CN116396350A - Tea saponin purification method - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 35
- 238000000746 purification Methods 0.000 title claims abstract description 12
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- 238000011084 recovery Methods 0.000 claims abstract description 53
- 238000000605 extraction Methods 0.000 claims abstract description 37
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 238000005303 weighing Methods 0.000 claims abstract description 12
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 238000001816 cooling Methods 0.000 claims description 14
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- 238000002835 absorbance Methods 0.000 claims description 9
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- 238000000926 separation method Methods 0.000 claims description 5
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- 239000012488 sample solution Substances 0.000 claims description 3
- 239000012043 crude product Substances 0.000 abstract description 11
- 238000006243 chemical reaction Methods 0.000 abstract description 5
- 238000011156 evaluation Methods 0.000 abstract description 3
- 239000000706 filtrate Substances 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- 238000010998 test method Methods 0.000 abstract description 3
- 238000002390 rotary evaporation Methods 0.000 abstract description 2
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- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 description 3
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 description 3
- 235000012141 vanillin Nutrition 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000011481 absorbance measurement Methods 0.000 description 2
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- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/256—Polyterpene radicals
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Abstract
The invention discloses a purification method of tea saponin, which comprises the following steps: weighing a small amount of crude tea saponin, and dissolving with 10ml of hot water to prepare a tea saponin solution with the mass fraction of 10%; the method is characterized in that crude tea saponin developed in a laboratory is used as a raw material, saturated n-butanol is used as an extractant, the recovery rate of the tea saponin is used as an evaluation index, the influence of each single factor on the recovery rate of the tea saponin is systematically researched, an orthogonal test method is adopted to optimize the conversion extraction to obtain the optimal process condition, when the dosage of the saturated n-butanol is 35mL, the pH is 4, the mass fraction of the crude product is 20%, the extraction times are 3 times, the filtrates of the 3 times are combined, rotary evaporation and drying are carried out to obtain a milky tea saponin product with the purity of 97% and the recovery rate of 53.25%, and compared with the prior art, the purity of the tea saponin can be improved by purifying the crude tea saponin, and the influence of the saturated n-butanol on the recovery rate of the purified tea saponin is reduced as much as possible.
Description
Technical Field
The invention relates to the technical field of tea saponin purification, in particular to a purification method of tea saponin.
Background
The tea saponin is extracted from tea pulp, and along with the gradual enhancement of environmental awareness of people and the shortage of acceleration of current energy sources, the harmful surfactant is continuously removed, for example, the dodecyl sulfate is forbidden to be used, and the tea saponin is taken as a natural nonionic surfactant, which is an environmental-friendly surfactant required in industrial production, so that not only can tea seed resources be fully utilized, but also the development of the environmental-friendly industry in China is promoted;
the traditional tea saponin extraction can be carried out by adopting extraction methods such as a water extraction method, an alcohol extraction method and the like, crushing tea seed meal, degreasing by petroleum ether to obtain defatted tea seed meal, extracting tea saponin in the defatted tea seed meal by any extraction method, obtaining tea saponin solution after centrifugal separation, and concentrating and drying to obtain crude tea saponin.
However, in the existing tea saponin extraction process, the extracted crude tea saponin contains impurities such as pigment, protein, flavone, tannin and the like, so that the crude tea saponin has a darker color and lower purity, and directly affects the quality of shampoo produced by the tea saponin, and therefore, a purification method of the tea saponin is proposed to solve the above problems.
Disclosure of Invention
In order to overcome the defects in the prior art and solve at least one of the problems, the invention provides a purification method of tea saponin.
