A microcapsule of a guggul triterpenoid compound, a preparation method and application thereof

Abstract

The application provides a microcapsule of triterpenoid compounds of Radix Morindae Officinalis and a preparation method thereof. The preparation method is as follows: taking Radix Morindae Officinalis powder as raw material, first extracting with ethanol under ultrasonic, then extracting with ethyl acetate to obtain triterpenoid compounds of Radix Morindae Officinalis; mixing malt dextrin and gum arabic, dissolving in water to obtain a wall material solution; adding the triterpenoid compounds of Radix Morindae Officinalis into the wall material solution, then adding glycerol monostearate, stirring to obtain an emulsion; spray drying the emulsion to obtain the microcapsule of triterpenoid compounds of Radix Morindae Officinalis. The application also provides application of the microcapsule in preparation of antibacterial drugs or antioxidant active drugs. The extracted triterpenoid compounds of Radix Morindae Officinalis and the microcapsule based on the compounds have good antibacterial and antioxidant activities; through the microencapsulation method, the microcapsule has good water solubility, can avoid being attacked by gastric acid, has the effect of being released fully in the small intestine, and has the advantages of high sustained-release stability and high effective component utilization rate.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, and particularly relates to a triterpenoid compound microcapsule of Liquidambaris, a preparation method and application thereof. BACKGROUND

[0002] Traditional Chinese medicine Liquidambaris is the dried mature inflorescence of Liquidambar formosana Hance in Hamamelidaceae, which contains various effective components, including triterpenoids, flavonoids, phenolic acids, volatile oils and the like. Among them, triterpenoids are important active substances in Liquidambaris, and have important application value in the fields of medicine, chemical industry, agriculture and the like.

[0003] Microencapsulation technology is an encapsulation technology of wrapping active ingredients in a small capsule shell, aiming to improve the stability and bioavailability of active ingredients. This technology can effectively control the drug release rate, prolong the half-life of the drug in the body, thereby increasing the efficacy of the drug and reducing side effects.

[0004] Accordingly, it is tried to develop a triterpenoid compound microcapsule of Liquidambaris for the field of biological medicine. SUMMARY

[0005] The technical problem to be solved by the present application is to provide a triterpenoid compound microcapsule of Liquidambaris and a preparation method and application thereof.

[0006] The present application solves the above technical problems by adopting the following technical solutions:

[0007] A preparation method of a triterpenoid compound microcapsule of Liquidambaris, comprising the following steps:

[0008] S1, using Liquidambaris powder as raw material, first extracting with ethanol by ultrasonic, and then extracting with ethyl acetate to obtain triterpenoid compounds of Liquidambaris;

[0009] S2, uniformly mixing maltodextrin and gum arabic, and then dissolving with water to obtain a wall material solution;

[0010] S3, adding the triterpenoid compounds of Liquidambaris obtained in step S1 to the wall material solution, and then adding glycerol monostearate, and stirring to obtain an emulsion;

[0011] S4, performing spray drying treatment on the emulsion to obtain the target triterpenoid compound microcapsule of Liquidambaris.

[0012] As one of the preferred modes of the present application, in step S1, the specific preparation method of the triterpenoid compounds of Liquidambaris is:

[0013] (1) taking the powder of road road access, adding ethanol, alcohol water ratio 80%, material liquid ratio 1:10 (g / mL), ultrasonic time 0.5h, ultrasonic temperature 60℃, obtaining ethanol extract, recovering ethanol under reduced pressure after filtration, obtaining ethanol crude extract;

[0014] (2) the ethanol crude extract is suspended with distilled water, then sequentially extracted with petroleum ether, dichloromethane, ethyl acetate and n-butanol, taking the ethyl acetate layer, then recovering ethyl acetate under reduced pressure, obtaining the ethyl acetate extract, which is the road road access triterpenoid compound.

[0015] As one of the preferred modes of the present application, the obtained road road access triterpenoid compound has antibacterial and antioxidant activity in the step S1.

[0016] As one of the preferred modes of the present application, the mixing mass ratio of malt dextrin and gum arabic is 1:1 (g / g) and the wall material solution concentration is 2-10% in the step S2.

[0017] As one of the preferred modes of the present application, the road road access triterpenoid compound is the core material, the malt dextrin and gum arabic are the wall materials, and the core-wall mass ratio is 1:(5-25) (g / g) in the step S3.

[0018] As one of the preferred modes of the present application, the added amount of glycerol monostearate is 1% in the step S3.

[0019] As one of the preferred modes of the present application, the inlet air temperature of spray drying is 160-200℃ in the step S4.

[0020] As one of the preferred modes of the present application, the wall material solution concentration, the core-wall mass ratio and the spray drying temperature are determined by single factor experiment combined with response surface analysis method and combined with actual operation; finally, the wall material solution concentration is 5.7%, the core-wall mass ratio is 1:12, and the inlet air temperature is 181℃.

[0021] A road road access triterpenoid compound microcapsule is prepared according to the preparation method.

[0022] The application of the above road road access triterpenoid compound microcapsule in preparing antibacterial drugs or antioxidant active drugs.

[0023] The present application has the following advantages compared with the prior art:

[0024] The road road access triterpenoid compound microcapsule prepared by the present application has good morphology, good fluidity, no caking and a special aromatic odor. The microcapsule meets the requirements of oral drugs in terms of solubility, bioavailability, targeting, biocompatibility, controlled release and the like.

