CN112168851A - Preparation method of nitraria tangutorum bobr extract - Google Patents
Preparation method of nitraria tangutorum bobr extract Download PDFInfo
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61K2236/30—Extraction of the material
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- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A61K2236/50—Methods involving additional extraction steps
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Abstract
The invention relates to a method for preparing nitraria tangutorum bobr extract, which comprises the steps of leaching nitraria tangutorum bobr with ethanol for 2-4 times, combining extracting solutions of each time, centrifuging, taking supernate and concentrating to obtain the nitraria tangutorum bobr extract. The nitraria tangutorum bobr extract prepared by the invention contains higher procyanidine, is extracted by ethanol, has mild extraction conditions and large extraction amount, and is suitable for industrial production; the inventor finds that the content of the procyanidine in the extract can be improved by performing ethanol extraction after the nitraria tangutorum bobr is treated, and the content of the procyanidine is finally improved because the effective active ingredients in the nitraria tangutorum bobr are fully extracted by the combined action of supercritical extraction and acetic acid in the pretreatment of the nitraria tangutorum bobr; the nitraria tangutorum bobr extract prepared by the invention has high content of effective active ingredients and better health-care effect, and can be prepared into health-care products with rich nutrition.
Description
Technical Field
The invention belongs to the technical field of plant extracts, and particularly relates to a preparation method of nitraria tangutorum bobr extract.
Background
The white thorn is rich in resources in China and widely distributed, and has excellent wild excellent shrubs for improving the natural environment, preventing wind, fixing sand and improving soil. The fruits of nitraria tangutorum bobr have rich nutrition, can be processed into wine, beverages and health care products, can also be used as medicines, and has very high edible and medicinal values. Its tender stem and leaf are good pasture, its fruit can be used for edible and medicinal purposes, and its kernel can be used for extracting oil. The nitraria tangutorum bobr not only contains abundant saccharides, fat, protein, vitamin C and organic acid, but also contains various physiologically active substances such as amino acid, macroelement, microelement and unsaturated fatty acid. Due to the restriction of various factors, the resource of the nitraria tangutorum bobr and the nitraria tangutorum bobr has not been well valued and developed in China all the time. At present, few large-scale industrial production of nitraria tangutorum bobr products exists, the pharmacological efficacy research foundation is weak, and the scientific basis of deep development is lacked. Therefore, scientific research and economic value excavation of the nitraria tangutorum bobr still need to be further deepened, so that the nutritional and medicinal values of the nitraria tangutorum bobr are fully utilized.
The nitraria tangutorum bobr fruit contains abundant natural red pigment, has bright rose color and can be used as a good natural food pigment. The extraction separation and the stability of the nitraria tangutorum bobr red pigment are researched, and the nitraria tangutorum bobr red pigment has a very wide application prospect for realizing industrial production of the nitraria tangutorum bobr red pigment.
The present invention has been made in view of the above circumstances.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a method for preparing nitraria tangutorum bobr extract, which has higher content of procyanidine, high content of other active ingredients and better health-care effect.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing Nitraria sibirica pall extract comprises extracting Nitraria sibirica pall with ethanol for 2-4 times, mixing extractive solutions, centrifuging, collecting supernatant, and concentrating to obtain Nitraria sibirica pall extract.
Further, the nitraria tangutorum bobr is firstly treated as follows: performing supercritical extraction on the nitraria tangutorum bobr, and adding acetic acid into the extract for standing treatment.
Furthermore, the pressure of the supercritical extraction is 40-45MPa, the temperature is 40-50 ℃, the flow rate of carbon dioxide is 0.4-0.8L/min, and the extraction time is 1.4-1.8 h.
Furthermore, the pressure of the supercritical extraction is 42.5MPa, the temperature is 45 ℃, the flow rate of carbon dioxide is 0.6L/min, and the extraction time is 1.6 h.
Furthermore, the mass ratio of the extract to the acetic acid is 1: 0.2-0.4.
Further, the volume fraction of the ethanol is 40-60%.
Further, the volume fraction of the ethanol is 50%.