A purification method of tea saponin, comprising the steps of:
s1: weighing a small amount of crude tea saponin, and dissolving with 10ml of hot water to prepare a tea saponin solution with the mass fraction of 10%;
s2: regulating the pH value of the tea saponin solution to be 5, and cooling for standby;
s3: adding n-butanol solution or saturated n-butanol solution into cooled tea saponin solution, and shaking uniformly;
s4: extracting the evenly-shaken tea saponin mixed solution to obtain supernatant;
s5: concentrating and drying the obtained supernatant to constant weight to obtain pure tea saponin;
s6: testing the recovery rate of pure tea saponin;
wherein, two parts of crude tea saponin are weighed, respectively dissolved by using 10Ml of hot water, pH is adjusted, and after cooling, n-butanol and saturated n-butanol with the same volume are respectively added, the steps S4 to S6 are repeated, and the extractant is determined to be n-butanol solution or saturated n-butanol solution.
Preferably, in the step S3, a saturated n-butanol solution is used as an extractant, and in order to determine the optimal amount of the saturated n-butanol solution, different volumes of saturated n-butanol solutions are added to the cooled tea saponin solution, and steps S4 to S6 are repeated to determine the optimal amount of the saturated n-butanol solution.
Preferably, in the step S2, in order to determine the optimal pH value, after the step S1, the pH values of different tea saponin solutions are prepared, and after cooling, the steps S4 to S6 are repeated to determine the optimal pH value.
Preferably, in the step S4, in order to determine the optimal extraction times, the extraction is performed for different times, different supernatants after extraction are obtained after separation, and steps S5 to S6 are repeated to determine the optimal extraction times.
Preferably, in the step S1, in order to determine the optimal mass fraction of crude tea saponin, after a small amount of crude tea saponin is dissolved with 10ml of hot water, the crude tea saponin is respectively prepared into tea saponin solutions with different mass fractions, and steps S2 to S6 are repeated to determine the optimal mass fraction of crude tea saponin.
Preferably, in the step S5, the obtained supernatant is concentrated to paste at the temperature of 70 ℃ under the vacuum degree of 0.09Mpa, and is continuously dried to constant weight to obtain the pure tea saponin.
Preferably, in the step S7, the calculation formula of the purity and recovery rate of the tea saponin is as follows:
wherein, C is the purity of tea saponin; c is the concentration mg/ml in the sample solution calculated by the regression equation; x is the dilution of the solution; v is the volume ml of the solution; y is the recovery rate of tea saponin; m is M 1 The amount mg of the tea saponin pure product is weighed for measuring absorbance; m is M 3 The amount of the tea saponin is mg.
Preferably, in the step S7, the absorbance measurement is specifically that the pure tea saponin prepared in the quantitative step S6 is weighed, dissolved in methanol and fixed in volume, developed by a concentrated sulfuric acid-vanillin method, and absorbance a is tested at a wavelength of 550nm, and substituted into a regression equation:
A＝8.0918c+0.0626。
the invention has the advantages that:
the invention takes crude tea saponin developed in a laboratory as a raw material, saturated n-butyl alcohol as an extractant and the recovery rate of tea saponin as an evaluation index, systematically researches the influence of each single factor on the recovery rate of tea saponin, adopts an orthogonal test method to optimize the conversion and extraction to obtain the optimal process condition, combines the filtrates of 3 times when the dosage of saturated n-butyl alcohol is 35mL, the pH is 4, the mass fraction of the crude product is 20 percent, the extraction times are 3 times, and carries out rotary evaporation and drying to obtain a milky tea saponin product with the purity of 97 percent and the recovery rate of 53.25 percent.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions of the prior art, the drawings which are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that other drawings can be obtained from these drawings without inventive faculty for a person skilled in the art.
