CN101301401A - Camellia effective ingredients and extracting method and use thereof - Google Patents
Camellia effective ingredients and extracting method and use thereof Download PDFInfo
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- CN101301401A CN101301401A CNA2008100118991A CN200810011899A CN101301401A CN 101301401 A CN101301401 A CN 101301401A CN A2008100118991 A CNA2008100118991 A CN A2008100118991A CN 200810011899 A CN200810011899 A CN 200810011899A CN 101301401 A CN101301401 A CN 101301401A
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Abstract
The invention provides an extracting method, comprising a water extraction method, an alcohol extraction, a macroporous resin purification method and a solvent extraction method. By adopting the method, the contents of main effective constituents of the obtained extractive are: 1 to 25 percent of TFA, 1 to 20 percent of total Saponins and 1 to 3 percent of phenethyl alcohol glycoside. The invention also provides a method for making the appropriate preparation formulation by using the camellia directly or mixing the solid or liquid of the preparation and the like pharmaceutically in general. The effective constituents of the camellia and the preparation thereof have the functions of reducing blood sugar and lipid and protecting the gastric mucosa, etc. The extracting method has simple implementation and low cost, extracts sufficiently and is suitable for the industrial production; and the extractive has evident activity, precise healing effect and does not have the toxic and side effect.
Description
Technical field:
The invention belongs to medical technical field, relate to effective ingredient and the extracting method and the purposes of Flos Camelliae Japonicae, and the production and processing method of camellia effective ingredients preparation.
Background technology:
Flos Camelliae Japonicae (Camaellia sinensis) is Theaceae, the flower of Camellia plant Flos Camelliae Japonicae.In China's southern area commerial growing, cultivation history is long.Contain abundant flavonoid (being mainly nimbecetin type flavone, Quercetin type flavone, flavane), saponins (theasaponin type) and phenethyl alcohol glycosides in the Flos Camelliae Japonicae.Pharmacological evaluation shows that Flos Camelliae Japonicae extract has effects such as good blood sugar lowering, blood fat reducing, protection gastric mucosa, antioxidation.And being used for blood sugar lowering, blood fat reducing, protection gastric mucosa, Flos Camelliae Japonicae extract yet there are no report.
Summary of the invention:
The object of the present invention is to provide the extracting method of camellia effective ingredients, and utilize its several preparations of effective ingredient production and processing, with it as treatment or improve the medicine and the functional food of symptoms such as hyperglycemia, hyperlipidemia, gastric mucosa injury.
The present invention seeks to be achieved by the following scheme:
The preparation method of camellia effective ingredients is divided three kinds:
Method 1: after getting dry Flos Camelliae Japonicae roguing, the ethanol that adds the 5%-95% of 15 to 30 times of amounts decocts and is no less than twice, each extraction time is no less than 1 hour, extracting solution is concentrated into 1/5th to 1/20th of medicine liquid volume, extracting solution carries out separation and purification through macroporous resin (HPD 100 or D101), respectively with water, 5%-20% ethanol, 30%-80% ethanol, eluting, 5%-20% ethanol elution thing is an effective ingredient group 1, and 30%-80% ethanol elution thing is an effective ingredient group 2, and both are combined as the Flos Camelliae Japonicae effective extract by amounting to the raw medicinal herbs amount at 1: 1.
Method 2: after getting dry Flos Camelliae Japonicae roguing, the ethanol that adds the 5%-95% of 15 to 30 times of amounts decocts and is no less than twice, each extraction time is no less than 1 hour, extracting solution use respectively twice of petroleum ether (or cyclohexane extraction, normal hexane), dichloromethane (or chloroform), ethyl acetate, n-butanol extraction above to extraction fully.Reclaim solvent, gained n-butanol extraction layer is through macroporous resin (HPD 100 or D101) purification, respectively with water, 60%-100% ethanol, ethanol elution, 60%-100% ethanol elution thing is an effective ingredient component 1, and the ethyl acetate extraction layer is an effective ingredient group 2, and the two is combined as the Flos Camelliae Japonicae effective extract by amounting to the raw medicinal herbs amount at 1: 1.
