CN115813964A - Extraction process and application of antioxidant centella asiatica extract - Google Patents
Extraction process and application of antioxidant centella asiatica extract Download PDFInfo
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- CN115813964A CN115813964A CN202211396450.8A CN202211396450A CN115813964A CN 115813964 A CN115813964 A CN 115813964A CN 202211396450 A CN202211396450 A CN 202211396450A CN 115813964 A CN115813964 A CN 115813964A
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- asiaticoside
- madecassoside
- centella asiatica
- ethanol
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- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 19
- 238000000605 extraction Methods 0.000 title claims abstract description 18
- 239000003963 antioxidant agent Substances 0.000 title claims abstract description 9
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- WYQVAPGDARQUBT-XCWYDTOWSA-N asiaticoside Natural products O=C(O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@H](O)[C@H](O)[C@H](O[C@H]3[C@H](O)[C@H](O)[C@@H](O)[C@H](C)O3)[C@@H](CO)O2)O1)[C@@]12[C@@H]([C@@H](C)[C@H](C)CC1)C=1[C@](C)([C@@]3(C)[C@@H]([C@@]4(C)[C@H]([C@@](CO)(C)[C@@H](O)[C@H](O)C4)CC3)CC=1)CC2 WYQVAPGDARQUBT-XCWYDTOWSA-N 0.000 claims abstract description 60
- WYQVAPGDARQUBT-FGWHUCSPSA-N Madecassol Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O)OC[C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)OC(=O)[C@]12CC[C@H]([C@@H]([C@H]1C=1[C@@]([C@@]3(CC[C@H]4[C@](C)(CO)[C@@H](O)[C@H](O)C[C@]4(C)[C@H]3CC=1)C)(C)CC2)C)C)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O WYQVAPGDARQUBT-FGWHUCSPSA-N 0.000 claims abstract description 57
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- BNMGUJRJUUDLHW-HLUHVYOBSA-N Madecassoside Natural products C[C@@H]1CC[C@@]2(CC[C@]3(C)C(=CC[C@@H]4[C@@]5(C)C[C@@H](O)[C@H](O)[C@@](C)(CO)[C@@H]5[C@H](O)C[C@@]34C)[C@@H]2[C@H]1C)C(=O)O[C@@H]6O[C@H](CO[C@@H]7O[C@H](CO)[C@@H](O[C@@H]8O[C@H](C)[C@H](O)[C@@H](O)[C@H]8O)[C@H](O)[C@H]7O)[C@@H](O)[C@H](O)[C@H]6O BNMGUJRJUUDLHW-HLUHVYOBSA-N 0.000 claims abstract description 37
- 229940090813 madecassoside Drugs 0.000 claims abstract description 37
- 239000002253 acid Substances 0.000 claims abstract description 5
- JXSVIVRDWWRQRT-UYDOISQJSA-N asiatic acid Chemical compound C1[C@@H](O)[C@H](O)[C@@](C)(CO)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C)[C@H](C)[C@H]5C4=CC[C@@H]3[C@]21C JXSVIVRDWWRQRT-UYDOISQJSA-N 0.000 claims abstract description 5
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- PRAUVHZJPXOEIF-AOLYGAPISA-N madecassic acid Chemical compound C1[C@@H](O)[C@H](O)[C@@](C)(CO)[C@@H]2[C@H](O)C[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C)[C@H](C)[C@H]5C4=CC[C@@H]3[C@]21C PRAUVHZJPXOEIF-AOLYGAPISA-N 0.000 claims abstract description 5
- 229940011656 madecassic acid Drugs 0.000 claims abstract description 5
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 51
- 239000007788 liquid Substances 0.000 claims description 16
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- 239000003814 drug Substances 0.000 claims description 13
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- 244000146462 Centella asiatica Species 0.000 claims description 10
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- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 4
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- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
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- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
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Abstract
The invention discloses an extraction process and application of an antioxidant centella extract, which comprises the application of the centella extract in preparing a product for removing free radicals; comprises madecassoside, asiaticoside and asiaticoside. Asiaticosides include asiatic acid, madecassic acid and triterpenic acids; the mass fractions of madecassoside, asiaticoside and asiaticoside are all 1.0% -100%. According to the invention, the extraction purity is high, the separation component is accurate, and the free radical scavenging capability is high.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine preparation, and particularly relates to an extraction process and application of an antioxidant centella extract.