Furthermore, the volume-mass ratio of the ethanol to the nitraria tangutorum bobr is 8-20ml to 1 g.
Further, the nitraria tangutorum bobr is extracted by ethanol for 3 times, and the extraction time is 1-4h each time.
Further, the centrifugation speed is 2800-.
Further, the speed of centrifugation is 3000r/min, and the centrifugation time is 30 min.
Compared with the prior art, the invention has the beneficial effects that:
(1) the nitraria tangutorum bobr extract prepared by the invention contains higher procyanidine, is extracted by ethanol, has mild extraction conditions and large extraction amount, and is suitable for industrial production;
(2) the inventor finds that the content of the procyanidine in the extract can be improved by performing ethanol extraction after the nitraria tangutorum bobr is treated, and the content of the procyanidine is finally improved because the effective active ingredients in the nitraria tangutorum bobr are fully extracted by the combined action of supercritical extraction and acetic acid in the pretreatment of the nitraria tangutorum bobr;
(3) the nitraria tangutorum bobr extract prepared by the invention has high content of effective active ingredients and better health-care effect, and can be prepared into health-care products with rich nutrition.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.
Example 1
The preparation method of nitraria tangutorum bobr extract of the embodiment includes the steps of leaching nitraria tangutorum bobr with 40% ethanol for 2 times, combining extracting solutions of each time, enabling the volume mass ratio of ethanol to the nitraria tangutorum bobr to be 8ml:1g, enabling the extracting time to be 1 hour each time, centrifuging at the speed of 2800r/min for 35min, taking supernate and concentrating to obtain the nitraria tangutorum bobr extract.
Example 2
The preparation method of nitraria tangutorum bobr extract of the embodiment includes the steps of leaching nitraria tangutorum bobr with 50% of ethanol for 3 times, combining extracting solutions of each time, enabling the volume mass ratio of the ethanol to the nitraria tangutorum bobr to be 14ml:1g, enabling the extracting time to be 2.5h, centrifuging at the speed of 3000r/min for 30min, and taking supernate to concentrate to obtain the nitraria tangutorum bobr extract.
Example 3
The preparation method of nitraria tangutorum bobr extract of the embodiment includes the steps of leaching nitraria tangutorum bobr with 60% ethanol for 4 times, combining extracting solutions of each time, enabling the volume mass ratio of the ethanol to the nitraria tangutorum bobr to be 20ml:1g, enabling the extracting time to be 4 hours each time, centrifuging at the speed of 3200r/min for 25 minutes, and taking supernate to concentrate to obtain the nitraria tangutorum bobr extract.
Example 4
The preparation method of the nitraria tangutorum bobr extract in the embodiment is the same as that in the embodiment 1, except that the nitraria tangutorum bobr is treated before ethanol extraction, and the specific method is as follows: performing supercritical extraction on the nitraria tangutorum bobr, wherein the pressure of the supercritical extraction is 40MPa, the temperature is 40 ℃, the flow rate of carbon dioxide is 0.4L/min, the extraction time is 1.4h, adding acetic acid into the extract, and standing, wherein the mass ratio of the extract to the acetic acid is 1: 0.2.
Example 5
The preparation method of the nitraria tangutorum bobr extract in the embodiment is the same as that in the embodiment 2, except that the nitraria tangutorum bobr is treated before ethanol extraction, and the specific method is as follows: carrying out supercritical extraction on the nitraria tangutorum bobr, wherein the pressure of the supercritical extraction is 42.5MPa, the temperature is 45 ℃, the flow rate of carbon dioxide is 0.6L/min, the extraction time is 1.6h, adding acetic acid into the extract, and standing, wherein the mass ratio of the extract to the acetic acid is 1: 0.3.
Example 6
The preparation method of the nitraria tangutorum bobr extract in the embodiment is the same as that in the embodiment 3, except that the nitraria tangutorum bobr is treated before ethanol extraction, and the specific method is as follows: performing supercritical extraction on the nitraria tangutorum bobr, wherein the pressure of the supercritical extraction is 45MPa, the temperature is 50 ℃, the carbon dioxide flow rate is 0.8L/min, the extraction time is 1.8h, adding acetic acid into the extract, and standing, wherein the mass ratio of the extract to the acetic acid is 1: 0.4.
