WO2015032212A1

WO2015032212A1 - Method for extracting chlorogenic acid from green coffee beans

Info

Publication number: WO2015032212A1
Application number: PCT/CN2014/077364
Authority: WO
Inventors: 罗明锋; 房杰; 何勇; 李改丽
Original assignee: 四川圣湖生物科技有限公司
Priority date: 2013-09-04
Filing date: 2014-05-13
Publication date: 2015-03-12
Also published as: CN103497106B; CN103497106A

Abstract

The invention relates to a method for extracting chlorogenic acid from green coffee beans, which solves the problems of the high energy consumption, difficulty in adsorbent resin cleaning, long extraction time, low extraction rate, low product purity, and high caffeine content in the existing methods. A raw material is first crushed and sieved with a 10-mesh sieve, and is then subjected to continuous ultrasonic counter-current extraction using a 0.1% sodium sulfite aqueous solution; the extract solution is filtered with a 100-mesh vibrating sieve, subjected to horizontal spiral centrifugation and disk centrifugation, and then directly enters an XAD-16 macroporous resin for adsorption; the unadsorbed liquid is adsorbed by an LX-28 macroporous resin; after the adsorption is saturated, the resins are rinsed with purified water, and eluted with 50%-80% ethanol, the eluents are concentrated, ethanol is recovered, and then spray drying is carried out. The eluted product from the XAD-16 macroporous resin elution is caffeine. The eluted product from the LX-28 macroporous resin is 50% chlorogenic acid, with a yield of approximately 10%. The 50% chlorogenic acid is dissolved by using 10 times the amount thereof of 80% ethanol, subjected to active carbon decolorization and filtration, concentrated to a specific weight of 1.15, and left for freeze crystallization; and after microwave drying, a product with a content of 90% is obtained with a yield of above 3.5%.

Description

Method for extracting chlorogenic acid from green coffee beans

The invention relates to a process for extracting chlorogenic acid from green coffee beans, in particular to a method for purifying chlorogenic acid from green coffee beans by ultrasonic countercurrent extraction and macroporous resin separation, and effectively removing caffeine .

Background technique

Green coffee beans are derived from the seeds of the Rubiaceae plant, small fruit coffee, medium fruit coffee and large fruit coffee. The green coffee bean extract contains more chlorogenic acid and is therefore often used as a source of effective chlorogenic acid.

Chlorogenic acid is a condensed acid composed of caffeic acid and quinic acid. Its chemical name is 3 - caffeoyl quinic acid. It is known as "plant gold" and is a manufacturer. Chasing the object of production. Chlorogenic acid is an important biologically active substance with antibacterial, antiviral, elevated white blood cells, liver and gallbladder, anti-tumor, blood pressure lowering, blood lipid lowering, free radical scavenging and excitatory central nervous system. Modern science on green The research on the bioactivity of ortho-acids has penetrated into many fields such as food, health care, medicine and daily chemical industry. The main source of chlorogenic acid is extracted from natural plants. It has been used as a raw material for honeysuckle and eucommia in China, and mainly as coffee beans. At present, the main methods for extracting chlorogenic acid are:

1) Water extraction method: The water extraction method is a method in which chlorogenic acid is easily dissolved in water and hot water is used as a leaching agent. Using 10 times the amount of water, decocted at 50 °C for 2 times, each time boiling for 1.5 h, the crude chlorogenic acid was obtained. The water extraction process is simple, low in cost and low in investment. However, the energy consumption is high, the production cycle is long, the amount of wastewater is large, and the purity of the product is low.

2) Organic solvent extraction method: Chlorogenic acid is easily soluble in polar solvents such as methanol, ethanol and acetone. The mixture was refluxed with 10 times 70% ethanol for 1 h to obtain crude chlorogenic acid. The organic solvent extraction process is simple, the investment is small, and it is suitable for industrial production, but the solvent recovery cost is increased due to the use of organic solvents.

3) Physical field intensification extraction method: Adding physical fields such as ultrasonic or microwave to traditional solvent extraction, strengthening extraction, reducing extraction time and reducing degradation of active substances. The chlorogenic acid was obtained under the conditions of microwave power 340W, radiation time 20min, tincture ethanol volume fraction 60%, ethanol dosage 9 times, and soaking time 30 min. Product. However, the current industrial production technology is still immature and needs further research.