[0025] The application researches and finds that the extracted triterpenoids of Evodia rutaecarpa have good antibacterial and antioxidant activities. The application uses the triterpenoids of Evodia rutaecarpa as active ingredients, so that the prepared microcapsules have antibacterial and antioxidant activities, and through the microencapsulation method, the microcapsules have good water solubility, avoid the invasion of gastric acid, achieve the effect of releasing in the small intestine, have the advantages of high sustained-release stability and high utilization rate of effective components, and can effectively reduce the use dosage and side effects. In addition, the embedding structure of the microcapsules of the application protects the antibacterial biological molecules, prevents the destruction of ultraviolet light to the internal antibacterial components, and improves the stability of the effective components. At the same time, the preparation process of the application is simple, has high automation degree, is easy to expand production, and has very good application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 It is a morphological structure diagram of the triterpenoids of Evodia rutaecarpa microcapsules under scanning electron microscope;

[0027] Figure 2 It is an infrared spectrum of the triterpenoids of Evodia rutaecarpa microcapsules;

[0028] Figure 3 It is a liquid chromatography-mass spectrometry analysis spectrum of the triterpenoids of Evodia rutaecarpa microcapsules (in the figure, A is a negative ion flow diagram; B is a positive ion flow diagram);

[0029] Figure 4 It is a particle size distribution diagram of the triterpenoids of Evodia rutaecarpa microcapsules;

[0030] Figure 5 It is a standard curve of oleanolic acid;

[0031] Figure 6 It is an in-vitro release state diagram of the triterpenoids of Evodia rutaecarpa microcapsules;

[0032] Figure 7 It is an influence diagram of different temperatures on the antibacterial activity of the components before and after microencapsulation;

[0033] Figure 8 It is an influence diagram of different processing times on the antibacterial activity of the components before and after microencapsulation;

[0034] Figure 9 It is an influence diagram of different ultraviolet irradiation times on the antibacterial activity of the components before and after microencapsulation;

[0035] Figure 10 It is a DPPH clearance rate determination result diagram of the triterpenoids of Evodia rutaecarpa microcapsules;

[0036] Figure 11 It is an ABTS clearance rate determination result diagram of the triterpenoids of Evodia rutaecarpa microcapsules;

[0037] Figure 12 Effect of wall material concentration on entrapment efficiency - Figure of clearance rate determination result;

[0038] Figure 13 Figure of effect of wall material concentration on entrapment efficiency;

[0039] Figure 14 Figure of effect of core / wall ratio on entrapment efficiency;

[0040] Figure 15 Figure of effect of inlet air temperature on entrapment efficiency

[0041] Figure 16 Figure of response surface (in the figure, A is the effect of interaction of wall material concentration and core / wall ratio on entrapment efficiency; B is the effect of interaction of wall material concentration and inlet air temperature on entrapment efficiency; C is the effect of interaction of core / wall ratio and inlet air temperature on entrapment efficiency). DETAILED DESCRIPTION

[0042] The following detailed description of the embodiments of the present application is given on the premise of the technical solution of the present application, and detailed implementation modes and specific operation processes are given, but the protection scope of the present application is not limited to the following embodiments. Meanwhile, the experimental methods used in the following embodiments are conventional methods unless otherwise specified, and the materials, reagents and the like used are available from commercial channels unless otherwise specified.

[0043] Example 1

[0044] Preparation of triterpenoid compounds of Radix Morindae Officinalis:

[0045] (1) Radix Morindae Officinalis powder passing through a 60-mesh sieve was added with ethanol, the alcohol / water ratio was 80%, the solid / liquid ratio was 1:10 (g / mL), the ultrasonic time was 0.5 h, and the ultrasonic temperature was 60°C, to obtain an ethanol extract, which was filtered and then the ethanol was recovered under reduced pressure to obtain an ethanol crude extract.

[0046] (2) The ethanol crude extract was suspended with distilled water in an amount of twice the mass of the ethanol crude extract, and then extracted with petroleum ether, dichloromethane, ethyl acetate and n-butanol in equal volumes in sequence, and then the petroleum ether layer, the dichloromethane layer, the ethyl acetate layer, the n-butanol layer and the water layer were recovered under reduced pressure to obtain a petroleum ether extract (yield 2.88±0.05%), a dichloromethane extract (yield 2.57±0.02%), an ethyl acetate extract (yield 0.99±0.03%), an n-butanol extract (yield 1.17±0.07%) and a water extract (yield 0.58±0.09%), wherein the ethyl acetate extract is the triterpenoid compound of Radix Morindae Officinalis.

[0047] Example 2

[0048] Preparation of microcapsule of triterpenoid compounds of Morinda officinalis How

[0049] (1) Maltodextrin and gum arabic were mixed uniformly at a mass ratio of 1:1 (g / g), and then dissolved in water to obtain a wall material solution with a concentration of 2 wt%.

[0050] (2) The triterpenoid compounds of Morinda officinalis How (core material) prepared in Example 1 were added to the wall material solution at a core-wall mass ratio of 1:5 (g / g), and glycerol monostearate was added to make its concentration 1 wt%, and then stirring treatment was performed, and an emulsion was formed by high-speed shearing.

[0051] (3) Spray drying was performed at an inlet air temperature of 160°C to obtain the triterpenoid compounds of Morinda officinalis How microcapsule.

[0052] Example 3

[0053] Preparation of microcapsule of triterpenoid compounds of Morinda officinalis How

[0054] (1) Maltodextrin and gum arabic were mixed uniformly at a mass ratio of 1:1 (g / g), and then dissolved in water to obtain a wall material solution with a concentration of 10 wt%.

[0055] (2) The triterpenoid compounds of Morinda officinalis How (core material) prepared in Example 1 were added to the wall material solution at a core-wall mass ratio of 1:25 (g / g), and glycerol monostearate was added to make its concentration 1 wt%, and then stirring treatment was performed, and an emulsion was formed by high-speed shearing.

