CN101804132B - Saponins of saponins and their preparation methods and applications - Google Patents
Saponins of saponins and their preparation methods and applications Download PDFInfo
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- CN101804132B CN101804132B CN2010101507867A CN201010150786A CN101804132B CN 101804132 B CN101804132 B CN 101804132B CN 2010101507867 A CN2010101507867 A CN 2010101507867A CN 201010150786 A CN201010150786 A CN 201010150786A CN 101804132 B CN101804132 B CN 101804132B
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Abstract
本发明公开了虎皮兰皂苷及其制备方法和应用，虎皮兰皂苷是将虎皮兰制成干粉，经水、乙醇或甲醇提取，石油醚或乙醚萃取，再经正丁醇或戊醇萃取制得的虎皮兰总皂苷浸膏；本发明一种虎皮兰皂苷的制备方法，优化了工艺条件，操作简单，费用低，工艺设备要求不高，可以减少溶剂的消耗，提高浸出效率。灭菌后的虎皮兰皂苷，对人白血病细胞系JurKet的生长有明显的抑制作用，对人宫颈癌细胞系HeLa、人胃癌细胞系BGC823无活性杀伤效果，对正常的中国仓鼠卵巢细胞CHO无伤害，细胞生长正常没有明显的变化。The invention discloses a saponin of tiger lily and a preparation method and application thereof. The saponin of tiger lily is prepared by making tiger lily into dry powder, extracting it with water, ethanol or methanol, extracting it with petroleum ether or ether, and then extracting it with n-butanol or amyl alcohol to obtain a total saponin extract of tiger lily; the preparation method of tiger lily saponin of the invention optimizes the process conditions, is simple to operate, has low cost, has low requirements for process equipment, can reduce the consumption of solvents, and improves the leaching efficiency. The sterilized tiger lily saponin has a significant inhibitory effect on the growth of human leukemia cell line JurKet, has no active killing effect on human cervical cancer cell line HeLa and human gastric cancer cell line BGC823, has no damage to normal Chinese hamster ovary cells CHO, and has normal cell growth without significant changes.
Description
Technical field
The present invention relates to a kind of extraction of vegetable active composition, specifically Sansevieria trifasciata saponin
Background technology
Budgerigar blue (Sansevieria trifasciata) has another name called Folium Sansevieriae Trifasclatae, Your Highness is blue, is the perennial meat herbaceous plant of Agavaceae, Folium Sansevieriae Trifasclatae genus.Originate in Africa, India, belong to desert plant, can restrain oneself harsh environmental conditions, strong to the adaptability of environment; Be prone to breeding, growth soon, the happiness temperature climate, cold-resistant; The preference temperature of growth is 20-30 ℃, when temperature is lower than 13 ℃, can stop growing, and the temperature of surviving the winter can not be lower than 8 ℃.The Budgerigar orchid has the root stock of crawling to grow, and blade directly grows from rhizome, and blade is strong upright.
At present, point out that the Budgerigar orchid has the effect that purifies the air of a room in a lot of reports.It can discharge oxygen in absorbing carbon dioxide, the negative ion concentration in the room air is increased.It is reported, indoor at 15 square metres, it is blue to put 2-3 basin Budgerigar, just can the interior harmful gas more than 80% of absorption chamber.It also has the ability of very strong absorption formaldehyde, therefore is called by people " natural street cleaner ".
The Budgerigar orchid also has effects such as heat-clearing and toxic substances removing, promoting flow of QI and blood, relaxing muscles and tendons and activating QI and blood in the collateral, and diseases such as cardiovascular, gastric ulcer, arteriosclerosis are had therapeutic efficiency.The Budgerigar orchid also has the effect of antiinflammatory, heat-clearing and toxic substances removing, in addition, and can also the lowering blood-fat and reducing weight blood pressure lowering.Not only do not suffer tremendous loss in strength or resources, can supplement the nutrients on the contrary, increase effects such as human body energy.
