Method for separating multiple components from raspberry fruits

Abstract

The invention provides a method for separating multiple components from raspberry fruits, and belongs to the technical field of biological extraction. The method comprises the following steps: adding water into raspberry fruits, pulping, heating, cooling, centrifuging, collecting centrifugate for later use, carrying out alcohol extraction on centrifugal residues, filtering, concentrating an alcohol extract until no alcohol smell exists, mixing the concentrated alcohol extract with the centrifugate, centrifuging again, filtering the centrifugate by a ceramic membrane, enabling filtrate to respectively pass through XDA-6 and 001 * 7 resin, and drying to obtain the raspberry extract. Concentrating the effluent to thick paste, adding high alcohol to carry out alcohol precipitation treatment to obtain a polysaccharide component as a precipitate, and concentrating and drying a supernatant part to obtain monosaccharide and inorganic salt; xDA-6 is eluted to obtain anthocyanin components, and 001 * 7 is eluted to obtain organic acid components. According to the method, a series of components in the raspberry fruits can be extracted, the extraction process is simple, convenient and feasible, the extraction efficiency is high, and a foundation is laid for development of downstream products of the raspberry fruits.

Description

Technical Field

[0001] The invention belongs to the technical field of biological extraction, and in particular relates to a method for separating multiple components from mountain berries. Background Art

[0002] Raspberry is a plant of the genus Rubus in the Rosaceae family. Its Latin name is Rubus corchorifolius Lf. It is commonly found on sunny slopes, stream banks, valleys, wastelands and moist places in dense bushes at an altitude of 200-2200 meters. The fruit is sweet and contains sugar, malic acid, citric acid and vitamin C. It can be eaten raw, made into jam and brewed wine. The fruit, roots and leaves are used as medicine, which has the effects of promoting blood circulation, detoxifying and stopping bleeding. The flowering period is from February to March, and the fruiting period is from April to June. Due to its wide distribution and thorny body, it has not been artificially introduced and cultivated on a large scale, except for a few transplants used for fence isolation.

[0003] A Chinese invention patent with publication number CN101628863A discloses a method for extracting luteolin and raspberry from fresh raspberry fruit, including extracting fresh raspberry fruit with alcohol, fermenting with Trichoderma, separating by adsorption with silica gel column and macroporous resin, eluting, concentrating and crystallizing to obtain pure luteolin and raspberry. A Chinese invention patent with publication number CN108078855A discloses a raspberry fruit extract and its extraction process, including washing, air-drying, crushing and filtering the fresh fruit, and then treating with alcohol and alkaline salt, concentrating and extracting with ethyl acetate to obtain high-content amino acids. Both patents extract and separate the components of raspberry fruit separately, but only study luteolin, raspberry and amino acids, and there is no relevant report on the systematic extraction and separation of raspberry fruit components. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a method for separating multiple components from raspberries, which has a simple process flow and can systematically separate anthocyanins, organic acids, polysaccharides, and monosaccharides / inorganic salts in raspberries. The components are classified and segmented, and the product is simple and easy to obtain.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0006] The invention provides a method for separating multiple components from raspberry fruit, comprising the following steps: beating the raspberry fruit into pulp, heating the pulp in a water bath, then cooling the pulp in an ice water bath to room temperature, centrifuging the cooled pulp to obtain a centrifuge liquid and centrifuge residues; mixing the centrifuge residues with ethanol, filtering to obtain an ethanol extract, concentrating, then mixing the concentrate with the centrifuge liquid, and centrifuging again to obtain a clarified centrifuge liquid; passing the clarified centrifuge liquid through XDA-6 and 001×7 series resin columns in sequence, eluting with water after complete adsorption, and collecting an effluent liquid and a water eluate; disconnecting the XDA-6 and 001×7 series resin columns, eluting the XDA-6 with ethanol, and removing the effluent liquid from the XDA-6 with ethanol. resin column, collecting ethanol eluate, eluting 001×7 resin column with ammonia water, collecting ammonia eluate; concentrating the effluent and water eluate, the ethanol eluate, and the ammonia eluate to obtain effluent and water eluate concentrates, ethanol eluate concentrates, and ammonia eluate concentrates respectively; mixing the effluent and water eluate concentrates with ethanol and standing, separating the supernatant and the precipitate, and concentrating the supernatant to obtain a supernatant concentrate; drying the ethanol eluate concentrate, the ammonia eluate concentrate, the supernatant concentrate, and the precipitate respectively to obtain anthocyanins, organic acids, monosaccharides / inorganic salts, and polysaccharides.