[0056] (3) Spray drying was performed at an inlet air temperature of 200°C to obtain the triterpenoid compounds of Morinda officinalis How microcapsule.

[0057] Example 4

[0058] Preparation of microcapsule of triterpenoid compounds of Morinda officinalis How

[0059] (1) Maltodextrin and gum arabic were mixed uniformly at a mass ratio of 1:1 (g / g), and then dissolved in water to obtain a wall material solution with a concentration of 6 wt%.

[0060] (2) The triterpenoid compounds of Morinda officinalis How (core material) prepared in Example 1 were added to the wall material solution at a core-wall mass ratio of 1:10 (g / g), and glycerol monostearate was added to make its concentration 1 wt%, and then stirring treatment was performed, and an emulsion was formed by high-speed shearing.

[0061] (3) Spray drying was performed at an inlet air temperature of 180°C to obtain the triterpenoid compounds of Morinda officinalis How microcapsule.

[0062] Example 5

[0063] Pharmacodynamic experiment of triterpenoid compounds of Morinda officinalis How:

[0064] I. Antibacterial test

[0065] Preparation of solutions required for the experiment:

[0066] LB (Luria-Bertain) medium: Tryptone 10 g, sodium chloride 10 g, yeast extract 5 g, distilled water to 1000 mL, pH adjusted to 7.0, sterilized.

[0067] Sample solution: 100 mg / L of triterpenoids of R. typhina (Example 1).

[0068] Diameter of the inhibition zone experiment: In a single clean bench, take 4 sterilized test tubes with stoppers, put 5 mL of LB liquid medium into each test tube, then take 20 μL of E. coli and S. aureus and put them into the test tubes, blow them evenly with a pipette gun, tightly cap with a rubber cap, wrap with a newspaper, and place in a table top constant temperature shaker for culture. When the culture of E. coli and S. aureus reaches OD 600 value of 0.5 ± 0.2, plate (take 50 μL of the cultured E. coli and S. aureus and spread on LB solid medium with a spreader). Place sterilized filter paper pieces evenly above, left and right lower parts of the culture dish, and add a small amount of drug, blank control and positive control (kanamycin sulfate) on the filter paper pieces for 30 μL, place in a 37°C incubator for one day, observe, take photos, and record the diameter of the inhibition zone.

[0069] The results are: the diameter of the inhibition zone of the triterpenoids of R. typhina drug group is 8.2 ± 0.37 (E. coli) and 8.1 ± 0.26 cm (S. aureus). The inhibition rates of the other groups are worse than that of the triterpenoids of R. typhina group.

[0070] II. Antioxidant activity test

[0071] The following antioxidant tests were performed on different concentrations (0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL) of triterpenoids of R. typhina sample solution (Example 1) (L-ascorbic acid solution as positive control).

[0072] (1) Determination of DPPH free radical scavenging ability

[0073] Take 3 mL of DPPH solution (anhydrous ethanol as solvent) (concentration 0.05 mg / mL), add 1 mL of sample solution of different concentrations, shake well, store at room temperature for 30 min in the dark, measure the absorbance at 517 nm, and record it as A S; another 3 mL DPPH solution, 1 mL of anhydrous ethanol was added, and the absorbance A was measured C ; 1 mL of sample solution of different concentrations was added to 3 mL of anhydrous ethanol solution, and the absorbance A was measured j The above test was repeated three times, and the average value was taken.

[0074] The DPPH free radical scavenging rate was calculated according to the following formula:

[0075]

[0076] A C : blank control absorbance; A j : background absorbance of different concentration solutions; A S : absorbance of different concentrations.

[0077] The results showed that the DPPH free radical scavenging ability of the triterpenoid compounds of Ruitong increased with the increase of concentration after embedding, and there was no significant difference in free radical scavenging rate before and after embedding at high concentration greater than 0.5 mg / mL (P>0.05).

[0078] (2) Determination of ABTS free radical scavenging ability

[0079] Preparation of ABTS: 5 mL of 7 mmol / L ABTS (3.84 mg / mL) was taken, 88 μL of 140 mmol / L potassium persulfate (3.77 mg / mL) was added, and the mixture was placed in the dark at room temperature for 12-16 h. The ABTS mother liquor was obtained. Anhydrous ethanol was mixed with the mother liquor, diluted according to the ratio of 1:60, adjusted to an absorbance of 0.700±0.020 at 734 nm, and the ABTS working solution was obtained. It was preheated at 30°C and ready for use.

[0080] 10 μL of sample solution of different concentrations was taken, 100 μL of ABTS working solution was added, mixed, and reacted in the dark at room temperature for 6 min. The absorbance A was measured at a wavelength of 734 nm C ; control group: 10 μL of anhydrous ethanol was taken, 100 μL of ABTS working solution was added, mixed, and reacted in the dark at room temperature for 6 min. The absorbance A was measured at a wavelength of 734 nm B . The test was repeated three times in parallel, and the average value was taken.

[0081] The ABTS free radical scavenging rate was calculated according to the following formula:

[0082]

[0083] The results showed that the ABTS free radical scavenging ability of Lulutong triterpenoid compounds increased slightly with the increase of concentration before and after encapsulation. Except for the significant decrease in free radical scavenging rate after encapsulation at a concentration of 1 mg / mL (P<0.05), there was almost no significant difference in free radical scavenging rate before and after encapsulation at other concentrations greater than 0.25 mg / mL (P>0.05).

