Preparation method and application of wood vinegar calcium preparation

Abstract

The invention provides a preparation method and application of a pyroligneous calcium preparation, the preparation method comprises the following steps: 1, adding calcium hydroxide powder into pyroligneous, and magnetically stirring for 1.5-3 hours, 2, dropwise adding an ammonia water solution into a product obtained in the step 1, and continuously stirring to obtain a mixture containing the pyroligneous calcium preparation; and step 3, performing vacuum suction filtration on the mixture obtained in the step 2, collecting the wood vinegar calcium preparation, washing with ethanol, and performing vacuum drying, thereby facilitating the preservation and transportation of the wood vinegar, being directly used for treatment, solving the problems that phenolic substances in the wood vinegar are relatively fast in failure and easy to oxidize, and being applicable to the fields of food, medicine, feed and health care, such as feed additives, feed additives and the like. And a calcium supplement preparation.

Description

Technical Field

[0001] The invention relates to the fields of medicine and animal husbandry, and in particular to a preparation method and application of a wood vinegar calcium preparation. Background Art

[0002] Wood vinegar is a reddish-brown acidic liquid, which mainly refers to the smoky-smelling liquid obtained by condensing and standing the gas mixture produced in the distillation process of agricultural and forestry wastes, residues and other biomass. The main components of wood vinegar include water, acid, and phenolic substances. Its composition is complex, so it has been widely used in many fields such as biology and medicine. For example, the phenolic substance isoeugenol in wood vinegar is the main antibacterial substance, and a certain amount of acetic acid can enhance its antibacterial properties; wood vinegar has anti-inflammatory properties both in vivo and in vitro, among which 2-methoxy-4-methylphenol is the main anti-inflammatory compound; wood vinegar prepared from the top of rhizophora plants, polyphenolic compounds are the main antioxidants, including 2,6-dimethoxyphenol. It can be seen that wood vinegar has strong antioxidant and antibacterial activities, and can be widely used as an antioxidant and antibacterial agent in medicine, food, agriculture, animal husbandry and other fields.

[0003] At present, the main active ingredients in wood vinegar, phenolic compounds, are easily oxidized and ineffective, which seriously restricts the application of wood vinegar. Summary of the invention

[0004] The purpose of the present invention is to provide a method for preparing a wood vinegar calcium preparation, which is beneficial to the storage and transportability of the wood vinegar, can be directly used for treatment, and solves the problem that the phenolic substances in the wood vinegar lose their effectiveness quickly and are easily oxidized.

[0005] The purpose of the present invention is achieved through the following technical solutions: A method for preparing a wood vinegar calcium preparation comprises the following steps: Step 1: Add calcium hydroxide powder to wood vinegar and stir magnetically for 1.5-3 hours. 2+ Introduced into bio-oil; The mass ratio of the calcium hydroxide powder to the wood vinegar is (1.8-3.2):40; Step 2, dripping an ammonia solution into the product obtained in step 1 and continuously stirring to obtain a mixture containing the wood vinegar calcium preparation; The concentration of the ammonia solution is 1-6 mol / L, and the volume mass ratio of the ammonia solution to the wood vinegar is 6:40 mL / g.

[0006] Step 3, vacuum filtering the mixture obtained in step 2, collecting the wood vinegar calcium preparation, washing it with ethanol and then vacuum drying it; As a more optimal technical solution of the present invention, the concentration of the hydrochloric acid solution in step 4 is 4 mol / L; and the volume mass ratio of the hydrochloric acid solution to the wood vinegar is 12:40 mL / g.

[0007] As a more optimal technical solution of the present invention, the stirring time in step 2 is 5-35 minutes.

[0008] As a more optimal technical solution of the present invention, the stirring time in step 2 is 25 minutes.

[0009] As a more preferred technical solution of the present invention, the reaction temperature in step 2 is 20°C-70°C.

[0010] As a more optimal technical solution of the present invention, the reaction temperature in step 2 is 60°C.

[0011] Another object of the present invention is to provide a wood vinegar calcium preparation prepared by the above method.

