Preparation method of a papaya mespilic acid component

Abstract

A preparation method of a papaya fruit acid component, comprising: mixing papaya powder with 0.1-0.5 mol / L inorganic acid aqueous solution according to a solid-liquid ratio of 1:10-1:30 to obtain a mixed solution A; extracting the mixed solution A at 50-100 DEG C for 1-6 h, then filtering and water washing to obtain an extraction residue and an extraction solution A; mixing the extraction residue with 0.1-0.5 mol / L inorganic base aqueous solution according to a solid-liquid ratio of 1:10-1:30 to obtain a mixed solution B; extracting the mixed solution B at 40-90 DEG C for 1-5 h, then filtering and water washing to obtain an extraction solution B; mixing the extraction solution A with the extraction solution B, carrying out acid-base neutralization, and concentrating to obtain an extraction solution C; carrying out macroporous adsorption resin purification on the extraction solution C, recovering inorganic salt and obtaining a fruit acid purified product. The acid-base two-step extraction method is adopted to realize simultaneous extraction of free fruit acid components and fruit acid components of combined glycosides in papaya, the preparation method is simple, the cost is low, the extraction rate is high, and the industrial implementation is easy.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of extraction and separation and purification of fruit acid components in Chaenomeles speciosa, and particularly relates to a preparation method of fruit acid components in Chaenomeles speciosa. BACKGROUND

[0002] Chaenomeles speciosa is the mature fruit of Chaenomeles speciosa in Rosaceae, and is widely distributed in Shaanxi, Hubei, Anhui, Jiangxi, Jiangsu and Shandong. The fruit of Chaenomeles speciosa contains a large amount of organic acid components, which makes the taste of the fruit sour and astringent, and the fruit is not suitable for raw food, and is mainly used for medicinal purposes. The fruit of Chaenomeles speciosa has the effects of relaxing muscles and meridians, relieving inflammation and pain, and stopping diarrhea, so that it is widely used.

[0003] The acid substances contained in the fruit of Chaenomeles speciosa are mainly triterpene acids, and common substances are oleanolic acid and ursolic acid, and the like. Oleanolic acid belongs to pentacyclic triterpenes, and is widely distributed in Chaenomeles speciosa, and usually exists in the form of free monomers and glycosides. Oleanolic acid is usually used as an auxiliary drug for liver diseases, and has the effects of improving liver function and treating hepatitis; and can be used for rheumatoid arthritis, soreness of waist and knees, acute and chronic hepatitis and the like. Oleanolic acid has the pharmacological properties of reducing liver damage, inhibiting liver fibroblast growth, and protecting chromosome damage. Ursolic acid is another pentacyclic triterpene compound that exists in a large amount in Chaenomeles speciosa, and mainly exists in the form of glycosides combined with sugar substances or free form. Ursolic acid has the functions of antibacterial, anti-inflammatory and anticancer.

[0004] It can be seen that the fruit acid components have significant biological activity and important application value, and therefore it is very important to establish a green, efficient, low-cost and easy-to-industrialize preparation method of fruit acid components. However, due to the great difference in physical and chemical properties such as polarity caused by the different molecular structures of glycosides and aglycones, the currently disclosed technology can only realize the single extraction of free fruit acid components or fruit acid glycosides in Chaenomeles speciosa, so that the extraction rate is low and a large amount of raw material resources are wasted, or a large amount of waste liquid is generated in the extraction process, so that it is difficult to be industrialized and popularized. In addition, there are a large amount of non-fruit acid components in the extract which need to be further removed, thereby increasing the cost. SUMMARY

[0005] One of the purposes of the present application is to provide a preparation method of fruit acid components in Chaenomeles speciosa, which realizes the simultaneous extraction of free fruit acid components and combined glycoside fruit acid components in Chaenomeles speciosa by adopting an acid-base two-step extraction method, and has the advantages of simple preparation method, low cost, high extraction rate and easy industrialization.

