Application of mangrove plant extract in preparation of bacteriostatic and antioxidant products
The roots, stems and leaves of mangrove plants are extracted through organic solvents to prepare mangrove plant extracts, which solves the problem of insufficient development and utilization of mangrove plant resources, improves the antibacterial and antioxidant effects, and provides a new application direction.
Patent Information
- Application Number
- CN202510629771.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-16
- Publication Date
- 2025-07-11
AI Technical Summary
In the prior art, the research and resource development and utilization of mangrove plants have been insufficient, especially the application in antibacterial and antioxidant has not been fully utilized.
Organic solvents (such as 78% ethanol solution) were used to extract the roots, stems and leaves of mangrove plants to prepare mangrove plant extracts, which were used to prepare antibacterial and antioxidant products, and the antibacterial effect of mangrove plant extraction complex was optimized.
The effect of mangrove plant extracts in antibacterial and antioxidant aspects has been improved, a new direction of utilization of mangrove plant resources has been provided, and the antibacterial effect has been optimized.
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Figure CN120285032A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical fields of drug development and antibacterial and antioxidant products, and particularly relates to the application of a mangrove plant extract in the preparation of antibacterial and antioxidant products. Background Art
[0002] China is rich in mangrove resources. Mangrove plants contain a large number of compounds with unique structures and remarkable activities. However, the research and development and utilization of the medicinal value resources of mangrove plants are insufficient. On the basis of protecting existing resources, this project focuses on the research of extracts and complexes of several mangrove plants and their applications in antibacterial and antioxidant activities. This study conducts research on the separation, extraction, antibacterial testing, and antioxidant testing of active substances from several mangrove plants. Previous studies mainly focused on the antitumor and anti-inflammatory activities of mangrove plant extracts. This study aims to study the antibacterial and antioxidant activities of extracts from several common mangrove plants, including the ethanol extracts and complexes of the roots, stems, and leaves of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia candel, Rhizophora apiculata, Avicennia marina, Xylocarpus granatum, Ceriops tagal, Rhizophora stylosa, and Acanthus ilicifolius, so as to provide data reference for the research on the antibacterial and antioxidant biological activities of mangrove plants. Summary of the Invention
[0003] The present invention provides an application of a mangrove plant extract in the preparation of an antibacterial product, wherein the mangrove plant is at least one of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia candel, Rhizophora apiculata, Avicennia marina, Xylocarpus granatum, Ceriops tagal, Acanthus ilicifolius, and Rhizophora stylosa, and the bacteria are Escherichia coli or Bacillus subtilis.
[0004] In one embodiment of the present invention, the mangrove plant extract is prepared by extraction with an organic solvent.
[0005] In one embodiment of the present invention, the organic solvent is an ethanol solution.
[0006] In one embodiment of the present invention, the ethanol solution is a 78% ethanol solution.
[0007] In one embodiment of the present invention, the mangrove plant extract is an extract of any one of the roots, stems, and leaves of the mangrove plant.
[0008] In one embodiment of the present invention, the mangrove plant extract is composed of extracts of Avicennia marina and Bruguiera gymnorrhiza.
[0009] In one embodiment of the present invention, the mass ratio of the extracts of Avicennia marina and Bruguiera gymnorrhiza is 1:1.
[0010] The present invention also provides an application of a mangrove plant extract in the preparation of an antioxidant product, wherein the mangrove plant is at least one of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia candel, Rhizophora apiculata, Xylocarpus granatum, Ceriops tagal, Acanthus ilicifolius leaves, and Rhizophora stylosa.
[0011] In one embodiment of the present invention, the mangrove plant extract is prepared by extraction with an organic solvent.
[0012] In one embodiment of the present invention, the organic solvent is a 78% ethanol solution.
[0013] Compared with the prior art, the present invention has the following beneficial effects:
[0014] The present invention applies the extract of mangrove plant active substances to antioxidant and antibacterial applications, and has found an optimized mangrove plant extraction complex, whose antibacterial effect is better than that of a single mangrove plant extract, providing a new direction for the utilization of mangrove plants. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] Figure 1 Shows the influence of three factors on the size of the antibacterial zone of the extract of Acanthus ilicifolius in Example 1.
[0016] Figure 2 Shows the scavenging effect of the crude extract solutions of the roots, stems and leaves of eight mangrove plants on DPPH in Example 1.
