CN106188213B - A kind of Simple Purification method of Tea Saponin - Google Patents

Abstract

The invention discloses a simple purification method of tea saponin, which belongs to the technical field of plant extraction. The technical scheme is as follows: 1) dissolving crude tea saponin in an alcohol solution with a volume fraction of 80%, centrifuging, and taking the supernatant 2) Put the supernatant in a Soxhlet extraction flask, take the cholesterol, wrap it with filter paper, transfer it to a Soxhlet extraction tube, heat and reflux for extraction for 6-7 hours, cool to room temperature, filter the extract, wash, Obtain the tea saponin-cholesterol complex; 3) dry the tea saponin-cholesterol complex to constant weight, wrap it with filter paper after cooling, put it into a Soxhlet extraction tube, add the extractant to the Soxhlet flask, and heat to reflux After extraction for 5-6 hours, after cooling, the purified tea saponin is obtained from the Soxhlet extraction tube with a purity of 95%-98%. The invention has the advantages of simple operation, low cost and high purification efficiency (purity is 95%-98%), and is suitable for industrial scale-up production.

Description

A kind of simple purification method of tea saponin

technical field

The invention belongs to the technical field of plant extraction, and in particular relates to a simple purification method of tea saponin.

Background technique

Tea saponin is a glycoside compound extracted from the seeds of the Theaceae plant. It belongs to saponins and is a natural non-ionic surfactant. It is a by-product of extracting tea seed oil---tea seed meal main active substance. Because tea saponin can combine with cholesterol in the cell membrane to form a complex, thereby changing the permeability of the cell membrane, destroying the structure of the cell membrane, causing the extravasation of the cytoplasm, and even death. Therefore, although tea seed meal contains a certain amount of active protein, polysaccharide, etc., because it contains a certain amount of tea saponin, tea seed meal can only be used as fertilizer.

With people's continuous understanding of the nutritional value of tea seed oil, tea seed oil, commonly known as "Oriental olive oil", has become a seasoning oil on people's tables, but because tea seeds are difficult to harvest and labor costs are high, tea Seed oil processing costs are rising. Therefore, actively developing and utilizing other active ingredients in tea seeds to realize the resource utilization of tea seeds has certain significance for reducing the production cost of tea seed oil and truly becoming the first-class oil on the public table.

Tea saponin is a natural non-ionic surfactant with good performance. Its excellent foaming and foam stabilizing properties make it widely used in light industry, chemical industry, pesticide, feed, breeding, textile, oil extraction, mining, building materials and highway construction. In the fields of detergents, pesticide auxiliaries, feed additives, protection agents for crab and shrimp farming, textile auxiliaries, oilfield foam agents, mining flotation agents, air-entrained concrete foam stabilizers and concrete admixtures - antifreeze is an important component. It is of practical significance to develop tea saponin resources and obtain high-purity tea saponin.

Because tea saponin is insoluble in anhydrous methanol and ethanol, insoluble in organic solvents such as ether, acetone, benzene, petroleum ether, etc., it is easily soluble in aqueous methanol, aqueous ethanol, glacial acetic acid, acetic anhydride, pyridine, etc. Therefore, tea saponin is generally extracted with aqueous methanol and ethanol as an extractant, and the tea saponin obtained by this method generally has a purity of 60% to 80%, and is a light yellow powder. Someone uses water or low-concentration ammonia water as an extractant to obtain tea saponin with a purity of 40-50% and a brownish-red powder. Someone uses water as the extractant and ethanol or methanol as the flocculant to obtain relatively high-purity tea saponin. Generally speaking, the lower the purity of tea saponin, the darker its color, and the narrower its application. Improving the purity of tea saponin is the main measure to broaden the application of tea saponin.

Contents of the invention

The purpose of the present invention is a simple purification method of tea saponin, which is simple in operation, strong in practicability, high in purification efficiency, high in purity of the prepared tea saponin and wide in application range.