Saponin (Saponins) is the glycosides compound that is present in a botanic class formation more complicated, and its aglycon belongs to mostly and has spirostane and similar steroid or the triterpenoid compound of source of students thereof.Can produce persistent foam after the jolting of most of saponin aqueous solution, so claim saponin.Up to the present, the saponins compound of finding in the world mainly is distributed in the plant of 15 sections, and wherein the shared quantity of monocotyledon is more, and 41 genus of 9 sections are arranged, and mainly concentrates in the subjects such as Dioscoreaceae, Agavaceae, Liliaceae.Chinese medicine that some are commonly used such as Radix Ginseng, Radix Bupleuri, Radix Notoginseng, Semen Ziziphi Spinosae, Herb Gynostemmae Pentaphylli etc. all contain a large amount of steroidal saponins.The saponin aboundresources of China, wherein more, than system with the report of saponin in the Araliaceae, use also relatively extensively, like Radix Ginseng, Radix Panacis Quinquefolii etc.In addition, in recent years the research report of saponin in Dioscoreaceae, strand Landtek, the Liliaceae is also day by day increased.
Saponin by sapogenin (Sapogenins) and sugar, alduronic acid, also might be to form with other organic acid.The common sugar of forming saponin has glucose, galactose, rhamnose, arabinose, wooden sugar and other pentose class.Common alduronic acid has galacturonic acid, glucuronic acid etc.These glucosides alduronic acid are the forms that are combined into sugar chain (oligosaccharide) earlier, link to each other with sapogenin again, and few cases is that the form with monosaccharide links to each other with sapogenin.
The saponin kind is numerous and diverse, and is different according to the chemical constitution of sapogenin, can saponin be divided into triterpene saponin (TriterpenoidSaponins) and steroidal saponin (Steroid Saponins).Up to now, from plant, just isolated more than 400 kind of steroidal saponin.But directly applying to clinical medicine, the single component saponin also do not have, and different with plant variety, and effect also is not quite similar.
The spiral sterin alkane saponin of taking from Folium Dracaenae cambodianae has anti-gram positive bacteria effect with the astersaponin of taking from Asterias amurensis Lutken.
Take from Radix Glycyrrhizae monopotassium glycyrrhizunate (a kind of amyrin saponin) but external concentration deactivation Measles virus and herpesvirus when 5 μ mol/L.Saikoside (a kind of β balsam saponin) can suppress multiple DNA and RNA viruses, irreversibly deactivation herpes simplex virus, and (human immunodeficiency virus HIV) also has inhibitory action to the HIV.
The Dodonoside A that obtains from aalii (Sapindaceae) can reduce mice pawl angle dish glue and cause edema 33%, and the Crossop tine B that obtains from Rubiaceae is better than aspirin and aescine reducing the effect that mice pawl angle dish glue causes aspect the edema.The saponin that from Quillaia saponaria (Rosaceae), obtains is studied for many years at world wide, and it is gone on the market as immune adjuvant.
Cardiac glycoside (lanatoside) is one type has the important compound of unique effect to heart, and they are all on the books in many medical books.At present, the steroidal saponin of single component directly applies to clinical medicine also not to be had, but total steroidal saponin is more in clinical practice as the medicine of treatment diseases of cardiovascular and cerebrovascular systems, and evident in efficacy.Such as: by the pure Chinese medicinal preparation that contains 8 kinds of steroidal saponins---the DIAOXINXUE KANG JIAONANG that extracts in the Dioscorea panthaica Prain et Burkill (Dioscoreapanthaica), preparation---the Xinnao Shutong of being developed by Fructus Tribuli (Tribulusterrestris) fruit total steroid saponin.Preparation by Rhizoma Dioscoreae Zingiberensis (Dioscoreazingiberensis) rhizome water-solubility saponin development---peltate leaf perhexiline and the main component of using clinically are the decoction etc. Radix Ophiopogonis of steroidal saponin.These medicines are at aspects such as the sequela of treating coronary heart disease, angina pectoris, myocardial ischemia, cerebral arteriosclerosis and cerebral thrombosis, chronic pulmonary heart diseases, and clinical effectiveness very significantly.
Chlorophytum malayense has potential cytotoxicity to tumor; Solanum nigrum is used for various treatment for cancer in China and Japan; The Herba Phyllanthi Urinariae also treats at the antineoplastic that is used among the people.