[0017] Figure 3 Shows the scavenging effect of the crude extract solution of the roots of eight mangrove plants on DPPH in Example 1.
[0018] Figure 4 Shows the scavenging effect of the crude extract solution of the stems of eight mangrove plants on DPPH in Example 1.
[0019] Figure 5 Shows the scavenging effect of the crude extract solution of the leaves of eight mangrove plants on DPPH in Example 1.
[0020] Figure 6 Shows the scavenging effect on DPPH of the best scavenging effect among the roots, stems and leaves of each mangrove plant in Example 1. DETAILED DESCRIPTION OF THE INVENTION
[0021] Example 1
[0022] Experimental materials and methods
[0023] Mangrove plants: the roots, stems and leaves of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia obovata, Rhizophora apiculata, Avicennia marina, Xylocarpus granatum, Ceriops tagal, Acanthus ilicifolius and Rhizophora stylosa.
[0024] Antibacterial test strains: Bacillus subtilis, Escherichia coli, Saccharomyces cerevisiae, Aspergillus flavus
[0025] Bacterial culture medium: NaCl (0.05 g), beef extract (0.03 g), peptone (1.0 g), agar (1.5 g), purified water (100 ml).
[0026] Yeast culture medium: yeast extract (0.3 g), glucose (0.1 g), peptone (0.8 g), agar (1.5 g), purified water (100 ml).
[0027] Fungal culture medium: potato powder (20 g), glucose (2 g), agar (1.5 g), purified water (100 ml).
[0028] 1. Ethanol extraction of roots, stems and leaves of mangrove plants
[0029] Collect the roots, stems and leaves of several mangroves including Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia obovata, Rhizophora apiculata, Avicennia marina, Xylocarpus granatum, Ceriops tagal, Acanthus ilicifolius and Rhizophora stylosa respectively. After washing and air-drying, they are crushed. The crushed roots, stems and leaves are placed in 78% ethanol solution according to the material-liquid ratio of 1:12 (mg / ml) respectively, and extracted for 24 h to obtain the extraction solutions of roots, stems and leaves of each mangrove. The extraction solutions are distilled under reduced pressure by a rotary evaporator at 50 - 160 r / min and 66 °C to obtain the crude extracts of roots, stems and leaves of each mangrove (each 250 mL of extraction solution needs to be distilled for 1 h).
[0030] 2. In vitro antibacterial effect of crude extracts of roots, stems and leaves of mangrove plants
[0031] The disc diffusion method is used to determine the antibacterial effects of extracts of roots, stems and leaves of several mangroves on Escherichia coli, Bacillus subtilis and Saccharomyces cerevisiae. The specific operations are as follows:
[0032] (1) Strain activation: Pick up a loop of bacteria from the slant-preserved culture and inoculate it into the liquid culture medium. Saccharomyces cerevisiae is cultured at 30 °C and 120 rpm for 48 h; Bacillus subtilis and Escherichia coli are cultured at 37 °C and 120 rpm for 24 h.
[0033] (2) Preparation of bacterial suspension: Under sterile conditions, the activated strains are adjusted to a bacterial liquid concentration of 10 -4 CFU / mL with sterile normal saline respectively.
[0034] (3) Antibacterial zone test with different extract concentrations: The extract is prepared into an acetone solution with a concentration of 0.08 g (powder raw material) / L. The acetone solution without extract is used as the blank control (CK). The solid culture medium sterilized by a high-pressure steam sterilizer at 121 °C for 20 min is poured into the petri dish to make a plate. Use a pipette to suck 10 -4200 μL of the bacterial suspension at CFU / ml was placed in a petri dish and then spread evenly with a sterilized spreading rod by burning. A 6-mm filter paper disk was attached to the center of the bacteria-containing petri dish with forceps, and then 20 μl, 60 μl, and 100 μl of the acetone solutions of the extracts from the roots, stems, and leaves of Acanthus ilicifolius were respectively pipetted onto the center of the filter paper disk (three parallel experiments were set for different volumes of the sample solutions and the blank group). Finally, the inoculated petri dish was placed in an incubator for cultivation (bacteria were cultivated at 37 °C for 24 h, and yeast was cultivated at 28 °C for 48 h). Observe whether an inhibition zone appears in the petri dish and measure the size and area of the inhibition zone (π * longest * widest / 4).