4) Macroporous resin adsorption method: Macroporous adsorption resin is a polymer adsorbent which has no exchange group and has a macroporous structure. It is also a lipophilic substance with good selectivity, high mechanical strength and convenient regeneration. It has the advantages of fast adsorption speed, simple operation, stable product quality and low production cost, so it is especially suitable for separating and purifying water-soluble compounds. CN 101704748A uses NKA-Π polar macroporous resin, the loading concentration is 0.26mg/mL, pH2.0, the sample volume/macroporous resin mass ratio is 12, the flow rate is 2mL/min, the eluent is 30% ethanol, The purity of the chlorogenic acid extract can reach 25.3%. However, this method takes a long time and it is difficult to clean the adsorbing resin.

In general, the above commonly used methods for extracting chlorogenic acid have their own drawbacks, such as: high energy consumption, long extraction time, low extraction rate, low product purity, and high caffeine content.

Summary of the invention

SUMMARY OF THE INVENTION The object of the present invention is to provide a method for extracting chlorogenic acid from green coffee beans with low energy consumption, easy cleaning of the adsorbing resin, low extraction time, high extraction rate, high product purity and low caffeine content.

The present invention is implemented as follows:

A method for extracting chlorogenic acid from green coffee beans, the extraction process comprising the following steps:

(1) First, the green coffee beans are crushed through a 10-mesh sieve to obtain a green coffee bean powder;

(2) Adding 10 times mass percent concentration of green coffee bean powder to 0.1% sodium sulfite aqueous solution, 4 (T60 ° C ultrasonic countercurrent continuous extraction, sodium sulfite aqueous solution flow rate is 5 times the mass of green coffee bean powder, obtaining green coffee bean extract ;

(3) filtering the extract in step (2) through a 100-mesh shaking sieve, centrifuging the snail, and centrifuging the disc to obtain a supernatant;

(4) The supernatant in step (3) is directly poured into XAD-16 macroporous resin. The XAD-16 macroporous resin acts to selectively adsorb coffee and thus does not adsorb chlorogenic acid. The ratio of resin adsorption to supernatant is 1:1; After the adsorption is saturated, rinse with purified water, elute with mass concentration of 5 (Γ80% ethanol, elution amount is 2 times the volume of the resin, collect the eluent;

(5) The XAD-16 macroporous resin loss solution in step (4) is poured into LX-28 macroporous resin, and the resin adsorption amount and the lost liquid ratio are 1:1; after the adsorption is saturated, it is rinsed with purified water, and the mass is used. The concentration was 5 (Γ80% ethanol eluted, the elution amount was 2 times the volume of the resin, and the eluate was collected;

(6) concentrating the eluate obtained in the step (4), recovering the ethanol, and spray-drying to obtain a caffeine by-product; (5) The eluate obtained is concentrated, the ethanol is recovered, and the chlorogenic acid product is spray-dried;

(7) The chlorogenic acid in step (6) is dissolved in 80% by mass of 80% by mass of ethanol, decolorized by activated carbon, and concentrated to a specific gravity of 1.15. Get chlorogenic acid products. The temperature of the crystallization is 5% of the mass of the chlorogenic acid, the temperature is 70 V, and the holding time is 20 min; the specific gravity of the crystal is 1. l (Tl. 15, the temperature is ( T4 ° C.

In step (1), the green coffee bean powder is required to pass through 10 mesh, and the range is 1 (Γ20 mesh, too large extraction is insufficient, and too fine extraction liquid is difficult to discharge;

The extract in the step (2) is 0.1% aqueous sodium sulfite solution, and the extract can ensure the weak alkalinity of the extract and increase the water solubility of the chlorogenic acid. The experiment shows that the enthalpy of the extract of the present invention is 7~7. 5, without adding a mass percent concentration of 0.1% sodium sulfite, the extract value is 5~6, and the solubility of chlorogenic acid is decreased by 16%. The second is to prevent the chlorogenic acid from being oxidized and to reduce the degradation of the chlorogenic acid. The experimental data proves that the degradation rate of the chlorogenic acid aqueous solution having a mass percentage of 5% in boiling water for 1 hour is 6.2%, and the degradation rate is 2 hours. 10.7%. The 5%, the degradation rate of the chlorogenic acid aqueous solution was 0.1%, and the degradation rate was 0.57%.

