肿瘤、降血糖、降血脂等。王草含有丰富的多糖成分，从王草中提取多糖，对拓宽王草利用渠道和提高种植王草的经济效益有重要意义。目前，据了解和检索，关于王草的植物多糖提取尚未有相关的报道和文献记载。Plant polysaccharides have a variety of biologically active functions, including immune regulation, anti-tumor, hypoglycemic, hypolipidemic, etc. King grass is rich in polysaccharides. Extracting polysaccharides from king grass is of great significance to broaden the utilization channels of king grass and improve the economic benefits of planting king grass. At present, as far as we know and retrieve, there are no relevant reports and literature records about the extraction of plant polysaccharides from King Grass.
发明内容Contents of the invention
为了克服以上技术存在的不足，本发明旨在提供一种王草多糖的提取方法。In order to overcome the deficiencies in the above technologies, the present invention aims to provide a method for extracting Wangcao polysaccharide.
本发明解决技术问题所采用的技术方案为：一种王草多糖的提取方法，该方法包括下列步骤：The technical scheme adopted by the present invention to solve technical problems is: a kind of extracting method of Wangcao polysaccharide, the method comprises the following steps:
（1）将新鲜王草茎叶的王草原料切成小段，在烘箱中烘烤46～50小时干燥至恒重，然后用粉碎机粉碎成粉末，过30～50目筛后备用；(1) Cut the raw material of fresh king grass stems and leaves into small pieces, bake in an oven for 46-50 hours and dry to constant weight, then crush it into powder with a pulverizer, pass through a 30-50 mesh sieve and set aside;
（2）在王草粉末中加入10～30倍重量的蒸馏水倒入圆底烧瓶容器中，放入恒温水浴锅中匀速振荡，提取温度为60～90℃，提取时间为20～50分钟，重复上述提取过程1～3次，提取完成后待溶液冷却，双层中速定性滤纸过滤，蒸馏水冲洗滤渣，合并滤液并定容，待测；(2) Add 10-30 times the weight of distilled water to the king grass powder, pour it into a round-bottomed flask container, put it into a constant temperature water bath and shake at a constant speed, the extraction temperature is 60-90°C, the extraction time is 20-50 minutes, repeat The above extraction process was carried out 1 to 3 times. After the extraction was completed, the solution was cooled, filtered with double-layer medium-speed qualitative filter paper, and the filter residue was rinsed with distilled water. The filtrate was combined and fixed to volume for testing;
（3）以葡萄糖标准品溶液为对照，采用苯酚硫酸法测定提取液中多糖的含量；(3) Using the glucose standard solution as a control, the content of polysaccharides in the extract was determined by the phenol-sulfuric acid method;
（4）向提取液中加入95%乙醇溶液并摇匀，在4℃冰箱中静置24小时，经过滤或离心后得到沉淀物，用丙酮充分洗涤，再次过滤或离心，得到的产物经冷冻干燥后为王草多糖。(4) Add 95% ethanol solution to the extract and shake it well, let it stand in a refrigerator at 4°C for 24 hours, obtain a precipitate after filtration or centrifugation, wash thoroughly with acetone, filter or centrifuge again, and freeze the obtained product After drying, it is king grass polysaccharide.
较佳地，所述的一种王草多糖的提取方法，其特征在于：Preferably, the extraction method of a polysaccharide from Wangcao Wangcao is characterized in that:
（1）所述王草原料为新鲜王草茎叶，切成约3cm小段，在65℃烘箱中烘烤48小时干燥至恒重，然后用粉碎机粉碎成粉末，过40目筛后备用；(1) The raw material of king grass is fresh stems and leaves of king grass, cut into small pieces of about 3 cm, baked in an oven at 65°C for 48 hours and dried to constant weight, then crushed into powder with a pulverizer, passed through a 40-mesh sieve for later use;
（2）在王草粉末中加入15倍重量的蒸馏水倒入圆底烧瓶容器中，放入恒温水浴锅中匀速振荡，提取温度为80℃，提取时间为30分钟，重复上述提取过程2次，提取完成后待溶液冷却，双层中速定性滤纸过滤，蒸馏水冲洗滤渣，合并滤液并定容，待测；(2) Add 15 times the weight of distilled water to the king grass powder, pour it into a round-bottomed flask container, put it into a constant temperature water bath and vibrate at a constant speed, the extraction temperature is 80°C, and the extraction time is 30 minutes. Repeat the above extraction process twice, After the extraction is completed, wait for the solution to cool down, filter with double-layer medium-speed qualitative filter paper, rinse the filter residue with distilled water, combine the filtrate and constant volume, and wait for the test;
（3）以葡萄糖标准品溶液为对照，采用苯酚硫酸法测定提取液中多糖的含量；(3) Using the glucose standard solution as a control, the content of polysaccharides in the extract was determined by the phenol-sulfuric acid method;
（4）按与提取液体积比为1:1加入95%乙醇溶液并摇匀，在4℃冰箱中静置24小时，经过滤或离心后得到沉淀物，用沉淀物2倍体积丙酮充分洗涤，再次过滤或离心，得到的沉淀物经冷冻干燥后为王草多糖。(4) Add 95% ethanol solution at a volume ratio of 1:1 to the extract and shake well, let it stand in a refrigerator at 4°C for 24 hours, obtain a precipitate after filtration or centrifugation, and wash thoroughly with acetone with 2 times the volume of the precipitate , filtered or centrifuged again, and the obtained precipitate was freeze-dried to be Wangcao polysaccharide.