FIG. 1 is a flow chart of a purification method according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of testing recovery rates using different extractants in one embodiment of the invention;
FIG. 3 is a schematic diagram showing test recovery rates of saturated n-butanol using different amounts in accordance with one embodiment of the present invention;
FIG. 4 is a schematic diagram of testing recovery rates for different pH values in accordance with one embodiment of the present invention;
FIG. 5 is a schematic diagram showing the selection of different extraction times for testing recovery rate in an embodiment of the present invention;
FIG. 6 is a schematic diagram showing the test recovery rate of crude tea saponin with different mass fractions according to one embodiment of the present invention;
FIG. 7 is a graph showing the relationship between the logarithm of concentration and the surface tension in an embodiment of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to fig. 1 to 7, a purification method of tea saponin comprises the following steps:
s1: weighing a small amount of crude tea saponin, and dissolving with 10ml of hot water to prepare a tea saponin solution with the mass fraction of 10%;
s2: regulating the pH value of the tea saponin solution to be 5, and cooling for standby;
s3: adding n-butanol solution or saturated n-butanol solution into cooled tea saponin solution, and shaking uniformly;
s4: extracting the evenly-shaken tea saponin mixed solution to obtain supernatant;
s5: concentrating and drying the obtained supernatant to constant weight to obtain pure tea saponin;
s6: testing the recovery rate of pure tea saponin;
weighing two parts of crude tea saponin, respectively dissolving with 10Ml of hot water, regulating pH, cooling, respectively adding n-butanol and saturated n-butanol with the same volume, repeating steps S4 to S6, and determining whether the extractant is n-butanol solution or saturated n-butanol solution;
specifically, determining that the extractant is n-butanol or saturated n-butanol solution, weighing 2 parts of laboratory self-made tea saponin crude products, and respectively numbering as l and 2; dissolving with 10mL of hot water, respectively adjusting pH value to 5, cooling, adding 30mL of n-butanol into I, adding 30mL of saturated n-butanol solution into II, extracting twice, testing the influence of different extractants on the recovery rate of tea saponin, calculating the recovery rate of tea saponin, and selecting saturated n-butanol solution as extractant, wherein the recovery rate of tea saponin is higher than that of the n-butanol solution, and the extraction rate of tea saponin by saturated n-butanol is obviously higher than that by n-butanol, as shown in figure 2. The method is characterized in that n-butanol and water are in a slightly-soluble relationship, and when the n-butanol is used as an extracting agent, liquid separation is often involved, so that a large number of emulsifying layers are generated, the emulsifying layers are difficult to separate, the emulsifying layers are greatly reduced after the saturated n-butanol is used, the solubility of the n-butanol is increased by the saturated n-butanol solution, and the method is suitable for deep extraction, so that the saturated n-butanol solution is used as the extracting agent.
In the step S3, a saturated n-butanol solution is used as an extractant, and in order to determine the optimal amount of the saturated n-butanol solution, different volumes of saturated n-butanol solutions are added into the cooled tea saponin solution respectively, and steps S4 to S6 are repeated to determine the optimal amount of the saturated n-butanol solution;
specifically, determining the dosage of the optimal saturated n-butanol solution, specifically weighing 6 parts of crude tea saponin, dissolving with 10mL of hot water to prepare a tea saponin solution with the mass fraction of 10%, adjusting the pH value to be 5, cooling for standby, respectively adding 10mL, 20mL, 30mL, 40mL, 50mL and 60mL of saturated n-butanol solution, shaking for 5min, extracting for l h for two times, examining the influence of the dosage of the saturated n-butanol solution on the recovery rate of the tea saponin, and calculating the recovery rate of the tea saponin; according to calculation, 30ml of saturated n-butanol solution is selected, and the recovery rate of tea saponin is relatively high; as shown in figure 3, along with the increase of the saturated n-butanol dosage, the recovery rate of the tea saponin is also increased sharply, when the saturated n-butanol dosage reaches 30mL, and then the extractant dosage is increased, so that the recovery rate of the tea saponin is not changed greatly, and from the aspect of saving cost, 30mL of saturated n-butanol is preferably selected to be suitable.