[0006] The second object of the present application is to provide a method for recycling salt components generated in the extraction of fruit acids, which realizes the recycling of the extraction waste liquid and the preparation of additional products through acid-base neutralization reaction, and is green, environmentally friendly, economical and efficient, and has good market application prospects.

[0007] To achieve the above object, the present application provides a preparation method of fruit acid components in papaya, which comprises the following steps:

[0008] 1) The papaya powder is mixed with 0.1-0.5 mol / L inorganic acid aqueous solution according to a solid-liquid ratio of 1:10-1:30 to obtain a mixed liquid A;

[0009] 2) The mixed liquid A is extracted at 50-100℃ for 1-6h by solvent extraction method, and then filtered and washed with water to obtain an extraction residue and an extraction liquid A;

[0010] 3) The extraction residue obtained in step 2) is mixed with 0.1-0.5 mol / L inorganic base aqueous solution according to a solid-liquid ratio of 1:10-1:30 to obtain a mixed liquid B;

[0011] 4) The mixed liquid B is extracted at 40-90℃ for 1-5h by solvent extraction method, and then filtered and washed with water to obtain an extraction liquid B;

[0012] 5) The extraction liquid A obtained in step 2) is mixed with the extraction liquid B obtained in step 4) to perform acid-base neutralization, and then concentrated to obtain an extraction liquid C;

[0013] 6) The extraction liquid C obtained in step 5) is purified by macroporous adsorption resin to recover inorganic salt and obtain a fruit acid purified product.

[0014] As a further preferred technical solution of the present application, the inorganic acid in the inorganic acid aqueous solution is one or more of sulfuric acid, hydrochloric acid, phosphoric acid and nitric acid.

[0015] As a further preferred technical solution of the present application, the inorganic base in the inorganic base aqueous solution is one or more of potassium hydroxide, calcium hydroxide, sodium hydroxide, potassium carbonate and sodium carbonate.

[0016] As a further preferred technical solution of the present application, the macroporous adsorption resin has a particle size of 0.3-1.25mm.

[0017] As a further preferred technical solution of the present application, the macroporous adsorption resin is D101, HP-20, HPD100, X-5, AB-8, LSA-40 or DM130.

[0018] As a further preferred technical solution of the present application, in step 6), the specific operation of macroporous adsorption resin purification is as follows:

[0019] The extract C is prepared into a sample solution, 1-5 BV of the sample solution is added into the macroporous resin column at a loading flow rate of 0.5-2 BV / h for adsorption, and 1-3 BV of water is added for elution to obtain recovered inorganic salts; then ethanol solution is used for desorption; and the desorption solution is concentrated and dried to obtain fruit acid purified product.

[0020] As a further preferred technical solution of the present application, the extract C is prepared into a sample solution with a concentration of 0.5-3 mg / mL.

[0021] As a further preferred technical solution of the present application, the ethanol solution used for desorption has a concentration of 30-80% and a dosage of 2-5 BV.

[0022] The preparation method of the fruit acid components in the papaya of the present application adopts the acid-base two-step extraction method, and can realize the simultaneous extraction of free fruit acid components and fruit acid components in the form of glycosides in the papaya, and has high extraction rate. In the specific method: in the first step, the fruit acid components in the papaya are extracted by using inorganic acid solution, which can not only realize the extraction of part of the free fruit acid components, but also can hydrolyze the fruit acid glycosides, so that the fruit acid glycosides are converted into free fruit acid components and extracted; in the second step, the extraction residue after the first step extraction is extracted again by using inorganic alkali solution, which can react with the fruit acid components to form salts, thereby increasing the water solubility and being more helpful for the dissolution of the fruit acid components; finally, the acid extraction liquid and the alkali extraction liquid are mixed and neutralized to obtain the free fruit acid component extract, and then the extract is purified by using the macroporous adsorption resin purification technology to recover the fruit acid purified product and inorganic salt components, so that the simple, efficient, green, and low-cost preparation of the fruit acid components in the papaya is realized. In addition, the sodium salt, potassium salt, calcium salt, phosphorus salt, and nitrate salt and other substances recovered after the neutralization of the acid extraction liquid and the alkali extraction liquid have great application value in the fields of biological medicine, agricultural fertilizer, and food additive. BRIEF DESCRIPTION OF DRAWINGS

[0023] The present application will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0024] Figure 1 The flow chart of the preparation method of the fruit acid components in the papaya of the present application.