本发明具体的有益效果为：考虑了多种因素对多糖提取的影响，采用正交试验方法对提取过程中提取剂水与王草粉末的比例、提取时间、提取次数和提取温度进行严格控制，整个提取过程操作简单、程序简化、提取准确、重复性好、提高了王草多糖的提取率。The specific beneficial effects of the present invention are: taking into account the influence of various factors on the extraction of polysaccharides, adopting an orthogonal test method to strictly control the ratio of extractant water and Wangcao powder, extraction time, extraction times and extraction temperature during the extraction process, The whole extraction process is simple in operation, simplified in procedure, accurate in extraction and good in repeatability, and improves the extraction rate of Wangcao polysaccharide.
具体的实施方式specific implementation
实施例一Embodiment one
一种王草多糖的提取方法，该方法包括下列步骤：A method for extracting Wangcao polysaccharide, the method comprising the following steps:
（1）王草原料为新鲜王草茎叶，切成约3cm小段，在65℃烘箱中烘烤48小时干燥至恒重，然后用粉碎机粉碎成粉末，过40目筛后备用；(1) The raw material of king grass is fresh stems and leaves of king grass, cut into small pieces of about 3cm, baked in an oven at 65°C for 48 hours and dried to constant weight, then crushed into powder with a grinder, passed through a 40-mesh sieve for later use;
（2）在王草粉末中加入15倍重量的蒸馏水倒入圆底烧瓶容器中，放入恒温水浴锅中匀速振荡，提取温度为80℃，提取时间为30分钟，重复上述提取过程2次，提取完成后待溶液冷却，双层中速定性滤纸过滤，蒸馏水冲洗滤渣，合并滤液并定容，待测；(2) Add 15 times the weight of distilled water to the king grass powder, pour it into a round-bottomed flask container, put it into a constant temperature water bath and vibrate at a constant speed, the extraction temperature is 80°C, and the extraction time is 30 minutes. Repeat the above extraction process twice, After the extraction is completed, wait for the solution to cool down, filter with double-layer medium-speed qualitative filter paper, rinse the filter residue with distilled water, combine the filtrate and constant volume, and wait for the test;
（3）以葡萄糖标准品溶液为对照，采用苯酚硫酸法测定提取液中多糖的含量，本实施例中多糖含量为9.20%；(3) Using the glucose standard solution as a control, the polysaccharide content in the extract was determined by the phenol-sulfuric acid method, and the polysaccharide content in this example was 9.20%;
（4）按与提取液体积比为1:1加入95%乙醇溶液并摇匀，在4℃冰箱中静置24小时，经过滤或离心后得到沉淀物，用沉淀物2倍体积丙酮充分洗涤，再次过滤或离心，得到的沉淀物经冷冻干燥后为王草多糖。(4) Add 95% ethanol solution at a volume ratio of 1:1 to the extract and shake well, let it stand in a refrigerator at 4°C for 24 hours, obtain a precipitate after filtration or centrifugation, and wash thoroughly with acetone with 2 times the volume of the precipitate , filtered or centrifuged again, and the obtained precipitate was freeze-dried to be Wangcao polysaccharide.