Comparative example 1
The preparation method of Nitraria sibirica pall extract in this example is the same as that of example 2, except that Nitraria sibirica pall is treated by standing without adding acetic acid.
Detection of procyanidins
1. Preparation reagent
(1) A: 1% vanillin solution (weighing 1.000g vanillin in methanol solution, and adding volume to 100 ml);
(2) b: 8% hydrochloric acid solution (8 ml of concentrated hydrochloric acid is dissolved in methanol, and the volume is fixed to 100 ml);
(3) color developing agent: a and B are 1:1, and are prepared as before.
2. And (3) measuring the total amount of procyanidine of nitraria tangutorum bobr: taking 1-2 g nitraria tangutorum bobr, adding 10ml of methanol, shaking for 20min, centrifuging at 3000r/min for 20min, taking 1ml of supernatant, and performing color development analysis (if the concentration is too high, the supernatant can be diluted again).
3. Measuring the content of procyanidine in the nitraria tangutorum bobr extract: 1ml of the supernatant after ethanol extraction is taken for color analysis (if the concentration is too high, the supernatant can be diluted again).
Measuring the content of procyanidine by spectrophotometry: 1ml of sampling liquid is added with 5ml of color developing agent and shaken up. Keeping away from light, and keeping in constant temperature water bath at 30 + -1 deg.C for 30 min. Taking out, measuring the absorbance under the wavelength of 500nm, and calculating the content of procyanidin in the sample according to the standard curve.
4. Drawing a standard curve: preparing procyanidin standard solution with concentration of 1.2 mg/ml. 1, 2, 3, 4 and 5ml are respectively taken and then the volume is up to 10 ml. 1ml of each reagent was added with 5ml of color developing agent. And (5) keeping the temperature and carrying out color comparison, wherein the operation is the same as 1.2.3, and drawing a standard curve.
5. Determination of content
The sample is measured according to a standard curve method, a reagent blank is taken as a reference, and the content of the sample is calculated according to a regression equation.
Test example 1 influence of ethanol concentration
Respectively weighing 50g of nitraria tangutorum bobr, putting the nitraria tangutorum bobr into a round-bottom flask, adding 12 times of ethanol with different concentrations, performing reflux extraction for 2 hours, and performing extraction once, wherein the content of procyanidine in an extracting solution and the cream yield are shown in table 1.
TABLE 1
| Ethanol volume concentration (%) | 30 | 40 | 50 | 60 | 70 |
| Procyanidins (mg/g) | 4.32 | 5.51 | 6.74 | 7.02 | 7.25 |
| Percentage of cream discharged (%) | 29.9 | 28.5 | 27.4 | 25.5 | 24.8 |
As can be seen from table 1, the extraction rate of procyanidin increases and the extraction yield decreases with the increase of ethanol concentration, and when the ethanol concentration increases to 50%, the increase of procyanidin is not obvious, and 50% ethanol is selected as the extraction solvent in comprehensive consideration.
Test example 2 influence of ethanol volume
Respectively weighing 50g of nitraria tangutorum bobr, respectively adding 50% ethanol in an amount which is 8, 10, 12, 15 and 20 times of ml of the nitraria tangutorum bobr, performing reflux extraction for 2 hours, and performing extraction once, wherein the content of procyanidine in an extracting solution and the cream yield are shown in table 2.
TABLE 2
| Volume multiple of ethanol | 8 | 10 | 12 | 15 | 20 |
| Procyanidins (mg/g) | 5.41 | 6.74 | 6.83 | 7.36 | 7.71 |
| Percentage of cream discharged (%) | 24.3 | 27.4 | 28.5 | 30.4 | 33.5 |
As can be seen from table 2, with the increase of the volume of ethanol, the procyanidin and the cream yield are both increased, the increase of the cream yield is obvious, and considering that the effect of a small amount of multiple extraction is better, 12 times of ethanol is selected as the extraction solvent.
