CN101549117A - Bagasse (leaf) extract with anti-tumor effect and its application - Google Patents
Bagasse (leaf) extract with anti-tumor effect and its application Download PDFInfo
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- CN101549117A CN101549117A CNA2009101140573A CN200910114057A CN101549117A CN 101549117 A CN101549117 A CN 101549117A CN A2009101140573 A CNA2009101140573 A CN A2009101140573A CN 200910114057 A CN200910114057 A CN 200910114057A CN 101549117 A CN101549117 A CN 101549117A
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Abstract
The invention relates to bagasse and sugarcane leaves extract, and an extracting method of the extract and the application of the extract. The inventive extract is the extract of bagasse (leaf), specifically the extract with anti-tumor effect. The application of this invention refers to the application of using the extract with anti-tumor effect to prepare cancer therapeutic agents or supplementary drugs, or the application for preparation of functional food, or the application for preparation of the raw material of aforementioned medicines and food.
Description
Technical field
The present invention relates to the extract of a kind of bagasse or leaf of Caulis Sacchari sinensis, particularly relate to a kind of extract and uses thereof with antitumor action.
Background technology:
Caulis Sacchari sinensis Saccharum sinensis Roxb. is the cane of grass, extensively plants in the temperate zone and the torrid areas, and all there is cultivation each province, China south.The traditional Chinese medical science thinks that the Caulis Sacchari sinensis sweet in the mouth is cool in nature, clearing away heat and promoting production of body fluid is arranged, moisturize effect such as relieve the effect of alcohol, and can be used for diseases such as calentura Tianjin wound, restlessness and thirst, regurgitation vomiting, xeropulmonary cough, constipation due to dry stool.Bagasse and leaf of Caulis Sacchari sinensis are the side-product behind the harvesting sugarcane, and bagasse its main uses at present has the paper pulp of manufacturing, is used for boiler oil and generating, produces wood-based plate and environmental protection tableware, makes xylitol, furfural, ethanol, compost etc.Leaf of Caulis Sacchari sinensis also contains number of chemical compositions such as vitamin C, equisetic acid, Caulis Sacchari sinensis polysaccharide except that containing a large amount of chlorophylls.Leaf of Caulis Sacchari sinensis output is very big, and each province, annual China south leaf of Caulis Sacchari sinensis output is up to tens million of tons.But a large amount of leaf of Caulis Sacchari sinensis also is underutilized, except that small part as the animal feed, most of by sugarcane grower's on-site incineration, cause the wasting of resources and serious environment pollution.Though aspect rationally the utilizing of bagasse and leaf of Caulis Sacchari sinensis, done some research work and obtained certain achievement in recent years, do not see its health care and medicinal aspect report.The inventor explores the application of leaf of Caulis Sacchari sinensis at medical health field in order to attempt to excavate the health care and the medical value of leaf of Caulis Sacchari sinensis, and having carried out is the functional food of main material and the research and development research work of medicine with the leaf of Caulis Sacchari sinensis.This further promotes the Caulis Sacchari sinensis industry development to have positive effect for the comprehensive utilization and the deep development of Caulis Sacchari sinensis resource.
Summary of the invention
The objective of the invention is from bagasse (leaf), to extract active substance, and with the raw material of these active substances as preparation functional food or medicine, be applied to the auxiliary treatment or the treatment of cancer, thereby prolong the Caulis Sacchari sinensis industrial chain, utilize the agricultural waste material resource of China better.
The inventor finds that by the research to bagasse and leaf of Caulis Sacchari sinensis the extract of bagasse (leaf) has obvious antineoplastic, in experiment in vitro, has obtained excellent curative.
Extract of the present invention, its chemical identification is as follows:
(1) get the extract sample for preparing under the preparation method of the present invention, be dissolved in ethanol, add a little magnesium powder jolting, drip several concentrated hydrochloric acid again, heating is 1-2 minute in the water-bath, takes out, i.e. displaing amaranth.
(2) get the extract sample for preparing under the preparation method of the present invention, be dissolved in ethanol, get the alcoholic solution point after (putting 1 time after doing) on the filter paper again, doing, the 1% aluminum chloride ethanol test solution of spraying is observed under ultra-violet lamp, presents yellow-green fluorescence.
The extract that the inventor extracts from Caulis Sacchari sinensis (slag) leaf in pharmacological research, has stronger specific aim, obtains preliminary effect.The present invention simultaneously also provides Preparation method and use.
Experimental study shows: bagasse of the present invention (leaf) extract has inhibitory action external to human hepatoma cell strain Hele 7404, cervical cancer cell strain Hela, human stomach cancer cell line SGC7901.Conclusion: bagasse (leaf) extract has obvious antineoplastic.