Background
Radicals are various atoms, molecules or ions containing an unpaired electron, and can be classified into oxygen radicals and non-oxygen radicals according to whether they contain an oxygen molecule or not. Free radicals can be produced by both endogenous and exogenous pathways. 90% of the endogenous free radicals in eukaryotic cells are generated by the escape of mitochondrial electrons, which are associated with O 2 Reaction to form O -2 Which is then converted to H 2 O 2 . The remaining 10% are produced in peroxisomes, plasma and nuclear membranes, etc. The exogenous free radicals are generated in a wide range of ways, external stimuli such as light, heat, ultrasound and radiation can cause redox reaction and electrolytic reaction to generate free radicals, and the external environment can increase psychological pressure and stress sensation and enable organisms to generate free radicals. The body can prevent the excessive accumulation of free radicals by the antioxidant defense system, balancing its destructive action. Scientific research on aging of human body caused by free radicals has been accepted by most people, and aging-causing mechanisms and forms are various, but are comprehensively expressed by various age-related stress reactions and inflammatory reactions.
Endogenous and exogenous stimuli produce free radicals upon acting on the body, which intervene and interfere with normal signal transduction pathways, but the ability to cause injury and even aging depends not only on the intensity of the stimuli, but also on the fitness of the body.
Asiaticoside (AS) is a triterpenoid compound in centella asiatica and has a molecular formula of C 48 H 78 O 19 And a molecular weight of 959.12.As one of the main pharmacological active ingredients of the centella asiatica, the AS has various pharmacological actions of resisting tumors and inflammation, protecting the function of the nervous system, promoting wound healing and the like.
Asiaticoside is prepared by combining asiatic acid, madecassic acid and triterpenic acid in asiatic pennywort herb extract, and has strong application value in removing free radicals and resisting oxidation of skin.
Madecassoside (MC) is the pentacyclic triterpene with highest content in herba Centellae, and has molecular formula of C 48 H 78 O 20 The crystal had a white appearance relative to a molecular mass of 975.13. Madecassoside is a saponin secondary plant metabolite synthesized by centella asiatica in the isoprene pathway. Naturally derived compounds generally have a wide range of biological activities in the biomedical field and also have a wide range of applications in clinical and everyday life.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide the extraction process and the application of the antioxidant centella asiatica extract, which have the advantages of high extraction purity, accurate separation component and high-efficiency free radical scavenging capability. To achieve the above objects and other advantages in accordance with the present invention, there are provided
Application of herba Centellae extract in preparing antioxidant product is provided.
Preferably, it comprises madecassoside, asiaticoside and asiaticoside.
Preferably, asiaticosides include asiatic acid, madecassic acid and triterpenic acids.
Preferably, the mass fractions of the madecassoside, the asiaticoside and the asiaticoside are all 1% -100%.
Preferably, the mass fractions of the madecassoside, the asiaticoside and the asiaticoside are 90-100%.
A process for extracting madecassoside for scavenging free radicals, comprising:
s1, smashing the whole centella asiatica, and stirring and extracting;
s2, performing filter pressing, adsorbing the liquid medicine on a resin column, and eluting effective components from the resin column by using ethanol;
s3, decoloring the liquid medicine by applying the liquid medicine to a decoloring resin column, and concentrating the decolored liquid into thick paste by a scraper concentrator;
s4, transferring to a microwave drying box of a D-level clean area for drying;
s5, grinding, sieving and mixing to obtain the asiaticoside;
s6, dissolving the ethanol to separate the asiaticoside to obtain a madecassoside crude product;
s7, dissolving and refining by 95% ethanol, drying, crushing and screening by using a screen to obtain the madecassoside.
Preferably, in step S1, the whole plant of centella is pulverized into 60 mesh fine powder, and the powder is put into an extraction pot, 900kg of the fine powder is added into 4.5 tons of boiling water per pot, and stirred and extracted for 2 hours.
Preferably, the screens in step S5 and step S7 are both 100 mesh.
Preferably, the ethanol in step S6 is 5 times 95% ethanol.
Application of madecassoside in preparing antiallergic product is provided.
Application of madecassoside in preparing chemical burn repairing product is provided.
Compared with the prior art, the invention has the beneficial effects that: the asiaticoside, the madecassoside and the asiaticoside extracted by the preparation process have high extraction purity and accurate separation component, and the main efficacy components of the asiaticoside, the madecassoside and the asiaticoside in the centella asiatica are high in free radical scavenging capability.