The present invention is achieved through the following technical solutions:

A kind of simple and easy purification method of tea saponin, comprises the following steps:

1) Dissolve the crude tea saponin in an alcohol solution with a volume fraction of 80% according to the dosage ratio of (10-15) g: 10 mL, centrifuge, and take the supernatant;

2) Put the supernatant in a Soxhlet extraction flask, take the cholesterol according to the mass ratio of crude tea saponin: cholesterol = (10-15): (7-12), wrap it with filter paper, and transfer it to Soxhlet extraction In the tube, heat and reflux for extraction for 6-7 hours, cool to room temperature, filter the extract, wash to obtain the tea saponin-cholesterol complex;

3) Dry the tea saponin-cholesterol complex to constant weight, wrap it with filter paper after cooling, put it into a Soxhlet extraction tube, add the extractant to the Soxhlet flask, heat and reflux for extraction for 5-6 hours, after cooling, extract from Purified tea saponin is obtained in the Soxhlet extraction tube with a purity of 95% to 98%.

In step 1), the centrifugation treatment is at 2500-3500r/min, and the centrifugation treatment is 8-12min.

In step 2), the heating and reflux speed is controlled at 7-8 min/time.

In step 2), washing is performed 2 to 4 times with ethanol.

In step 3), the tea saponin-cholesterol complex is dried at 55-60°C.

Step 3) The extractant used is ether, toluene or sherwood oil.

In step 3), the heating and reflux speed is controlled at 8-10 min/time.

Step 3) also includes the step of recovering the intermediate product for reuse, specifically:

Collect the extract extracted by heating and reflux, concentrate the volume to 1/3-2/3, then cool to obtain cholesterol crystals, filter and dry to obtain cholesterol.

Compared with the prior art, the present invention has the following beneficial technical effects:

The simple purification method of tea saponin disclosed by the invention, because the tea saponin is easily soluble in aqueous methanol/ethanol, it is dissolved in 80v/v ethanol and loaded into a Soxhlet flask. Although cholesterol is soluble in ethanol, its solubility does not change much with temperature at 20-50°C, but when it exceeds 50°C, its solubility suddenly increases. The Soxhlet purification method is based on this principle. Tea saponin is easily combined with cholesterol in the liquid phase to form a tea saponin-cholesterol complex. The tea saponin-cholesterol complex is insoluble in ethanol/methanol containing water and is easily separated from the extractant (ethanol/methanol containing water). Because tea saponin is insoluble in organic solvents such as ether/toluene/petroleum ether, and cholesterol is easily soluble in organic solvents such as ether/toluene/petroleum ether, tea saponin and cholesterol can be separated by using this as an extraction agent. The cholesterol used in the method of the present invention has no loss in the purification process of tea saponin, can be used repeatedly, and will not significantly increase the purification cost of tea saponin. The purification cost of tea saponin mainly comes from the energy consumption of the extraction process, and can also solve the problem There is the problem of difficult solvent recovery in the technology, the operation is simple, the cost is low, the purification efficiency is high (purity is 95%-98%), and it is suitable for industrial scale-up production.

Detailed ways

The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

The reaction principle of the present invention is: tea saponin is easily combined with cholesterol in liquid phase to form tea saponin-cholesterol complex, and tea saponin-cholesterol complex is insoluble in water-containing ethanol/methanol, is easy to be mixed with extractant (water-containing ethanol/methanol) separation. And because tea saponin is insoluble in organic solvents such as ether/toluene/petroleum ether, and cholesterol is easily soluble in organic solvents such as ether/toluene/petroleum ether, it is an extraction agent in turn, and tea saponin and cholesterol can be separated.

Take 80v/v methanol/ethanol as the solvent, dissolve 10-15g of crude tea saponin in 100mL methanol/ethanol with a volume fraction of 80v/v, centrifuge at 3000 rpm for 10min, and put the supernatant into In a 150mL Soxhlet extraction flask, weigh 7.0-12g of cholesterol, wrap it with filter paper, transfer it to a Soxhlet extraction tube, install a Soxhlet extraction device, turn on the cooling water switch, turn on the heating device for heating, and control the reflux rate to 1 time/7~8min, continuous extraction for 6~7h, turn off the power switch, after cooling to room temperature, remove the Soxhlet extraction device. The extract was centrifuged and filtered to obtain the tea saponin-cholesterol complex, which was washed three times with 50 mL of methanol to obtain a white tea saponin complex.