[0035] (4) Antibacterial tests of several tree species: The extracts from the roots, stems, and leaves of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia candel, Rhizophora apiculata, Avicennia marina, Sonneratia alba, Ceriops tagal, Acanthus ilicifolius, and Rhizophora stylosa were respectively prepared into acetone solutions with a concentration of 0.08 g (powder raw material) / L. The extracts of Avicennia marina and Bruguiera gymnorrhiza were mixed in a volume ratio of 1:1 to prepare an acetone solution with a concentration of 0.08 g (powder raw material) / L (the extracts from the roots, stems, and leaves were mixed separately), and the acetone solution without the extract was used as the blank control; 10 -4 200 μL of the bacterial suspension at CFU / mL was placed in a petri dish and spread evenly with a spreading rod; then a 6-mm sterile filter paper disk was placed flat on the center of the bacteria-containing petri dish, and then 60 μL of the acetone solutions of the extracts from the roots, stems, and leaves of each mangrove tree were respectively pipetted onto the center of the filter paper disk, and three parallels were set respectively; it was incubated upside down in an incubator (yeast was cultivated at 30 °C for 48 h, and bacteria were cultivated at 37 °C for 24 h); observe whether an inhibition zone appears in the petri dish and measure the size of the inhibition zone. Judgment of the inhibition zone test: An inhibition zone diameter > 20 mm is extremely sensitive; 15 - 20 mm is highly sensitive; 10 - 15 mm is moderately sensitive; 7 - 9 mm is lowly sensitive; < 7 mm is insensitive.
[0036] 3. Scavenging effect of the crude extracts from the roots, stems, and leaves of mangroves on DPPH
[0037] The DPPH reaction method was used to explore the antioxidant capacity of the crude extracts from the roots, stems, and leaves of different mangroves. The specific operations are as follows:
[0038] (1) Preparation of DPPH standard solution: Prepare a DPPH anhydrous ethanol solution with a concentration of 3.5×10 -2 mg / mL, and note that it should be prepared freshly and stored in the dark.
[0039] (2) Detection and calculation of clearance rate: Prepare a series of concentration gradients of the mangrove extract alcohol solution. In a 10 mL colorimetric tube, add the extract solution and the DPPH anhydrous ethanol solution at a volume ratio of 1:4. After placing it in the dark for 30 min, measure the absorbance at a wavelength of 517 nm, denoted as Aj. The blank group is a mixture of anhydrous ethanol solution and the extract solution, and its absorbance is denoted as Ai. The control group is a mixture of the DPPH anhydrous ethanol solution and anhydrous ethanol, and its absorbance is denoted as Ac. Use the anhydrous ethanol solution as the reference solution.
[0040] Calculate the clearance rate of each crude mangrove extract solution against DPPH according to the following formula:
[0041] S% = [1 - (Ai - Aj) / Ac] × 100%.
[0042] Preparation of DPPH clearance rate curve: Make a DPPH clearance rate curve based on the DPPH radical clearance rate of different concentrations of the mangrove extract alcohol solution.
[0043] Results
[0044] 1. Extraction rate of ethanol from the roots, stems and leaves of several mangrove species
[0045] As can be seen from Table 1, the alcohol extraction efficiencies of the roots, stems and leaves of different mangrove species are different. The overall extraction rates of Aegiceras corniculatum, Ceriops tagal and Rhizophora stylosa are relatively high. For the same mangrove species, the extraction rates of stems and leaves are mostly higher than those of roots. Among them, the leaf extraction rate of Aegiceras corniculatum is the highest, reaching 30.94%; while the root and stem extraction rates of Bruguiera gymnorrhiza are relatively low, 4.71% and 4.07% respectively. Generally speaking, the difference in extraction rates between tree species is not very large, but the difference between different tree species reaches a significant level. However, the difference in extracts between different parts shows such a pattern: leaf > stem > root.
[0046] Table 1 Extraction rates of the roots, stems and leaves of several mangrove species
[0047]
[0048]
[0049] 2. Antibacterial test analysis of extracts from three parts of Acanthus ilicifolius at different concentrations
[0050] At the same time, the antibacterial tests on extracts from three parts of Acanthus ilicifolius at different concentrations found that as the concentration of the extract increased, the size of the inhibition zone increased rapidly. The extract from the stem of Acanthus ilicifolius had good antibacterial effects on 3 kinds of microorganisms at 100 μL, especially the largest antibacterial effect on Bacillus.