Drawings
FIG. 1 is a process flow diagram of the extraction process and application of centella asiatica extract having antioxidant activity according to the present invention;
FIG. 2 is a graph of 90% concentration of asiaticoside versus ABTS + free radical clearance for the extraction process and application of centella asiatica extract having antioxidant properties according to the present invention;
FIG. 3 is a graph of 90% asiaticoside concentration versus ABTS + free radical clearance for the extraction process and application of an antioxidant centella asiatica extract according to the present invention;
FIG. 4 is a graph of 90% madecassoside concentration versus ABTS + free radical scavenging rate for the extraction process and application of centella asiatica extract having antioxidant properties according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-4, the use of centella asiatica extract in the preparation of antioxidant products.
Further, it comprises madecassoside, asiaticoside and asiaticoside.
Further, asiaticosides include asiatic acid, madecassic acid and triterpenic acids.
Furthermore, the mass fractions of the madecassoside, the asiaticoside and the asiaticoside are all 1% -100%.
Furthermore, the mass fractions of the madecassoside, the asiaticoside and the asiaticoside are all 90-100%.
A process for extracting madecassoside for scavenging free radicals, comprising: s1, smashing the whole centella asiatica, and stirring and extracting;
s2, performing filter pressing, adsorbing the liquid medicine on a resin column, and eluting effective components from the resin column by using ethanol;
s3, decoloring the liquid medicine by passing the liquid medicine through a decoloring resin column, and concentrating the decolored liquid into thick paste through a scraper concentrator;
s4, transferring to a microwave drying box of the D-level clean area for drying;
s5, grinding, sieving and totally mixing to obtain the asiaticoside;
s6, dissolving the ethanol to separate the asiaticoside to obtain a madecassoside crude product;
s7, dissolving and refining by 95% ethanol, drying, crushing and screening by using a screen to obtain the madecassoside.
A process for extracting asiaticoside to remove free radicals includes such steps as pulverizing the whole plant of asiatic centella to 60 meshes, loading it in extracting tank, adding 4.5 tons of boiling water to 900kg of fine powder, stirring for 2 hr, filter pressing, adsorbing by resin column, eluting with 60-75% alcohol, decoloring by resin column, concentrating by scraper concentrator (55-75 deg.C and vacuum more than 0.06 MPa) to obtain thick paste (relative density 1.05-1.15 (75 deg.C), baking in microwave drying box (30-65 deg.C and vacuum more than 0.06 MPa), pulverizing, sieving by 100 meshes, and mixing. Dissolving asiaticoside with 5 times of 95% ethanol to obtain asiaticoside crude product, dissolving and refining with 95% ethanol, oven drying, pulverizing, and sieving with 100 mesh sieve to obtain herba Centellae.
An extraction process of asiaticoside for removing free radicals comprises the steps of crushing whole centella into fine powder of 60 meshes, putting the fine powder into an extraction tank, adding 4.5 tons of boiling water into each tank of 900kg of fine powder, stirring and extracting for 2 hours, carrying out filter pressing, adsorbing liquid medicine on a resin column, eluting effective components from the resin column by using ethanol (60% -75%), then putting the liquid medicine on a decolorizing resin column, decolorizing the liquid medicine, concentrating the decolorized liquid into thick paste (relative density is 1.05-1.15 (75 ℃) by using a scraper concentrator (temperature is 55-75 ℃ and vacuum is more than 0.06 MPa), transferring to a microwave drying box (temperature is 30-65 ℃ and vacuum is more than 0.06 MPa) of a D-grade clean area, drying, crushing, sieving by using a sieve of 100 meshes, and carrying out total mixing to obtain the asiaticoside. Hydrolyzing, standing for precipitation, discarding precipitate, filtering the supernatant with plate-and-frame filter, refining, oven drying, pulverizing, and sieving with 100 mesh sieve to obtain asiaticoside.
ABTS + free radical scavenging test
Purpose of detection
The samples to be detected were detected as three samples: the antioxidation of 90% of asiaticoside, 90% of asiaticoside and 90% of madecassoside is the ability to scavenge free radicals.
Detection instrument and apparatus
Analyzing a precision balance: weighing 0.1mg;
volumetric flask: 10mL, 20mL, 50mL, 100mL;
adjustable pipette: 1.00mL;
an enzyme-linked immunoassay detector: UV-1800 in the ultraviolet-visible spectrophotometer.
Reagent
All reagents were analytically pure except where otherwise stated, and water was first grade water in accordance with GB/T6682.