Dry the tea saponin complex in an oven at 55-60°C to constant weight, wrap it with filter paper after cooling, put it into a Soxhlet extraction tube, and transfer 100-150 mL of the extraction agent ether/toluene/petroleum ether into the Soxhlet flask During the process, turn on the cooling water system, turn on the power supply to heat, control the reflux rate to 1 time/8~10min, and extract continuously for 5~6h. After cooling, dismantle the Soxhlet extraction device, and the purified tea is removed from the Soxhlet tube. Saponin, its purity is 95～98%, and color and luster is pure and white, and its aqueous solution looks bright. The extract contains cholesterol, and 1/3 to 2/3 of the ether/toluene/petroleum ether extract is recovered, cooled to obtain cholesterol crystals, filtered, and dried to obtain cholesterol.

The solubility of cholesterol in methanol/ethanol is 10-12g/100mL, but it changes with temperature, especially the solubility of tea saponin in ethanol before 55°C does not change much with temperature, but when the temperature rises to 55-65°C When , the solubility increases sharply, almost more than twice the solubility at room temperature. The present invention just utilizes this property of cholesterol. Cholesterol is placed in a Soxhlet extraction tube. The methanol/ethanol vaporized by heat in the Soxhlet extraction device is cooled by a condenser, and its temperature is mostly above 55°C. At this time, the cholesterol The solubility of the product rises rapidly, and the cholesterol dissolved in methanol/ethanol is combined with the tea saponin in the Soxhlet flask through reflux to form a cholesterol-tea saponin complex. The method simplifies the tea saponin purification process and avoids the technical problem of difficult recovery of mixed solvents.

Example 1

A kind of simple and easy purification method of tea saponin, comprises the following steps:

1) Dissolve 10 g of crude tea saponin in 100 mL of alcohol solution with a volume fraction of 80%, centrifuge at 3000 rpm for 10 min, and take the supernatant;

2) Put the supernatant in a Soxhlet extraction flask, take 7g of cholesterol, wrap it with filter paper, transfer it to a Soxhlet extraction tube, install the Soxhlet extraction device, turn on the cooling water switch, turn on the heating device for heating, and control the reflux The speed is 1 time/7-8min. After heating and reflux extraction for 6-7 hours, turn off the power switch, cool to room temperature, filter the extract, and divide the lotion with 50mL methanol for 3 times to obtain the tea saponin-cholesterol complex;

3) Dry the tea saponin-cholesterol complex in an oven at 55°C to constant weight, wrap it with filter paper after cooling, put it into a Soxhlet extraction tube, add 100mL petroleum ether into the Soxhlet flask, and turn on the cooling water system , turn on the power to heat, control the reflux rate to 1 time/8-10min, heat and reflux extraction for 5-6h, after cooling, obtain purified tea saponin from the Soxhlet extraction tube with a purity of 97%. The separated tea saponin has a white color, and its aqueous solution has a bright color. The extract contains cholesterol, and 1/3 to 2/3 of the petroleum ether extract is recovered, cooled to obtain cholesterol crystals, filtered, and dried to obtain cholesterol.

Example 2

A kind of simple and easy purification method of tea saponin, comprises the following steps:

1) Dissolve 15 g of crude tea saponin in 100 mL of alcohol solution with a volume fraction of 80%, centrifuge at 2500 rpm for 12 min, and take the supernatant;

2) Put the supernatant in a Soxhlet extraction flask, take 10g of cholesterol, wrap it with filter paper, transfer it to a Soxhlet extraction tube, install the Soxhlet extraction device, turn on the cooling water switch, turn on the heating device for heating, and control the reflux The speed is 1 time/7-8min. After heating and reflux extraction for 6-7 hours, turn off the power switch, cool to room temperature, filter the extract, and divide the lotion twice with 50mL of methanol to obtain the tea saponin-cholesterol complex;

3) Dry the tea saponin-cholesterol complex in an oven at 60°C to constant weight, wrap it with filter paper after cooling, put it into a Soxhlet extraction tube, add 150mL ether to the Soxhlet flask, and turn on the cooling water system, Turn on the power supply to heat, control the reflux rate to 1 time/8-10min, heat and reflux extraction for 5-6h, after cooling, obtain purified tea saponin from the Soxhlet extraction tube with a purity of 95%. The separated tea saponin has a white color, and its aqueous solution has a bright color. The extract contains cholesterol, recovering 1/3 to 2/3 of the ether extract, cooling to obtain cholesterol crystals, filtering, and drying to obtain cholesterol.