[0084] (3) Hydroxyl radical scavenging ability

[0085] The Fenton method was used to determine the ultrasonic extraction of volatile oil. · OH scavenging ability:

[0086] Pipette 10 μL of 4.5mmol / L ferrous sulfate and 10 μL of 4.5mmol / L salicylic acid-ethanol solution, mix them, add 15 μL of sample solution and 105 μL of distilled water, and finally add 10 μL of 10mmol / L hydrogen peroxide; heat in a 37℃ water bath for 30 minutes; measure the absorbance at 510nm, which is recorded as A X Same as above, but 10 μL of distilled water is used instead of 10 μL of 10 mmol / L hydrogen peroxide, which is recorded as A. X0 Set the blank control variable, replace the sample solution with 15 μL distilled water and record it as A0, and calculate according to the following formula:

[0087]

[0088] The results showed that the triterpenoids of Lulutong had a significant effect on OH before and after embedding. - The free radical scavenging ability increased with the increase of concentration. When the concentration was 2 mg / mL, there was no significant difference in the free radical scavenging rate before and after embedding (P>0.05). When the concentration was lower than this, the free radical scavenging rate decreased slightly after embedding (P<0.05).

[0089] Example 6

[0090] Determination of Lulutong triterpenoid microcapsules:

[0091] The microcapsules of the triterpenoid compound of Lulutong prepared in the present invention (Example 4) were subjected to scanning electron microscopy, infrared spectroscopy, liquid chromatography-mass spectrometry analysis and particle size analysis, and the results were as follows:

[0092] 1. Scanning electron microscopy analysis of microcapsule morphology

[0093] Scanning electron microscopy images of microcapsules of triterpenoid compounds of Lulutong Figure 1 shown.

[0094] 2. Infrared spectroscopy analysis of microcapsules

[0095] The infrared spectrum of the microcapsules of triterpenoid compounds of Lulutong is shown in Figure 2. Figure 2As shown in the infrared spectrum of the core material, 3428cm -1 The OH bond stretching vibration absorption peak is at 2935cm, which indicates the widespread existence of hydroxyl groups in the extract of triterpenoid compounds of P. lulutong. These hydroxyl groups are important functional groups in the structure of triterpenoid compounds and are closely related to their water solubility and biological activity. -1 The stretching vibration of the C-H bond should be at 1704 cm, which reflects the presence of alkyl components in triterpenoids and indicates that the molecule contains a long carbon chain structure; -1 The stretching vibration peak of carbonyl C=O indicates the presence of possible keto or acidic carbonyl functional groups in the structure of triterpenoid compounds, which is an important functional group in triterpenoid compounds; 1614cm -1 The stretching vibration peak of C=C double bond is at 1515cm, especially in aromatic ring or unsaturated fatty chain; -1 and 1459cm -1 The 1380 cm-1 region is the bending vibration of the C-H bond, especially in alkyl and aromatic rings; -1 The bending vibration of the CH bond, especially in the methyl (CH3) group, indicates the presence of methyl groups in the triterpenoid compounds (core material) of Lulutong; 1208cm -1 and 1121cm -1 The peak at 1037 cm is the stretching vibration peak of the CO bond, especially in ether, alcohol or phenol, indicating the presence of an oxidized functional group such as an ether or alcohol group; -1 The stretching vibration peak of the COC bridge bond is at 586 cm, indicating a possible ether connection or sugar connection, which is a typical structural feature of triterpenoids. -1 The difference may be related to some special environment or impurities of the sample.

[0096] From the infrared spectrum of the wall material, we can see that 3425cm -1 The OH bond stretching vibration absorption peak is at 2924cm. In maltodextrin and gum arabic, hydroxyl groups are abundant, which is part of their structure and is responsible for forming hydrogen bonds and maintaining water solubility. -1 The stretching vibration of the C-H bond should be the result, indicating that there are long carbon chains or alkyl components in maltodextrin and gum arabic; 1642 cm -1 The peak at 1423 cm is the stretching vibration peak of C=O, indicating the existence of amide bond, which may come from the protein component in gum arabic. -1 The bending vibration of the C-H bond reflects the presence of the alkyl component; 1158 cm -1 The stretching vibration of the C-O-C bond is at 1023 cm, which may represent the glycosidic bond in the polysaccharide structure in maltodextrin and gum arabic. -1The stretching vibration peak of CO bond is at 854 cm, especially in polyols and polysaccharides, indicating the rich carbohydrate structure in maltodextrin and gum arabic; -1 The stretching vibration peak of the COC bond is at 577 cm, especially the characteristic vibration of the α-glycosidic bond, which indicates the ring structure of the sugar molecule; -1 These may be related to impurities or specific structural features of a particular sample.

[0097] From the infrared spectrum of the microcapsules, we can see that 3425 cm -1 The OH bond stretching vibration absorption peak is at 2921cm, which may be the hydroxyl vibration of triterpenoid compounds or other phenolic acid compounds; -1 The stretching vibration of the C-H bond should be present, indicating that the microcapsules contain aliphatic organic compounds; 1735 cm -1 The peak at 1631 cm is the stretching vibration peak of carbonyl C=O, which may indicate the degree of esterification in the microcapsule material or the ketone group in the triterpenoid compound; -1 The peak at 1420 cm is the stretching vibration peak of the C=C double bond, which is common in unsaturated fatty acids or aromatic rings. This may reflect the unsaturated nature of triterpenoids or the presence of other aromatic compounds. -1 The bending vibration of the C-H bond may indicate the methyl and methylene groups in the microcapsule wall material or the components of the passivation; 1251 cm -1 The peak at 1025 cm is the stretching vibration peak of CO bond, which may indicate the degree of glycosylation in maltodextrin and gum arabic and the presence of triterpenoids; 1 The stretching vibration peak of the C-O-C bond at 577 cm suggests the presence of polysaccharide or carbohydrate structure, which is characteristic of maltodextrin and gum arabic. -1 The presence of these impurities may be related to the impurities or specific structural features of the specific sample. For Lulutong microcapsules, this may point to the special structures or impurities formed during the microcapsule preparation process.