[0012] Another object of the present invention is to provide a wood vinegar calcium preparation for use as a medicine for treating bacterial infection or improving appetite and relieving gastrointestinal discomfort.

[0013] Another object of the present invention is to provide a wood vinegar calcium preparation for use as a food additive.

[0014] Beneficial effects: The preparation method provided by the present invention studies a new extraction technology for separating phenolic compounds from wood vinegar, which has low cost, high extraction rate of phenolic compounds, no environmental pollutant discharge and the residual wood vinegar can be utilized. Specifically, an appropriate amount of calcium hydroxide is added to the wood vinegar, and first reacts with acetic acid present in the wood vinegar. The production of calcium acetate means that Ca 2+ Then, the phenolic compounds and Ca in the wood vinegar were adjusted with an ammonia solution. 2+ The complex is filtered and dried, and then dissolved with hydrochloric acid; high-purity phenolic compounds are collected from the dissolved solution by extracting ethyl acetate. Since only a small amount of ammonia solution is introduced into the wood vinegar, the residual wood vinegar can be further utilized. The residual wood vinegar after filtration is mainly composed of calcium acetate, which can be used as a snow melting agent.

[0015] Calcium wood vinegar preparations can be divided into two parts: complex and calcium acetate. The part that forms a complex with phenols has many advantages and is conducive to further application. The complex can obtain phenolic compounds under acidic conditions, and the pH of gastric acid is 0.9-1.5. Therefore, calcium wood vinegar preparations can be directly used for treatment, solving the problem that phenolic substances in wood vinegar lose their effectiveness quickly and are easily oxidized. 2+After coordination, there is no smoky smell, because phenolic compounds are the main source of the smoky smell of wood vinegar. It is helpful for the application of wood vinegar calcium preparations in the fields of food, animal husbandry, etc. The other part of calcium acetate can be used for the preparation of environmentally friendly snow melting agents. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is the extraction rate of phenolic compounds under different calcium hydroxide addition amounts in the preparation method of the present invention.

[0017] Figure 2 It is the extraction rate of phenolic compounds under different concentrations of ammonia solution in the preparation method of the present invention.

[0018] Figure 3 It is the extraction rate of phenolic compounds at different reaction temperatures in the preparation method of the present invention.

[0019] Figure 4 It is the extraction rate of phenolic compounds under different reaction times of the preparation method of the present invention.

[0020] Figure 5 It is the infrared spectrum of calcium hydroxide, wood vinegar and the complex.

[0021] Figure 6 is the standard curve of EGCG.

[0022] Figure 7 The release of phenolic substances from calcium wood vinegar preparation in simulated gastric fluid within 270 minutes. DETAILED DESCRIPTION

[0023] The present invention is further described below in conjunction with embodiments, but is not intended to limit the present invention thereto.

[0024] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. If no specific techniques or conditions are specified in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used that do not specify the manufacturer are all conventional products that can be obtained commercially.

[0025] The calcium wood vinegar preparation prepared by the preparation method provided in the following embodiment is conducive to the storage and transportation of wood vinegar. Under the optimal reaction conditions (2.8g calcium hydroxide, 1mol / L ammonia solution, 60°C reaction temperature and 25min reaction time), the phenol extraction rate is 80.6%. In addition, an in vitro digestion simulation experiment was carried out to verify the feasibility of direct use of the calcium wood vinegar preparation. The main component of the wood vinegar remaining after the preparation of the calcium wood vinegar preparation is calcium acetate, which can be used to make an environmentally friendly snow melting agent. The calcium wood vinegar preparation can be used in medicine and animal husbandry, for example, as a feed additive.

[0026] Fourier transform infrared spectroscopy (FT-IR) analysis was performed by a Nicolet 330 FT-IR spectrometer to determine the functional groups of guaiacol and the obtained functional groups. The solid complex was mixed and ground with potassium bromide at a mass ratio of 1:100, and then 60 mg of the mixture was pressed into a tablet. The mixture tablet was placed in a sample holder and scanned from 4000-450 cm-1 at a resolution of 4 cm-1.