In the step S2, in order to determine the optimal pH value, after the step S1, different pH values of the tea saponin solution are prepared, and after cooling, the steps S4 to S6 are repeated to determine the optimal pH value;
specifically, determining an optimal pH value, specifically weighing 6 parts of crude tea saponin, dissolving with 10mL of hot water to prepare a tea saponin solution with the mass fraction of 10%, adjusting the pH values to be about 4, 5, 6, 7, 8 and 9 respectively, cooling for standby, adding 30mL of saturated n-butanol solution, shaking for 5min, extracting for 1h respectively, examining the influence of the pH value on the recovery rate of the tea saponin, and calculating the recovery rate of the tea saponin; the calculation shows that when the pH value is adjusted to 5, the purity and the recovery rate of the tea saponin are relatively high; as shown in figure 4, the recovery rate of tea saponin is continuously increased at the pH value of 4 to 5, and the recovery rate is continuously reduced from 5, which indicates that the recovery rate of tea saponin reaches the maximum value at the pH value of 5; the reason is probably because the tea saponin is easily dissolved in the weak alkaline solution, so that the tea saponin is more difficult to extract from the aqueous solution during the extraction, and therefore, the pH value is preferably 5.
In S4, in order to determine the optimal extraction times, the different times are extracted respectively, the supernatants after different extractions are obtained after separation, and steps S5 to S6 are repeated to determine the optimal extraction times;
specifically, determining optimal extraction times, namely weighing 5 parts of crude tea saponin, dissolving the crude tea saponin with 10mL of hot water to prepare a tea saponin solution with the mass fraction of 10%, adjusting the pH value to be 5, cooling for standby, adding 30mL of saturated n-butanol solution, shaking for 5min, extracting 1, 2, 3, 4 and 5 times respectively, extracting for 1h, examining the influence of the extraction times on the recovery of the tea saponin, and calculating the recovery rate of the tea saponin; the purity and recovery rate of the tea saponin are relatively high when the extraction times are three times; as shown in figure 5, when the tea saponin is extracted once by saturated n-butanol, the recovery rate of the tea saponin is very low, the extraction times are continuously increased, the recovery rate of the tea saponin is continuously increased, but the extraction times are selected to be three times after reasonable consideration is given to the factors of resource waste, excessive energy consumption, solvent waste and the like caused by increasing the extraction times.
In the step S1, in order to determine the optimal crude tea saponin mass fraction, a small amount of crude tea saponin is dissolved in 10ml of hot water and then is respectively prepared into tea saponin solutions with different mass fractions, and the steps S2 to S6 are repeated to determine the optimal crude tea saponin mass fraction;
specifically, determining the mass fraction of the optimal crude tea saponin, specifically weighing 6 parts of crude tea saponin, dissolving with 10mL of hot water to prepare tea saponin solutions with mass fractions of 5%, 10%, 15%, 20%, 25%, 30% and 35%, adjusting the pH value to be 5, cooling for standby, adding 30mL of saturated n-butanol solution, shaking for 5min, extracting for 1h, extracting for two times, examining the influence of the mass fraction of the crude tea saponin on the recovery rate of the tea saponin, and calculating the recovery rate of the tea saponin; the calculation shows that the purity and recovery rate of the crude tea saponin are relatively high when the mass fraction of the crude tea saponin is 25%; as shown in figure 6, the recovery rate is continuously increased from 5% by mass to 25% by mass of the crude product, and the recovery rate of tea saponin is maximized. At this time, the mass fraction was increased again and the recovery rate decreased with the increase, indicating that 30mL of saturated n-butanol had been extracted from the crude product to a maximum value, so the mass fraction of the crude product was selected to be 25%.
In the step S5, the obtained supernatant is concentrated to paste at the temperature of 70 ℃ under the vacuum degree of 0.09Mpa, and is continuously dried to constant weight to obtain the pure tea saponin.