Method 3: after getting dry Flos Camelliae Japonicae roguing, adding 15 to 30 times of water gagings decoctions is no less than twice, each extraction time is no less than 1 hour, extracting solution is concentrated into 1/5th to 1/20th of medicine liquid volume, add ethanol precipitation 1-2 time, concentration of alcohol is at 50%-90% in the control extracting solution, complete at 0-10 ℃ of following stand at low temperature 12-48 hour to precipitation, filter once more, filtrate recycling ethanol is not distinguished the flavor of to there being alcohol, the gained supernatant carries out separation and purification through macroporous resin (HPD 100 or D101), respectively with water, 5%-20% ethanol, 30%-60% ethanol elution, 5%-20% ethanol elution thing is an effective ingredient group 1, and 30%-60% ethanol elution thing is an effective ingredient group 2.Both are by amounting to the raw medicinal herbs amount 1: 1: be combined as the Flos Camelliae Japonicae effective extract.
In the gained Flos Camelliae Japonicae effective component extracts, total flavones 1%-25%, total saponins 1%-20%, phenethyl alcohol glycoside 1%-3%.
By said method as seen, the Flos Camelliae Japonicae effective component extracts is made up of total flavones, total saponins and phenethyl alcohol glycoside, can prepare by above-mentioned three kinds of methods, can sketch to be method one: alcohol extraction-macroporous resin segmentation purification; Method two: alcohol extraction-solvent stage extraction-purification by macroporous resin; Method three: water is carried-precipitate with ethanol-macroporous resin segmentation purification.
Extract of the present invention has blood sugar lowering, blood fat reducing, protection gastric mucosa, antioxidation isoreactivity, can be used for treatment or improves symptoms such as hyperglycemia, hyperlipidemia, gastric mucosa injury.
Advantage of the present invention is as follows:
(1) this preparation method gained Flos Camelliae Japonicae extract is formed clearly, is made up of three kinds of dissimilar natural products, has the low advantage of many target spots synergism and toxic and side effects.
(2) easy, the easy row of preparation method extracts fully, is suitable for commercial production
(3) the camellia effective ingredients extract is significantly active, and determined curative effect can effectively reduce the Chinese medicine dose.
(4) camellia effective ingredients extract action range, the low functional food that can be used as of toxicity.
The specific embodiment:
Embodiment 1
According to effective ingredient preparation method 1
(1) gets dry Flos Camelliae Japonicae 10kg, remove impurity, dust.
(2) 70% ethanol of adding medical material and 20 times of amounts in the Chinese medicine extraction jar, heating and refluxing extraction three times was respectively 2 hours, and 1.5 hours, 1 hour.
(3) merge three extracting solution, be concentrated into 20L, last sample is distinguished water (5V to the non-polar macroporous resin post
R), 10% ethanol (4V
R), 70% ethanol (8V
R), 95% ethanol (4V
R) gradient elution, thing be must wash and 10% ethanol elution thing, 70% ethanol elution thing discarded according to amounting to the crude drug amount 1: 1 mixed, get the Flos Camelliae Japonicae effective extract, wherein general flavone content is 21%, and total saponin content is 16%, and phenethyl alcohol glycoside content is 1.5%.
Embodiment 2
According to effective ingredient preparation method 2
(1) gets dry walking fern herb 6kg, remove impurity, dust
(2) 75% ethanol of adding medical material and 25 times of amounts in the Chinese medicine extraction jar, heating and refluxing extraction three times was respectively 2 hours, and 2 hours, 1 hour.