Summary of the invention
The object of the invention provides Sansevieria trifasciata saponin, and it is that the Budgerigar orchid is processed dry powder, extracts through saponin extract solution, and petroleum ether or extracted with diethyl ether make through n-butyl alcohol or amylalcohol extraction again;
Described saponin extract solution is water, ethanol or methanol;
Another object of the present invention is that a kind of method for preparing of Sansevieria trifasciata saponin is provided:
1) the blue dry powder of Budgerigar, with 70% soak with ethanol 24h,
2) supersonic generator is transferred to 40 ℃ of temperature, solid-to-liquid ratio is 1: 15, carries out ultrasound wave lixiviate 50min;
3) be evaporated to no ethanol, with petroleum ether extraction 3 times, discard ether layer.
4) the saturated n-butanol extraction of reuse is 3 times, and evaporation volatilizes, and gets total saponins bullion extractum.
Another technical scheme of the method for preparing of Sansevieria trifasciata saponin of the present invention is:
1), gets the blue dry powder of Budgerigar, after spending the night with 90% soak with ethanol;
2), in normal thermostat water bath, be heated to 75 ℃, the solid-liquid ratio is 1: 12, reflux, extract, 4 times, each 4h;
3), merge extractive liquid,, filter, evaporation and concentration is to there not being ethanol;
4), the extracting solution after will concentrating is with petroleum ether extraction 3 times, discard ether layer;
5), the saturated n-butanol extraction of reuse 3 times, reclaim n-butanol layer after, evaporation volatilizes, both the blue total saponins extractum of Budgerigar;
The blue dry powder of the described Budgerigar of step 1), through behind the multigelation, reuse is alcohol-pickled.
Another object of the present invention is the application of Sansevieria trifasciata saponin at the acute T Lymphocytic leukemia medicine of preparation treatment.
Sansevieria trifasciata saponin of the present invention is that the Budgerigar orchid is processed dry powder, extracts through saponin extract solution, and petroleum ether or extracted with diethyl ether extract the blue total saponins extractum of the Budgerigar that makes through n-butyl alcohol or amylalcohol again; The method for preparing of a kind of Sansevieria trifasciata saponin of the present invention has been optimized process conditions, and is simple to operate, and expense is low, and process equipment is less demanding, can reduce the consumption of solvent, improves leaching efficiency.Sansevieria trifasciata saponin after the sterilization; The growth that is JurKat to acute T lymphocytic leukemia cell has the obvious suppression effect; To human cervical carcinoma cell is that HeLa, SGC-7901 BGC823, Chinese hamster ovary cell CHO do not have injury, and the cell growth does not normally have obvious variation.
Description of drawings
Fig. 1 is Sansevieria trifasciata saponin of the present invention scanning spectra under ultraviolet spectrophotometer.
Fig. 2 is Sansevieria trifasciata saponin cell toxicity test result of the present invention, a left side: acute T HTLV cell line JurKat application of sample group; Right: acute T lymphocytic leukemia cell is the JurKat matched group.
The specific embodiment
Embodiment 1
1) getting the blue sample powder 10g of exsiccant Budgerigar, is that extractant soaks 24h with 70% ethanol,
2) with KQ-3200E supersonic generator (city of Kunshan's ultrasonic instrument company limited), transfer to 40 ℃ of temperature, solid-to-liquid ratio is under 1: 15 the condition, to carry out ultrasound wave lixiviate 50min;
3) do not have alcohol and distinguish the flavor of through extractant being evaporated into RE-52A Rotary Evaporators (Shanghai Yarong Biochemical Instrument Plant), with last solution with petroleum ether extraction 3 times, discard ether layer.
4) the saturated n-butanol extraction of reuse is 3 times, then n-butanol layer is volatilized n-butyl alcohol with Rotary Evaporators, gets total saponins bullion extractum.
Embodiment 2
1), gets the blue powder of exsiccant Budgerigar, after spending the night with 90% soak with ethanol;
2), reach in the HH-S6 type digital display thermostat water bath that the instruments used for education company limited produces in Changzhou HuaPu, be heated to 75 ℃, the solid-liquid ratio is 1: 12, reflux, extract, 4 times, each 4h;
3), merge extractive liquid,, filter, do not distinguish the flavor of to there being alcohol with RE-52A rotary evaporator (Shanghai Yarong Biochemical Instrument Plant) evaporation and concentration.