Test example 3 Effect of extraction frequency
Weighing 50g of nitraria tangutorum bobr, putting the nitraria tangutorum bobr into a round-bottom flask, adding 50% ethanol with the volume being 12 times that of the nitraria tangutorum bobr, performing reflux extraction for 2 hours, and performing extraction for four times, wherein the procyanidine and the cream yield in the extracting solution are shown in table 3.
TABLE 3
| Number of times of extraction | 1 time of | 2 times (one time) | 3 times of | 4 times (twice) |
| Procyanidins (mg/g) | 6.74 | 3.55 | 0.81 | 0.32 |
| Percentage of cream discharged (%) | 27.4 | 10.3 | 5.6 | 2.3 |
As can be seen from table 3, with the increase of the extraction times, the procyanidins which can be further extracted and the extraction rate are gradually reduced, and after two extractions, the extraction is basically complete, so two extractions are selected.
Test example 4 Effect of extraction time
Weighing 50g of nitraria tangutorum bobr, putting the nitraria tangutorum bobr into a round-bottom flask, adding 50% ethanol with the volume being 12 times that of the nitraria tangutorum bobr, performing reflux extraction for 1 hour, 2 hours, 3 hours and 4 hours, and extracting for 1 time, wherein the procyanidine and the cream yield in the extracting solution are shown in table 4.
TABLE 4
| Extraction time (h) | 1 | 2 | 3 | 4 |
| Procyanidins (mg/g) | 3.88 | 6.74 | 7.12 | 7.25 |
| Percentage of cream discharged (%) | 20.8 | 27.4 | 29.8 | 31.2 |
As can be seen from table 4, the procyanidin and the extraction yield both increased with the increase of the extraction time, and the increase of procyanidin was not significant after 2 hours of extraction, so 2 hours was selected as the extraction time.
Experimental example 5 orthogonal experiment
From the results of test examples 1 to 4, ethanol concentration, ethanol volume, and extraction time were selected as factors of the orthogonal test by the above one-factor test, and the levels of the factors of the orthogonal test are shown in Table 5 below.
TABLE 5 orthogonal experiment factor table
Performing tests according to an orthogonal design, and respectively calculating the extraction rate of procyanidine and the extraction rate of extractum under each factor level, wherein the extraction rate of procyanidine accounts for 60 percent, and the extraction rate of extractum accounts for 40 percent; the total score is procyanidin extraction rate 60+ extractum extraction rate 40, the orthogonal test arrangement and result analysis are shown in table 6, and the orthogonal test result variance analysis is shown in table 7.
TABLE 6
| Factors of the fact | Concentration of ethanol | Volume of ethanol | Number of times of extraction | Extraction time | Composite score |
| Experiment 1 | 1(40％) | 1(10) | 1(1) | 1(2) | 61.66 |
| Experiment 2 | 1(40％) | 2(12) | 2(2) | 2(3) | 72.20 |
| Experiment 3 | 1(40％) | 3(15) | 3(3) | 3(4) | 75.82 |
| Experiment 4 | 2(50％) | 1(10) | 2(2) | 3(4) | 72.58 |
| Experiment 5 | 2(50％) | 2(12) | 3(3) | 1(2) | 78.12 |
| Experiment 6 | 2(50％) | 3(15) | 1(1) | 2(3) | 72.78 |
| Experiment 7 | 3(60％) | 1(10) | 3(3) | 2(3) | 76.68 |
| Experiment 8 | 3(60％) | 2(12) | 1(1) | 3(4) | 68.02 |
| Experiment 9 | 3(60％) | 3(15) | 2(2) | 1(2) | 76.28 |
| Mean value 1 | 69.893 | 70.307 | 67.487 | 72.020 | - |
| Mean value 2 | 74.493 | 72.780 | 73.687 | 73.887 | - |
| Mean value 3 | 73.660 | 74.960 | 76.873 | 72.140 | - |
| Extreme difference | 4.600 | 4.653 | 9.386 | 1.867 | - |
TABLE 7
| Factors of the fact | Sum of squares of deviation | Degree of freedom | F ratio | Critical value of F | Significance of |
| Concentration of ethanol | 36.042 | 2 | 0.681 | 4.460 | - |
| Volume of ethanol | 32.523 | 2 | 0.614 | 4.460 | - |
| Number of times of extraction | 136.704 | 2 | 2.582 | 4.460 | - |
| Extraction time | 6.550 | 2 | 0.124 | 4.460 | - |
| Error of the measurement | 211.82 | 8 | - | - | - |
As can be seen from tables 6 and 7, the analysis of K1, K2 and K3 combined with the results of the level of each factor in the orthogonal test preliminarily determined that the extraction process is A2B3C3D2Namely ethanol concentration of 50%, solvent volume of 15 times of water, extraction times of 3 times, and extraction time of 2 hours each time. According to the comparison of the range R, the sequence of the influencing factors of the extraction process is C > B > A > D, namely the extraction times > ethanol volume > ethanol concentration > extraction time. In conclusion, the optimal extraction process is finally determined to be 50% of ethanol concentration, 15 times of ethanol volume of water, 3 times of extraction and 2 hours of extraction time each time.