[0027] The total phenol method was used to quantitatively analyze the phenolic compounds of the acidified calcium wood vinegar preparation. The extraction rate of phenolic compounds was obtained by comparing the total phenol content after extracting 40g of wood vinegar with 8mL of ethyl acetate. Extraction Rate (%) = C / C0 × 100%, where C is the concentration of the dissolved complex and C0 is the concentration of the wood vinegar after extraction.

[0028] Example 1 2.8 g of calcium hydroxide powder was added to 40 g of wood vinegar at room temperature and magnetically stirred for 2 h. The acetic acid in the wood vinegar reacted with calcium hydroxide to form calcium acetate, that is, the complex agent Ca2+ was introduced into the bio-oil. 6 mL of a 1 mol / L ammonia solution was dropped into the bio-oil, the reaction temperature was 60 ° C, and the stirring was continued for 25 min; the wood vinegar calcium preparation gradually appeared in the solution. The mixture was filtered using a Buchner funnel in a vacuum pump. The wood vinegar calcium preparation on the filter paper was then collected, washed three times with ethanol, and then further dried in a vacuum drying oven at 105 ° C for 4 hours to remove traces, and weighed. The dried complex was dissolved in 12 mL of hydrochloric acid at a concentration of 4 mol / L for 20 min. The phenolic compounds in the dissolved solution were extracted with 8 mL of ethyl acetate.

[0029] Example 2 The difference between this embodiment and embodiment 1 is only that: the calcium hydroxide powder addition amount is 2.0g.

[0030] Example 3 The difference between the present embodiment and embodiment 1 is only that: the calcium hydroxide powder addition amount is 2.2g.

[0031] Example 4 The difference between the present embodiment and embodiment 1 is only that: the calcium hydroxide powder addition amount is 2.4g.

[0032] Example 5 The difference between the present embodiment and embodiment 1 is only that: the calcium hydroxide powder addition amount is 2.6g.

[0033] Example 6 The difference between this embodiment and embodiment 1 is only that: the calcium hydroxide powder addition amount is 1.8g.

[0034] Example 7 The difference between this embodiment and embodiment 1 is only that: the calcium hydroxide powder addition amount is 3.0g.

[0035] Example 8 The difference between the present embodiment and embodiment 1 is only that: the calcium hydroxide powder addition amount is 3.2g.

[0036] like Figure 1 As shown in the figure, the effect of calcium hydroxide addition in the range of 1.8-3.2g on the extraction rate of phenols was tested, while the concentration of ammonia solution, reaction temperature and reaction time were fixed at 1mol / L, 20℃ and 25min respectively. As the addition of calcium hydroxide increased from 1.8g to 2.8g, the extraction rate of phenolic compounds increased from 9.8% to 64.6%, and the extraction rate decreased slightly when it was higher than 2.8g. During the experiment, it was found that when the amount of calcium hydroxide added was too much, some of it was insoluble and too much precipitation was formed, the fluidity of the solution became worse, and the difficulty of the separation process increased.

[0037] Example 9 The only difference between this embodiment and embodiment 1 is that the concentration of the aqueous ammonia solution is 1 mol / L.

[0038] Example 10 The only difference between this embodiment and embodiment 1 is that the concentration of the ammonia solution is 21 mol / L.

[0039] Embodiment 11 The only difference between this embodiment and embodiment 1 is that the concentration of the ammonia solution is 3 mol / L.

[0040] Example 12 The only difference between this embodiment and embodiment 1 is that the concentration of the aqueous ammonia solution is 4 mol / L.

[0041] Embodiment 13 The only difference between this embodiment and embodiment 1 is that the concentration of the ammonia solution is 5 mol / L.

[0042] Embodiment 14 The only difference between this embodiment and embodiment 1 is that the concentration of the aqueous ammonia solution is 6 mol / L.

