阅读说明：本技术 一种连续从白刺种子中提取油脂、黄酮和多糖的方法 (Method for continuously extracting grease, flavone and polysaccharide from nitraria tangutorum bobr seeds ) 是由 赵振 马烨 安庆莲 赵婌君 于 2020-08-22 设计创作，主要内容包括：本发明公开了一种连续从白刺种子中提取油脂、黄酮和多糖的方法,包括如下步骤：S1、滤渣滤液分离；S2、提取油脂；S3、提取黄酮；S4、提取多糖；本发明能够连续从白刺种子中提取出油脂、黄酮和多糖类化合物,提取工艺简单可行,设备投入低,仅需要一套提取设备。并且在提取不同成分时不需要将物料从提取罐中取出,节约了时间和人力,避免了物料在转移过程中的损耗。在本发明的优选方案下,油脂、黄酮和多糖三种活性成分的得率均达到较高值,提高了白刺种子的利用率。(The invention discloses a method for continuously extracting grease, flavone and polysaccharide from nitraria tangutorum bobr seeds, which comprises the following steps: s1, separating filter residue and filtrate; s2, extracting grease; s3, extracting flavone; s4, extracting polysaccharide; the method can continuously extract the grease, the flavone and the polysaccharide compounds from the nitraria tangutorum bobr seeds, has simple and feasible extraction process and low equipment investment, and only needs one set of extraction equipment. And need not take out the material from extracting the jar when extracting different composition, practiced thrift time and manpower, avoided the loss of material in the transfer process. Under the preferred scheme of the invention, the yield of the three active ingredients of the grease, the flavone and the polysaccharide reaches a higher value, and the utilization rate of the nitraria tangutorum bobr seeds is improved.)

1. A method for continuously extracting grease, flavone and polysaccharide from nitraria tangutorum bobr seeds is characterized by comprising the following steps:

s1, separating filter residue and filtrate: cleaning, drying and crushing nitraria tangutorum bobr seeds to obtain nitraria tangutorum bobr seed powder; adding ethanol into the nitraria seed powder, mixing, and heating and extracting for 1-3 times; filtering, and mixing the filtrates to obtain filter residue I and filtrate I;

s2, extracting oil: concentrating the filtrate I in the step S1 under reduced pressure, recovering ethanol, adding purified water for dilution to obtain a concentrated solution I, extracting with petroleum ether, and separating to obtain a petroleum ether layer and a water layer; concentrating the petroleum ether layer under reduced pressure, and recovering petroleum ether to obtain Nitraria seed oil;

s3, extracting flavone: adding purified water into the filter residue I in the step S1, and heating and extracting for 1-3 times; after extraction, performing suction filtration to obtain a filtrate II, and performing reduced pressure concentration on the filtrate II to obtain a concentrated solution II; adding 95% ethanol into the concentrated solution II, standing for precipitation, and centrifugally separating supernatant from precipitate; concentrating the supernatant under reduced pressure to recover ethanol, and mixing

S2, mixing the middle water layers, preparing a sample loading liquid with corresponding concentration, adjusting the pH value, and loading the sample into a macroporous adsorption resin column; eluting with clear water until the effluent is colorless, eluting with ethanol solution until the eluate is colorless and transparent, collecting ethanol eluate, concentrating, and lyophilizing to obtain total flavone;

s4, extracting polysaccharide: and (4) freeze-drying the precipitate in the step S3 to obtain the polysaccharide.

2. The method for continuously extracting oil, flavone and polysaccharide from nitraria tangutorum bobr seeds of claim 1, wherein in step S1, nitraria tangutorum bobr seeds are dried in the sun until the moisture content is less than or equal to 5% before being crushed; crushing nitraria tangutorum bobr seeds to 40-60 meshes; adding 55-95% ethanol into the nitraria tangutorum bobr seed powder according to the liquid-material ratio of 4: 1-14: 1 mL/g; heating to 45-85 deg.C, extracting for 2 times, each time for 1-3 hr.