[0098] 3. Microcapsule liquid chromatography-mass spectrometry analysis spectrum

[0099] The ingredients of the microcapsules of triterpenoid compounds of Lulutong Figure 3As shown, first, the data file is opened using Xcalibur software, the mass spectrum is viewed to obtain the mass-to-charge ratio and relative abundance of the compound, then, the characteristic peak is selected for secondary mass spectrometry analysis, the compound structure is inferred from the fragment ions, and the compound name is finally determined by matching with relevant literature and databases such as HMDB and PubChem, and the output results include key data such as mass-to-charge ratio, retention time, response value and metabolite information, etc. The responses of triterpenoids (oleanolic acid, Cimifugin, Picraflavone, etc.) and phenolic acids (gallic acid, ellagic acid, etc.) in the microcapsules of Radix Cynanchi Wilfordii triterpenoids are higher, in addition, there are flavonoids (liquiritin), anthraquinones (Catenarin) and esters (cinnamyl cinnamate, picralactone) and other active substances.

[0100] 4. Microcapsule particle size analysis

[0101] The particle size distribution of the microcapsules of Radix Cynanchi Wilfordii triterpenoids prepared by the present application is in the range of 1.61 μm to 20.09 μm, as shown in Figure 4 .

[0102] 5. Microcapsule embedding rate

[0103] The embedding rate of the microcapsules of Radix Cynanchi Wilfordii triterpenoids prepared by the present application is 89.87% ± 0.23%.

[0104] 6. Microcapsule simulated in vitro gastrointestinal digestion and sustained release test

[0105] Preparation of simulated gastric juice: 8.5 mL of concentrated hydrochloric acid with a mass concentration of 36-38% is taken by a pipette, diluted with distilled water to 1000 mL to obtain 0.1 mol / L hydrochloric acid (pH = 1.2).

[0106] Preparation of simulated intestinal juice: 6.8 g of potassium dihydrogen phosphate is taken and dissolved in 250 mL of distilled water, 190 mL of 0.2 mol / L NaOH and 400 mL of distilled water are added, and the pH is adjusted to 7.5, and then water is added to make up to 1000 mL.

[0107] To further explore the relationship between the in vitro sustained release after embedding and the triterpenoid components, oleanolic acid was taken as an index, and the ultraviolet spectrophotometry was used to determine the absorbance at a wavelength of 552 nm. The absorbance corresponding to the oleanolic acid concentration in the range of 0.01, 0.04, 0.07, 0.10, 0.12, 0.15 (mg / mL) was 0.056, 0.135, 0.234, 0.339, 0.411, 0.494, respectively, and the results showed that the oleanolic acid concentration in the range had a good linear relationship. The regression equation result is as shown in Figure 5 , which is y = 3.41079x - 0.00595, r 2 = 0.99546.

[0108] From Figure 6 It can be seen that in artificial gastric juice, the release rate is stable within 10%, indicating that the microcapsule has good protection for active substances under strong acidic conditions and is not easily damaged. Under artificial intestinal juice digestion, the release rate increases first and then tends to be flat, and after 2.5h, the microcapsule release rate increases to more than 20%, indicating that the active substance is more easily released under weak acidic conditions.

[0109] 7. Analysis of the antibacterial activity of microcapsules

[0110] The turbidity observation of the triterpenoid compounds of Radix Rhei (a) and the microencapsulated triterpenoid compounds of Radix Rhei (b) in a 96-well plate is shown in Table 1. Through turbidity observation, both the embedded and unembedded drugs have better effects on S. aureus, and the antibacterial rate increases with the increase of drug concentration. The minimum inhibitory concentration of the triterpenoid compounds of Radix Rhei on E. coli is 25 mg / mL, and the minimum inhibitory concentration on S. aureus is 3.13 mg / mL. The minimum inhibitory concentration of the microencapsulated triterpenoid compounds of Radix Rhei on E. coli is 25 mg / mL, and the minimum inhibitory concentration on S. aureus is 6.25 mg / mL, indicating that the antibacterial activity before and after embedding does not change much, and the active antibacterial components before embedding still exist after embedding.

[0111] Table 1. Turbidity observation of drug concentration and antibacterial situation before and after embedding

[0112]

[0113] Note: (a) indicates triterpenoid compounds of Radix Rhei; (b) indicates microencapsulated triterpenoid compounds of Radix Rhei; "-" indicates no turbidity; "+" indicates turbidity.

[0114] The specific antibacterial rate is shown in Table 2. The specific antibacterial data before and after embedding can be obtained, and the specific antibacterial effect of the microcapsule after embedding is slightly lower than that before embedding at the same mass concentration, but overall, there is a certain antibacterial effect at the same concentration. We need to focus on protecting the stability of these substances and controlling their release behavior, rather than directly enhancing the antibacterial effect through technical processing. It is worth noting that the embedding technology does not change the essential properties of the antibacterial substance, i.e. its antibacterial effect mainly depends on the chemical structure and mechanism of the antibacterial substance itself, and the embedding process only provides a more stable and controllable environment for the antibacterial substance to maintain its durability and stability.

[0115] Table 2. Antibacterial rate of samples before and after embedding

[0116]

[0117] Note: (a) indicates the olibanum triterpenoids; (b) indicates the olibanum triterpenoids microcapsules; inhibition rate / %.