[0051] From Figure 1It can be seen that the combination of three factors: A - strain, B - part, and C - level has an impact on the antibacterial effect in the antibacterial test. It can be seen that as the concentration of the extract increases, the size of the antibacterial zone increases rapidly. The extract from the stem of Acanthus ilicifolius has good antibacterial effects on three kinds of microorganisms at 100 μL, especially the largest antibacterial effect on Bacillus sp., corresponding to Figure 1 the highest blue column in
[0052] 3. Antibacterial effects in vitro of crude extracts from the roots, stems, and leaves of mangroves
[0053] As can be seen from Table 2, each mangrove extract has obvious antibacterial effects on the three tested bacteria. Different mangroves have different antibacterial effects on the same strain, and the same mangrove also has different antibacterial effects on different strains.
[0054] The antibacterial zones of the root and stem extracts of Bruguiera gymnorrhiza against Bacillus subtilis, Escherichia coli, and Saccharomyces cerevisiae are all greater than 20 mm. The three bacteria are extremely sensitive to its root and stem extracts, while the sensitivity to the leaves is relatively low. The roots, stems, and leaves of four mangroves, Kandelia obovata, Rhizophora apiculata, Avicennia marina, and Xylocarpus granatum, all have extremely strong inhibitory effects on Escherichia coli. Except that the roots of Kandelia obovata have no obvious inhibitory effect on Saccharomyces cerevisiae, these four mangroves as a whole have good antibacterial activities. The extracts from the roots, stems, and leaves of Aegiceras corniculatum have relatively good antibacterial effects on Bacillus subtilis and Escherichia coli, and relatively weak antibacterial effects on Saccharomyces cerevisiae.
[0055] By comparing the diameters of the antibacterial zones of several mangroves against the three bacteria, it can be obtained that the antibacterial effect sizes of the extracts of each mangrove are in the order of Avicennia marina > Bruguiera gymnorrhiza > Rhizophora apiculata > Xylocarpus granatum > Rhizophora stylosa > Ceriops tagal > Kandelia obovata > Aegiceras corniculatum; by comparing the diameters of the antibacterial zones of mangroves against different bacteria, it can be obtained that the antibacterial effect of the extracts of each mangrove on Escherichia coli is the best, followed by Bacillus subtilis, and the worst on Saccharomyces cerevisiae. The combined extract of Avicennia marina and Bruguiera gymnorrhiza has the best antibacterial effects on Escherichia coli, Bacillus cereus, and Saccharomyces cerevisiae. The extract of Acanthus ilicifolius has no antibacterial effect on Aspergillus flavus, and the extract of Bruguiera gymnorrhiza has no antibacterial effect on Aspergillus flavus, but the combined extract of Acanthus ilicifolius + Bruguiera gymnorrhiza has obvious antibacterial effects on Aspergillus flavus.
[0056] Table 2 Antibacterial zone diameters (mm) of extracts from several mangrove plants against three bacteria
[0057]
[0058]
[0059] Note: -: indicates that the antibacterial zone diameter is less than 7 mm, with no obvious inhibitory effect.
[0060] 4. Scavenging effects of crude extracts from the roots, stems, and leaves of several mangroves on DPPH
[0061] From Figure 3 K1-K8 among them, it can be seen that the scavenging rates of the extracts from the roots, stems and leaves of each mangrove on DPPH free radicals increase with the increase of concentration within the experimental concentration range, and show a good linear relationship (R 2 > 0.95). The scavenging effects of the extracts from the roots, stems and leaves of the same mangrove on DPPH free radicals are different. Among them, the extracts from the stems of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia candel, Rhizophora apiculata and Rhizophora stylosa have the strongest free radical scavenging ability, the extracts from the leaves of Avicennia marina and Ceriops tagal have the strongest DPPH free radical scavenging ability. The difference in the DPPH free radical scavenging ability between the stems and leaves of Ceriops tagal is not significant, but both are better than the roots.