The extraction temperature in step (2) is 4 (T6 (TC, the temperature is too low, the extract is insufficient, too high material is easy to paste, which is not conducive to liquid discharge;

The extraction device in the step (2) is an ultrasonic continuous countercurrent extraction device. The advantage of the device is that the extraction temperature is not high, the flow of the material and the extract are opposite, the extraction rate is increased, and the oxidation of the chlorogenic acid is effectively reduced. Compared with the conventional multi-functional extraction tank, the extraction rate of chlorogenic acid increased from 87% to 96%, and the extraction effect increased by more than 10%. The filtration conditions in the step (3) are oscillating sieve filtration, horizontal snail centrifugation, and disc centrifugation to ensure clarification of the extract, and the macroporous resin is not blocked in the post-treatment process;

The adsorbing resin in the step (4) is a XAD-16 macroporous resin, and the resin selectively adsorbs caffeine in green coffee beans without adsorbing chlorogenic acid. The content of caffeine in 50% chlorogenic acid products in the market is generally above 5%. Since caffeine can increase nerve excitability, most customers hope that the lower the caffeine in 50% chlorogenic acid products, the better, in order to reduce caffeine. The content is currently mainly tetrachloromethane, and the lethal dose of tetrachloromethane is 4 ml. The XAD-16 macroporous resin of the present invention can solve the problem well, and the caffeine content of the 50% chlorogenic acid product treated by XAD-16 is less than 0.5%.

The adsorbing resin in the step (5) is a LX-28 macroporous resin, and the resin is a weakly polar resin, which can selectively adsorb chlorogenic acid, and the LX-28 resin has a large adsorption amount compared with other macroporous resins. , the adsorption is very concentrated. The eluent content is above 50%. The product eluted by LX-28 macroporous resin is chlorogenic acid with a content of 50%, and the yield is about 10% (that is, 100 kg of product per ton of raw material).

The method of the invention is a process technology for extracting and purifying chlorogenic acid from green coffee beans by using ultrasonic countercurrent extraction and two macroporous resin adsorption methods. The invention pulverizes the raw material through 10 mesh, extracts by ultrasonic wave, and filters by shaking sieve. The horizontal snail centrifugation and the disc centrifugation solve the problem that the resin cleaning is difficult in the macroporous resin adsorption method alone; the invention adopts the XAD-16 macroporous resin to selectively adsorb coffee and thus does not adsorb chlorogenic acid, thereby effectively reducing the green genus. The caffeine content in the acid product solves the problem of high caffeine content in the chlorogenic acid product during the general resin adsorption process; at the same time, the caffeine can be recycled; the solvent in the invention only uses purified water and ethanol, is safe and environmentally friendly, and The purity of chlorogenic acid obtained after alcohol washing and water washing and purification is as high as 90%. The conversion rate of chlorogenic acid is high, and the content of chlorogenic acid in the product is high. It has high extraction efficiency, simple operation, low production cost, easy to be used in large production, solvent is only used in purified water and ethanol, safe and environmentally friendly, and 50% of chlorogenic acid products have low caffeine content.

DRAWINGS

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

detailed description

Example 1

The technical method of the present invention mainly includes the following steps:

1. Material preparation: Crush the green coffee beans into a 15 mesh powder;

2. Ultrasonic extraction: 8 times the amount of sodium sulfite aqueous solution (mass concentration of 0.1%) 50 °C ultrasonic countercurrent continuous extraction, water flow is 5 times the mass of the material, get green coffee bean extract;

3. Centrifugation: The ultrasonic extract is filtered through a 100-mesh sieve, centrifuged at a snail, and centrifuged at a disc;

4. XAD-16 macroporous resin adsorption caffeine: The supernatant after centrifugation is adsorbed by XAD-16 macroporous resin. The ratio of resin adsorption to raw material is 1:1 (volume/mass, 1 cubic resin corresponds to 1 ton of green coffee) Extract of bean raw material);