Test example 7
The contents of procyanidins in the supernatants of Nitraria tangutorum fruits prepared in examples 1 to 6 and comparative example were measured, respectively, and the results are shown in Table 8.
TABLE 8
As can be seen from table 8, the procyanidin content of the extractive solution can be increased after the nitraria tangutorum bobr is processed before ethanol extraction, because the effective active ingredients in the nitraria tangutorum bobr are fully extracted through the combined action of supercritical extraction and acetic acid in the pretreatment of the nitraria tangutorum bobr, and the procyanidin content is finally increased.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (10)
1. A method for preparing nitraria tangutorum bobr extract is characterized in that nitraria tangutorum bobr extract is obtained by leaching nitraria tangutorum bobr with ethanol for 2-4 times, combining extracting solutions of each time, centrifuging, and concentrating supernate.
2. The method for preparing Nitraria tangutorum bobr extract according to claim 1, wherein the Nitraria tangutorum bobr is first treated as follows: performing supercritical extraction on the nitraria tangutorum bobr, and adding acetic acid into the extract for standing treatment.
3. The method for preparing nitraria tangutorum bobr extract according to claim 2, wherein the pressure of supercritical extraction is 40-45MPa, the temperature is 40-50 ℃, the carbon dioxide flow rate is 0.4-0.8L/min, and the extraction time is 1.4-1.8 h.
4. The method for preparing nitraria tangutorum bobr extract according to claim 3, wherein the pressure of supercritical extraction is 42.5MPa, the temperature is 45 ℃, the flow rate of carbon dioxide is 0.6L/min, and the extraction time is 1.6 h.
5. The method for preparing nitraria tangutorum bobr extract according to claim 2, wherein the mass ratio of the extract to the acetic acid is 1: 0.2-0.4.
6. The method for preparing nitraria tangutorum bobr extract according to claim 1, wherein the volume fraction of the ethanol is 40-60%, preferably 50%.
7. The preparation method of nitraria tangutorum bobr extract as claimed in claim 6, wherein the volume-mass ratio of the ethanol to the nitraria tangutorum bobr is 8-20ml:1 g.
8. The method for preparing nitraria tangutorum bobr extract according to claim 1, wherein nitraria tangutorum bobr is extracted with ethanol for 3 times, and the extraction time is 1-4 hours each time.
9. The method for preparing Nitraria tangutorum bobr extract as claimed in claim 1, wherein the centrifugation speed is 2800-.
10. The method for preparing Nitraria sibirica pall extract according to claim 9, wherein the centrifugation speed is 3000r/min, and the centrifugation time is 30 min.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105054070A (en) * | 2015-09-07 | 2015-11-18 | 中国科学院西北高原生物研究所 | Sophora davidii anthocyanin crude extract and microcapsule thereof |
| CN107684575A (en) * | 2016-08-04 | 2018-02-13 | 北京铂鑫天然生物技术有限公司 | The preparation method and purposes of white bur activity extract |
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Application publication date: 20210105 |