[0043] like Figure 2As shown in the figure, the experiment was carried out under the conditions of 2.8g calcium hydroxide, reaction temperature of 20℃ and reaction time of 25min. The concentration of ammonia solution has a significant effect on the extraction rate of phenolic compounds. The lower the concentration, the more favorable it is to the extraction rate of phenolic compounds. When the concentration of ammonia solution increases from 1mol / L to 3mol / L, the extraction rate of phenolic compounds decreases slightly, from 77% to 65.8%. When the concentration of ammonia solution is higher than 3mol / L, the extraction rate of phenolic compounds decreases significantly to 49.9%. Because the pH after the reaction has reached 12 when no ammonia solution is added, the addition of ammonia solution does not have much beneficial effect on the extraction rate of phenolic compounds. However, when 1mol / L ammonia solution is added, the extraction rate of phenolic compounds is higher than that without addition.

[0044] Embodiment 15 The only difference between this embodiment and embodiment 1 is: 20°C reaction temperature.

[0045] Example 16 The only difference between this embodiment and embodiment 1 is: 30°C reaction temperature.

[0046] Embodiment 17 The only difference between this embodiment and embodiment 1 is: 40°C reaction temperature.

[0047] Embodiment 18 The only difference between this embodiment and embodiment 1 is: 50°C reaction temperature.

[0048] Embodiment 19 The only difference between this embodiment and embodiment 1 is: 70°C reaction temperature.

[0049] like Figure 3 As shown in the figure, when the amount of calcium hydroxide added, the concentration of ammonia solution and the reaction time were fixed at 2.8g, 1mol / L and 25min, the effect of reaction temperature on the extraction rate of phenolic compounds was studied. The higher the temperature, the higher the extraction rate of phenolic compounds. When the reaction temperature increased from 20℃ to 60℃, the extraction rate of phenolic compounds increased significantly, from 65.8% to 79.1%. High temperature promotes the reaction, but the solubility of the complex increases at high temperature, which is not conducive to the separation of the complex. In the experiment, the solution was first cooled to room temperature and then filtered for separation, which is conducive to the full precipitation and complete separation of the complex.

[0050] Embodiment 20 The only difference between this embodiment and embodiment 1 is that stirring is continued for 5 minutes.

[0051] Embodiment 21 The only difference between this embodiment and embodiment 1 is that stirring is continued for 10 min.

[0052] Embodiment 22 The only difference between this embodiment and embodiment 1 is that stirring is continued for 15 min.

[0053] Embodiment 23 The only difference between this embodiment and embodiment 1 is that stirring is continued for 20 min.

[0054] Embodiment 24 The only difference between this embodiment and embodiment 1 is that stirring is continued for 30 min.

[0055] Embodiment 25 The only difference between this embodiment and embodiment 1 is that stirring is continued for 35 min.

[0056] like Figure 4 As shown in the figure, under the conditions of 2.8g calcium hydroxide addition, 1mol / L ammonia solution concentration and 20℃ reaction temperature, the effect of reaction time after adding ammonia solution on the extraction rate of phenolic compounds was studied. When the reaction time was greater than 25min, the extraction rate of guaiacol was still high, then decreased slightly, and decreased slightly again when the reaction time was less than 25min. This indicates that Ca 2+ The complexation reaction with the phenolic compound requires sufficient time.

[0057] like Figure 5 The infrared spectra of calcium hydroxide, wood vinegar and the prepared complex are shown. Under the optimal conditions, a complex with a good phenol extraction rate was prepared. The liquid wood vinegar and the solid complex have similar characteristic peaks. The transmittance of the phenolic hydroxyl group in the complex is significantly lower than that of guaiacol. At the same time, the fingerprint peaks between guaiacol and the complex are also different, indicating that the complex has a different chemical structure from wood vinegar. The complex and calcium hydroxide have a strong affinity at 3600cm -1 There are free hydroxyl peaks everywhere, which is due to the addition of excess calcium hydroxide during the preparation process. 2+ If the reaction generates calcium phenolate, no phenolic hydroxyl group should be detected. -1 The peak at indicates the presence of phenolic hydroxyl groups in the complex, indicating that the complex formed during the preparation process is not phenolic calcium salt.

[0058] like Figure 6 As shown, the total phenol content of the calcium wood vinegar preparation after dissolution was calculated to be 6641µg / mL. Similarly, the total phenol content of 40g of wood vinegar extracted with 8mL of ethyl acetate was 8240µg / mL. The phenol extraction rate under the optimal conditions was calculated to be 80.6%. The total phenol content measures the biological activity of wood vinegar. From this study, it can be seen that the calcium wood vinegar preparation can extract and preserve most of the active ingredients in wood vinegar. It is helpful for its application in the medical field.