5. XAD-16 macroporous resin analysis: After adsorption saturation, rinse with purified water, the elution amount is 2 times the volume of the resin; collect the eluent, concentrate and dry, get the crude caffeine

6. LX-28 macroporous resin enriched with chlorogenic acid: The leaching solution is adsorbed by macroporous resin. The ratio of resin adsorption to raw material is 1:1 (volume/mass, 1 cubic resin corresponds to 1 ton of green coffee bean raw material extraction) liquid);

7. LX-28 macroporous resin analysis (elution): After the adsorption is saturated, rinse with purified water, and use high quality 70% ethanol elution, the elution amount is twice the volume of the resin;

8. Concentration and drying: The eluate is concentrated, and the ethanol is recovered and spray-dried to obtain a chlorogenic acid product having a content of 50%;

9. Decolorization and crystallization: The chlorogenic acid with a content of 50% is dissolved in 10 times of 80% by mass of ethanol, decolorized by activated carbon, concentrated to a specific gravity of 1.15, and then placed in a frozen crystal. 5%以上。 The product having a purity of more than 3.5%.

Example 2.

The method for extracting chlorogenic acid includes the following steps:

(1) First, the green coffee beans are crushed through a 10-mesh sieve to obtain green coffee bean powder; 1 kg is taken for extraction, and the raw material chlorogenic acid content is 6.4%;

(2) adding 10 times the mass percent concentration of 0.1% aqueous solution of sodium sulfite, and continuously extracting by ultrasonic countercurrent at 40 ° C, the water flow rate is 5 times of the mass of the material, and the crude extract of green coffee beans is obtained;

(3) The extract in (2) is filtered through a 100-mesh sieve, centrifuged in a snail, and centrifuged to obtain a supernatant; the supernatant is poured into a resin column containing 7 liters of XAD-16 resin, and then used. Rinse twice with column volume of purified water, collect the leaching solution and the washing solution separately, adsorb 1 liter of LX-28 macroporous resin on the leaching solution, elute with 80% ethanol by mass concentration, concentrate the eluent, spray dry . 6%。 The purity of the caffeine was 73.6%.

5%。 The chlorogenic acid content was 53.3%, the yield was 3.6%, the purity was 90.5%. The purity (content) of caffeine was 73.6%. Example 3

(2) adding 10 times of purified water, and continuously extracting by ultrasonic countercurrent at 40 °C, the water flow rate is 5 times of the mass of the material, and the crude extract of green coffee beans is obtained;

(3) The extract in (2) is filtered through a 100-mesh sieve, centrifuged in a snail, and centrifuged to obtain a supernatant; the supernatant is poured into a resin column containing 7 liters of XAD-16 resin, and then used. Rinse with 2 column volumes of purified water, collect the leaching solution and the washing solution separately, and wash 1 liter of LX-28 macroporous resin on the leaching solution, elute with 80% ethanol, concentrate the eluent, and spray dry. 6%。 The purity of the caffeine was 73.6%.

8%的纯度。 83%。 The chlorogenic acid, the yield was 2.8%, the purity was 83%. The caffeine purity (content) was 73.6%. Example 4

The method for extracting chlorogenic acid includes the following steps:

(1) First, the green coffee beans are crushed through a 10-mesh sieve to obtain a green coffee bean powder; 7 kg is taken for extraction, and the raw material chlorogenic acid content is 6.4%;

(2) adding 10 times the mass percent concentration of 0.1% sodium sulfite aqueous solution, continuously extracting under ultrasonic conditions at 40 ° C, the water flow is 5 times the mass of the material, and obtaining the crude extract of green coffee beans;

(3) The extract in (2) is filtered through a 100-mesh sieve, centrifuged in a snail, and centrifuged to obtain a supernatant; the supernatant is poured into a resin column containing 7 liters of XAD-16 resin, and then used. Rinse with 2 column volumes of purified water to collect the effluent and water wash. Combine and bisect 7 parts, respectively, 1 liter of LX-28, DM130, D101, HZ816, AB-8, SPD600, HP-20 resin, elute with 80% ethanol by mass concentration, concentrate the eluent, spray dry .