3. The method for continuously extracting oil, flavone and polysaccharide from nitraria tangutorum bobr seeds as claimed in claim 1, wherein in the step S2, the volume ratio of the added purified water to the concentrated solution I is 0: 1-1: 1; the volume ratio of the added petroleum ether to the diluted concentrated solution I is 1: 1-3: 1; the petroleum ether extraction time is 10-60 min; the number of times of petroleum ether extraction is 1-4.

4. The method for continuously extracting the grease, the flavone and the polysaccharide from the nitraria tangutorum bobr seeds as claimed in claim 1, wherein in the step S3, purified water is added into the filter residue I according to the liquid-material ratio of 5: 1-20: 1 mL/g; heating reflux extraction at 60-100 ℃; extracting for 1 time, 1-3 h each time; according to the weight percentage of ethanol: and (3) adding 95% ethanol into the concentrated solution II according to the volume ratio of 2: 1-5: 1 of the concentrated solution II, and standing and precipitating for 12 hours at 4 ℃.

5. The method for continuously extracting oil, flavone and polysaccharide from nitraria tangutorum bobr seeds of claim 1, wherein in step S3, the macroporous absorption resin column material is AB-8 type macroporous resin; the concentration of flavone in the sample loading solution is 0.5-2.5 mg/mL; the pH value of the sample liquid is 3-7; the sample loading liquid is loaded into the macroporous adsorption resin column at the flow speed of 0.5-2.5 BV/h; the volume fraction of the ethanol solution for elution is 15-95 percent; the elution flow rate of the ethanol solution is 0.5-2.5 BV/h.

Technical Field

The invention relates to a method for continuously extracting grease, flavone and polysaccharide from nitraria tangutorum bobr seeds.

Background

The white thorn is multi-branch, salt and alkali resistant and xerophyte type short shrubs of Zygophyllaceae, and has the physiological characteristics of drought resistance, high temperature resistance, strong water retention capacity and the like. The nitraria tangutorum bobr has rich resources and wide distribution, and is wild in the northwest desert, the north China and the northeast coastal areas, Zhangjiakou dam, Tianjin, Cangzhou and the like. According to statistics, the nitraria tangutorum bobr basins are distributed in pollution-free areas with the altitude of more than 3000 m, 6-10 ten thousand tons of nitraria tangutorum bobr can be produced every year, and the method has great development value.

The nitraria tangutorum bobr fruits have higher economic and medicinal values and are well researched, developed and applied. In the process of utilizing the nitraria tangutorum bobr, a large amount of nitraria tangutorum bobr seeds are produced. The seed oil of Nitraria tangutorum has been reported to contain high content of unsaturated fatty acids, wherein the content of linoleic acid is 65.85% (Soxhlet, Gaohang, Wang-Cynanchum Paniculatum, Tanggute Nitraria, China: research on chemical components of seed oil by supercritical carbon dioxide extraction (English) [ J)]Natural product research and development, 2004(01):16-18.), is a functional oil rich in unsaturated fatty acids which is rarely found in nature. Some methods for extracting Nitraria tangutorum bobr seed oil have been disclosed, for example, in the report of the text "Natural products research and development" No. 16 volume No. 1 of 2004, "research on chemical compositions of Nitraria tangutorum bobr seed oil by supercritical carbon dioxide extraction (English)" in supercritical CO₂The fluid technology is carried out at the pressure of 22.0MPa, the temperature of 42 ℃ and CO₂The tangut nitraria tangutorum bobr seeds are extracted for 8 hours under the condition of the flow rate of 46L/h, and the oil yield is 11.6 percent; the same supercritical CO is used in the national patent CN200510041879₂The fluid technology is carried out at the pressure of 15-45 MPa, the temperature of 25-50 ℃ and CO₂Extracting the Nitraria tangutorum bobr seeds for 6 hours under the condition of the flow rate of 45-250L/h, wherein the oil yield is 10.13%; in "CN 201110374709", a method for extracting nitraria seed oil by using subcritical fluid is described, and the yield of oil is about 12%. And "supercritical CO" in "food research and development" 2015 3, 36 th volume, 6 th₂The best technology for extracting Siberian Nitraria seed oil is disclosed as the research article, wherein the optimal technology is that the extraction pressure is 40MPa, the extraction temperature is 55 ℃, and CO is adopted₂The flow is 10L/min, the yield is 4.63% under the condition, and the difference of the grease yield caused by different varieties of the nitraria tangutorum bobr is possible. The method disclosed above has high yield and high oil qualityGood and the like, but all need complicated and expensive extraction equipment, which is not beneficial to industrial popularization.

