阅读说明：本技术 基于亚临界萃取工艺的绿原酸提取方法 (Chlorogenic acid extraction method based on subcritical extraction process ) 是由 陈维 邹俊辉 黄德春 于 2021-09-30 设计创作，主要内容包括：本发明公开基于亚临界萃取工艺的绿原酸提取方法,本方法以金银花为原料,以亚临界水为溶剂萃取绿原酸,收集提取液。本发明使绿原酸提取率从24.941mg/g提高至32.195mg/g,最高增加了30％,有效解决了传统提取绿原酸的提取率较低、大量使用有机溶剂的问题。(The invention discloses a chlorogenic acid extraction method based on a subcritical extraction process. The method improves the extraction rate of the chlorogenic acid from 24.941mg/g to 32.195mg/g, increases the extraction rate by 30 percent at most, and effectively solves the problems of low extraction rate and large use of organic solvents in the traditional extraction of the chlorogenic acid.)

1. A chlorogenic acid extraction method based on a subcritical extraction process is characterized by comprising the following steps: the method comprises the following steps:

(1) preprocessing honeysuckle;

(2) subcritical extraction: putting pretreated honeysuckle into an extraction tank, adding pure water, turning on a heating switch, stopping heating after reaching a certain temperature, keeping the temperature and pressure in the kettle at a certain pressure, extracting for a period of time, turning on a condensed water circulating pump after extraction is finished, cooling steam discharged from the extraction tank, further cooling the steam after passing through a low-temperature cooling tank, and finally feeding extract into a receiving tank to collect the extract.

2. The method for extracting chlorogenic acid based on subcritical extraction process according to claim 1, characterized in that: the liquid-material ratio in the step (2), namely the ratio of pure water to honeysuckle flower is 70: 1-100: 1.

3. The method for extracting chlorogenic acid based on subcritical extraction process according to claim 1, characterized in that: in the step (2), the extraction temperature is 105-130 ℃.

4. The method for extracting chlorogenic acid based on subcritical extraction process according to claim 1, characterized in that: in the step (2), the extraction pressure is 0.05-0.24 Mpa.

5. The method for extracting chlorogenic acid based on subcritical extraction process according to claim 1, characterized in that: in the step (2), the extraction time is 20-50 min.

Technical Field

The invention belongs to a chlorogenic acid extraction method, and particularly relates to a chlorogenic acid extraction method by utilizing a subcritical water extraction process.

Background

Chlorogenic acid (Chlorogenic acid) is a phenylpropanoid compound produced by plants in the aerobic respiration process through the shikimic acid pathway. Chlorogenic acid is widely present in plants of Eucommiaceae and Caprifoliaceae, but the number of plants with higher content is not large, and is mainly distributed in leaves of eucommia ulmoides, dried flower buds of Lonicera japonica, and the like. As one of the main effective components of the Chinese herbal medicines such as honeysuckle, oriental wormwood, eucommia bark and the like, chlorogenic acid has multiple effects of protecting the cardiovascular system, resisting oxidation, reducing blood sugar, regulating immunity, resisting bacteria and the like. Chlorogenic acid has various biological activities, and can inhibit tumor (in vitro) growth, including colorectal cancer, liver cancer and larynx cancer; prevention of type 2 diabetes; and has antihypertensive, antiviral, antibacterial, and antifungal activities. Meanwhile, the chlorogenic acid has relatively low toxicity and no side effect. These properties of chlorogenic acid have been used in the manufacture of food additives and cosmetics.

Subcritical extraction is a novel extraction and separation technology which utilizes subcritical fluid as an extractant, transfers fat-soluble components in solid materials to a liquid extractant through a molecular diffusion process of the extraction materials and the extractant in a soaking process according to the principle that organic matters are similar and soluble mutually in a closed, oxygen-free and low-pressure container, and separates the extractant from a target product through a reduced pressure evaporation process to finally obtain the target product. The subcritical water extraction method is a green and environment-friendly extraction technology which is popular in recent years, and subcritical water is used as a high-efficiency extractant to extract polar and nonpolar substances. The extraction rate of the current subcritical extraction is not ideal enough, and chlorogenic acid can be efficiently extracted from honeysuckle by controlling the temperature and the pressure of subcritical water.

Disclosure of Invention

The purpose of the invention is as follows: the invention aims to provide a chlorogenic acid extraction method based on a subcritical extraction process, which has high extraction rate.

The technical scheme is as follows: the invention relates to a chlorogenic acid extraction method based on a subcritical extraction process, which comprises the following steps:

1) preprocessing honeysuckle;

2) subcritical extraction: putting pretreated honeysuckle into an extraction tank, adding pure water, turning on a heating switch, stopping heating after reaching a certain temperature, keeping the temperature and pressure in the kettle at a certain pressure, extracting for a period of time, turning on a condensed water circulating pump after extraction is finished, cooling steam discharged from the extraction tank, further cooling the steam after passing through a low-temperature cooling tank, and finally feeding extract into a receiving tank to collect the extract.

Further, the liquid-material ratio, i.e. the ratio of pure water to honeysuckle flower is 70: 1-100: 1(mL/g), the extraction temperature is 105-130 ℃, the extraction time is 20-50 min, and the extraction liquid is collected, wherein the extraction pressure is 0.05-0.24 MPa.