Experimental Example 1 : Chlorogenic acid content and conversion rate of LX-28 macroporous resin and other resin final products The experimental comparison results are shown in Table 1 below:

Table 1. Comparison of experimental results of different resin models

Experimental example 2

(1) First, the green coffee beans are crushed through a 10-mesh sieve to obtain a green coffee bean powder; 2 kg is taken for extraction, the raw material content is 6.04%. The caffeine content is 1. 2%.

(2) Adding 10 times the amount of 0.1% aqueous solution of sodium sulfite, continuously extracting by ultrasonic countercurrent at 40 °C, the water flow rate is 5 times of the mass of the material, and the crude extract of green coffee beans is 18 liters, and the average is divided into 2 parts (per The content of chlorogenic acid is 18.39%, caffeine content 1.89%). One liter of XAD-16 resin is as follows: the upper column flow rate is 3 column volumes, and after the upper column is finished, it is rinsed with 2 volumes of purified water, combined with the lost liquid and water washing solution, and concentrated by spray drying. The other one was tetrachloromethane, used in an amount of 27 liters, extracted 3 times, 9 liters each time, and the combined aqueous phases were concentrated and dried. The results are shown in Table 2:

Table 2. Effect of resin method and carbon tetrachloride on caffeine removal and conversion to chlorogenic acid Item The present invention The present invention absorbs tetrachloromethane tetrachloromethanetetrahydromethane extract and absorbs adsorption conversion rate 垸 extraction 垸 extraction conversion rate before and after weight (g) 314. 7 232. 5 314. 7 266 . 8

Chlorogenic acid content (%) 18. 39 23. 65 95. 01% 18. 39 19. 52 89. 93% Caffeine content (%) 1. 89 0. 14 5. 47% 1. 89 0. 27 12 11% As can be seen from the above table, the removal of caffeine by the resin method is obviously superior to the currently used tetrachloromethane extraction method, and the conversion rate of chlorogenic acid is also significantly improved.

Claims

claims

1. A method for extracting chlorogenic acid from green coffee beans, characterized in that the extraction process includes the following steps:

(1) First, crush the green coffee beans and pass them through a 10-mesh sieve to obtain green coffee bean powder;

(2) Add 10 times the mass percentage of green coffee bean powder to a 0.1% sodium sulfite aqueous solution, conduct continuous ultrasonic countercurrent extraction at 40 to 60°C, and the sodium sulfite aqueous solution flow rate is 5 times the mass of the green coffee bean powder to obtain a green coffee bean extract. ;

(3) Filter the extract in (2) through a 100-mesh vibrating sieve, decanter centrifuge, and disc centrifuge to obtain the supernatant;

(4) Pour the supernatant in (3) directly into XAD-16 macroporous resin. The function of XAD-16 macroporous resin is to selectively adsorb caffeine without adsorbing chlorogenic acid. The ratio of resin adsorption capacity to supernatant is 1 :1; After the adsorption is saturated, rinse it with purified water, and elute with ethanol with a mass concentration of 50 to 80%. The elution volume is 2 times the volume of the resin, and the eluent is collected;

(5) Pour the XAD-16 macroporous resin loss liquid in (4) into the LX-28 macroporous resin. The ratio of the resin adsorption capacity to the loss liquid is 1:1; after the adsorption is saturated, rinse it with purified water and use mass percent Elute with ethanol with a concentration of 50% to 80%, the elution volume is 2 times the resin volume, and collect the eluent;

(6) Concentrate the eluent obtained in (4), recover ethanol, and spray-dry it into caffeine by-products with a content greater than 70%; Concentrate the eluent obtained in (5), recover ethanol, and spray-dry it into a caffeine by-product with a content greater than 50%. Chlorogenic acid products;

(7) Dissolve the chlorogenic acid in (6) with 8-10 times the amount of ethanol with a mass concentration of 80%, decolorize and filter with activated carbon, concentrate to a specific gravity of 1.15, freeze and crystallize, and dry with microwave to obtain chlorogenic acid. acid products.

2. The method according to claim 1, the decolorizing agent in step (6) is medical needle-type activated carbon, the dosage is 5% of the mass of chlorogenic acid, the temperature is 70°C, the holding time is 20min; the specific gravity of the crystallization is 1. l(Tl.15, the temperature is (T4°C.