Preparation method of salicornia europaea biologic salt

Abstract

The invention provides a preparation method of salicornia europaea bio-salt. The preparation method comprises the following steps: step 1, pretreating raw materials: preparing a culture solution from salicornia europaea raw materials; step 2, mixing and extracting; step 3, sterilization; step 4, inoculating bacillus subtilis; step 5, fermenting; step 6, centrifuging; step 7, carrying out solid-liquid separation; 8, clarifying: removing impurities from the liquid obtained after solid-liquid separation in the step 7 by using a natural clarifying agent, so that the solution is clear and transparent; and 9, crystallizing to obtain the salicornia europaea biologic salt. According to the method disclosed by the invention, the bio-salt is prepared by coupling bacillus subtilis fermentation with ZTC 1 + 1 natural clarifying agent clarification, so that zero-emission, zero-pollution and zero-load environment-friendly manufacturing is realized.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of plant salt preparation, and particularly relates to a preparation method of biological salt of Salicornia. BACKGROUND

[0002] Current treatment measures for saline-alkali soil are physical methods (drainage, flushing, sanding, and alkali pressing), chemical methods (gypsum, phosphogypsum, and desulfurization gypsum), and microbial methods (salt-tolerant microbial strains), all of which face high costs, limited effects, and possible secondary pollution, and cannot effectively treat severe saline-alkali soil. The present application intends to use Salicornia to convert inorganic salt in the soil into biological salt (organic mineral salt) through "biological conversion".

[0003] Biological salt has a significant positive impact on human health, environmental protection, sustainable development, and the development and utilization of agricultural resources. In the field of biological salt preparation, the existing methods mainly use Suaeda as raw material, and the main technical process of preparation is: preparation, raw material crushing, complex enzyme hydrolysis, filtration, fermentation, extraction, flocculation and decolorization, purification, and drying and concentration. The raw materials for preparing biological salt mainly include Suaeda, Nitraria, West Coast Sea Pine, Sea Fennel, Russian hemp, and Alternanthera philoxeroides. Referring to the patent application files with the application numbers 201911400266.4, 201210051265.5, 201711041401.1, 201711015459.9, 201811651347.7, 202010672993.2, and the like; and Salicornia species-derived salt substitutes (application number: CN200880013938.X), the main preparation process is: pretreatment, extraction, ultrafiltration, and drying. However, there are problems such as incomplete impurity removal, low quality, possible loss of nutrients, and low extraction rate of biological salt. No mature patent application or publication has been found for preparing biological salt using Salicornia as raw material. The existing biological salt preparation still has many drawbacks, mainly in the purity of biological salt, consumer acceptance, complex production process, large functional differences, and the like. It is necessary to further optimize the production process, improve the product purity, enrich the product function, strengthen the market promotion, and pay attention to ecological protection. Based on the above considerations, the preparation process of Salicornia biological salt is invented.

[0004] Salicornia is rich in organic nutrients such as amino acids and enzymes, but there is no technical solution for preparing biological salt using Salicornia as raw material at present. Therefore, the present application intends to design a scheme for efficiently preparing Salicornia biological salt using Salicornia as raw material, i.e., a method for preparing biological salt by using Bacillus subtilis fermentation coupled with ZTC 1+1 natural clarifying agent clarification. SUMMARY

[0005] In view of the defects of low extraction rate, low purity, low productivity and low activity of other biological salt preparation technical solutions in the prior art, and the blank of the salt sedge biological salt preparation technical solution, the biological salt is prepared by utilizing Bacillus subtilis fermentation coupled with ZTC 1+1 natural clarifying agent clarification, so that the effects of natural pollution-free, improved biological activity, enhanced nutrition, efficient clarification and effective component protection can be achieved.

[0006] The technical solutions adopted by the present application to solve the problems in the prior art are as follows: A preparation method of salt sedge biological salt, comprising the following steps: Step 1, raw material pretreatment: collecting mature salt sedge raw materials for cleaning, then airing, crushing and sieving under natural conditions to obtain salt sedge powder samples, and dissolving the powder in distilled water to prepare a culture solution; Step 2, mixed extraction: weighing the culture solution in step 1 and placing it into a double water bath constant temperature oscillator for mixed extraction reaction; Step 3, sterilization: placing the culture solution obtained in step 2 into a sterilization pot for sterilization; Step 4, inoculation of Bacillus subtilis: inoculating Bacillus subtilis into the culture solution after sterilization in step 3, and controlling the inoculation amount; Step 5, fermentation: placing the sample solution obtained in step 4 into a two-layer oscillation incubator for fermentation, and controlling the fermentation time and temperature; Step 6, centrifugation: placing the fermentation solution prepared in step 5 into a centrifuge for centrifugation to obtain a centrifugal solution; Step 7, solid-liquid separation: performing solid-liquid separation on the impurity-containing centrifugal solution obtained in step 6 by vacuum filtration in a vacuum pump; Step 8, clarification: using a natural clarifying agent to remove impurities from the liquid prepared after solid-liquid separation in step 7, so that the solution is clear and transparent; Step 9, crystallization: placing the clear solution obtained in step 8 under vacuum conditions, gradually evaporating the solvent in the solution, and gradually removing the crystallization water in the sample, so that the solute in the sample reaches a saturated state, thereby precipitating crystals, and obtaining salt sedge biological salt.

[0007] The sieving condition in step 1 is: 60 mesh sieve, and the solid-liquid ratio of salt sedge powder to distilled water is 1:20-1:30.

[0008] The oscillation speed of the oscillator in the mixed extraction process in step 2 is 180 rpm.

[0009] The sterilization time in step 3 is 30 min, and the sterilization temperature is 121 DEG C.