(3) merge three extracting solution, be concentrated into 10L, equal-volume cyclohexane extraction, dichloromethane, ethyl acetate, n-butanol extraction four times.Get cyclohexane extraction extract 60 grams, dichloromethane extract 55g, acetic acid ethyl ester extract 170 grams, n-butyl alcohol extract 740 grams.Reclaim solvent, the gained n-butyl alcohol extract is further through purification by macroporous resin, and respectively with water, 75% ethanol elution, reclaim under reduced pressure solvent get 75% ethanol elution thing, 340 grams.75% ethanol elution thing and acetic acid ethyl ester extract by amounting to crude drug than 1: 1 mixed, are obtained the camellia effective ingredients extract.Get the Flos Camelliae Japonicae effective extract, wherein general flavone content is 22%, and total saponin content is 19%, and phenethyl alcohol glycoside content is 1.1%.
Embodiment 3
According to effective ingredient preparation method 3
(1) gets dry Flos Camelliae Japonicae 5kg, remove impurity, dust
(2) water of adding medical material and 15 times of amounts in the Chinese medicine extraction jar, heated and boiled extracts twice, each 1.5 hours.
(3) merge extracted twice liquid, be concentrated into 10L, add ethanol, regulate determining alcohol to 50%, extremely precipitated fully in 24 hours 10 ℃ of following stand at low temperature, filter once more, filtrate recycling ethanol is to the pure flavor of nothing.
(4) sample is distinguished water (5V to the non-polar macroporous resin post on the supernatant
R), 15% ethanol (4V
R), 60% ethanol (6V
R), 90% ethanol (4V
R) gradient elution, must wash thing and discard, 20% ethanol elution thing, 60% ethanol elution thing get the Flos Camelliae Japonicae effective extract according to amounting to the crude drug amount 1: 1 mixed, and wherein general flavone content is 20%, and total saponin content is 15%, and phenethyl alcohol glycoside content is 1%.
Embodiment 4 Flos Camelliae Japonicae extracts are to the influence of glucose load rat blood sugar
Laboratory animal: male Wistar rat, body weight 130～150g is available from Japanese KiwaLaboratory Animal Co.Ltd. (Wakayama).Animal constant temperature (23 ± 2 ℃) is raised, and feedstuff is available from Japanese MF.Oriental Yeast Co.Ltd. (Tokyo).The animal fasting is 20～24 hours before the experiment, freely fetches water.
Experimental technique: get the male Wistar rat of 130～150g, fasting was irritated stomach and is given and the Flos Camelliae Japonicae extract after 20～24 hours, 0.5 after hour, irritate stomach and give sucrose (1.0g/kg), after 0.5 hour, blood is got on the optical fundus, adopts the determination of glucose oxidase blood sugar level behind the centrifugal separation plasma.
Experimental result sees Table 1, and the Flos Camelliae Japonicae extract can significantly reduce glucose load rat blood sugar level at 250～1000mg/kg dosage.
Table 1 Flos Camelliae Japonicae extract is to the influence of glucose load rat blood sugar
**p＜0.01
Embodiment 5 Flos Camelliae Japonicae extracts are to the influence of triglyceride levels in the oil load rat plasma
Laboratory animal: male DDY mice, body weight 25～30g is available from Japanese Kiwa LaboratoryAnimal Co.Ltd. (Wakayama).Animal constant temperature (23 ± 2 ℃) is raised, and feedstuff is available from Japanese MF.Oriental Yeast Co.Ltd. (Tokyo).The animal fasting is 20～24 hours before the experiment, freely fetches water.
Experimental technique: get the male DDY mice of 25～30g, fasting was irritated stomach and is given and the Flos Camelliae Japonicae extract after 20～24 hours, 0.5 after hour, irritate stomach and give olive oil (5mL/kg), after 2 hours, blood is got on the optical fundus, adopts triglyceride levels in the WAKO kit measurement blood plasma behind the centrifugal separation plasma.
Experimental result sees Table 2, and the Flos Camelliae Japonicae extract is to the influence of triglyceride levels in the oil load mice plasma, and Flos Camelliae Japonicae extract acetic acid ethyl ester extract and n-butyl alcohol extract can significantly reduce triglyceride levels in the oil load mice plasma at 500mg/kg dosage.
Table 2 Flos Camelliae Japonicae extract is to the influence of triglyceride levels in the oil load rat plasma
**p＜0.01
The gastric mucosa injury that the protection of embodiment 6 Flos Camelliae Japonicae extracts is alcohol induced
Laboratory animal: male SD rat, body weight 230～250g is available from Japanese Kiwa LaboratoryAnimal Co.Ltd. (Wakayama).Animal constant temperature (23 ± 2 ℃) is raised, and feedstuff is available from Japanese MF.Oriental Yeast Co.Ltd. (Tokyo).The animal fasting is 24～26 hours before the experiment, freely fetches water.
Experimental technique: get the male SD rat of 230～250g, fasting was irritated stomach and is given and Flos Camelliae Japonicae extract (5% arabic gum suspendible) after 24～26 hours, after 1 hour, the filling stomach causes gastric mucosa injury to 99.5% ethanol (1.5mL), and after 1 hour, etherization is put to death.Take out the rat stomach, behind the extraction content, the formalin solution of injection 10mL 1.5% is coat of the stomach fixedly, and with flattening after the flushing with clean water, computer scanning calculates the gastric mucosa injury rate repeatedly.
Experimental result: the result is shown in Table 3, and the Flos Camelliae Japonicae extract can significantly reduce the gastric mucosa injury that ethanol causes at 50～200mg/kg dosage; Flos Camelliae Japonicae extract n-butanol extraction layer can significantly reduce the gastric mucosa injury that ethanol causes at 25～200mg/kg dosage.
The gastric mucosa injury that the protection of table 3 Flos Camelliae Japonicae extract is alcohol induced
*p＜0.05，
**p＜0.01
The inductive gastric mucosa injury of embodiment 7 Flos Camelliae Japonicae extracts protection indomethacin
Laboratory animal: male SD rat, body weight 230～250g is available from Japanese Kiwa LaboratoryAnimal Co.Ltd. (Wakayama).Animal constant temperature (23 ± 2 ℃) is raised, and feedstuff is available from Japanese MF.Oriental Yeast Co.Ltd. (Tokyo).The animal fasting is 24～26 hours before the experiment, freely fetches water.
Experimental technique: the male SD rat of getting 230～250g, after the fasting 24～26 hours, irritating stomach gives and Flos Camelliae Japonicae extract (5% arabic gum suspendible), after 1 hour, (the 20mg/kg indomethacin is dissolved in 5% sodium bicarbonate solution, behind the distilled water diluting to indomethacin to irritate stomach, add the neutralization of 0.2M hydrochloric acid, 1.5ml/rat) cause gastric mucosa injury, after 4 hours, etherization is put to death.Take out the rat stomach, behind the extraction content, the formalin solution of injection 10mL 1.5% is coat of the stomach fixedly, and with flattening after the flushing with clean water, the gastric mucosa injury rate is calculated in scanning repeatedly.
Experimental result: see Table shown in 4, the Flos Camelliae Japonicae extract can significantly reduce the gastric mucosa injury that indomethacin causes at 50～200mg/kg dosage.Flos Camelliae Japonicae extract n-butanol extraction layer can significantly reduce the gastric mucosa injury that indomethacin causes at 50～200mg/kg dosage.
The inductive gastric mucosa injury of table 4 Flos Camelliae Japonicae extract protection indomethacin
*p＜0.05，
**p＜0.01
Embodiment 8
Get Flos Camelliae Japonicae effective extract (preparation method such as embodiment 1) dry, pulverize, cross 80 mesh sieves, directly or add suitable diluents (dextrin, microcrystalline cellulose etc.) fill capsule.
Embodiment 9
Get Flos Camelliae Japonicae effective extract (preparation method such as embodiment 2) and add diluent (dextrin, starch etc.), dry, pulverizing, granulation, tabletting or pill agent in 1: 1～1: 4 ratio.
Claims (7)
1, camellia effective ingredients is characterized in that: comprise and contain abundant flavonoid, total saponins class and phenethyl alcohol glycosides in the Flos Camelliae Japonicae.
2, camellia effective ingredients according to claim 1 is characterized in that: described flavones ingredient is nimbecetin type flavone, Quercetin type flavone, flavane, and saponin component is a theasaponin type saponin.
3, a kind of extracting method of camellia effective ingredients as claimed in claim 1 is characterized in that: its extracting method is as follows:
After getting dry Flos Camelliae Japonicae roguing, the ethanol that adds the 5%-95% of 15 to 30 times of amounts decocts and is no less than twice, each extraction time is no less than 1 hour, extracting solution is concentrated into 1/5th to 1/20th of medicine liquid volume, extracting solution carries out separation and purification through HPD 100 or D101 macroporous resin, respectively with water, 5%-20% ethanol, 30%-80% ethanol, eluting, 5%-20% ethanol elution thing is an effective ingredient group 1, and 30%-80% ethanol elution thing is an effective ingredient group 2, and both are combined as the Flos Camelliae Japonicae effective extract by amounting to the raw medicinal herbs amount at 1: 1;
Or after getting dry Flos Camelliae Japonicae roguing, the ethanol that adds the 5%-95% of 15 to 30 times of amounts decocts and is no less than twice, each extraction time is no less than 1 hour, extracting solution respectively with twice of petroleum ether or cyclohexane extraction or normal hexane, dichloromethane or chloroform, ethyl acetate, n-butanol extraction above to extraction fully.Reclaim solvent, gained n-butanol extraction layer is through HPD 100 or D101 purification by macroporous resin, respectively with water, 60%-100% ethanol, ethanol elution, 60%-100% ethanol elution thing is an effective ingredient component 1, and the ethyl acetate extraction layer is an effective ingredient group 2, and the two is combined as the Flos Camelliae Japonicae effective extract by amounting to the raw medicinal herbs amount at 1: 1;
Or after getting dry Flos Camelliae Japonicae roguing, adding 15 to 30 times of water gagings decoctions is no less than twice, each extraction time is no less than 1 hour, extracting solution is concentrated into 1/5th to 1/20th of medicine liquid volume, add ethanol precipitation 1-2 time, concentration of alcohol is at 50%-90% in the control extracting solution, complete at 0-10 ℃ of following stand at low temperature 12-48 hour to precipitation, filter once more, filtrate recycling ethanol is to there not being the alcohol flavor, the gained supernatant carries out separation and purification through HPD 100 or D101 macroporous resin, respectively with water, and 5%-20% ethanol, the 30%-60% ethanol elution, 5%-20% ethanol elution thing is an effective ingredient group 1, and 30%-60% ethanol elution thing is an effective ingredient group 2, and both are by amounting to the raw medicinal herbs amount 1: 1: be combined as the Flos Camelliae Japonicae effective extract.
4, the extracting method of camellia effective ingredients according to claim 3 is characterized in that: in the described effective ingredient group: total flavones 1%-25%, total saponins 1%-20%, phenethyl alcohol glycoside 1%-3%.
5, the extracting method of camellia effective ingredients according to claim 2 is characterized in that: the spray-dried pulverulent solids of making of camellia effective ingredients group extracting solution can also be extracted total active constituent content＞40% in the powder.
6, the extracting method of camellia effective ingredients according to claim 5 is characterized in that: its spray drying condition is: the air preheat to 280 that sprays into ± 20 ℃, the air pressure that sprays into is 3-6kg/cm
2, dry indoor temperature remains on 120 ± 20 ℃.
7, camellia effective ingredients is in preparation treatment hyperglycemia, hyperlipidemia, the medicine of gastric mucosa injury or the application in the functional food.
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| CN101974063A (en) * | 2010-11-09 | 2011-02-16 | 苏州派腾生物医药科技有限公司 | Method for preparing camellia saponin B2 |
| CN103082051A (en) * | 2013-01-11 | 2013-05-08 | 李滋星 | Natural plant combined pinocytosis and preparation method thereof |
| CN103182004A (en) * | 2011-12-30 | 2013-07-03 | 株式会社爱茉莉太平洋 | Composition for preventing or improving metabolic syndrome comprising tea llant leaf, flower and seed extract |
| CN103689740A (en) * | 2013-12-12 | 2014-04-02 | 华南农业大学 | Extractive of camellia oleifera flowers and application of extractive in preparation of health-care drink |
| CN105194102A (en) * | 2015-10-30 | 2015-12-30 | 中国科学院昆明植物研究所 | Camellia sinensis var.assamica extract, preparation method thereof and application of Camellia sinensis var.assamica extract in cosmetics |
| CN105287909A (en) * | 2015-10-30 | 2016-02-03 | 中国科学院昆明植物研究所 | Camellia yuhsienensis extract and preparation method and application thereof |
| CN109793073A (en) * | 2019-03-21 | 2019-05-24 | 苏州美稀生物科技有限公司 | The extracting method at Ilex paraguarensis drink Lipid-lowering activities position and hypoglycemic activity position |
| CN115252489A (en) * | 2022-07-13 | 2022-11-01 | 上海林清轩生物科技有限公司 | Preparation method and application of camellia japonica flower and leaf extract |
| CN117224453A (en) * | 2023-11-15 | 2023-12-15 | 北京小鹿科技有限公司 | Antibacterial skin-friendly wet tissue containing camellia and preparation method thereof |
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| CN1775249A (en) * | 2005-11-18 | 2006-05-24 | 北京天新园医药科技开发有限公司 | Camellia-flower total flavone medicinal preparation, its preparing method and use |
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| CN101974063B (en) * | 2010-11-09 | 2012-10-03 | 苏州派腾生物医药科技有限公司 | Method for preparing camellia saponin B2 |
| CN101974063A (en) * | 2010-11-09 | 2011-02-16 | 苏州派腾生物医药科技有限公司 | Method for preparing camellia saponin B2 |
| CN103182004B (en) * | 2011-12-30 | 2019-03-15 | 株式会社爱茉莉太平洋 | Containing tealeaves, camellia and tea seed extract, for preventing or improving the composition of metabolic syndrome |
| CN103182004A (en) * | 2011-12-30 | 2013-07-03 | 株式会社爱茉莉太平洋 | Composition for preventing or improving metabolic syndrome comprising tea llant leaf, flower and seed extract |
| CN103082051A (en) * | 2013-01-11 | 2013-05-08 | 李滋星 | Natural plant combined pinocytosis and preparation method thereof |
| CN103082051B (en) * | 2013-01-11 | 2014-06-04 | 李滋星 | Natural plant combined pinocytosis and preparation method thereof |
| CN103689740A (en) * | 2013-12-12 | 2014-04-02 | 华南农业大学 | Extractive of camellia oleifera flowers and application of extractive in preparation of health-care drink |
| CN105194102A (en) * | 2015-10-30 | 2015-12-30 | 中国科学院昆明植物研究所 | Camellia sinensis var.assamica extract, preparation method thereof and application of Camellia sinensis var.assamica extract in cosmetics |
| CN105287909A (en) * | 2015-10-30 | 2016-02-03 | 中国科学院昆明植物研究所 | Camellia yuhsienensis extract and preparation method and application thereof |
| CN109793073A (en) * | 2019-03-21 | 2019-05-24 | 苏州美稀生物科技有限公司 | The extracting method at Ilex paraguarensis drink Lipid-lowering activities position and hypoglycemic activity position |
| CN115252489A (en) * | 2022-07-13 | 2022-11-01 | 上海林清轩生物科技有限公司 | Preparation method and application of camellia japonica flower and leaf extract |
| CN117224453A (en) * | 2023-11-15 | 2023-12-15 | 北京小鹿科技有限公司 | Antibacterial skin-friendly wet tissue containing camellia and preparation method thereof |
| CN117224453B (en) * | 2023-11-15 | 2024-02-06 | 北京小鹿科技有限公司 | Antibacterial skin-friendly wet tissue containing camellia and preparation method thereof |
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