Has the advantages that: compared with the prior art, the invention has the following remarkable advantages:

the extraction method does not use any organic solvent, and the result shows that the extraction rate of the chlorogenic acid is improved from 24.941mg/g to 32.195mg/g, the extraction rate is increased by 30 percent, and the high-efficiency extraction of the chlorogenic acid in the honeysuckle is realized.

Drawings

FIG. 1 is a standard curve of chlorogenic acid by HPLC;

FIG. 2 is a graph comparing the extraction rate of chlorogenic acid with the conventional hot water extraction rate in example 11.

Detailed Description

Fitting by using high performance liquid chromatography with peak area standard curve measured by chlorogenic acid standard as reference to obtain a relation between peak area and concentration: and (4) calculating the content of the extracted chlorogenic acid by using the formula, wherein the A (peak area) is 0.6098C (concentration) -0.1762.

Example 1

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 105 ℃, keeping the pressure in the kettle at 0.05Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 27.737mg/g by using high performance liquid chromatography measurement.

Example 2

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 110 ℃, keeping the pressure in the kettle at 0.1Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 30.925mg/g by using high performance liquid chromatography measurement.

Example 3

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 120 ℃, keeping the pressure in the kettle at 0.2Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 27.872mg/g by using high performance liquid chromatography measurement.

Example 4

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 130 ℃, keeping the pressure in the kettle at 0.24Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 25.131mg/g by using high performance liquid chromatography measurement.

The extraction data for examples 1-4 are shown in FIG. 1.

TABLE 1

As can be seen from the data in the comparison table, the extraction rate of chlorogenic acid is increased as a whole with the increase of the temperature, because the molecular diffusion rate is increased with the increase of the temperature, and the extraction efficiency of the solvent to the chlorogenic acid is also increased. When the temperature reaches 110 ℃, the extraction rate reaches the highest value of 30.925mg/g, and when the temperature is increased continuously, partial chlorogenic acid is decomposed due to the overhigh temperature, so that the extraction rate of the chlorogenic acid is reduced. Therefore, the optimum extraction temperature is selected as 110 ℃, and the extraction rate is the maximum.

Experiments prove that the subcritical extraction time has a great influence on the extraction rate of chlorogenic acid, and the specific implementation process is shown in examples 5-8.

Example 5

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 110 ℃, keeping the pressure in the kettle at 0.1Mpa, and extracting for 20 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 27.219mg/g by using high performance liquid chromatography measurement.

Example 6

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 110 ℃, keeping the pressure in the kettle at 0.1Mpa, and extracting for 30 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 27.592mg/g by using high performance liquid chromatography measurement.

Example 7

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 110 ℃, keeping the pressure in the kettle at 0.1Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 30.925mg/g by using high performance liquid chromatography measurement.

Example 8

Accurately weighing 20g of pretreated honeysuckle, placing the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 110 ℃, keeping the pressure in the kettle at 0.1Mpa, and extracting for 50 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 30.892mg/g by using high performance liquid chromatography measurement.

The extraction data for examples 5-8 are shown in FIG. 2.

TABLE 2

As can be seen from the data in the comparison table, the extraction rate of chlorogenic acid increases with time, and particularly when the extraction time is prolonged from 30min to 40min, the extraction rate increases significantly. When the extraction time is prolonged to 50min, the extraction rate is not obviously improved any more, and the conclusion can be drawn that the subcritical water under the temperature and the pressure has the best extraction effect when the extraction time is 40 min.

Meanwhile, the liquid-material ratio of the subcritical extraction has a certain influence on the extraction rate of chlorogenic acid, and the specific implementation process of the relationship is researched, which is shown in examples 9-12.

Example 9

Accurately weighing 22.86g of pretreated honeysuckle (liquid-material ratio is 70: 1), putting the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, turning on a heater, heating to 110 ℃, keeping the pressure in the tank at 0.1Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 25.266mg/g by using high performance liquid chromatography measurement.

Example 10

Accurately weighing 20g of pretreated honeysuckle (liquid-material ratio is 80: 1), putting the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 110 ℃, keeping the pressure in the tank at 0.1Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 30.925mg/g by using high performance liquid chromatography measurement.

Example 11

Accurately weighing 17.78g (liquid-material ratio is 90: 1) of pretreated honeysuckle, putting the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, turning on a heater, heating to 110 ℃, keeping the pressure in the tank at 0.1Mpa, and extracting for 40 min. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 32.195mg/g by using high performance liquid chromatography measurement.

Example 12

Accurately weighing 16g of pretreated honeysuckle (liquid-material ratio is 100: 1), putting the honeysuckle into an extraction tank, adding 1600mL of pure water, sealing the tank body, starting a stirrer, adjusting to 120r/min, starting a heater, heating to 110 ℃, keeping the pressure in the tank at 0.1Mpa, and extracting for 40min under the condition of heat preservation and pressure maintaining. And after extraction is finished, a circulating pump is started, steam discharged from the extraction tank is cooled by cooling water, the steam is further cooled after passing through a low-temperature cooling tank, and finally the extract enters a receiving tank, and the extraction rate is 30.921mg/g by using high performance liquid chromatography measurement.

The extraction data for examples 9-12 are shown in FIG. 3.

TABLE 3

As can be seen from the data in the comparison table, the extraction rate of the chlorogenic acid increases along with the increase of the liquid-material ratio, and the extraction rate of the chlorogenic acid reaches the highest point when the liquid-material ratio is 90: 1. Therefore, the optimum liquid-to-material ratio can be selected as the condition of 90: 1.