实施例二Embodiment two
一种王草多糖的提取方法，该方法包括下列步骤：A method for extracting Wangcao polysaccharide, the method comprising the following steps:
（1）王草原料为新鲜王草茎叶，切成约3cm小段，在65℃烘箱中烘烤48小时干燥至恒重，然后用粉碎机粉碎成粉末，过40目筛后备用；(1) The raw material of king grass is fresh stems and leaves of king grass, cut into small pieces of about 3cm, baked in an oven at 65°C for 48 hours and dried to constant weight, then crushed into powder with a grinder, passed through a 40-mesh sieve for later use;
（2）在王草粉末中加入20倍重量的蒸馏水倒入圆底烧瓶容器中，放入恒温水浴锅中匀速振荡，提取温度为70℃，提取时间为20分钟，重复上述提取过程1次，提取完成后待溶液冷却，双层中速定性滤纸过滤，蒸馏水冲洗滤渣，合并滤液并定容，待测；(2) Add 20 times the weight of distilled water to the king grass powder, pour it into a round bottom flask container, put it into a constant temperature water bath and shake it at a constant speed, the extraction temperature is 70°C, the extraction time is 20 minutes, repeat the above extraction process once, After the extraction is completed, wait for the solution to cool down, filter with double-layer medium-speed qualitative filter paper, rinse the filter residue with distilled water, combine the filtrate and constant volume, and wait for the test;
（3）以葡萄糖标准品溶液为对照，采用苯酚硫酸法测定提取液中多糖的含量，本实施例中多糖含量为6.32%；(3) Using the glucose standard solution as a control, the polysaccharide content in the extract was determined by the phenol-sulfuric acid method, and the polysaccharide content in this example was 6.32%;
（4）按与提取液体积比为1:1加入95%乙醇溶液并摇匀，在4℃冰箱中静置24小时，经过滤或离心后得到沉淀物，用沉淀物2倍体积丙酮充分洗涤，再次过滤或离心，得到的沉淀物经冷冻干燥后为王草多糖。(4) Add 95% ethanol solution at a volume ratio of 1:1 to the extract and shake well, let it stand in a refrigerator at 4°C for 24 hours, obtain a precipitate after filtration or centrifugation, and wash thoroughly with acetone with 2 times the volume of the precipitate , filtered or centrifuged again, and the obtained precipitate was freeze-dried to be Wangcao polysaccharide.
实施例三Embodiment Three
一种王草多糖的提取方法，该方法包括下列步骤：A method for extracting Wangcao polysaccharide, the method comprising the following steps:
（1）王草原料为新鲜王草茎叶，切成约3cm小段，在65℃烘箱中烘烤48小时干燥至恒重，然后用粉碎机粉碎成粉末，过40目筛后备用；(1) The raw material of king grass is fresh stems and leaves of king grass, cut into small pieces of about 3cm, baked in an oven at 65°C for 48 hours and dried to constant weight, then crushed into powder with a grinder, passed through a 40-mesh sieve for later use;
（2）在王草粉末中加入25倍重量的蒸馏水倒入圆底烧瓶容器中，放入恒温水浴锅中匀速振荡，提取温度为90℃，提取时间为40分钟，重复上述提取过程3次，提取完成后待溶液冷却，双层中速定性滤纸过滤，蒸馏水冲洗滤渣，合并滤液并定容，待测；(2) Add 25 times the weight of distilled water to the king grass powder, pour it into a round bottom flask container, put it into a constant temperature water bath and vibrate at a constant speed, the extraction temperature is 90°C, the extraction time is 40 minutes, repeat the above extraction process 3 times, After the extraction is completed, wait for the solution to cool down, filter with double-layer medium-speed qualitative filter paper, rinse the filter residue with distilled water, combine the filtrate and constant volume, and wait for the test;
（3）以葡萄糖标准品溶液为对照，采用苯酚硫酸法测定提取液中多糖的含量，本实施例中多糖含量为6.78%；(3) Using the glucose standard solution as a control, the polysaccharide content in the extract was determined by the phenol-sulfuric acid method, and the polysaccharide content in this example was 6.78%;
（4）按与提取液体积比为1:1加入95%乙醇溶液并摇匀，在4℃冰箱中静置24小时，经过滤或离心后得到沉淀物，用沉淀物2倍体积丙酮充分洗涤，再次过滤或离心，得到的沉淀物经冷冻干燥后为王草多糖。(4) Add 95% ethanol solution at a volume ratio of 1:1 to the extract and shake well, let it stand in a refrigerator at 4°C for 24 hours, obtain a precipitate after filtration or centrifugation, and wash thoroughly with acetone with 2 times the volume of the precipitate , filtered or centrifuged again, and the obtained precipitate was freeze-dried to be Wangcao polysaccharide.
以上所揭露的仅为本发明的较佳实施例而已，当然不能以此来限定本发明之权利范围，因此依本发明权利要求所作的等同变化，仍属于本发明所涵盖的范围。The above disclosures are only preferred embodiments of the present invention, and certainly cannot limit the scope of rights of the present invention. Therefore, equivalent changes made according to the claims of the present invention still fall within the scope of the present invention.
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