Below be preparation method of extract of the present invention:
1, preparation method:
(1) get bagasse (leaf), extracting in water 2-4 time filters, and merges the water extract, concentrates, and gets bagasse (leaf) extract cream.
(2) get bagasse (leaf), add alcohol reflux and extract 2-4 time, filtration merges alcohol extract, concentrates and reclaims alcohol, gets bagasse (leaf) extract cream.
(3) get bagasse (leaf), extracting in water 2-4 time filters, and merges the water extract, being concentrated into relative density is 1.05-1.15, the ethanol that adds 80%-95% in the concentrated solution contains alcohol to medicinal liquid and measures 40%-75%, leaves standstill 12-72 hour, filter, filtrate recycling ethanol does not extremely have the alcohol flavor, concentrates, and gets bagasse (leaf) extract cream.
(4) get bagasse (leaf), add ethyl acetate extraction 2-4 time, filter, the combined ethyl acetate extracting solution concentrates, and gets bagasse (leaf) extract cream.
The explanation of preparation method:
1) in the above-mentioned preparation method, amount of water or add pure consumption be bagasse (leaf) weight 6-12 doubly
2) alcohol in the preparation method (2) comprises ethanol, methanol, propanol, n-butyl alcohol for the used alcohol of conventional extraction.
3) the leaf of Caulis Sacchari sinensis cream after above-mentioned preparation method concentrates can be prepared into the clear paste that relative density is 1.20-1.35, and being used for routinely, formulation method prepares liquid preparation; Also the needs of formulation method routinely are dried to dried cream, and it is standby to beat powder, is used to make solid preparation.
2, functional food and medicament preparation method embodiment:
Embodiment 1.
Raw material: bagasse (leaf)
Method for making: get bagasse (leaf), decoct with water and extract 4 times, each 6 times of water gagings filter, and merge the water extract, are condensed into the clear paste that relative density is 1.20-1.35, and 80 ℃ of drying under reduced pressure are pulverized, and add right amount of auxiliary materials, make tablet.
Embodiment 2.
Raw material: bagasse (leaf)
Method for making: get bagasse (leaf), decoct with water and extract 2 times, each 12 times of water gagings filter, and merge the water extract, are condensed into the clear paste that relative density is 1.20-1.35, and drying is pulverized, and adds right amount of auxiliary materials, makes granule.
Embodiment 3.
Raw material: bagasse (leaf)
Method for making: bagasse (leaf) is ground into coarse powder, decocts with water to extract 3 times, and each 8 times of water gagings filter, and merge the water extract, are condensed into the clear paste that relative density is 1.20-1.35, and drying is pulverized, and adds right amount of auxiliary materials, makes powder.
Embodiment 4.
Raw material: bagasse (leaf)
Method for making: get bagasse (leaf), add alcohol reflux 2 times, each 8 times of amount ethanol, filter, merge alcohol extract, reclaim ethanol and be condensed into the clear paste that relative density is 1.2-1.35 (60 ℃ of surveys), behind glyceryl monostearate heating and melting mixing, splash in the not miscible condensed fluid, shrink condensation and make sphere or the spheric preparation of class, packing promptly gets drop pill.
Embodiment 5.
Raw material: bagasse (leaf)
Method for making: get bagasse (leaf), decoct with water and extract each 6 times of water gagings 4 times, filter, merge the water extract, be condensed into the clear paste that relative density is 1.05-1.15, add 95% ethanol and transfer to and contain alcohol amount 70%, stir, left standstill 12 hours, filter, decompression filtrate recycling ethanol, and the simmer down to relative density is the clear paste (60 ℃) of 1.2-1.35,80 ℃ of drying under reduced pressure are pulverized, and add adjuvant, mixing, briquetting or adorn tea bag or fry in shallow oil in the bag then, packing promptly gets medicinal tea.
Embodiment 6.
Raw material: bagasse (leaf)
Method for making: bagasse (leaf), be ground into coarse powder, decoct with water and extract 2 times, each 8 times of water gagings filter, and merge the water extract, be condensed into the clear paste that relative density is 1.05-1.15, add 95% ethanol and transfer to and contain alcohol amount 60%, stir, left standstill 72 hours, and filtered decompression filtrate recycling ethanol, and the simmer down to relative density is the clear paste (60 ℃) of 1.2-1.35, add refined honey and receive cream, add appropriate amount of auxiliary materials, packing promptly gets soft extract.
Embodiment 7.