[0118] The results of the effect of different temperatures on the antibacterial activity of the microencapsulated olibanum triterpenoids are shown in Table 3. Figure 7 It can be seen that with the increase of the treatment temperature, the antibacterial effect of the embedded olibanum triterpenoids is more stable than that of the unembedded olibanum triterpenoids. When the temperature reaches 80℃, it can be seen that the antibacterial activity of the unembedded ingredients begins to decrease obviously, and when the temperature reaches 100℃, the antibacterial effect of the unembedded ingredients is greatly weakened, indicating that it is sensitive to high temperature and does not show antibacterial effect under high temperature conditions. While the antibacterial effect of the embedded olibanum triterpenoids is relatively flat with the increase of the temperature, indicating that it can still well protect the active ingredients under high temperature conditions, so as to maintain high antibacterial capacity.

[0119] The results of the effect of different times on the antibacterial activity of the microencapsulated olibanum triterpenoids are shown in Table 4. Figure 8 The samples were placed in an oven at 25℃ for different times, and it can be observed that with the increase of the treatment time, the antibacterial capacity of the unembedded olibanum triterpenoids samples is in a state of gradual weakening, which indicates that the unembedded samples are more susceptible to the influence of external environmental temperature and humidity, resulting in the gradual degradation or inactivation of the active ingredients (such as volatile active ingredients) therein, while the embedded samples can well maintain the balance, indicating that the embedding technology effectively protects the active ingredients in the samples from the influence of the external environment.

[0120] The results of the effect of ultraviolet irradiation on the antibacterial activity of the microencapsulated olibanum triterpenoids are shown in Table 5. Figure 9 The samples were irradiated under an ultraviolet lamp for different lengths of time, and it can be observed that with the increase of the treatment time, the antibacterial capacity of the unembedded olibanum triterpenoids samples is in a state of obvious weakening, which may be because the ultraviolet light has a destructive effect on some antibacterial ingredients in the samples, resulting in the loss or weakening of the antibacterial capacity of these ingredients. Since ultraviolet light is a high-energy electromagnetic wave, it can destroy the structure of biological molecules, including those with antibacterial function. While the antibacterial capacity of the microcapsules of the embedded olibanum triterpenoids remains stable, indicating that the embedding structure of the microcapsules plays a protective role and prevents the destruction of the internal antibacterial ingredients by ultraviolet light.

[0121] 8. Analysis of the antioxidant activity of the microcapsules

[0122] From Figure 10 , Figure 11 , Figure 12It can be seen that the DPPH scavenging rate of the Chinese herbal medicine Rhizoma Lulutong extract increases with increasing concentration. The positive control ascorbic acid can achieve a good antioxidant effect at a low concentration. The microcapsules and Rhizoma Lulutong triterpenoids have good scavenging capabilities when the concentration reaches 1 mg / mL. The ABTS scavenging rate of the Chinese herbal medicine Rhizoma Lulutong extract slowly increases with increasing concentration. The positive control ascorbic acid begins to have a good antioxidant effect at 0.5 mg / mL. The microcapsules and Rhizoma Lulutong triterpenoids have weak overall ABTS scavenging capabilities. The hydroxyl radical scavenging rate of the Chinese herbal medicine Rhizoma Lulutong extract increases with increasing concentration. The positive control ascorbic acid has a strong antioxidant effect at a low concentration. The concentration of microcapsules and Rhizoma Lulutong triterpenoids needs to reach 1 mg / mL to have a good hydroxyl radical scavenging effect. In summary, a comprehensive comparison of the microcapsules encapsulating active ingredients and the unencapsulated Rulutong triterpenoid compounds shows that the antioxidant scavenging rates of both increase with increasing concentrations. The clearance rates of microcapsules and Rulutong triterpenoid compounds are relatively close at high concentrations, and the microcapsules are slightly inferior to Rulutong triterpenoid compounds at low concentrations. This may be due to the particle size distribution problem of the microcapsules, which causes their performance to fluctuate. It may also be because the encapsulated microcapsules have not been completely and effectively released, and the release time is not sufficient, resulting in low antioxidant efficiency. In the future, we can further study the encapsulation material and reaction time to verify the relationship between the two in order to achieve the best antioxidant effect.

[0123] Example 7

[0124] Experiment on the influence of various factors on the performance of microcapsules:

[0125] 1. Effect of wall material concentration on microcapsule embedding efficiency

[0126] A wall material, maltodextrin, was mixed with gum arabic in a ratio of 1:1 (g / g). Water was added and stirred to fully dissolve the mixture. Wall material concentrations were set at 2%, 4%, 6%, 8%, and 10%. 1% glyceryl monostearate was added and stirred for a preferred duration of 6 hours. The drug was then accurately weighed and added to the wall material at a core-to-wall ratio of 1:15 (g / g). Stirring was continued using a magnetic stirrer for 6 hours to obtain a sample solution. The sample solution was placed in a spray dryer at an inlet air temperature of 180°C for spray drying. The triterpenoid compounds were microencapsulated. The microcapsulation efficiency was measured under different wall material concentrations, and the effects of these conditions on the encapsulation efficiency were analyzed. Three replicates were performed, and the results were averaged.

[0127] The embedding efficiency of triterpenoid compounds in microcapsules of Lulutong was analyzed by using different wall material concentrations as the measurement index. Figure 13 As shown. Figure 13It can be seen that when the wall material concentration reaches 6%, the encapsulation rate is the highest, and the encapsulation rate trend increases first and then decreases with the increase of the wall material concentration. When the wall material concentration is low, the encapsulation rate also gradually increases with the increase of the wall material concentration, because appropriate increase of the wall material concentration can provide more wall material molecules, which can help to form more complete and dense encapsulation structure, thereby enhancing the encapsulation effect of the triterpenoid compounds in road rut. However, when the wall material concentration is too high, the encapsulation rate will decrease instead.