Example 3

A kind of simple and easy purification method of tea saponin, comprises the following steps:

1) Dissolve 13g of crude tea saponin in 100mL of alcohol solution with a volume fraction of 80%, centrifuge at 3500 rpm for 8min, and take the supernatant;

2) Put the supernatant in a Soxhlet extraction flask, take 12g of cholesterol, wrap it with filter paper, transfer it to a Soxhlet extraction tube, install the Soxhlet extraction device, turn on the cooling water switch, turn on the heating device for heating, and control the reflux The speed is 1 time/7-8min. After heating and reflux extraction for 6-7 hours, turn off the power switch, cool to room temperature, filter the extract, and divide the lotion with 50mL methanol for 4 times to obtain the tea saponin-cholesterol complex;

3) Dry the tea saponin-cholesterol complex in an oven at 60°C to constant weight, wrap it with filter paper after cooling, put it into a Soxhlet extraction tube, add 130mL toluene to the Soxhlet flask, and turn on the cooling water system, Turn on the power to heat, control the reflux rate to 1 time/8-10min, heat and reflux extraction for 5-6h, after cooling, obtain purified tea saponin from the Soxhlet extraction tube with a purity of 98%. The separated tea saponin has a white color, and its aqueous solution has a bright color. The extract contains cholesterol, and 1/3 to 2/3 of the toluene extract is recovered, cooled to obtain cholesterol crystals, filtered, and dried to obtain cholesterol.

In summary, the method of the present invention is simple to operate, low in cost, high in purification efficiency, cholesterol is not lost in the purification process of tea saponin, can be used repeatedly, can not significantly increase the purification cost of tea saponin, and the purification of tea saponin The cost mainly comes from the energy consumption of the extraction process.

Claims (6)

1. a kind of Simple Purification method of Tea Saponin, it is characterised in that comprise the following steps：

1) (10~15) g is pressed：100mL amount ratio, Tea Saponin crude product is dissolved in the alcoholic solution that volume fraction is 80%, centrifugation Processing, takes supernatant；

2) supernatant is placed in surname extraction flask, by Tea Saponin crude product：Cholesterol=(10~15)：The quality of (7~12) Than taking cholesterol, being wrapped with filter paper, is transferred in soxhlet extraction pipe, after being heated to reflux 6~7h of extraction, be cooled to room temperature, will extract Liquid filters, and washing, obtains Tea Saponin-cholesterin complex；Wherein, speed control is heated to reflux at 7~8min/ times；

3) by Tea Saponin-cholesterin complex drying to constant weight, wrapped, be fitted into soxhlet extraction pipe with filter paper after cooling, to rope Extractant is added in family name's flask, is heated to reflux 5~6h of extraction, after cooling, is obtained from soxhlet extraction pipe by purification process Tea Saponin, purity are 95%~98%；Wherein, speed control is heated to reflux at 8~10min/ times.

2. the Simple Purification method of Tea Saponin according to claim 1, it is characterised in that in step 1), centrifugal treating is Under 2500~3500r/min, 8~12min of centrifugal treating.

3. the Simple Purification method of Tea Saponin according to claim 1, it is characterised in that in step 2), washing is to use second Alcohol cleans 2~4 times.

4. the Simple Purification method of Tea Saponin according to claim 1, it is characterised in that in step 3), be by Tea Saponin- Cholesterin complex is dried at 55~60 DEG C.

5. the Simple Purification method of Tea Saponin according to claim 1, it is characterised in that extractant used in step 3) is second Ether, toluene or petroleum ether.

6. the Simple Purification method of Tea Saponin according to claim 1, it is characterised in that also include after step 3) in recovery Between product recycle the step of, be specially：

The extract for being heated to reflux extraction is collected, concentration volume to 1/3~2/3, is then cooled down, obtains cholesterol crystal, filter, Drying, obtains cholesterol.