Besides oil, flavonoids are also an important active ingredient in nitraria tangutorum bobr seeds. The total flavone content in the nitraria tangutorum bobr seeds is reported to be about 1.1-1.9%, and the flavones have the effects of clearing free radicals, repairing vascular endothelial cell injury and the like. In the published extraction method of the flavones of the nitraria tangutorum bobr seeds, such as CN200510041880, ethanol is used as a solvent for hot reflux extraction, and the extract is purified by a nonpolar macroporous resin tower. However, it is known that active ingredients in plants are complex and various, and various effective ingredients exist in extracts, so that the total flavone is obtained only from the nitraria tangutorum bobr seed extract, and the development and utilization of the nitraria tangutorum bobr seeds are not facilitated.

Polysaccharides are also an important active ingredient in plants, and many plant polysaccharides have antioxidant and anti-inflammatory effects. The nitraria tangutorum bobr polysaccharide research is mainly focused on nitraria tangutorum bobr fruits, and no nitraria tangutorum bobr seed polysaccharide extraction report is found, but in our research, the polysaccharide compound content in the nitraria tangutorum bobr seeds is more than 6%, so that the polysaccharide extraction is expected to become a new way for comprehensively utilizing and developing the nitraria tangutorum bobr seeds.

In industry, different extraction equipment or extraction processes are often required for extracting different target components from plants, so that manufacturers need to invest more equipment, labor, time and bear more risks, and therefore, a method which is simple, rapid and can simultaneously or continuously extract different effective components from plants is urgently needed to be developed.

Disclosure of Invention

Aiming at the defects of the prior art, the invention solves the problems that: provides a process for quickly, conveniently and continuously extracting grease, flavone and polysaccharide from nitraria tangutorum bobr seeds.

In order to solve the problems, the technical scheme adopted by the invention is as follows:

a method for continuously extracting grease, flavone and polysaccharide from nitraria tangutorum bobr seeds comprises the following steps:

s1, separating filter residue and filtrate: cleaning, drying and crushing nitraria tangutorum bobr seeds to obtain nitraria tangutorum bobr seed powder; adding ethanol into the nitraria seed powder, mixing, and heating and extracting for 1-3 times; filtering, and mixing the filtrates to obtain filter residue I and filtrate I;

s2, extracting oil: concentrating the filtrate I in the step S1 under reduced pressure, recovering ethanol, adding purified water for dilution to obtain a concentrated solution I, extracting with petroleum ether, and separating to obtain a petroleum ether layer and a water layer; concentrating the petroleum ether layer under reduced pressure, and recovering petroleum ether to obtain Nitraria seed oil;

s3, extracting flavone: adding purified water into the filter residue I in the step S1, and heating and extracting for 1-3 times; after extraction, performing suction filtration to obtain a filtrate II, and performing reduced pressure concentration on the filtrate II to obtain a concentrated solution II; adding 95% ethanol into the concentrated solution II, standing for precipitation, and centrifugally separating supernatant from precipitate; concentrating the supernatant under reduced pressure to recover ethanol, mixing with the water layer in step S2, preparing into a sample solution with corresponding concentration, adjusting pH, and loading into a macroporous adsorbent resin column; eluting with clear water until the effluent is colorless, eluting with ethanol solution until the eluate is colorless and transparent, collecting ethanol eluate, concentrating, and lyophilizing to obtain total flavone;

s4, extracting polysaccharide: and (4) freeze-drying the precipitate in the step S3 to obtain the polysaccharide.