In S7, the calculation formula of the purity and recovery rate of tea saponin is as follows:
wherein, C is the purity of tea saponin; c is the concentration mg/ml in the sample solution calculated by the regression equation; x is the dilution of the solution; v is the volume ml of the solution; y is the recovery rate of tea saponin; m is M 1 The amount mg of the tea saponin pure product is weighed for measuring absorbance; m is M 3 The amount of the tea saponin is mg;
specifically, after the extractant is saturated n-butanol, in order to determine the influence of the synergistic effect of a plurality of single factors on the recovery rate of the tea saponin, a four-factor three-level method is adopted to design an orthogonal test, and the dosage (A), the pH (B), the mass fraction (C) of the crude product and the extraction frequency (D) of the extractant are selected as investigation factors so as to further optimize the extraction conditions. The results of the orthogonal test design factor level are shown in table 1;
TABLE 1 orthogonal test design factor level
Obtaining an L9 (43) 9 group test according to the single-factor test result and the designed four-factor three-level table, wherein the test result is shown in a table 2;
TABLE 2 results of orthogonal experiments
| Test number | A | B | C | D | Recovery/% |
| 1 | 1 | 1 | 1 | 1 | 33.91 |
| 2 | 1 | 2 | 2 | 2 | 39.18 |
| 3 | 1 | 3 | 3 | 3 | 41.01 |
| 4 | 2 | 1 | 2 | 3 | 51.21 |
| 5 | 2 | 2 | 3 | 1 | 29.36 |
| 6 | 2 | 3 | 1 | 2 | 39.73 |
| 7 | 3 | 1 | 3 | 2 | 43.48 |
| 8 | 3 | 2 | 1 | 3 | 53.76 |
| 9 | 3 | 3 | 2 | 1 | 28.35 |
| Ⅰ | 114.10 | 128.60 | 127.40 | 91.61 | |
| Ⅱ | 120.30 | 122.30 | 118.73 | 122.64 | |
| Ⅲ | 125.58 | 109.08 | 113.85 | 145.98 | |
| K1 | 38.03 | 42.46 | 42.46 | 30.54 | |
| K2 | 40.10 | 40.76 | 39.57 | 40.88 | |
| K3 | 41.86 | 36.36 | 37.95 | 48.66 | |
| R | 3.83 | 6.5 | 4.51 | 18.12 |
As can be seen from Table 2, the four selected factors, namely saturated n-butanol dosage (A), pH (B), crude product mass fraction (C), extraction times (D), influence the recovery rate of tea saponin in the order of D > B > C > A, namely extraction times > pH > crude product mass fraction > saturated n-butanol dosage; the best extraction optimization process conditions are A3B1C1D3, namely: the dosage of saturated n-butanol is 35mL, the pH is 4, the mass fraction of the crude product is 20%, and the extraction times are 3.
In order to verify the accuracy of the obtained result, the test is verified that the optimal optimized process condition is that the saturated n-butanol dosage is 35mL, the pH is 4, the mass fraction of the crude product is 20%, the extraction times are 3, the purity is 97%, and the recovery rate is 53.25%.
In S7, the absorbance measurement is specifically that the pure tea saponin prepared in the quantitative step S6 is weighed, dissolved in methanol and fixed in volume, developed by concentrated sulfuric acid-vanillin method, and absorbance a is tested at 550nm wavelength, and substituted into regression equation:
A＝8.0918c+0.0626(R 2 ＝0.989)
specifically, the color development principle of the concentrated sulfuric acid-vanillin method is that strong oxidizing property of the concentrated sulfuric acid is utilized, triterpenoid saponin is dehydrogenated under the action of strong oxidizing acid, and a colored substance is generated by adding vanillin after oxidation, and the colored substance is red. The method is simple and convenient to operate, sensitive in reaction, easy to control reaction conditions and stable in color development, and is a better determination method;
the determination of λmax is specifically: precisely weighing 41.4mg of tea saponin standard substance dried to constant weight, dissolving with 70% methanol, fixing the volume to a 50ML volumetric flask, shaking uniformly, sucking 0.2ML of the standard solution into a test tube, adding 0.3ML of solvent (70% methanol), sucking 5ML of the now prepared 8% vanillin solution (2.0 g vanillin and a certain amount of absolute ethyl alcohol are dissolved, fixing the volume to a 25ML volumetric flask), sucking 4ML of 77% concentrated sulfuric acid under an ice bath environment, shaking uniformly, reacting the test tube in a water bath at 60 ℃ for 18min, then placing the test tube in an ice bath for 10min, taking out the test tube to return to room temperature, and measuring absorbance A at 480-600nm by using reagent blank as a reference to obtain a lambda max of 550nm;
working principle: taking crude tea saponin developed in a laboratory as a raw material, taking saturated n-butanol as an extractant, taking the recovery rate of the tea saponin as an evaluation index, systematically researching the influence of each single factor on the recovery rate of the tea saponin, adopting an orthogonal test method to optimize the conversion extraction to obtain the optimal process condition, when the dosage of the saturated n-butanol is 35mL, the pH is 4, the mass fraction of the crude tea saponin is 20%, the extraction times are 3, merging the filtrates of the 3 times, rotary evaporating, and drying to obtain a milky tea saponin product with the purity of 97% and the recovery rate of 53.25%;
qualitative analysis is carried out on tea saponin prepared under the optimal process conditions:
foam performance testing, specifically, preparing aqueous solutions with tea saponin concentration of 0.0l%, O.05%, 0.1%, O.5%, 1.00% and 2.00%, respectively transferring 1mL of the aqueous solutions into graduated test tubes, shaking up and down by hand for 1min, standing, reading the concave liquid surface of the test tubes in a head-up manner, recording the foam height mark as H, and starting recording the time T required for complete foam fading.
