CN1528773A - Method for extracting naringin from pomelo - Google Patents

Abstract

The invention relates to a method for extracting naringin monomer from pomelo (such as Shatian pomelo, etc.). The method is as follows: slice or pulverize pomelo, extract with water one or more times, filter, and combine the filtrate; the filtrate is concentrated into an extract, the extract is alcohol-precipitated, and then put on a column or directly put on a column for column separation and eluted with an organic solvent; The eluate was evaporated to dry the solvent to obtain the crude product; after repeated recrystallization, naringin crystals were obtained. By adopting the method of the invention, naringin is extracted from Shatian pomelo, and the extraction yield is all above 85%, and the content of naringin monomer in the obtained naringin sample reaches above 98%. Therefore, the method of the present invention has the characteristics of simple extraction process, high extraction rate, and high purity of the extracted naringin monomer.

Description

Method for extracting naringin from pomelo

technical field

The invention relates to a method for extracting naringin monomer from pomelo Citus grandis (L) Osbeck (such as Shatian pomelo, etc.).

Background technique

Naringin is a flavonoid compound widely present in various Chinese herbal medicines, such as Citrus aurantium, Citrus aurantium, citrus, lemon, grapefruit, tangerine, orange, etc. Naringin is also contained in grapefruit Citus grandis (L) Osbeck, and the content is relatively high. Pomelo Citus grandis (L) Osbeck has many varieties, including Wendan pomelo Citus grandis (L) Osbeck var.wentanyu Hort., Shatian pomelo Citus grandis (L) Osbeckv var.shatianyu Hort., Pingshan pomelo Citus grandis (L) Osbeck var. Hort., Citus grandis (L) Osbeck var. szechipow Hort. in four seasons, Citus grandis (L) Osbeck var. tahungpow in big red. Naringin is one of the main active ingredients of various medicinal materials. Studies have proved that naringin has antibacterial and anti-inflammatory effects, relieves cough and reduces phlegm, reduces plasma cholesterol concentration, and improves vascular circulation. It has been used as a bittering agent, sweating agent, and lowering cholesterol.

The obtaining route of naringin, except Rosenmund (Rosenmund, Ber., 61,2608 (1958)) and Zemlen, the method described by Bognar (Ber., 75,648 (1942)) and other chemical methods are synthesized and obtained, also It can be obtained by extracting from medicinal materials. Many medicinal materials such as pomelo and citrus aurantium have a high content of naringin in their medicinal materials. The content of naringin in the fresh fruit of Meizhou Shatian pomelo is about 2%. However, medicinal materials contain various flavonoids, polysaccharides, tannins, alkaloids, anthraquinones and other substances, and flavonoids also contain many substances with similar structures. To obtain naringin monomer, it is necessary to separate naringin from other substances. Previous studies failed to provide a feasible extraction method to obtain naringin monomer. The method for extracting naringin monomer from medicinal materials (especially pomelo) has not yet been reported.

The structural formula of naringin:

Contents of the invention

The purpose of this invention is to provide a kind of method that extracts naringin monomer from pomelo Citus grandis (L) Osbeck.

The steps of the naringin extraction method of the present invention comprise:

① Slice or crush pomelo, extract with water, filter, and combine the filtrate;

②The filtrate is concentrated into an extract, and the extract is alcohol-precipitated and put on the column or directly on the column for column separation and organic solvent elution;

③ The eluent was evaporated to dry the solvent to obtain the crude naringin product: after repeated recrystallization, the naringin monomer (pure product) was obtained.

The grapefruit mentioned in the above-mentioned method of the present invention can be the young fruit or the peel or the whole fruit of Shatian pomelo or the fruit of the same family and genus; it can be fresh or dried medicinal materials; for example: Shatian pomelo Citus grandis (L) Osbeckvvar.shatianyu Hort., Wendanyu Citus grandis(L)Osbeck var.wentanyu Hort., Pingshanyou Citus grandis(L)Osbeck var.Hort., Four Seasons Citus grandis(L)Osbeckvar.szechipow Hort. or Dahongdao Citus grandis (L) Osbeck var. tahungpow. and other varieties of pomelo.