Raw material: bagasse (leaf)
Method for making: get bagasse (leaf), decoct with water and extract 2 times, 8 times of amount of water, filter by 6 times, merge the water extract, be condensed into the clear paste that relative density is 1.05-1.15, add 85% ethanol and transfer to and contain alcohol amount 55%, stir, left standstill 48 hours, filter, decompression filtrate recycling ethanol, and the simmer down to relative density is the clear paste (60 ℃) of 1.2-1.35,80 ℃ of drying under reduced pressure, pulverize, add right amount of auxiliary materials, mixing, be divided in Capsules or be sealed in the soft capsule material, packing promptly gets hard capsule product or soft capsule product.
Embodiment 8.
Raw material: bagasse (leaf)
Method for making: get bagasse (leaf), decoct with water and extract each 8 times of water gagings 2 times, filter, merge the water extract, be condensed into the clear paste that relative density is 1.05-1.15, add 90% ethanol and transfer to the alcohol amount 65% that contains, stir, left standstill 24 hours, filter, decompression filtrate recycling ethanol, and the simmer down to relative density is the clear paste (60 ℃) of 1.2-1.35, adds right amount of auxiliary materials, makes mixture.
Embodiment 10.
Raw material: bagasse (leaf)
Method for making: get bagasse (leaf), decoct with water and extract each 8 times of water gagings 2 times, filter, merge the water extract, be condensed into the clear paste that relative density is 1.05-1.15, add 90% ethanol and transfer to the alcohol amount 60% that contains, stir, left standstill 36 hours, filter, decompression filtrate recycling ethanol, and the simmer down to relative density is the clear paste (60 ℃) of 1.2-1.35, adds right amount of auxiliary materials, makes syrup.
3, pharmacology embodiment
Embodiment 1:
Leaf of Caulis Sacchari sinensis extract mtt assay is measured: every hole adds the RPMI1640 culture fluid of 100 μ l (containing three kinds of tumor cells of 5000/ml) 10%NBS (new-born calf serum) in 96 well culture plates (NUNC).Cell puts 37%, cultivate 24h in the 5%CO2 incubator after, experimental group adds four kinds of different extract culture fluid 100 μ l that concentration is 50 μ g/ml, 25 μ g/ml, 12.5 μ g/ml, 6.25 μ g/ml respectively, blank group adds the culture fluid of equal-volume solvent, and repeat 3 times in every group 3 hole.Put 37 ℃, 5%CO
2Incubator was cultivated after 3 days, abandoning supernatant, and every hole adds the serum-free medium of the freshly prepared 5mg/ml of the containing MTT of 20 μ l, 37 ℃, 5%CO
2Continue cultivating 4h, abandon supernatant, add 150 μ l dimethyl sulfoxide (DMSO), behind the vibration mixing, is 550nm with the wavelength on microplate reader, and reference wavelength is that 450nm measures optical density (OD) value, result of calculation.Be calculated as follows the suppression ratio of medicine: growth of tumour cell suppression ratio=(the average OD value of the 1-experimental group/average OD value of blank group) * 100% to growth of tumour cell.The result shows, the leaf of Caulis Sacchari sinensis extract has inhibitory action to 7901 when 1mg/ml, 31.25 μ g/ml, 15.63 μ g/ml, 7.81 μ g/ml concentration, but P＞0.05; The leaf of Caulis Sacchari sinensis ethyl acetate extract all has inhibitory action in each concentration, and suppression ratio reaches 10.17 (P＜0.01) when 3.91 μ g/ml.
Embodiment 2:
The bagasse extract mtt assay is measured: the trophophase cell of taking the logarithm, behind 0.25% trypsinization, be inoculated on the 96 porocyte culture plates (3 * 104cells/ml), every hole 100 μ l.Put 37 ℃, 5%CO
2After cultivating 24h in the incubator, experimental group adds the culture fluid that final concentration is 250,125,62.5,31.25 μ g/ml extracts respectively, and matched group then adds the culture fluid of equal-volume solvent, and repeat 3 times in every group 3 hole.Establish blank hole (only add RPMI1640 culture fluid and medicine, do not add cell) simultaneously.At 37 ℃, 5%CO
2Cultivate 3d in the incubator, add MTT (5mg/ml) 20 μ l and continue to cultivate 4 hours, abandon supernatant, add DMSO150 μ l, concussion 15min surveys each hole absorbance (OD) with microplate reader at the 490nm place, deduct blank during mensuration, be calculated as follows the suppression ratio of medicine growth of tumour cell.The result shows that in 31.25～250 μ g/ml concentration ranges, all there is the obvious suppression effect at the ethyl acetate extraction position to 3 kinds of tumor cell lines, and along with the increase of extract concentrations, its suppression ratio is more and more higher, has dose dependent.When concentration is 250 μ g/ml, suppression ratio to 3 kinds of tumor cells such as SGC7901, Hela, BEL7404 is respectively 45.54%, 52.11% and 49.10%, calculates its half-inhibition concentration (IC50) by the improvement karber's method and is respectively 228.03,204.30 and 190.22 μ g/ml.Also there is certain inhibitory action at water extract and alcohol extraction position to 3 kinds of tumor cells.