[0062] From Figures 3 - 6 it can be seen that among several mangroves, the free radical scavenging ability of the roots, stems and leaves of Avicennia marina is the weakest. The trend lines of the scavenging rates of its roots, stems and leaves are significantly deviated from those of other mangrove plants. While the DPPH free radical scavenging rates corresponding to the extracts from the roots, stems and leaves of the remaining mangroves are close when the concentrations are the same, and the scavenging abilities are not much different. And the extracts of the seven mangroves also show a relatively high DPPH free radical scavenging rate at low concentrations, and the scavenging effect is good.
[0063] From Figure 2 it can be seen that Ceriops tagal and Ceriops tagal have the strongest DPPH free radical scavenging ability, and the free radical scavenging abilities of the two are similar. Kandelia candel and Rhizophora stylosa are the second, followed by Bruguiera gymnorrhiza, Aegiceras corniculatum and Rhizophora apiculata, and the scavenging abilities of the three are also close to each other. Obviously, Avicennia marina is the one with the weakest free radical scavenging ability among the 8 mangrove plants.
[0064] By measuring the absorbance value and its corresponding DPPH free radical scavenging rate, simulating the curve equation, and thus calculating the half scavenging concentration IC50 of each sample. The smaller the IC50 value, the stronger the free radical scavenging ability of the sample. It can be seen from Table 4 that except for the stems, the extracts of Acanthus ilicifolius have certain antioxidant abilities. Especially the leaf extracts have antioxidant abilities much higher than those of gallic acid (flavonoids) and catechin (polyphenols). Generally, it shows that: Acanthus ilicifolius leaves > roots > stems.
[0065] Table 3 DPPH analysis of different extracts of Acanthus ilicifolius
[0066]
[0067]
[0068] Table 4 DPPH analysis of different extracts of 9 mangrove plants
[0069]
[0070]
[0071] Furthermore, DPPH antioxidant activity regression curves were established for the crude extracts of the roots, stems and leaves of the above 9 mangrove tree species (Table 4), and their antioxidant activities were comprehensively evaluated based on the IC50 values. The DPPH values measured for the three repeated samples of the extracts in the tree species in Table 5 all had antioxidant capabilities. In particular, the DPPH antioxidant ability of the leaf extracts was stronger than that of the stems and roots, and the roots were slightly stronger than the stems, but the difference between the two was not very large. Comparing from the tree species, when comparing the antioxidant activities of the roots, stems and leaves with equal weights, the order of antioxidant activity from strong to weak was: *Rhizophora stylosa* > *Ceriops tagal* > *Rhizophora apiculata* > *Kandelia obovata* > *Bruguiera gymnorrhiza* > *Lumnitzera racemosa* > *Aegiceras corniculatum* > *Avicennia marina* > *Acanthus ilicifolius*.
[0072] The embodiments described above are only descriptions of the preferred embodiments of the present invention and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.
Claims
1. Application of a mangrove plant extract in the preparation of an antibacterial product, characterized in that, The mangrove plant is at least one of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia candel, Rhizophora apiculata, Avicennia marina, Xylocarpus granatum, Ceriops tagal, Acanthus ilicifolius and Rhizophora stylosa, and the bacterium is Escherichia coli or Bacillus subtilis.
2. The application according to claim 1, wherein The mangrove plant extract is prepared by extraction with an organic solvent.
3. The application according to claim 2, wherein The organic solvent is an ethanol solution.
4. The application according to claim 3, characterized in that The ethanol solution is a 78% ethanol solution.
5. The application according to any one of claims 1 to 4, characterized in that, The mangrove plant extract is an extract of any one of the roots, stems and leaves of the mangrove plant.
6. The application according to claim 5, wherein The mangrove plant extract is composed of an extract of Avicennia marina and Bruguiera gymnorrhiza.
7. The application according to claim 6, characterized in that, The mass ratio of the extract of Avicennia marina and Bruguiera gymnorrhiza is 1:
1.
8. Use of a mangrove plant extract in the preparation of an antioxidant product, characterized in that, The mangrove plant is at least one of Bruguiera gymnorrhiza, Aegiceras corniculatum, Kandelia candel, Rhizophora apiculata, Xylocarpus granatum, Ceriops tagal, the leaves of Acanthus ilicifolius and Rhizophora stylosa.
9. The application according to claim 8, wherein The mangrove plant extract is prepared by extraction with an organic solvent.
10. The application according to claim 9, wherein The organic solvent is a 78% ethanol solution.