The water extraction in the method of the present invention can be carried out at room temperature to 100°C or/and under ultrasonic conditions; the number of extractions is generally one to three times, and the number of times can be increased without limitation. The amount of water is that the water can at least cover the medicinal materials, and there is no limit to the amount of water used too much. The extraction time is generally 0.1-24 hours, and the time extension is not limited. Extraction under heating or/and ultrasonic conditions can shorten the extraction time.

The ethanol used for the alcohol precipitation of the extract in the method of the present invention can be absolute ethanol or ethanol of various concentrations; after adding ethanol for precipitation, the ethanol final concentration of the mixed liquid can be various concentrations of 5% to 959%.

For the column separation in the method of the present invention, the column fillers used can be various fillers such as silica gel, polyamide, macroporous resin, and dextran gel.

The organic solvent elution in the method of the present invention, organic solvent can adopt various common organic solvents such as ethanol, methyl alcohol, ethyl acetate. The concentration of the organic solvent can be used in various concentrations, including a concentration of 100% organic solvent.

For the recrystallization in the method of the present invention, the solvent can be water or an organic solvent, and the organic solvent can be various common organic solvents such as ethanol, methanol, and ethyl acetate. The number of times of recrystallization can be from one to ten times, and the number of times can be increased without limitation.

By adopting the method of the present invention, naringin (pure product) is extracted from pomelo Citus grandis (L) Osbeck and its variant Shatian pomelo, etc., and the extraction yield is all above 85%, and the extraction yield is high.

Adopt the naringin monomer (pure product) that the inventive method obtains, through " Pharmacopoeia of the People's Republic of China " 2000 edition high-performance liquid chromatography is measured, carry out with the naringin reference substance provided by Chinese medicine and biological product test The external standard method is used for quantification to obtain the content of naringin monomer in the extracted naringin sample. The results showed that the content of naringin monomer in the sample was higher than 95%. Show that the process of the present invention is feasible.

The method for extracting naringin of the present invention has the characteristics of simple extraction process, high extraction rate, high purity of extracted naringin monomer, and the like.

specific implementation

Below in conjunction with embodiment the present invention will be further described.

The percentages of solids in solid mixtures, liquids in liquids, and solids in liquids referred to in each example are calculated by wt/wt, vol/vol, and wt/vol, respectively, unless otherwise specified.

Example 1

Cut the young fruit of Shatian pomelo into decoction pieces, add 100°C water to submerge the medicinal materials, extract three times with water, one hour each time, and filter; the filtrate is concentrated into an extract, passed through a macroporous resin column, washed with water, and then washed with 70% ethanol Elution: collect the ethanol eluent, recover ethanol from the eluent, obtain a precipitate, recrystallize the precipitate three times with water, and spray dry to obtain naringin monomer. The yield was 88.6%.

Example 2

Pulverize the pomelo peel of Shatian pomelo, do not sieve, add 100°C water to submerge the sample, extract three times, each time for 1 hour, filter, combine the filtrate; concentrate the filtrate, add absolute ethanol until the ethanol concentration is 50%, let stand overnight, filter , and the supernatant was recovered with ethanol. Pass the extract through a polyamide column, elute with water first, and then elute with 30% ethanol; collect the ethanol eluate, recover ethanol from the eluate, obtain a precipitate, dry the precipitate, and recrystallize five times with absolute ethanol to obtain pomelo dermoside monomer. The yield was 86.3%.

Example 3

The young fruit of Shatian pomelo is cut into decoction pieces, extracted twice by 70°C water for 3 hours each time, filtered, and the filtrate is combined; the filtrate is concentrated into an extract, and the extract is added with 95% ethanol until the ethanol concentration is 60%, and left to stand Overnight, a precipitate was produced, filtered, and the precipitate was removed. The filtrate recovered ethanol, and the obtained liquid aluminum peroxide column was eluted with water first, and then 95% ethanol; the ethanol eluate was collected, recovered and concentrated, and the precipitate was obtained. Recrystallize four times to get naringin monomer. The yield was 85.8%.