Embodiment 3:
The bagasse extract colony forming method is measured: the trophophase cell of taking the logarithm behind 0.25% trypsinization, is inoculated on the 24 porocyte culture plates (100cells/ml) every hole 1ml.Shake up rearmounted 37 ℃, 5%CO
2Cultivate 24h in the incubator, experimental group adds the culture fluid that final concentration is 250,125,62.5,31.25 μ g/ml extracts respectively, and matched group then adds the culture fluid of equal-volume solvent, every group 3 hole, every hole 1ml.Then 24 well culture plates are moved into 37 ℃, 5%CO
2Leave standstill 7d in the incubator, abandon culture fluid, (0.01mol/L pH7.4) carefully embathes 1 time, and methanol is 10min fixedly, abandons fixative, contains the above cell colony of 50 cells with counting behind Rui Shi-Giemsa staining with PBS.The ethyl acetate extraction position forms 3 kinds of tumor cell line colonies all the obvious suppression effect, and presents certain dose dependent in the test concentrations scope.When concentration is 250 μ g/ml, the colony of SGC7901, BEL7404 and Hela cell strain is formed suppression ratio reach 100%, 96.77% and 100%.Also there is certain inhibitory action at water extract and alcohol extraction position to 3 kinds of tumor cells, and gained result and mtt assay are identical substantially.
Claims (8)
1, a kind of extract with antitumor action is characterized in that: this extract is to be refined by bagasse or leaf of Caulis Sacchari sinensis to process.
2, as right 1 described preparation method of extract, it is characterized in that: get bagasse (leaf), extracting in water 2-4 time filters, and merges the water extract, concentrates, and gets bagasse (leaf) extract cream.
3, as right 1 described preparation method of extract, it is characterized in that: get bagasse (leaf), add alcohol reflux and extract 2-4 time, filtration merges alcohol extract, concentrates and reclaims alcohol, gets bagasse (leaf) extract cream.
4, as right 1 described preparation method of extract, it is characterized in that: get bagasse (leaf), extracting in water 2-4 time, filter, merge the water extract, being concentrated into relative density is 1.05-1.15, adds the ethanol of 80%-95% in the concentrated solution, contain alcohol to medicinal liquid and measure 55%-70%, left standstill 12-72 hour, and filtered, filtrate recycling ethanol is to there not being the alcohol flavor, concentrate, get bagasse (leaf) extract cream.
5, as right 1 described preparation method of extract, it is characterized in that: get bagasse (leaf), add ethyl acetate extraction 2-4 time, filter, the combined ethyl acetate extracting solution concentrates, and gets bagasse (leaf) extract cream.
6, the purposes of extract as claimed in claim 1 is characterized in that: this extract is in the application of producing functional food and pharmaceutical preparations.
7, the purposes of extract as claimed in claim 1 is characterized in that: this extract is in the application of treatment cancer.
8, the purposes of Chinese medicine preparation as claimed in claim 1 is characterized in that: the application of this extract in auxiliary for treating cancer.
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2009101140573A CN101549117A (en) | 2009-05-13 | 2009-05-13 | Bagasse (leaf) extract with anti-tumor effect and its application |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007090A (en) * | 2013-01-16 | 2013-04-03 | 广西中医药大学 | Preparation and application of sugarcane leaf extractive for preventing and curing decsyed teeth and preparations |
| CN103549252A (en) * | 2013-10-22 | 2014-02-05 | 广西金麦克生物科技有限公司 | Preparation method and application of sugarcane polysaccharide extract |
| CN104082704A (en) * | 2014-06-11 | 2014-10-08 | 南通昊友食品添加剂有限公司 | Composite sweetening agent |
-
2009
- 2009-05-13 CN CNA2009101140573A patent/CN101549117A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103007090A (en) * | 2013-01-16 | 2013-04-03 | 广西中医药大学 | Preparation and application of sugarcane leaf extractive for preventing and curing decsyed teeth and preparations |
| CN103549252A (en) * | 2013-10-22 | 2014-02-05 | 广西金麦克生物科技有限公司 | Preparation method and application of sugarcane polysaccharide extract |
| CN104082704A (en) * | 2014-06-11 | 2014-10-08 | 南通昊友食品添加剂有限公司 | Composite sweetening agent |
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Open date: 20091007 |