[0007] Preferably, during the process of beating the wild berries into pulp, the mass ratio of the wild berries to water is 1:0.5-2.5.

[0008] Preferably, the water bath heating temperature is 65-72°C and the heating time is 25-35 minutes.

[0009] Preferably, the cooling pulp is centrifuged at a speed of 3500-4500 rpm.

[0010] Preferably, during the mixing process of the centrifugal residue and ethanol, the volume percentage of ethanol is 90-98% ethanol aqueous solution, and the mixing and stirring time is 1.5-2.5 hours.

[0011] Preferably, the flow rate of the clarified centrifuge liquid through the XDA-6 and 001×7 resin columns connected in series is 3-5 L / h.

[0012] Preferably, the ethanol used to elute the XDA-6 resin column is a 45-55% by volume ethanol aqueous solution.

[0013] Preferably, the concentration of aqueous ammonia used for eluting the 001×7 resin column is 2-6% by volume.

[0014] Preferably, the parameters during the standing of the effluent and the concentrated water eluate with ethanol include: the ethanol is 92-98% ethanol aqueous solution by volume, and the standing time is 10-14 hours.

[0015] Preferably, the drying is spray drying or vacuum drying.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention proposes for the first time a method for systematically extracting anthocyanins, organic acids, polysaccharides and monosaccharides / inorganic salts from raspberries, which can obtain anthocyanins, organic acids, polysaccharides and monosaccharides / inorganic salts with relatively high yield and high content. The extraction process of the present invention is simple, the components are classified and segmented, and the product is simple and easy to obtain. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a process diagram for separating multiple components from raspberries. DETAILED DESCRIPTION

[0019] The invention provides a method for separating multiple components from raspberry fruit, comprising the following steps: (1) beating the raspberry fruit into pulp, heating the pulp in a water bath, then cooling the pulp in an ice water bath to room temperature, centrifuging the cooled pulp to obtain a centrifuge liquid and centrifuge residues; mixing the centrifuge residues with ethanol, filtering to obtain an ethanol extract, concentrating the centrifuge residues, then mixing the concentrate with the centrifuge liquid, and centrifuging again to obtain a clarified centrifuge liquid; (2) passing the clarified centrifuge liquid through XDA-6 and 001×7 series resin columns in sequence, eluting with water after complete adsorption, and collecting the effluent and the water eluate; (3) disconnecting the XDA-6 and 001×7 series resin columns, eluting the XDA-6 with ethanol, and removing the ethanol extract from the XDA-6 with ethanol. Resin column, collect ethanol eluate, elute 001×7 resin column with ammonia water, collect ammonia eluate; (4) concentrate the effluent and water eluate, the ethanol eluate, and the ammonia eluate to obtain effluent and water eluate concentrate, ethanol eluate concentrate, and ammonia eluate concentrate; (5) mix the effluent and water eluate concentrate with ethanol and let stand, separate the supernatant and the precipitate, and concentrate the supernatant to obtain a supernatant concentrate; (6) dry the ethanol eluate concentrate, the ammonia eluate concentrate, the supernatant concentrate, and the precipitate to obtain anthocyanin, organic acid, monosaccharide / inorganic salt, and polysaccharide. The present invention uses fresh raspberries picked, selects and removes impurities, and washes with deionized water 1-3 times. The raspberry of the present invention is an immature fresh fruit of the plant raspberry of the genus Rubus of the Rosaceae family, with a color of yellow to orange to red and firm flesh. The resin column of the present invention can be purchased through commercial channels.

[0020] In the present invention, during the process of beating the raspberries into pulp, the mass ratio of the raspberries to water is 1:0.5-2.5, preferably 1:1. The pulping of the present invention can be carried out using a conventional food processor.

[0021] In the present invention, the water bath heating temperature is 65-72°C, the time is 25-35min; preferably the temperature is 68°C, the time is 30min. The water bath heating time of the present invention starts when the temperature rises to 62-68°C, preferably starts at 65°C. The purpose of the water bath heating of the present invention is to kill enzymes and sterilize, and at the same time facilitate the extraction of effective ingredients.

[0022] In the present invention, the speed of the centrifugal cooling of the pulp is 3500-4500rpm, preferably 4000rpm. The present invention centrifuges the cooled pulp to obtain centrifugal residue and centrifugal liquid. In the process of mixing the centrifugal residue with ethanol, the volume percentage of ethanol is 90-98% ethanol aqueous solution, preferably 95%, and the mixing and stirring time is 1.5-2.5h, preferably 2h. The present invention concentrates the ethanol extract after mixing and filtering the centrifugal residue with ethanol until there is no alcohol taste.