4), the extracting solution after will concentrating is with petroleum ether extraction 3 times, discard ether layer;
5), the saturated n-butanol extraction of reuse 3 times, reclaim be concentrated into rotary evaporator behind the n-butanol layer dried, both the blue total saponins extractum of Budgerigar.
Embodiment 3
1) gets the blue powder of exsiccant Budgerigar, by adding water, multigelation 3 times, drying at 1: 2;
2) spend the night with 90% soak with ethanol;
3) reach in the HH-S6 type digital display thermostat water bath of instruments used for education company limited production in Changzhou HuaPu, be heated to 75 ℃, the solid-liquid ratio is 1: 12, reflux, extract, 4 times, each 4h;
4) merge extractive liquid, filters, and does not distinguish the flavor of to there being alcohol with RE-52A rotary evaporator (Shanghai Yarong Biochemical Instrument Plant) evaporation and concentration.
5) extracting solution after will concentrating is with petroleum ether extraction 3 times, discard ether layer;
6) the saturated n-butanol extraction of reuse is 3 times, reclaim be concentrated into rotary evaporator behind the n-butanol layer dried, both the blue total saponins extractum of Budgerigar.
With the blue total saponins extractum of Budgerigar, dilute with methanol; With ultraviolet-uisible spectrophotometer (DU-7500 ultraviolet spectrophotometer, U.S. Beckman company produces) scanning, the wavelength of scanning is observed its absorption spectrum characteristic in the 200-800nm scope with the solution after the dilution.The methanol diluent of Sansevieria trifasciata saponin bullion has maximum absorption peak at the 325nm place, see Fig. 2.
Embodiment 5
With human cervical carcinoma cell is HeLa, human leukemia cell line JurKet, SGC-7901 BGC823, Chinese hamster ovary cell CHO, with containing 10% calf serum, 1% pair of anti-RPMI-1640 culture medium, at 37 ℃, 5%CO
2The isoperibol condition under the cultivation of going down to posterity;
The take the logarithm cell of trophophase, through the digestion of 0.5 ‰ trypsin solutions, piping and druming is dispersed into single cell suspension, joins in the 96 porocyte culture plates with the amount of every hole 50 μ l, regulates cell number to 2 * 10
4Individual/hole, put into 37 ℃ of 5%CO
22h makes it adherent in the incubator.
The blue total saponins extractum of Budgerigar is used water dissolution, and making original solution concentration is 2mg/ml, through the filtering bacterium; Add respectively in the different cell holes; First hole adds 10 μ g/ml (parallel 3 holes), below each hole add 20 μ g/ml (parallel 3 holes) successively, add not commensurability culture medium then; Making it cumulative volume is 100 μ L, and each gradient is established three parallel holes; Blank (adding isopyknic empty mycoprotein and culture medium) is established in experiment.37 ℃, 5%CO
2Cultivate 48h under the condition, take out and be put in observed result under the inverted microscope.
The result finds that the growth that the blue total saponins of Budgerigar is JurKat to acute T lymphocytic leukemia cell has the obvious suppression effect.Along with the increase of the blue total saponins concentration of Budgerigar, the mortality of cell also increases gradually, and the most of change circle of experimental group cell, color are secretly, refractivity is poor even cracking dead; To human cervical carcinoma cell is that HeLa, SGC-7901 BGC823 do not have fragmentation effect, and normal Chinese hamster ovary cell CHO is not had injury, and the cell growth does not normally have obvious variation.See Fig. 2.
The blue total saponins of Budgerigar is to each cell line exercising result
Annotate: "+" expression effect is remarkable, " ± " expression DeGrain, and "-" expression is invalid.
Claims (4)
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| CN103965284A (en) * | 2014-05-07 | 2014-08-06 | 吉林农业大学 | Spindle tree fruit saponin and preparing method and application thereof |
| CN110897989B (en) * | 2019-12-24 | 2021-11-26 | 广州蜜妆生物科技有限公司 | Sensitive skin repair emulsion |
| CN114208857B (en) * | 2021-12-31 | 2022-08-30 | 青岛农业大学 | Application of sansevieria trifasciata crude extract in preparing bactericide for preventing and treating plant diseases caused by downy mildew |
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| CN101062045A (en) * | 2006-04-29 | 2007-10-31 | 华北制药集团新药研究开发有限责任公司 | Novel function of triterpene saponin compounds |
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