95% ethanol;
2,2' -biazoie-bis-3-ethylbenzthiazoline-6-sulfonic acid (Sigma);
positive control: vitamin E with purity not less than 96%.
Sample information
TABLE 4-1 sample essential information
Experimental procedure
5.1ABTS + mother liquor: equal volumes of 7.4mM ABTS aqueous solution and 2.6mM potassium persulfate aqueous solution are measured, mixed uniformly and reacted for 12-16h under the condition of room temperature and darkness. Preparing with brown bottle, wrapping with tinfoil, and storing at low temperature;
5.2ABTS + working solution: diluting ABTS + mother liquor with 95% ethanol until absorbance is 0.7 + -0.05, and keeping out of the sun;
5.3 Positive controls were diluted with 95% ethanol solution to: the test system was verified by using a series of concentration gradients of 0.08mg/mL, 0.04mg/mL, 0.02mg/mL, and 0.01 mg/mL.
5.4 treatment of the sample to be detected: the test substance is diluted by 95% ethanol to be a multi-concentration sample.
Referring to Table 5.4-1, 10mL test tubes were used to set up sample tubes (T), sample background (T0), ABTS + tubes (C) and solvent background (C0), 3 parallel tubes were set up for each sample concentration of sample tubes (T), and 3 parallel tubes were set up for ABTS + tubes (C).
1mL of the same concentration of sample solution was added to each of the sample tube (T) and sample background (T0).
5.5 in all test tubes (T, T, C, C) supplement solvent, the sample to be tested is added with 95% ethanol, make up for 3mL, and mix well.
5.6 adding 1mL of ABTS + ethanol solution into the sample tube (T) and the ABTS + tube (C), replacing the sample background (T0) and the solvent background (C0) with 95% ethanol, shaking up gently, and standing for 5 minutes at room temperature.
5.7 transfer each reaction solution into a 1cm cuvette and measure the absorbance at 734 nm.
TABLE 5.4-1 sample addition requirements
| T-sample tube | T0-sample background | C-ABTS + Pipe | C0-solvent background | |
| Sample solution (mL) | 1 | 1 | - | - |
| 95% ethanol solvent (mL) | 2 | 2 | 3 | 3 |
| ABTS + Working solution (mL) | 1 | - | 1 | - |
| 95% ethanol (mL) | - | 1 | - | 1 |
| Number of |
3/ |
1/ |
3/ |
1/test |
6. Results of the experiment
6.1 ABTS + free radical scavenging results for the test samples.
6.2 calculate ABTS + free radical clearance:
in the formula:
t-sample tube light absorption value, and the light absorption value of the solution after the sample reacts with ABTS +;
t0-sample background light absorption value;
C-ABTS + tube light absorption value is averaged for 3 times, namely the ABTS + solution light absorption value when no sample is added;
c0-background absorbance of solvent.
6.3 test results the elimination result of ABTS + free radical by 90% asiaticoside in the sample to be tested.
TABLE 6.3-1 asiaticoside 90% vs ABTS + Detection result of radical scavenging
* The data in the table are mean ± relative deviation.
* IC50 is the concentration tested for the corresponding sample at which 50% clearance was achieved.
6.4 test results the elimination result of ABTS + free radical by asiaticoside 90% in a sample to be tested
TABLE 6.4-1 Asiaticoside 90% vs ABTS + Detection results of radical scavenging
* The data in the table are mean ± relative deviation.
* IC50 is the concentration tested for the corresponding sample at which 50% clearance was achieved.
6.5 detection result the elimination result of 90% of madecassoside to ABTS + free radical of the sample to be detected.
TABLE 6.5-1 Madecassoside 90% vs. ABTS + Detection results of radical scavenging
* The data in the table are mean ± relative deviation.
* IC50 is the concentration tested for the corresponding sample at which 50% clearance is achieved.
Test with positive control, IC of vitamin E 50 The tested concentration of the sample (corresponding to the removal effect of 50 percent) is 0.02mg/mL-0.06mg/mL, and all batch test systems of the project are effective.