Further, in the step S1, before crushing, the nitraria tangutorum bobr seeds are dried in the sun until the moisture content is less than or equal to 5%; crushing nitraria tangutorum bobr seeds to 40-60 meshes; adding 55-95% ethanol into the nitraria tangutorum bobr seed powder according to the liquid-material ratio of 4: 1-14: 1 mL/g; heating to 45-85 deg.C, extracting for 2 times, each time for 1-3 hr.

Further, in the step S2, the volume ratio of the added purified water to the concentrated solution I is 0: 1-1: 1; the volume ratio of the added petroleum ether to the diluted concentrated solution I is 1: 1-3: 1; the petroleum ether extraction time is 10-60 min; the number of times of petroleum ether extraction is 1-4.

Further, in the step S3, adding purified water into the filter residue I according to the liquid-material ratio of 5: 1-20: 1 mL/g; heating reflux extraction at 60-100 ℃; extracting for 1 time, 1-3 h each time; according to the weight percentage of ethanol: and (3) adding 95% ethanol into the concentrated solution II according to the volume ratio of 2: 1-5: 1 of the concentrated solution II, and standing and precipitating for 12 hours at 4 ℃.

Further, in the step S3, the macroporous adsorbent resin column material is AB-8 type macroporous resin; the concentration of flavone in the sample loading solution is 0.5-2.5 mg/mL; the pH value of the sample liquid is 3-7; the sample loading liquid is loaded into the macroporous adsorption resin column at the flow speed of 0.5-2.5 BV/h; the volume fraction of the ethanol solution for elution is 15-95 percent; the elution flow rate of the ethanol solution is 0.5-2.5 BV/h.

The invention has the advantages of

1. The nitraria seed powder is subjected to two times of ethanol separation and extraction essentially, the two times of extraction filtrates are mixed to be used as carrier liquid for extracting flavone, and meanwhile, the first-time separated filtrate provides a carrier for oil separation, so that the process design is ingenious, efficient and convenient.

2. The method can continuously extract the grease, the flavone and the polysaccharide compounds from the nitraria tangutorum bobr seeds, has simple and feasible extraction process and low equipment investment, and only needs one set of extraction equipment. And need not take out the material from extracting the jar when extracting different composition, practiced thrift time and manpower, avoided the loss of material in the transfer process. Under the preferred scheme of the invention, the yield of the three active ingredients of the grease, the flavone and the polysaccharide reaches a higher value, and the utilization rate of the nitraria tangutorum bobr seeds is improved.

3. The invention researches the influence of factors such as material-liquid ratio, ethanol volume fraction, extraction temperature, petroleum ether extraction time, petroleum ether dosage, petroleum ether extraction frequency and the like on the extraction of the nitraria seed oil or total flavonoids, and determines the optimal process for extracting the nitraria seed flavonoids by ethanol. When the polysaccharide in the nitraria tangutorum bobr seeds is extracted by using water, the influence of factors such as a material-liquid ratio, an extraction temperature, extraction time, extraction times and the like on the yield of the polysaccharide is also researched, and the optimal process for extracting the polysaccharide is determined. In addition, the purification process of the nitraria seed flavone is optimized, the optimal process conditions such as the concentration of the sample loading liquid, the pH of the sample loading liquid, the flow rate of the sample loading liquid, the volume fraction of the eluent, the flow rate of the eluent and the like are determined, and the purity of the nitraria seed flavone is obviously improved.

Drawings

FIG. 1 is a process flow diagram of the present invention.

FIG. 2 is a standard curve of rutin as a reference substance for determining the content of total flavonoids in the method of the present invention.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings.