CN1237893C - Water-soluble propolis and its extraction method - Google Patents

Abstract

The invention provides a water-soluble propolis and an extraction method. The extraction method uses solubilization, ultrasonic technology, and controls reasonable temperature and heating time to extract active components in the propolis. The method extracts the active ingredients completely, can reduce the destruction of the active ingredients, and can avoid the use of alcohol, which is one of the allergens. The total flavonoid content of the propolis liquid extracted by this method is as high as 12.765 mg/ml, which has good anti-inflammatory immunity, lowering blood fat, lowering blood sugar and preventing diabetic complications, and has little influence on the adrenal cortex system. Because the water extraction preparation is easily absorbed by the body, there is no side effect caused by solvent residue, and it is easy to season, so it has great development potential and has a good application prospect in health care products, cosmetics and food additives.

Description

A kind of extraction method and application of water-soluble propolis

technical field

The invention belongs to the field of deep processing of bee products, and relates to a propolis extraction method, in particular to a water extraction method of water-soluble propolis. The propolis raw material prepared by the invention is especially suitable for health care products, cosmetics and food additives.

Background technique

At present, the active ingredients of propolis products at home and abroad are basically obtained through alcohol extraction. The traditional propolis alcohol extraction method has an unacceptable irritating taste, and people who are allergic to alcohol are prone to allergic reactions when used, which increases allergens. It is very restrictive as a raw material for cosmetics and oral health products, and propolis alcohol extraction The liquid is not easily soluble in water, the absorption and utilization rate is low, and some of the water-soluble active ingredients have not been extracted. Other extraction methods that have been reported generally include propolis chloroform, ethyl acetate, dichloromethane, glycerin extraction, oil extraction and supercritical extraction, etc., which are costly and the active ingredients are not fully extracted. It is reported that propolis water extraction also adopts cold water extraction or heat reflux, the extraction efficiency is low, and the degree of destruction of biologically active components is large.

For a long time, the water extraction technology of propolis routinely adopts cold water extraction, and the obtained propolis water extract contains only 1.914 mg/ml of flavonoids, although the propolis alcohol extract has the highest total flavonoid content under corresponding conditions. 51.36mg/ml, but when dissolved in water, 100ml of water can only dissolve 4-6ml of propolis alcohol extract. At present, there is a preparation method of water-soluble propolis using glycerin soaking and heating, but this physiological activity has not been tested, and it is a method of dissolving the alcohol extract in water instead of extracting it with water. Therefore, the water extraction technology of propolis has not obtained substantive breakthrough, has seriously restricted the further development of product. Since propolis is insoluble in water, most liquid preparations of propolis are extracted by ethanol. The smell of the product is strong and irritating, which is difficult for most consumers to accept and is not suitable for consumption. Alcohol-extracted propolis solid dosage form products are not irritating, but the dosage form of the product is limited, and its biological activity and human absorption rate are not as ideal as those of water-extracted propolis. At present, there is also a small amount of water-soluble propolis that is alkalized and treated with alcohol-extracted propolis on the market in China. Because the alkaline environment has a certain interference effect on the activity of propolis, the biological effect is affected to a certain extent. Therefore, the ideal water extraction technology is a hot and difficult point in the current research on the deep processing of propolis products.

Contents of the invention

An object of the present invention is to provide a kind of water-soluble propolis and extraction method, obtain the total flavonoid content in the water-soluble propolis water extract up to 12.765mg/ml, this extraction method mainly comprises: get propolis raw material (hair glue), water is Solvent, add a small amount of natural green surfactant, heat under temperature control, undergo ultrasonic treatment, and filter to prepare propolis water extract.

The invention provides the surfactant added in the propolis extraction method, including lecithin, soybean lecithin, polyethylene glycol 6000, Tween 80, glyceride and the like.

In the propolis extraction method provided by the invention, the heating temperature is 60-80°C.

In the propolis extraction method provided by the invention, the heating time is 10-16 hours.

In the propolis extraction method provided by the invention, the ultrasonic treatment is performed for 15 to 45 minutes.

Another object of the present invention is to provide various products prepared by using the propolis water extract extracted by the method of the present invention as raw material.

The propolis aqueous extract that adopts the inventive method to extract has the following advantages:

(1) Low toxicity. Acute toxicity experiments were performed by gavage to mice at 400 times and 600 times the normal human dose, and there was no death within a week, indicating that the toxicity is very low;

(2) The effect is good. The active ingredient content of the extraction is high. Compared with the conventional alcohol extraction method, the dosage is only half of the alcohol extraction to achieve the same effect, or even better;

(3) Little irritation. It reduces allergen components and expands the scope of application, especially in reducing alcohol allergy caused by traditional alcohol extraction;

(4) The dosage form is easy to be absorbed by the body and easy to flavor. Especially suitable for health products, cosmetics and food additives.

Description of drawings

Fig. 1 is a simple process flow chart of the inventive method.

Detailed ways

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

Embodiment 1 An example of the inventive method

Take propolis raw material (hair glue), use water as a solvent, add 1% lecithin, the solution is ultrasonically treated for 15-45 minutes, the temperature is controlled at 60-80°C, and the heating time is 10-16 hours, extracted and filtered to obtain propolis water extraction. liquid.

See accompanying drawing 1 for the simple technological process.

By adopting the method of the invention, the propolis extract with a total flavonoid content as high as 12.765 mg/ml can be extracted. However, the content of flavonoids in propolis water extract obtained by conventional cold water extraction is only 1.914mg/ml. Although the total flavonoid content of propolis alcohol extract is up to 51.36mg/ml under corresponding conditions, when dissolved in water, 100ml of water can only dissolve 4-6ml of propolis alcohol extract. The extraction rate of flavonoids in water-soluble propolis is much higher than that of traditional methods.

It is confirmed by animal experiments that the propolis liquid extracted by the method of the invention has good pharmacological activities, including anti-inflammation and immune regulation, anti-diabetic and blood-lipid-lowering effects.

Example 2 The anti-inflammatory immune effect of water-extracted propolis liquid

Four animal models of acute inflammation were used (rat paw swelling model induced by carrageenan, mouse abdominal capillary permeability increase model induced by acetic acid, rat pleurisy model induced by carrageenan, and acute rat model induced by oleic acid plus LPS). Lung injury model) and mouse cotton ball granuloma chronic inflammation model, complete Freund's adjuvant-induced immune inflammation model in rats, a total of 6 batches of whole animal experiments have observed the anti-inflammatory effect of propolis water extract, on this basis from The anti-inflammatory effect and mechanism of the propolis water extract extracted by this method were studied in many aspects and levels, such as the generation and release of inflammatory mediators, lipid peroxide content and SOD activity, cytokines, adhesion molecules, and transcriptional regulators. Studies have shown that the extract extracted by this method has a good anti-inflammatory effect, can inhibit the migration of white blood cells in the acute phase of inflammation, and reduce the degree of inflammation. The NO and PGE in the acute pleurisy inflammatory exudate of rats induced by carrageenan The production of ₂ has obvious inhibitory effect, reduces the interaction between NO and prostaglandins, reduces the activation effect of various proteases, and thus reduces the degree of inflammatory response. Moreover, it can resist the weight loss during the modeling period of adjuvant arthritis rats, has a good preventive effect on chronic inflammation and immune inflammation, and can prevent the occurrence of diabetic nephropathy, hyperlipidemia and hypertension in diabetic rats. Can control the rise of blood sugar to a certain extent. In particular, it exhibited low-dose, high-activity effects in diabetic models. Through immunohistochemistry and in situ hybridization, it was found that the water extract of propolis could inhibit the activation of NF-kBp65 and the expression of intercellular adhesion molecule-1 (CD54) in tissues, indicating that the water extract of propolis exerted anti-inflammatory effect through this pathway is the key One of the mechanisms of action. Links related to its anti-inflammatory effect include: reducing the generation of PGE ₂ and MDA at the inflammatory site; reducing the content of NO in the pleural effusion of rats with pleurisy; preventing the abnormal increase of IL-6 in the process of inflammatory response; inhibiting the acute lung injury Expression of CD54 in mouse bronchial epithelial cells; inhibits the activation of NF-kB p65 and the increase of p65 mRNA transcription in cells during acute inflammation, and controls the central link of inflammatory response.

Example 3 The effect of water-extracted propolis liquid on diabetes

The propolis liquid extracted by the method of the invention has a good preventive effect on diabetes under the condition of restricted feeding, and the effect is better than that of the propolis alcohol extract of the same dose. Propolis water dike extraction can reduce the blood sugar level in diabetic rats: after 2 weeks of administration, blood sugar in the water extract group decreased by 22.3% (P<0.05) compared with the model group, while the alcohol extract group decreased by 12.8%; After one week, compared with the model group, the water extract group decreased by 22.2% (P<0.01), while the alcohol extract group decreased by 15.2% (P<0.01); after 4 weeks of administration, compared with the model group, the water extract group decreased 17.9% (P＜0.05), while the alcohol extract group decreased by 6.8%; after administration for 5 weeks, the water extract group decreased by 27.3% (P＜0.01), while the alcohol extract group decreased by 18.6% (P＜0.05 ); after 6 weeks of administration, it decreased by 33.8% (P＜0.01), while the alcohol extract group decreased by 22.3% (P＜0.01); after 7 weeks of administration, the water extract group decreased by 30.4% (P＜0.01 ), while the alcohol extract group decreased by 21.7% (P<0.01); with the prolongation of time, the effect of controlling blood sugar tended to increase. Regulate lipid metabolism and protein metabolism in vivo, reduce protein consumption: compared with the model group, the total cholesterol in the water extract group decreased by 18.4% (P<0.05), while the alcohol extract group decreased by 15.8% (P<0.05); Very low-density lipoprotein cholesterol water extract group decreased by 34.4% (P<0.01), while alcohol extract group decreased by 24.5% (P<0.05); total protein water extract group increased by 4.8%, while alcohol extract group The water extract group increased by 5.4%, the albumin content in the water extract group increased by 32.1%, and the alcohol extract group increased by 9.5%. It has a certain protective effect on the kidney: compared with the model group, uric acid in the water extract group decreased by 12.5%, while that in the alcohol extract group decreased by 12.3%; creatinine in the water extract group decreased by 7.4%, while that in the alcohol extract group decreased 4.9%; and can inhibit kidney enlargement. The blood sugar control of propolis water extract on diabetes is to achieve the purpose of lowering blood sugar by inhibiting α-glucosidase.

Example 4 The effect of water-extracted propolis liquid on hyperlipidemia

Experimental studies have proved that the extract has a good therapeutic effect on hyperlipidemia. The extract extracted by the water extraction method can effectively inhibit the increase of serum triglyceride (TG), total cholesterol (TC) and low-density lipoprotein (LDL-C) in SD rats with hyperlipidemia; TC, TG value, and can significantly reduce the concentration of serum alanine aminotransferase (GOT) and aspartate aminotransferase (GPT); can significantly increase the activity of superoxide dismutase (SOD). It shows that the propolis liquid extracted by this method can significantly improve the fat metabolism of hyperlipidemia rats, and has a certain antioxidant effect.

Example 5 The content of total flavonoids in propolis liquid by different extraction methods and its effect on the growth of _S180 tumor in mice

The propolis liquid obtained by different extraction methods was compared to the content of total flavonoids and its effect on the growth of _S180 tumor in mice. The results are shown in Table 1.

Table 1 propolis liquid Concentration of total flavonoids in propolis extract (mg/ml) Tumor inhibition rate (%) 75% propolis tincture 27.099 48.76 80% propolis tincture 51.36 67.36 95% propolis tincture 22.644 55.75 Ordinary water extract 1.914 53.48 The inventive method water extract 12.765 80.78

Some references involved in the present invention:

(1) Guo Hua, Ye Tunhao, Li Cili, etc. Research on the influence of microwave on the extraction rate of propolis flavonoids. Jiangsu Food and Fermentation, 2001, 105(2): 10-11.

(2) Gu Qing, Zhang Yanping, Zhong Liren. Research on the Extraction Technology of Active Components in Propolis. Zhejiang Agricultural Journal, 2001, 13(3): 161-164.

(3) Xu Qian, Guo Hua, Ye Huaiyi. Study on Two-step Alcoholic Extraction of Active Components of Propolis. Applied Science and Technology, 2002, 29(7): 75-77.

(4) Chen Hefeng, Han Wenhui, Li Bing, etc. Comparison of antibacterial effects of various solvent extracts of propolis. Food Research and Development, 2001, 22 (Supplement): 18-19.

(5) Hu Fuliang. Extraction method of propolis. Beekeeping Science and Technology, 2000, (5): 28-31.

(6) Ye Luxing, Hu Fuliang, Xuan Hongzhuan, etc. Extraction methods and influencing factors of propolis. Bee Magazine, 2002, (7): 16-17.

(7) Krell R. Value-added products from beekeeping. FAO Bulletin 124, Rome, 1996.

(8) Vassya S, Bankova V. Propolis: Rencent advances in chemistry and plant origin. Apidologie. 2000, 31: 3-15.

(9) Wu Xiaowen, Wu Jiasen. Supercritical CO ₂ fluid extraction of propolis has bright prospects. Beekeeping Science and Technology, 2002,

(1): 11-12.

(10) Zhang Wei. Propolis Extraction Technology and Application of Extract. Chinese Beekeeping, 2002, 53(5): 28-29.

(11) You Hai, Chen Cen, Gao Yinyu, etc. Optimization of extraction process parameters of propolis flavonoids. Food Science, 2002, 23(8): 173-174.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (2)

1. A water-soluble propolis extraction method is characterized in that it is realized by the following steps: get propolis raw material, use water as solvent, add lecithin as surfactant, control heating temperature at 60～80 ℃, heating time control at 10～ After 16 hours, undergo ultrasonic treatment, the ultrasonic treatment time is controlled at 15-45 minutes, and filter to obtain propolis water extract. 2. the application of the propolis aqueous extract prepared by the water-soluble propolis extraction method according to claim 1 in propolis products.

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