[0059] The release of calcium wood vinegar preparation in the stomach was evaluated by simulating gastric digestion conditions in vitro. 800 mL of water was added to 16.4 mL of dilute hydrochloric acid (diluted from 23.4 mL of hydrochloric acid to 100 mL), mixed with 10 g of pepsin, and diluted with water to 1000 mL to obtain simulated gastric fluid. Weigh 50 mg of calcium wood vinegar preparation, add 5 mL of simulated gastric fluid, and incubate at 37°C for 5, 15, 30, 60, 120, 180, and 270 minutes, respectively. Then, shake the mixture evenly, filter it with a filter membrane, and determine the total phenol content in the solution. Figure 7 As shown in the figure, the phenolic compounds in the calcium wood vinegar preparation were rapidly released in the simulated gastric fluid in the first 60 minutes, and the release amount exceeded 80%. After that, the release of phenolic compounds began to slow down. At 180 minutes, about 90% of the total phenol in the simulated gastric fluid was released, and there was no significant change in the release after 120 minutes. The release of phenolic compounds in the calcium wood vinegar preparation was mainly due to the acidic environment that caused the coordination bonds to break and the formation of phenolic hydroxyl groups. This shows that the calcium wood vinegar preparation can quickly and efficiently release phenolic compounds in gastric acid.

[0060] The invention provides a preparation method of a calcium wood vinegar preparation, wherein the phenol extraction rate under optimal conditions is 80.6%. Infrared spectrum shows that a complex rather than calcium phenol salt is formed during the preparation process. The main component of the filtrate remaining after filtration during the preparation process is calcium acetate, which can be used for the preparation of an environmentally friendly snow-melting agent. In an in vitro digestion simulation experiment, the phenolic compounds in the calcium wood vinegar preparation can be quickly released, and the release amount is as high as 90%, indicating that the calcium wood vinegar preparation has the possibility of direct use.

[0061] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, and they should be covered by the protection scope of the present invention.

Claims

1. A method for preparing a wood vinegar calcium preparation, characterized in that: The steps include: Step 1, adding calcium hydroxide powder to wood vinegar and magnetically stirring for 1.5-3h; the mass ratio of the calcium hydroxide powder to the wood vinegar is (1.8-3.2):40; Step 2: dripping an ammonia solution into the product obtained in step 1 and continuously stirring to obtain a mixture containing a wood vinegar calcium preparation; the concentration of the ammonia solution is 1-6 mol / L, and the volume mass ratio of the ammonia solution to the wood vinegar is 6:40 mL / g; Step 3: vacuum filter the mixture obtained in step 2, collect the wood vinegar calcium preparation, wash it with ethanol and then vacuum dry it.

2. The method for preparing the wood vinegar calcium preparation as claimed in claim 1, wherein The mass ratio of the calcium hydroxide powder and the wood vinegar in step one is 2.8:

40.

3. The preparation method of the wood vinegar calcium preparation as claimed in claim 1, characterized in that, The stirring time in step 2 is 5-35 min.

4. (Stirring time after adding ammonia) The method for preparing the calcium wood vinegar preparation according to claim 2, characterized in that: The stirring time in step 2 is 25 min.

5. The method for preparing the wood vinegar calcium preparation as claimed in claim 1, characterized in that, The reaction temperature in step 2 is 20°C-70°C.

6. The method for preparing the calcium wood vinegar preparation according to claim 4, wherein: The reaction temperature in step 2 is 60°C.

7. A wood vinegar calcium preparation, characterized in that: The method is prepared by the preparation method of the wood vinegar calcium preparation according to any one of claims 1 to 6.

8. The wood vinegar calcium preparation according to claim 7, characterized in that The wood vinegar calcium preparation is used as a medicine for treating bacterial infection or improving appetite and relieving gastrointestinal discomfort.

9. The wood vinegar calcium preparation according to claim 7, characterized in that: The wood vinegar calcium preparation is used as a food additive.

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