[0025] The purpose implementation, functional characteristics, and advantages of the present application will be further described with reference to the embodiments and the accompanying drawings. DETAILED DESCRIPTION

[0026] The specific embodiments of the present application will be described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present application, and are not used to limit the present application.

[0027] Unless otherwise defined, the technical terms used in the following examples have the same meanings as commonly understood by those skilled in the art to which the present invention belongs. The test reagents used in the following examples are all conventional biochemical reagents unless otherwise specified; the experimental methods used are all conventional methods unless otherwise specified.

[0028] The following extraction processes all use solvent extraction method.

[0029] Example 1

[0030] The preparation method is described in the flowchart shown in Figure 1

[0031] 1. Take 100 g of papaya powder, add 3000 mL of 0.1 mol / L sulfuric acid aqueous solution, mix uniformly, extract at 100℃ for 1 h, filter and wash with water after extraction is completed, to obtain extraction residue and extraction liquid A (also known as acid extraction liquid);

[0032] 2. Take 3000 mL of 0.1 mol / L sodium hydroxide aqueous solution, add to the above extraction residue, mix thoroughly, extract at 60℃ for 4 h, then filter and wash to obtain extraction liquid B (also known as alkali extraction liquid);

[0033] 3. Combine extraction liquid A and extraction liquid B in equal volume, perform acid-base neutralization, and then perform vacuum concentration by rotary evaporator to obtain extraction liquid C;

[0034] 4. Prepare the extraction liquid C into a sample solution with a concentration of 1.5 mg / mL, load 2 BV of the sample solution into the D101 macroporous resin column at a loading flow rate of 1.0 BV / h. Then load 1 BV of water for elution, perform vacuum distillation on the eluate to obtain recovered inorganic salts. Then use 3 BV of 50% ethanol solution for desorption, dry the desorption liquid to obtain fruit acid purified product.

[0035] Example 2

[0036] 1. Take 100 g of papaya powder, add 2000 mL of 0.15 mol / L hydrochloric acid aqueous solution, mix uniformly, extract at 80℃ for 2 h, filter and wash with water after extraction is completed, to obtain extraction residue and extraction liquid A;

[0037] 2. Take 2000 mL of 0.15 mol / L potassium hydroxide aqueous solution, add to the above extraction residue, mix thoroughly, extract at 70℃ for 3 h, then filter and wash to obtain extraction liquid B;

[0038] 3. Combine extraction liquid A and extraction liquid B in equal volume, perform acid-base neutralization, and then perform vacuum concentration by rotary evaporator to obtain extraction liquid C;

[0039] ​4. The extract C was prepared into a sample solution with a concentration of 1 mg / mL, and 1 BV of the sample solution was added into the HP-20 macroporous resin column for adsorption at a loading flow rate of 1.5 BV / h. Then 1 BV of water was added for elution, and the eluate was subjected to reduced pressure distillation to recover the inorganic salts. Then 2 BV of 60% ethanol solution was used for desorption, and the desorption solution was concentrated and dried to obtain the fruit acid purified product.

[0040] Example 3

[0041] 1. 100 g of the papaya powder was mixed with 1500 mL of 0.2 mol / L nitric acid aqueous solution, and then extracted at 70°C for 4 h. After extraction, the extract residue and extract A were obtained by filtration and water washing;

[0042] 2. 1500 mL of 0.2 mol / L potassium hydroxide aqueous solution was added to the extract residue, and then extracted at 80°C for 2 h after mixing. Then the extract B was obtained by filtration and water washing;

[0043] 3. The extract A and the extract B were mixed in equal volume, and then subjected to acid-base neutralization and reduced pressure concentration by a rotary evaporator to obtain the extract C;

[0044] 4. The extract C was prepared into a sample solution with a concentration of 2 mg / mL, and 1 BV of the sample solution was added into the X-5 macroporous adsorption resin column for adsorption at a loading flow rate of 2 BV / h. Then 2 BV of water was added for elution, and the eluate was subjected to reduced pressure distillation to recover the inorganic salts. Then 3 BV of 70% ethanol solution was used for desorption, and the desorption solution was concentrated and dried to obtain the fruit acid purified product.

