CN1066055C - Production method for extracting flavonoid compound extract or powder from bamboo leaf - Google Patents

Abstract

The invention belongs to a production method for extracting flavonoid compounds from plants, in particular extracting flavonoid compound extract or powder from bamboo leaves. Using bamboo leaves as raw materials, the products obtained through extraction, concentration, separation, refining, or re-drying contain a large amount of bamboo leaf flavonoid glycosides, and compounds such as plant phenolic acids, anthraquinones, and terpene lactones. The preparation has excellent anti-free radical, anti-oxidation, anti-aging and hypolipidemic effects, and has a typical bamboo aroma, and can be widely used in various functional foods for the elderly and medical and health products. The preparation is extracted by heat reflux with low-concentration lower alcohol as solvent, and refined by liquid-liquid extraction with middle alcohol. The production process is safe, simple, pollution-free, easy to control, suitable for large-scale industrial production, and also has the characteristics of high flavonoid extraction rate and good purity.

Description

Production method for extracting flavonoids extract or powder from bamboo leaves

The invention relates to a production method for extracting flavonoids from plants, in particular to a production method for extracting flavonoids extract or powder from bamboo leaves.

Bioflavonoids widely exist in the plant kingdom and are the active ingredients of many Chinese herbal medicines. In addition to extracting flavonoids from ginkgo leaves, French pine bark, and Japanese eucommia, kudzu root, licorice, hawthorn, tea, persimmon leaves, masson pine needles and pine nuts, cinnamon bark, acacia bark, wolfberry root bark stems and leaves, dendrobium, cordyceps, The bioflavonoids contained in Shouwu, orange peel, Schisandra chinensis, dates, peanut shells, rice roots, eggplant stems, etc. all have development value. Taking ginkgo flavonoids, which is produced and used on the largest scale today, as an example, its production process roughly falls into the following three types:

(1) Steam distillation method: after drying and pulverizing Ginkgo biloba leaves, steam distillation at 100-120°C, after the distillate is cooled, adsorb with non-polar macroporous resin (or other adsorbents), and then elute with ethanol, Evaporate to dryness to obtain powdery crystals. This method has simple equipment, but the extraction rate is low, so it is rarely used now.

(2) Organic solvent extraction: this is the most widely used method at home and abroad, and there are many patents on its extraction process [method for extracting flavonoids from ginkgo leaves, Li Zhaolong, Chinese herbal medicine, 1992, 14 (6): 5 -6], which can be summarized into two forms. One is to prepare the crude extract: dry and pulverize the ginkgo leaves, extract with an organic solvent (such as ethanol or acetone), filter, concentrate under reduced pressure and recover the solvent to obtain the ginkgo extract. This method has simple production equipment and high product yield (about 12-14%), but contains a large amount of impurities, usually brownish black, and the total flavonoid content of the product is 7-10%. The other is to prepare the fine extract: this type of method is usually further refined on the basis of the crude extract, and the commonly used refining methods are: liquid-liquid extraction (such as degreasing with dichloromethane, chloroform, petroleum ether or benzene; or using Butanone, acetone-butanone or ethyl acetate extraction), precipitation method (precipitation with ammonia water or lead hydroxide) and adsorption-analysis method (adsorption with adsorbents such as macroporous resin, diatomaceous earth or silica gel, ethanol elution) . After drying with anhydrous sodium sulfate, filtering, and distilling off the solvent under reduced pressure, the fine extract can be obtained. It is generally yellowish-brown powder, the yield is about 1-3%, and the total flavonoid content is about 20-26%.

(3) Supercritical fluid extraction: In the past 10 years, more and more attention has been paid to using this technology to extract medicinal active ingredients in natural plants. Compared with the above-mentioned processes, it has high extraction efficiency, no solvent residue, and the active ingredients are not easily extracted Advantages such as decomposition and destruction. European patent EP0308675 (1988) reported a method for extracting active ingredients in ginkgo leaves with supercritical CO2. A lot of work has been done in this area in China in recent years, but most of them are in the stage of laboratory research, and there are no reports of industrial applications. .