[0023] In the present invention, the flow rate of the clarified centrifuge liquid through the XDA-6 and 001×7 series resin columns is 3-5 L / h, preferably 4 L / h. After complete adsorption, it is eluted with deionized water, and the effluent and the water eluate are collected.

[0024] In the present invention, the ethanol used to elute the XDA-6 resin column is a 45-55% ethanol aqueous solution by volume, preferably a 50% ethanol aqueous solution. The flow rate of the 50% ethanol aqueous solution used to elute the XDA-6 resin column is 3-5 L / h, preferably 4 L / h.

[0025] In the present invention, the concentration of ammonia water used to elute the 001×7 resin column is 2-6% by volume, preferably 4%. The flow rate of 4% ammonia water used to elute the 001×7 resin column is 3-5 L / h, preferably 4 L / h.

[0026] In the present invention, the parameters of the process of mixing the effluent and the concentrated water eluate with ethanol and standing still include: ethanol is 92-98% ethanol aqueous solution by volume, and the standing time is 10-14h. The effluent and the concentrated water eluate of the present invention are mixed and stirred with ethanol for 8-20min and then stood still.

[0027] In the present invention, the drying is spray drying or vacuum drying. The precipitate of the present invention is directly vacuum dried or dissolved and then spray dried to obtain polysaccharides. The ethanol eluate concentrate, ammonia eluate concentrate and supernatant concentrate of the present invention are spray dried or vacuum dried to obtain anthocyanins, organic acids, monosaccharides and polysaccharides / inorganic salts.

[0028] In the present invention, unless otherwise specified, all components or reagents are commercially available products well known to those skilled in the art.

[0029] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] Example 1

[0031] 1. Raw material extraction: 5 kg of fresh raspberry fruit after impurities were removed, washed twice with deionized water, and beaten into pulp with an equal mass of deionized water using a fruit and vegetable processor in batches, 9.8 kg of pulp was weighed, transferred to a 20L round-bottom flask, heated in a water bath at 68°C, and the timing started when the temperature in the bottle reached 65°C. After 30 minutes, the round-bottom flask was taken out and the pulp was poured into a stainless steel barrel, placed in an ice water bath and cooled to room temperature. The cooled pulp was centrifuged (4000 rpm) to obtain 7.5 kg of centrifugal liquid and 2.2 kg of centrifugal residue; 6 kg of 95% (v / v%) ethanol aqueous solution was added to the centrifugal residue, stirred for 2 hours, filtered to obtain an ethanol extract, concentrated until there was no alcohol taste, added to the centrifugal liquid, stirred and dispersed for 30 minutes, and centrifuged again to obtain 8.6 kg of clarified centrifugal liquid, and both the extraction residue and the centrifugal residue were discarded.

[0032] 2. Elution from resin column: Pass the clarified centrifuge liquid through XDA-6 (1 kg) and 001×7 (1 kg) series resin columns at a flow rate of 4 L / h. After complete adsorption, elute with 4 L of deionized water, and collect the effluent and water eluate. Disconnect the series resin columns, elute the XDA-6 resin column with 4 L of 50% (v / v%) ethanol aqueous solution at a flow rate of 4 L / h, and collect 4.2 L of ethanol eluate; elute the 001×7 resin column with 6 L of 4% (v / v%) ammonia water at a flow rate of 4 L / h, and collect 6.1 L of ammonia water eluate.

[0033] 3. Concentration: The XDA-6 ethanol eluate was concentrated (the concentration temperature was less than 55°C and the vacuum degree was less than -0.095 MPa) to obtain 95 g of ethanol eluate concentrate; the 001×7 ammonia eluate was concentrated (the concentration temperature was less than 55°C and the vacuum degree was less than -0.095 MPa) to obtain 113 g of ammonia eluate concentrate; the effluent and water eluate were concentrated (the concentration temperature was less than 55°C and the vacuum degree was less than -0.095 MPa) to obtain 431 g of effluent and water eluate concentrate.

[0034] 4. Alcohol precipitation: add 431g of the effluent and the concentrated water washing liquid to 1500ml of 95% (v / v%) ethanol aqueous solution, stir and disperse for 10min, let stand for 12h, separate the supernatant and the bottom precipitate, and concentrate the supernatant (concentration temperature is less than 55°C, vacuum degree is less than -0.095Mpa) to obtain 169g of supernatant concentrated thick paste (supernatant concentrate).