[0045] The yield and purity of the fruit acid purified products prepared in Examples 1-3 were detected, and the specific results are shown in Table 1. The yield and purity of the fruit acid were calculated by high performance liquid chromatography, and were calculated based on the amount of oleanolic acid and ursolic acid. The liquid chromatography detection reference conditions were as follows: Shimadzu LC-10ATvp high performance liquid chromatograph, Kromasil-C18 (4.6 mm x 200 mm, 5 μm) chromatographic column, methanol-0.2% acetic acid (75:25) as the mobile phase, sample volume 20 μL, detection wavelength 210 nm, flow rate 1.0 mL / min, column temperature 25°C.

[0046] Table 1. Yield and purity of oleanolic acid and ursolic acid

[0047]

[0048] Although the specific embodiments of the present application have been described above, it is understood by those skilled in the art that these are merely illustrative and various changes or modifications can be made to the present embodiments without departing from the principles and the spirit of the present application, and the scope of protection of the present application is defined only by the appended claims.

Claims

1. A method for preparing a papaya mespilic component, characterized by, The method comprises the following steps: 1) mixing the wood apple powder with 0.1-0.5 mol / L aqueous solution of inorganic acid at a solid-liquid ratio of 1:10-1:30 to obtain a mixture A; 2) extracting the mixture A at 50-100 ℃ for 1-6 h by solvent extraction, and then filtering and washing with water to obtain an extraction residue and an extraction liquid A; 3) mixing the extraction residue obtained in step 2) with 0.1-0.5 mol / L aqueous solution of inorganic base at a solid-liquid ratio of 1:10-1:30 to obtain a mixture B; 4) extracting the mixture B at 40-90 ℃ for 1-5 h by solvent extraction, and then filtering and washing with water to obtain an extraction liquid B; 5) mixing the extraction liquid A obtained in step 2) with the extraction liquid B obtained in step 4), and then performing acid-base neutralization and concentration to obtain an extraction liquid C; 6) purifying the extraction liquid C obtained in step 5) by macroporous adsorption resin, and recovering inorganic salt to obtain a fruit acid purified product.

2. The method of claim 1, wherein the preparation of the papaya mesocarpic acid components is characterized by, The inorganic acid in the aqueous solution of inorganic acid is one or more of sulfuric acid, hydrochloric acid, phosphoric acid and nitric acid.

3. The method of claim 1, wherein the preparation of the papaya mesocarpic acid components is characterized by, The inorganic base in the aqueous solution of inorganic base is one or more of potassium hydroxide, calcium hydroxide, sodium hydroxide, potassium carbonate and sodium carbonate.

4. The method of claim 1, wherein the preparation of the papaya mesocarpic acid components is characterized by, The macroporous adsorption resin has a particle size of 0.3-1.25 mm.

5. The method of claim 1, wherein the preparation of the papaya mesocarpic acid components is characterized by, The macroporous adsorption resin is one or more of D101, HP-20, HPD100, X-5, AB-8, LSA-40 and DM130.

6. The method of claim 1, wherein the preparation of the papaya mesocarpic acid components is characterized by, In step 6), the specific operation of macroporous adsorption resin purification is as follows: The extraction liquid C is prepared into a sample solution with a concentration of 0.5-3 mg / mL, and 1-5 BV of the sample solution is added to the macroporous resin column for adsorption at a sample loading flow rate of 0.5-2 BV / h, and 1-3 BV of water is added for elution to recover inorganic salt; then 30%-80% ethanol solution is used for desorption, the amount of ethanol is 2-5 BV, and the desorption liquid is concentrated and dried to obtain the fruit acid purified product.

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