7 detection conclusion
7.1 As can be seen from Table 6.3-1 and FIG. 2, the clearance of asiaticoside 90% to ABTS + free radical is not obvious in the anti-oxidation effect within the tested concentration range of 0.25-2.5mg/mL, and the clearance does not reach 50% (i.e. IC) 50 )；
7.2 As can be seen from Table 6.4-1 and FIG. 3, the 90% sample of asiaticoside has ABTS + free radical scavenging efficiencyThe test concentration range is 0.025-0.25mg/mL, and the sample concentration and clearance rate are in linear relationship and reach 50% clearance rate (IC) 50 ) The concentration is about 0.1651mg/mL;
7.3 As can be seen from Table 6.5-1 and FIG. 4, the clearance of 90% madecassoside sample to ABTS + free radicals is in the range of 0.25-2.5mg/mL of the tested concentration;
7.4 using ABTS + free radical scavenging detection method, the result is that asiaticoside and madecassoside both have certain antioxidant effect, but asiaticoside antioxidant effect is not obvious, antioxidant effect; asiaticoside > madecassoside > asiaticoside.
The number of devices and the scale of the processes described herein are intended to simplify the description of the invention, and applications, modifications and variations of the invention will be apparent to those skilled in the art.
While embodiments of the invention have been described above, it is not intended to be limited to the details shown, described and illustrated herein, but is to be accorded the widest scope consistent with the principles and novel features herein disclosed, and to such extent that such modifications are readily available to those skilled in the art, and it is not intended to be limited to the details shown and described herein without departing from the general concept as defined by the appended claims and their equivalents.
Claims (9)
1. Application of herba Centellae extract in preparing antioxidant product is provided.
2. Centella asiatica extract with antioxidant properties according to claim 1, comprising madecassoside, asiaticoside and asiaticoside.
3. The extract of centella asiatica having antioxidant properties according to claim 1, wherein the asiaticoside comprises asiatic acid, madecassic acid and triterpenic acid.
4. The process for extracting centella asiatica extract with antioxidant effect according to claim 1, wherein the mass fractions of madecassoside, asiaticoside and asiaticoside are 1.0-100%.
5. The extraction process of centella asiatica extract with antioxidant activity as claimed in claim 1, wherein the mass fractions of madecassoside, asiaticoside and asiaticoside are 90% -100%.
6. The process for extracting madecassoside scavenging free radicals as claimed in claim 1, comprising the following steps:
s1, smashing the whole centella asiatica, and stirring and extracting;
s2, performing filter pressing, adsorbing the liquid medicine on a resin column, and eluting effective components from the resin column by using ethanol;
s3, decoloring the liquid medicine by passing the liquid medicine through a decoloring resin column, and concentrating the decolored liquid into thick paste through a scraper concentrator;
s4, transferring to a microwave drying box of the D-level clean area for drying;
s5, grinding, sieving and totally mixing to obtain the asiaticoside;
s6, dissolving the ethanol to separate the asiaticoside to obtain a madecassoside crude product;
s7, dissolving and refining by 95% ethanol, drying, crushing and screening by using a screen to obtain the madecassoside.
7. The process for extracting madecassoside for scavenging free radicals as claimed in claim 1, wherein in step S1, the whole plant of centella asiatica is pulverized into fine powder of 60 meshes, and the fine powder is put into extraction tanks, each tank containing 900kg of fine powder, and 4.5 tons of boiling water are added for stirring and extraction for 2 hours.
8. The process for extracting madecassoside for scavenging free radicals as claimed in claim 1, wherein the sieves of step S5 and step S7 are 100 mesh.
9. The process for extracting madecassoside for scavenging free radicals as claimed in claim 1, wherein the ethanol used in step S6 is 5 times 95% ethanol.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1491656A (en) * | 2002-10-22 | 2004-04-28 | 上海海天医药科技开发有限公司 | Medicinal use of asiatic centella extract containing asiaticoside and hydroxy asiaticoside |
| CN103124560A (en) * | 2010-09-09 | 2013-05-29 | 纳图瑞克斯有限公司 | Method for preparing an extract of centella asiatica |
| CN104892719A (en) * | 2014-12-04 | 2015-09-09 | 陕西君碧莎制药有限公司 | Novel efficient preparation method for asiaticoside |
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| CN1491656A (en) * | 2002-10-22 | 2004-04-28 | 上海海天医药科技开发有限公司 | Medicinal use of asiatic centella extract containing asiaticoside and hydroxy asiaticoside |
| CN103124560A (en) * | 2010-09-09 | 2013-05-29 | 纳图瑞克斯有限公司 | Method for preparing an extract of centella asiatica |
| CN104892719A (en) * | 2014-12-04 | 2015-09-09 | 陕西君碧莎制药有限公司 | Novel efficient preparation method for asiaticoside |
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