The results were as follows: the foam performance test results are detailed in table 3 below:
TABLE 3 foam Properties of tea saponin
| Tea saponin concentration/% | 0.01 | 0.05 | 0.1 | 0.5 | 1.00 | 2.00 |
| Foam height/ |
8 | 19 | 32 | 49 | 62 | 76 |
| Time T/h of bubble stabilization | 1 | 2.5 | 4 | 9 | 24 | For more than 24 hours |
As can be seen from Table 3, at room temperature, the foam height increases linearly with increasing concentration of tea saponin, and the foam duration T also increases continuously with increasing foam height, which means that tea saponin is a surfactant with good foam performance, and according to the special property, tea saponin can be applied to products such as detergents, cosmetics, shampoo and the like.
The surface tension test is specifically to determine the surface tension of tea saponin according to the pull-off method, and when the surface tension of the solution reaches the minimum value, the concentration of the obtained solution is called Critical Micelle Concentration (CMC).
Tea saponin solutions with concentrations of O.001%, 0.002%, 0.004%, O.02%, 0.04%, 0.2%, 0.4%, l%, 2%, 4% and the like were prepared respectively, and the surface tension of distilled water was measured at 20 ℃ (the temperature difference was within 0.5 ℃) to be 70.06mN/m, and then the tea saponin solutions with different concentrations were used to measure the surface tension of tea saponin, and the measurement results are shown in FIG. 7. According to fig. 7, a Y-lgC diagram is drawn by taking the logarithm of the solution concentration as the abscissa and the measured surface tension value as the ordinate, Y rapidly slides down with the increase of lgC, and slowly rises or becomes gentle after reaching a certain value, a tangent line of the two curves is made, the value corresponding to the tangent line intersection point is an IgC value, and the converted C (concentration) value is CMC (critical micelle concentration) of tea saponin;
as shown in figure 7, the surface tension of the tea saponin solution is drastically reduced along with the continuous increase of the concentration logarithm, when the concentration logarithm is about 0.850%, that is, the concentration of the tea saponin is about O.142%, the surface tension is reduced to the minimum value of 34.86mN/m, and then the concentration is increased again, the surface tension is not reduced, and the tea saponin solution basically tends to be stable, so that a specific 'micelle' structure is formed, and at the moment, the concentration of the tea saponin is the Critical Micelle Concentration (CMC) of the tea saponin is O.142%.
In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing has shown and described the basic principles, principal features and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined in the appended claims.