[0128] 2. Effect of core-wall ratio on microcapsule embedding rate

[0129] The wall material maltodextrin was mixed with gum arabic at a ratio of 1:1 (g / g), water was added and stirred to make it fully dissolved. The wall material concentration was fixed at 6%, 1% glyceryl monostearate was added, and the stirring time was set to 6 hours. Then, the drug was accurately weighed and added to the wall material. The core-wall ratio was set to 1:5, 1:10, 1:15, 1:20 and 1:25 (g / g). The mixture was continuously stirred by a magnetic stirrer for 6 hours to obtain a sample solution. The sample solution was placed in a spray dryer, and the inlet temperature was set to 180°C for spray drying. The triterpenoid compounds in road rut were microencapsulated. Under different core-wall ratios, the microcapsule embedding rate was measured and analyzed to study the effect of the conditions on the embedding rate. This experiment was repeated three times, and the results were averaged.

[0130] By comparing the concentrations of different core-wall ratios, the triterpenoid compounds in road rut were prepared into microcapsules, and their embedding rates were evaluated and analyzed as shown in Table 1. Figure 14

[0131] When the core-wall ratio reaches 1:10 (g / g), the encapsulation rate is the highest, and the encapsulation rate trend increases first and then decreases with the increase of the wall material. When the wall material gradually increases (i.e. the core-wall ratio gradually decreases), the encapsulation rate also increases, because the wall material can better wrap the core material. However, when the wall material increases to a certain extent, further increase of the wall material amount will lead to a decrease in the encapsulation rate, which may be because excessive wall material will accumulate, overlap or form unnecessary structures when forming microcapsules, thereby affecting the overall quality and encapsulation effect of the microcapsules. When the core-wall ratio is 1:10 (g / g), the ratio of wall material to core material reaches a relatively ideal balance. The wall material can fully wrap the core material without wasting due to excessive amount, thereby achieving the highest encapsulation rate.

[0132] 3. Effect of inlet temperature on microcapsule embedding rate

[0133] ​The wall material maltodextrin is mixed with gum arabic at 1:1 (g / g), water is added and stirred to make it fully dissolved, the concentration of the fixed wall material is 6%, 1% glyceryl monostearate is added, the stirring time is 6h, the drug is accurately weighed and added to the wall material, the core-wall ratio is 1:15 (g / g), the magnetic stirrer is continuously stirred for 6h, the sample solution is obtained, the sample solution is placed in a spray dryer, the inlet temperature is set at 160℃, 170℃, 180℃, 190℃ and 200℃ for spray drying, and the triterpenoid compounds in Radix Sophorae Tonkinensis are microencapsulated. Under different inlet temperatures, the microcapsule encapsulation efficiency is measured and analyzed to study the effect of the conditions on the encapsulation efficiency. This experiment is repeated 3 times, and the results are averaged. The encapsulation efficiency formula is as follows:

[0134] Encapsulation efficiency % = m A / m0 (referring to the ratio of the mass of the loaded drug to the total drug input).

[0135] The microcapsules of triterpenoid compounds in Radix Sophorae Tonkinensis are prepared, the embedding rates at different inlet temperatures are compared, and the analysis is as shown in Figure 15 When the inlet temperature reaches 190℃, the encapsulation efficiency is the highest, and the encapsulation efficiency trend first increases and then decreases with the increase of the inlet temperature. Therefore, the inlet temperature in this response surface experiment is selected as 180℃, 190℃ and 200℃. With the increase of the inlet temperature, the drying speed increases, which helps to complete the drying process in a short time, thereby avoiding the degradation or denaturation of the core material during the drying process. When the inlet temperature is moderate (such as 190℃), the drying speed is fast enough to ensure that the wall material of the microcapsule forms a complete and dense network film structure before the core material degrades, thereby achieving a high encapsulation efficiency, promoting the improvement of the drying speed and the complete formation of the microcapsule wall material. However, when the inlet temperature is too high, the thermal effect begins to dominate, leading to the deformation or rupture of the microcapsule wall material, and thus reducing the encapsulation efficiency.

[0136] 4. Optimization of preparation process by response surface experiment

[0137] Based on the results of the single-factor experiment, the wall material concentration of 2%, 4% and 6%, the inlet temperature of 180℃, 190℃ and 200℃, and the core-wall ratio of 1:5 (g / g), 1:10 (g / g) and 1:15 (g / g) are finally selected, and the encapsulation efficiency of Radix Sophorae Tonkinensis triterpenoid compounds is used as an index to screen the best embedding process. The response surface factor level table is designed, as shown in Table 3.

[0138] Table 3 Response surface factor level table

[0139]

[0140] The Box-Behnken Design software is used for design, and the optimized results are obtained by software analysis.

[0141] Based on the single factor results, the response surface methodology with three factors and three levels was used to optimize the process. Seventeen experimental schemes were designed by Box-Behnken Design, and the final equation expressed by coded factors was obtained by the response surface software. The encapsulation efficiency of the active components of Rukong was equal to 72.23 + 9.29A + 19.65B - 2.36C + 1.57AB + 0.87AC - 1.74BC + 2.84A 2 -20.88B 2 + 3.04C 2 The absolute value of the coefficient indicated the significant influence of the factor on the response variable, the square term indicated the nonlinear effect of the factor, and the interaction term indicated the influence of the interaction between different factors on the response variable. Therefore, the experimental results showed that the core-wall ratio was the most significant factor affecting the response variable.