Example 4

The processed Shatian pomelo is cut into decoction pieces, add 100℃ water to submerge the medicinal materials, extract twice, each time for 2 hours, filter, and combine the filtrate; the filtrate is concentrated to obtain extract, the extract is passed through a polyamide column, and firstly washed with water , and then eluted with 60% ethanol; the ethanol eluate was collected, ethanol was recovered from the eluate, the obtained liquid was left overnight, filtered to obtain a precipitate, and the precipitate was recrystallized four times with water to obtain naringin monomer. The yield was 89.4%.

Example 5

Purity Determination Experiment of Naringin Monomer

Take an appropriate amount of naringin monomer prepared by the above-mentioned extraction method, dissolve it in methanol and settle to volume in a volumetric flask, filter it with a microporous membrane (0.45um), inject it into a high-performance liquid chromatograph, and test it with the Chinese pharmaceutical and biological product test method. The provided naringin reference substance is quantified by the external standard method to obtain the content of naringin monomer in the extracted naringin sample. The results are shown in Table 1.

Chromatographic conditions: Agilentl100 high performance liquid chromatography (autosampler, vacuum degasser, quaternary pump, column thermostat, diode array detector); chromatographic column: MARKER ODS column (5um, 4.0×250mm); mobile phase : methanol-water-glacial acetic acid (35:61:4); detection wavelength: 283nm; column temperature: 30°C; flow rate: 1ml/min; injection volume: 5μl.

Table 1: Sample content determination test results

                                                              

                                                                            Measured naringin content

sample number

Peak area

(mg/ml) (μl)

Concentration (mg/ml) (%)

Naringin

0.1004 5 815.75 0.1004 100

Reference substance

Sample 1 0.1004 5 798.85 0.0983 98.0

Sample 2 0.1192 5 963.64 0.1186 99.5

It can be seen from the results in Table 1 that the content of naringin monomers in the extracted samples is all higher than 98%. It shows that the naringin sample obtained by the extraction method of the present invention has high purity.

In summary, it shows that the naringin extraction method of the present invention can effectively extract naringin monomer from pomelo, and the obtained naringin samples have high purity, and the content is higher than 95%. The process is simple and feasible.

Claims (8)

1. method of extracting naringin from shaddock may further comprise the steps successively:

1. with shaddock section or pulverizing,, filter merging filtrate through water extraction;

2. filtrate is condensed into medicinal extract, and upper prop or direct upper prop carry out post and separate the organic solvent wash-out behind the medicinal extract alcohol precipitation;

3. elutriant volatilizes solvent, gets the naringin crude product; Again through recrystallization repeatedly, the naringin monomer.

2. in accordance with the method for claim 1, it is characterized in that said water extraction is or/and carry out under the ultrasonic wave condition under room temperature to 100 ℃ condition; Extraction time is one to three time.

3. in accordance with the method for claim 2, the extraction time that it is characterized in that said water extraction is 0.1-24 hour.

4. in accordance with the method for claim 1, it is characterized in that the used ethanol of said medicinal extract alcohol precipitation is the ethanol of dehydrated alcohol or various concentration; After adding ethanol sedimentation, the ethanol ultimate density of mixing liquid is 5% to 95%.

5. in accordance with the method for claim 1, it is characterized in that it is silica gel that said post separates the column packing that adopts, polymeric amide, macroporous resin or dextrane gel filler.

6. in accordance with the method for claim 1, it is characterized in that the used organic solvent of said organic solvent wash-out is to comprise that concentration is the ethanol or the methyl alcohol of 100%6 various concentration, ethyl acetate.

7. in accordance with the method for claim 1, it is characterized in that the used solvent of said recrystallization is water or organic solvent commonly used, organic solvent commonly used is an ethanol, methyl alcohol, ethyl acetate; The number of times of recrystallization is for once to ten times.

8. according to the described method of one of claim 1 to 7, it is characterized in that described shaddock is the young fruit of shatian pomelo or its equal genus kind fruit or the pericarp or the full fruit of mature fruit.