Bamboo is a perennial evergreen plant of the Poaceae (Poaceae) bamboo subfamily (Bambusoideae), and is a popular medicine and food dual-purpose plant. Bamboo is widely distributed in Southeast Asia. my country is one of the central production areas of bamboo in the world. There are more than 300 kinds of bamboo species and more than 60 million mu of bamboo forest area. The medical and health care function of bamboo has long been recognized by the working people in our country. "Materia Medica Seeking Truth" says: "Bamboo leaves cool the heart and relieve the spleen, clear phlegm and quench thirst. , vomiting, vomiting blood, and all strokes and convulsions, etc., are nothing more than because lightness can relieve the upper body, pungentness can relieve depression, sweetness can relieve the spleen, coolness can enter the heart, and coldness can heal heat.” Danzhuye and Danzhuli are traditional Chinese medicine for clearing away heat and detoxifying. The whole body of bamboo is a treasure, and it is one of the most valuable plants in the world today. However, in the comprehensive utilization of bamboo, the research and development of bamboo leaves obviously lag behind, except for a very small amount of medicinal and edible, it is basically in the original state. In recent years, the inventors have conducted systematic research on the active ingredients of bamboo leaves and their biological activities, and found that bamboo leaves are rich in flavonoids and other phenolic compounds, anthraquinones, terpenoid lactones, immune active polysaccharides, special amino acids (δ-hydroxylysine) and trace elements such as manganese, zinc, iron, selenium, among which flavonoids are its main functional components. The flavonoids contained in bamboo leaves are basically flavone glycosides, and a considerable part of them are carbon glycoside flavonoids, such as orientin xyloside, isoorientin, 4'-methoxy-vitexin, etc.

Studies have shown that bamboo leaf extract (Ebl) has excellent anti-free radical, anti-oxidation and anti-aging activities. A large number of studies have shown that Ebl has a strong ability to scavenge active oxygen free radicals and block nitrosation reactions; it can significantly induce the activity of endogenous antioxidant enzymes (SOD and GSH-Px) in aged mice, inhibit lipid Peroxidation and removal of peroxidation products; can significantly enhance the anti-fatigue and anti-stress ability of mice; Ebl can significantly reduce the concentration of rat blood triglyceride and total cholesterol, increase the concentration of HDL-cholesterol, reduce the concentration of LDL-cholesterol, Inhibition of weight gain in rats. The results of animal experiments showed that the effect of Ebl on reducing blood lipid and inducing endogenous SOD activity was the same as that of ginkgo biloba extract (Egb), and the effect on inhibiting lipid peroxidation and inducing GSH-Px activity was significantly better than that of Egb. In addition, no harmful components or anti-nutritional factors have been detected from bamboo leaves. Acute toxicity test shows that the oral LD ₅₀ of Ebl > 10g/kg body weight can be regarded as practically non-toxic; Ames test shows that Ebl has no mutagenicity. Therefore, Ebl is a safe and efficient bioflavonoid health factor, which can be widely used in the prevention and treatment of functional foods and various senile degenerative diseases.

The purpose of the present invention is to utilize bamboo leaves which are very rich in resources in my country and have not yet been effectively developed and utilized, to provide a kind of extraction from bamboo leaves with simple process, high extraction rate, easy control of the process, no pollution, and suitable for large-scale industrial production. The production method of flavonoid compound extract or powder.

The production method of extracting flavonoids extract or powder from bamboo leaves is characterized in that: bamboo leaves are used as raw materials, the bamboo leaves are washed, dried, and broken, and a lower concentration (20-40%, volume percent) is used Concentration) of lower alcohol (methanol or ethanol) as solvent, with 1: 10-15 solid-liquid ratio (weight percent) hot reflux extraction 1-2 hours, after the extract is concentrated, physically remove impurities, and then use intermediate alcohol (butanol or amyl alcohol) liquid-liquid extraction, the extract is concentrated under reduced pressure to recover the solvent, then add a small amount of water to dissolve it, and form a flavonoid compound extract, or vacuum or spray drying to obtain a flavonoid compound powder.

The present invention is achieved like this:

Bamboo leaves→washing→drying→crushing→ethanol～water solution hot reflux extraction→filtering→decompression concentration→static flocculation→separation and impurity removal→n-butanol step-by-step extraction→decompression concentration recovery solvent→hot water transfer→reduction Compression concentration→(extract)→vacuum or spray drying→bamboo leaf flavonoid powder

The technical points of this invention are based on the composition characteristics and physical and chemical properties (good hydrophilicity and thermal stability) of bamboo leaf flavonoids, using ethanol aqueous solution for one-time heat reflux extraction; according to the flocculation and precipitation characteristics of impurities under certain conditions , use physical methods to separate and remove impurities; then use n-butanol to extract step by step, recover the solvent, and obtain the extract after being dissolved in water, and then dry to obtain the powder. The process is simple and easy to control; less investment in equipment, good separation effect, high extraction rate; no organic solvent residue, good quality of finished products; the whole process does not use toxic, harmful reagents and strong acid and alkali, basically no "three wastes" discharge, no pollution environmental pollution. The process of the invention is suitable for continuous mass production.

The invention makes the long-term discarded bamboo leaves be deeply developed, not only realizes the utilization of the whole bamboo plant, but also provides a new bioflavonoid resource with a wide source, high quality and low price for the society, which is effective for human aging, tumors and cardiovascular diseases. The prevention and treatment of such degenerative diseases are of great significance.

The key points of the process for the preparation of total flavonoids from bamboo leaves are as follows:

(1) Bamboo leaf raw materials can be dried green leaves of scattered bamboos or clustered bamboos, such as bamboos of the genus Phyllostachys osmanthus (such as Phyllostachys pubescens, Osmanthus osmanthus, Phyllostachys osmanthus, Phyllostachys osmanthus, Bamboo japonica, Bamboo japonica, Bamboo sargassum, Purple bamboo, Bamboo mosaic, etc. ) is better, and the ideal harvesting period is autumn and winter (September to December).

(2) One-time heat reflux extraction. Take the fragments of dried green leaves, and use 1:10-15 solid-liquid ratio, 20-40% concentration of ethanol solution) to extract by hot reflux for 1-2 hours, the cycle is short, the equipment occupancy rate is low, and the extraction is relatively complete. In view of the extensiveness and low price of bamboo leaf raw materials, this one-time extraction method is the most economical and reasonable method.

(3) Physical method to remove impurities. Bamboo leaf extract is concentrated to a volume of about 1:1 (that is, 1 kg of dry leaves gets about 1 kg of concentrated liquid). During the static process of natural cooling, the contained macromolecular impurities and some polyphenols will flocculate and settle naturally. It can be removed by centrifugation, pressure filtration or vacuum filtration, which has a good clarification effect. Compared with conventional alcohol precipitation or other chemical methods, this physical decontamination method greatly saves production costs and avoids possible adverse effects of chemical reagents on the quality of the extract.

(4) The refining of bamboo leaf flavonoids adopts n-butanol liquid-liquid extraction. Because n-butanol has good solubility and selectivity to bamboo leaf flavonoid glycosides, in addition, it has a high boiling point, is not volatile at room temperature, has a small specific gravity, is easy to separate and recover, and is non-toxic and harmless. Using n-butanol for effective times of extraction, the extraction rate of flavonoids is high, and the color of the finished product is golden.

(5) After the n-butanol extract is concentrated under reduced pressure and the solvent is recovered, the extract is transferred into a flavonoid extract with a small amount of hot water, and then vacuum-dried or spray-dried to obtain a flavonoid powder.

The bamboo leaf flavonoid compound powder prepared by the technology of the present invention is yellow powder or yellow crystalline granules, with obvious bamboo leaf aroma, and the total flavonoid content is 24-30%, most of which are flavone glycosides, which contain a considerable amount of Carbon glycoside flavonoids, such as orientin xyloside, isoorientin and 4'-methoxy-vitexin, etc. Other ingredients include plant phenolic acids, anthraquinones, terpenoid lactones, nitrogenous compounds, soluble sugars and minerals.

Embodiment 1: dry green leaf 1kg of Phyllostachy nigra var.henonis (Bean) Stapf exRendle), be broken into about 10 order coarse pieces with crusher and put into the extraction tank of 20L after, add 30% ethanol 12kg , Soak for 1hr, heat to reflux for 1.5hr. The crude extract (about 10kg) is filtered and concentrated in a vacuum concentrator under reduced pressure (recovering the solvent at the same time). When concentrated to a volume of about 1L, the material is discharged, rested for 8 hours, and vacuum filtered to remove impurities. Extract with butanol three times (pre-saturated n-butanol with water, extract at 40-50°C with heat preservation, and stir for 0.5-1 hr each time), combine the n-butanol phase into a vacuum concentrator to concentrate under reduced pressure to recover the solvent, and transfer the extract to hot water , concentrated under reduced pressure to a certain concentration to form an extract, or spray-dried to obtain 25g yellow powder, the total flavonoid content of which is 28%.

Embodiment 2: the dried green leaf of 2kg sweet-scented bamboo (Phyllostachy bamboosoides Sieb.Et Zucc), be broken into the thick sheet of about 10 orders and put into the extraction jar of 50L, add the ethanol 30kg of 35%, soak 0.5hr, reflux 1hr, After the extract is filtered, put it into a storage tank, concentrate in batches with a vacuum concentrator to a final volume of about 2L, let it stand for 6 hours, and centrifuge to remove impurities. The clear liquid is extracted twice with 2 times the volume of n-butanol, and the n-butanol phase, concentrated under reduced pressure to recover the solvent, and after the extract was trans-dissolved in hot water, concentrated under reduced pressure until the specific gravity was greater than 1.12 to form an extractum, or dried in vacuo to obtain 45g yellow powder with a total flavonoid content of 27.6%.

Claims (1)

1, from Folium Bambusae, extracts the production method of flavonoid compound extract or powder, it is characterized in that: be raw material with the Folium Bambusae, with Folium Bambusae clean, oven dry, fragmentation, methanol or ethanol with 20～40% concentration expressed in percentage by volumes are solvent, solid-liquid percentage by weight hot reflux with 1: 10～15 was extracted 1～2 hour, extracting solution concentrates back physics roguing, reuse butanols or amylalcohol liquid～liquid extraction, after the extract concentrating under reduced pressure reclaims solvent, add low amounts of water and change molten, become flavonoid compound extract, or, promptly make the flavone compound powder again through vacuum or spray drying.