[0035] 5. Drying: The ethanol elution concentrate (spray drying: inlet air temperature 180-200°C, outlet air temperature 95-105°C, tower temperature 90-95°C), the ammonia elution concentrate (spray drying: inlet air temperature 180-200°C, outlet air temperature 95-105°C, tower temperature 90-95°C), the supernatant concentrate (spray drying: inlet air temperature 180-200°C, outlet air temperature 95-105°C, tower temperature 90-95°C) and the precipitate (vacuum drying, temperature 55-60°C, vacuum degree -0.098 MPa) were dried respectively to obtain 32.2 g of anthocyanins as the elution component of XDA-6, 46.6 g of organic acids as the elution component of 001×7, 121.1 g of the monosaccharide / inorganic salt portion of the supernatant, and 48.5 g of the polysaccharide portion of the precipitate.

[0036] Example 2

[0037] 1. Raw material extraction: Take 200 kg of fresh raspberry fruit after impurities are removed, wash it twice with deionized water, add 200 kg of water to squeeze the juice and beat it into pulp, transfer it into a 1000L multifunctional tank, steam heat, control the pulp temperature to 65-70°C, heat for 30 minutes (start timing after reaching the temperature), close the steam valve, open the condensation water to cool it to room temperature. Centrifuge to obtain centrifuge liquid, add 100 kg of 95% ethanol aqueous solution to 57 kg of centrifuge residue, stir for 2 hours, filter to obtain ethanol extract, concentrate until there is no alcohol taste, add it to the centrifuge liquid, stir and disperse for 30 minutes, and then obtain clarified centrifuge liquid by high-speed centrifugation, and discard the extract residue and centrifuge residue.

[0038] 2. Resin column elution: pass the clarified centrifuge liquid through DXA-6 (50kg) and 001×7 (50kg) resin columns in series at a flow rate of 2BV / h (2 column volumes per hour). After complete adsorption, elute with 2BV of deionized water, collect and combine the effluent and water eluate; elute the XDA-6 resin column with 2BV of 50% ethanol aqueous solution, and collect the eluate; elute the 001×7 resin column with 3BV of 4% ammonia water, and collect the eluate.

[0039] 3. Concentration: The XDA-6 ethanol eluate is concentrated (the concentration temperature is less than 55°C, and the vacuum degree is less than -0.095 MPa) to obtain an ethanol eluate concentrate; the 001×7 ammonia eluate is concentrated (the concentration temperature is less than 55°C, and the vacuum degree is less than -0.095 MPa) to obtain an ammonia eluate concentrate; the effluent and the water eluate are concentrated (the concentration temperature is less than 55°C, and the vacuum degree is less than -0.095 MPa) to obtain an effluent and water eluate concentrate.

[0040] 4. Alcohol precipitation: 21 kg of effluent and water eluate were concentrated into extract, 80 kg of 95% ethanol aqueous solution was added, stirred for 10 min, allowed to stand overnight, and the supernatant was sucked into a concentrator for concentration to obtain 8.3 kg of supernatant concentrated thick paste (supernatant concentrate), and the bottom precipitate weighed 3.7 kg.

[0041] 5. Drying: Dry the ethanol elution concentrate (spray drying: inlet air temperature 180-200°C, outlet air temperature 95-105°C, tower temperature 90-95°C), ammonia elution concentrate (spray drying: inlet air temperature 180-200°C, outlet air temperature 95-105°C, tower temperature 90-95°C), supernatant concentrate (spray drying: inlet air temperature 180-200°C, outlet air temperature 95-105°C, tower temperature 90-95°C) and precipitate (vacuum drying, temperature 55-60°C, vacuum degree -0.098Mpa) respectively to obtain 0.87kg of anthocyanins in the XDA-6 ethanol elution part, 1.64kg of organic acid in the 001×7 eluate part, 5.36kg of monosaccharide / inorganic salt part in the alcohol precipitation supernatant, and 1.25kg of polysaccharide part in the alcohol precipitation.

[0042] Comparative Example 1

[0043] 1. Raw material extraction: Take 5 kg of ripe raspberry fruits, wash them twice with deionized water, and beat them into pulp in batches with a fruit and vegetable processor with an equal mass of deionized water. Weigh and obtain 9.8 kg of pulp, transfer it into a 20L round-bottom flask, heat it in a 68°C water bath, and start timing when the temperature in the bottle reaches 65°C. After 30 minutes, take out the round-bottom flask and pour the pulp into a stainless steel barrel, place it in an ice water bath and cool it to room temperature. Centrifuge the cooled pulp (4000 rpm) to obtain 7.5 kg of centrifuge, and then pass the centrifuge through a high-speed centrifuge to obtain clarified centrifuge.