[0142] The 3D response surface graph was obtained by the response surface software, as shown in Figure 16 The interaction between the wall material concentration and the inlet air temperature had a small effect on the slope of the surface, which meant that these two parameters could be adjusted independently during the optimization process without worrying about the significant influence of their interaction on the encapsulation efficiency. The interaction between the wall material concentration and the core-wall ratio had a large effect, and their interaction would significantly affect the performance of the microcapsules when both of them changed simultaneously. For example, increasing the wall material concentration while keeping the core-wall ratio unchanged could improve the stability and encapsulation efficiency of the microcapsules, but if the wall material concentration was increased while the core-wall ratio was decreased, it might lead to insufficient drug loading and reduce the encapsulation efficiency. The interaction between the core-wall ratio and the inlet air temperature also had a large effect, mainly on the drying process and the stability of the active ingredients. For example, increasing the core-wall ratio while keeping the inlet air temperature unchanged could increase the drug content in the microcapsules, but it might also increase the difficulty of the drying process and the risk of drug degradation. However, decreasing the inlet air temperature while increasing the core-wall ratio could reduce the risk of drug degradation, but it might prolong the drying time. Therefore, by adjusting these parameters and their interactions reasonably, better encapsulation efficiency could be achieved to ensure the stability and effectiveness of the active ingredients of traditional Chinese medicine.

[0143] Table 4 Variance analysis of the response surface model for the encapsulation efficiency of the microcapsules

[0144]

[0145]

[0146] Note: R 2 = 0.9905; Radj 2 = 0.9783

[0147] Table 5 Microcapsule encapsulation rate model credibility analysis

[0148] Mean / % Standard Deviation [R 2 ]]> adjusted R 2 ]]> C.V. / % Precision 65.18 2.80 0.9905 0.9783 4.30 28.909

[0149] In combination with the results in Table 4 and Table 5, it is known that the wall material concentration and the core wall ratio have a significant effect, and the core wall ratio has the greatest effect. The influencing factors are verified by variance analysis, and the final optimized conditions are obtained as follows: the wall material concentration is 5.75%, the core wall ratio is 1:11.947, and the inlet air temperature is 180.81°C, and the embedding rate is 90.96%. For the convenience of the test, the wall material concentration is adjusted to 5.7%, the core wall ratio is 1:12, and the inlet air temperature is 181°C. The adjusted optimal process conditions are verified by three parallel verification experiments, and the microencapsulation embedding rate of the active substance of the road rutting is 89.87%.

[0150] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, and improvement within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing microcapsules of triterpenoid compounds of Lentulacus root, characterized in that: The steps include: S1. Using P. lupulus powder as raw material, first ultrasonic extraction with ethanol, then extraction with ethyl acetate to obtain P. lupulus triterpenoid compounds. The specific preparation method is as follows: (1) Take the powder of Rhizoma Lulutong, add ethanol, the alcohol-water ratio is 80%, the material-liquid ratio is 1:10, ultrasonic time is 0.5h, and the ultrasonic temperature is 60℃ to obtain ethanol extract. After filtering, reduce the pressure and recover ethanol to obtain ethanol crude extract; (2) suspending the crude ethanol extract with distilled water, and then extracting with petroleum ether, dichloromethane, ethyl acetate, and n-butanol in sequence, taking the ethyl acetate layer, and then recovering the ethyl acetate under reduced pressure to obtain the ethyl acetate extract, which is the triterpenoid compound of the Rhizoma Lulutongae; S2. Evenly mix maltodextrin and gum arabic, and then add water to dissolve them to obtain a wall material solution; S3, adding the L-type triterpenoid compound obtained in step S1 to the wall material solution, and then adding glyceryl monostearate, stirring to obtain an emulsion; S4. The emulsion is subjected to spray drying treatment to obtain the desired Lulutong triterpenoid compound microcapsules.

2. The method for preparing the Lulutong triterpenoid compound microcapsules according to claim 1, wherein In the step S1, the obtained Lulutong triterpenoid compounds have antibacterial and antioxidant activities.

3. The method for preparing the Lulutong triterpenoid compound microcapsules according to claim 1, wherein In step S2, the mixing mass ratio of maltodextrin to gum arabic is 1:1, and the concentration of the wall material solution is 2-10%.

4. The method for preparing the Lulutong triterpenoid compound microcapsules according to claim 1, wherein In step S3, the triterpenoid compound of Lulutong is used as the core material, maltodextrin and gum arabic are used as the wall material, and the core-to-wall mass ratio is 1:(5-25).

5. The method for preparing the Lulutong triterpenoid compound microcapsules according to claim 1, wherein In step S3, the amount of glyceryl monostearate added is such that the final concentration reaches 1%.

6. The method for preparing the Lentulatus triterpenoid compound microcapsules according to claim 1, wherein: In step S4, the inlet air temperature of the spray drying is 160-200°C.

7. The method for preparing the Lulutong triterpenoid compound microcapsules according to any one of claims 1 to 6, characterized in that: The wall material solution concentration, core-to-wall mass ratio, and spray drying temperature were all determined by single-factor experiments combined with response surface analysis and actual operations; the final wall material solution concentration was 5.7%, the core-to-wall mass ratio was 1:12, and the inlet air temperature was 181°C.

8. A microcapsule of triterpenoid compounds of Lentulacus root, characterized in that: It is prepared according to the preparation method according to any one of claims 1 to 7.

9. A use of the Lulutong triterpenoid compound microcapsules according to claim 8 in the preparation of antibacterial drugs, characterized in that: The bacteria are Escherichia coli and Staphylococcus aureus.

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