Claims (8)
1. A purification method of tea saponin is characterized in that: the purification method comprises the following steps:
s1: weighing a small amount of crude tea saponin, and dissolving with 10ml of hot water to prepare a tea saponin solution with the mass fraction of 10%;
s2: regulating the pH value of the tea saponin solution to be 5, and cooling for standby;
s3: adding n-butanol solution or saturated n-butanol solution into cooled tea saponin solution, and shaking uniformly;
s4: extracting the evenly-shaken tea saponin mixed solution to obtain supernatant;
s5: concentrating and drying the obtained supernatant to constant weight to obtain pure tea saponin;
s6: testing the recovery rate of pure tea saponin;
wherein, two parts of crude tea saponin are weighed, respectively dissolved by using 10Ml of hot water, pH is adjusted, and after cooling, n-butanol and saturated n-butanol with the same volume are respectively added, the steps S4 to S6 are repeated, and the extractant is determined to be n-butanol solution or saturated n-butanol solution.
2. The method for purifying tea saponin according to claim 1, wherein: in the step S3, the saturated n-butanol solution is used as an extractant, and in order to determine the optimal use amount of the saturated n-butanol solution, the saturated n-butanol solutions with different volumes are respectively added into the cooled tea saponin solution, and the steps S4 to S6 are repeated to determine the optimal use amount of the saturated n-butanol solution.
3. The method for purifying tea saponin according to claim 2, wherein: in the step S2, in order to determine the optimal pH value, after the step S1, the pH values of different tea saponin solutions are prepared, and after cooling, the steps S4 to S6 are repeated to determine the optimal pH value.
4. A method for purifying tea saponin according to claim 3, wherein: in the step S4, in order to determine the optimal extraction times, different times are extracted respectively, different extracted supernatants are obtained after separation, and the steps S5 to S6 are repeated to determine the optimal extraction times.
5. The method for purifying tea saponin according to claim 4, wherein: in the step S1, in order to determine the optimal crude tea saponin mass fraction, a small amount of crude tea saponin is dissolved by 10ml of hot water and then is respectively prepared into tea saponin solutions with different mass fractions, and the steps S2 to S6 are repeated to determine the optimal crude tea saponin mass fraction.
6. The method for purifying tea saponin according to claim 5, wherein: and in the step S5, the obtained supernatant is placed in a vacuum degree of 0.09Mpa and a temperature of 70 ℃ to be concentrated into paste, and the paste is continuously dried to constant weight to obtain the pure tea saponin.
7. The method for purifying tea saponin according to claim 6, wherein: in the step S7, the calculation formula of the purity and the recovery rate of the tea saponin is as follows:
wherein, C is the purity of tea saponin; c is the concentration in the sample solution calculated by the regression equationDegree mg/ml; x is the dilution of the solution; v is the volume ml of the solution; y is the recovery rate of tea saponin; m is M 1 The amount mg of the tea saponin pure product is weighed for measuring absorbance; m is M 3 The amount of the tea saponin is mg.
8. The method for purifying tea saponin according to claim 7, wherein: in the step S7, the absorbance is measured specifically by weighing the pure tea saponin prepared in the quantitative step S6, dissolving the pure tea saponin in methanol, fixing the volume, developing the color by a concentrated sulfuric acid-vanillin method, testing the absorbance A at the wavelength of 550nm, and substituting the absorbance A into a regression equation:
A＝8.0918c+0.0626。
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102372761A (en) * | 2011-10-10 | 2012-03-14 | 安徽农业大学 | Method for extracting tea saponin from sasanglla cake |
| CN110078782A (en) * | 2019-05-28 | 2019-08-02 | 中国科学院青岛生物能源与过程研究所 | A kind of Tea Saponin extraction and purification process |
| CN113024629A (en) * | 2021-01-05 | 2021-06-25 | 贵州乔盛生物科技有限公司 | Tea saponin purification method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102372761A (en) * | 2011-10-10 | 2012-03-14 | 安徽农业大学 | Method for extracting tea saponin from sasanglla cake |
| CN110078782A (en) * | 2019-05-28 | 2019-08-02 | 中国科学院青岛生物能源与过程研究所 | A kind of Tea Saponin extraction and purification process |
| CN113024629A (en) * | 2021-01-05 | 2021-06-25 | 贵州乔盛生物科技有限公司 | Tea saponin purification method |
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