[0044] 2. Elution: Pass the clarified centrifuge liquid through a series of D101 (1kg) and 001×7 (1kg) resin columns, elute with 4L of deionized water after complete adsorption, and collect the effluent and water eluate. Disconnect the series of resin columns, elute the D101 resin column with 4L of 50% ethanol, and collect 4.3L of eluate; elute the 001×7 resin column with 6L of 4% ammonia water, and collect 6.1L of eluate.

[0045] 3. Concentration: The D101 ethanol eluate was concentrated to obtain 100 g of concentrated solution; the 001×7 eluate was concentrated to obtain 122 g of concentrated solution; the effluent and water washing solution were concentrated to obtain 457 g of concentrated solution.

[0046] 4. Alcohol precipitation: add 457g of the effluent and the concentrated solution of the water washing solution into 1500ml of 95% ethanol, stir and disperse for 10min, let stand for 12h, separate the supernatant and the bottom precipitate 122g, and concentrate the supernatant to obtain 140g of thick paste.

[0047] 5. Drying: Dry the ethanol elution concentrate, ammonia elution concentrate, supernatant paste and precipitate separately to obtain 48.6 g of D101 part, 32.4 g of 001×7 part, 98.7 g of supernatant part and 38.5 g of precipitate part respectively.

[0048] Experimental Example 1

[0049] The yield of each extracted component and the content of each dried component were statistically analyzed. The content was detected using a universal UV detection method. The results are shown in Table 1.

[0050] Table 1 Yield and content of each extracted component

[0051]

[0052] As shown in Table 1, the yield and content of each extracted component obtained by the method proposed in the present invention are higher than those in Comparative Example 1, and the selected XDA-6 and 001×7 resin columns can obtain high content of anthocyanins, organic acids and polysaccharides.

[0053] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for separating multiple components from raspberries, characterized in that: The steps include: The raspberry fruit is beaten into pulp, the pulp is heated in a water bath, and then placed in an ice water bath to cool to room temperature, and the cooled pulp is centrifuged to obtain a centrifuge liquid and centrifuge residues; the centrifuge residues are mixed with ethanol, filtered to obtain an ethanol extract, concentrated, and then the concentrate is mixed with the centrifuge liquid, and centrifuged again to obtain a clarified centrifuge liquid; The clarified centrifuge liquid was passed through XDA-6 and 001×7 series resin columns in sequence, and after complete adsorption, it was eluted with water, and the effluent and water eluate were collected; Disconnect the XDA-6 and 001×7 resin columns connected in series, elute the XDA-6 resin column with ethanol, collect the ethanol eluate, elute the 001×7 resin column with ammonia water, and collect the ammonia water eluate; The effluent and water eluent, the ethanol eluent, and the ammonia eluent are respectively concentrated to obtain an effluent and water eluent concentrate, an ethanol eluent concentrate, and an ammonia eluent concentrate; The effluent and the concentrated solution of the water eluate are mixed with ethanol and allowed to stand, the supernatant and the precipitate are separated, and the supernatant is concentrated to obtain a supernatant concentrate; The ethanol eluate concentrate, the ammonia eluate concentrate, the supernatant concentrate and the precipitate are dried respectively to obtain anthocyanins, organic acids, monosaccharides / inorganic salts and polysaccharides.

2. The method according to claim 1, characterized in that In the process of beating the wild berries into pulp, the mass ratio of the wild berries to water is 1:0.5-2.

5.

3. The method according to claim 1, characterized in that The water bath heating temperature is 65-72°C and the heating time is 25-35 minutes.

4. The method according to claim 1, characterized in that The rotation speed of the cooling pulp centrifuge is 3500-4500rpm.

5. The method according to claim 1, characterized in that During the mixing process of the centrifugal residue and ethanol, the volume percentage of ethanol is 90-98% ethanol aqueous solution, and the mixing and stirring time is 1.5-2.5 hours.

6. The method according to claim 1, characterized in that The flow rate of the clarified centrifuge liquid through the XDA-6 and 001×7 series resin columns is 3-5 L / h.

7. The method according to claim 1, characterized in that The ethanol used to elute the XDA-6 resin column is a 45-55% by volume ethanol aqueous solution.

8. The method according to claim 1, characterized in that The concentration of ammonia water used to elute the 001×7 resin column is 2-6% by volume.

9. The method according to claim 1, characterized in that The parameters of the process of mixing the effluent and the concentrated solution of the water eluate with ethanol and standing still include: the ethanol is 92-98% ethanol aqueous solution by volume, and the standing time is 10-14 hours.

10. The method according to claim 1, characterized in that The drying is spray drying or vacuum drying.

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