[0010] The inoculation amount in step 4 is controlled to be 8 % to 12 % of the sample volume.

[0011] The fermentation time in step 5 is 12 hours to 36 hours, and the fermentation temperature is 30° C. to 40° C.

[0012] In step 6, the centrifuge speed is 8000 rpm and the centrifugation is performed for 15 minutes.

[0013] In step 8, the natural clarifier is ZTC 1+1 clarifier. 80 ml of the solution obtained in step 7 is measured and dissolved in distilled water at a ratio of 1:99, and the mixture is swelled for 12 hours with stirring. Component B is dissolved in 1% acetic acid at a ratio of 1:99, and the mixture is swelled for 12 hours with stirring. Component B is added to the solution first, followed by component A, with the addition ratio being B:A = 2:1. After adding component B, fine suspended matter can be seen to aggregate after about 5 minutes of treatment. Component A is then added for auxiliary treatment for 30 minutes. The temperature is maintained at 60°C throughout the process. The total amount of clarifier added to A+B is 6% to 10% of the volume of the measured solution.

[0014] In step 9, the clarification time is 70 min to 90 min.

[0015] Compared with the prior art, the present invention has the following significant improvements: (1) The present invention has outstanding economic and social value: The raw materials used in the present invention come from Salicornia herba grown on heavily saline-alkali land. According to incomplete statistics, saline-alkali land in the Yellow River S-shaped Bend Basin accounts for about 69% of the country's saline-alkali land area. The present invention can not only improve saline-alkali land but also use Salicornia herba to prepare biosalt. As a new type of edible salt, it has great potential in the market. With people's increasing attention to healthy diet, Salicornia herba biosalt is expected to become an important substitute for traditional table salt, meeting consumers' demand for health, low sodium, and balanced nutrition. The production of Salicornia herba biosalt involves multiple links, including raw material planting, processing, sales, etc., which can drive the development of related industrial chains; through continuous research and development and innovation, its quality and added value can be continuously improved to meet the diversified and personalized needs of the market.

[0016] (2) The present invention is highly feasible: This invention uses Salicornia herba, a salt-storing plant, as a raw material to produce biosalt. This not only reduces dependence on sea salt but also alleviates the burden on salt lakes and mineral salts. It also removes salt from saline-alkali land, providing a resource recycling method for saline-alkali land. The inventors developed a process for producing biosalt using Salicornia herba as a raw material through Bacillus subtilis fermentation coupled with a ZTC 1+1 natural clarifier. The process successfully produced biosalt from Salicornia herba with simple operation, high yield, low cost, and ease of large-scale production.

[0017] The preparation method of the present invention has the following advantages: ①Using Bacillus subtilis fermentation coupled with ZTC 1+1 natural clarifier to clarify the preparation of biological salt, realizing zero emission, zero pollution, zero load on the environment of green biological manufacturing.

[0018] ②Using Bacillus subtilis to increase active substances and promote the growth of beneficial bacteria.

[0019] ③Using ZTC 1+1 natural clarifier to clarify efficiently, safely and non-toxicly, and reduce production cost.

[0020] ④Bacillus subtilis tolerates high salt environment, and when coupled with ZTC 1+1 clarifier to ferment high-salt fermentation broth, it can still metabolize and remove impurities efficiently. DETAILED DESCRIPTION

[0021] The technical solutions of the present application are further specifically described below through examples.

[0022] Example 1 Matured Salicornia bigelovii was collected, washed, dried, crushed, and sieved through a 60-mesh sieve, and then dissolved in distilled water to prepare a 1:20 culture solution. The culture solution was placed in a double number water bath constant temperature shaker and shaken at a shaking speed of 180 rpm, and heated to 80℃ for 8h. The prepared sample was placed in a 121℃ sterilization pot, sterilized for 30 min, and then 12% Bacillus subtilis was added. The sample was cultured in a two-layer shaking incubator at 30℃ for 12h to obtain a fermentation broth. The number of bacteria colonies in the fermentation broth was 1.57×10 9 CFU / mL. The fermentation broth was centrifuged in an 8000 rpm centrifuge for 15 min, and then the centrifuged liquid was filtered under a circulating water vacuum pump. A φ50 mm microporous filter membrane was used to separate the solid and liquid, 80 ml of liquid was measured, 10% ZTC 1+1 clarifier was added, and the liquid was clarified for 80 min. Crystallization was performed to obtain Salicornia bigelovii biological salt. The removal rate of impurities in this process was 6.77%. The yield of the biological salt obtained according to the above process was 87.13%, and the ion content is shown in Table 1.

[0023] Example 2 Matured Salicornia bigelovii was collected, washed, dried, crushed, and sieved through a 60-mesh sieve, and then dissolved in distilled water to prepare a 1:20 culture solution. The culture solution was placed in a double number water bath constant temperature shaker and shaken at a shaking speed of 180 rpm, and heated to 80℃ for 8h. The prepared sample was placed in a 121℃ sterilization pot, sterilized for 30 min, and then 12% Bacillus subtilis was added. The sample was cultured in a two-layer shaking incubator at 30℃ for 12h to obtain a fermentation broth. The number of bacteria colonies in the fermentation broth was 1.57×10 9CFU / mL. Centrifuged in a centrifuge at 8000 rpm for 15 min, then the centrifugate was filtered under a circulating water vacuum pump, and solid-liquid separation was achieved using a φ50 mm microporous filter membrane. 80 ml of liquid was measured, 6% ZTC 1+1 clarifier was added, and clarification was performed for 70 min. Saltwort biological salt was obtained by crystallization, and the impurity removal rate in this process reached 5.77%. The yield of biological salt obtained according to the above process was 77.75%, and the ion content is shown in Table 1.

[0024] Example 3 Mature saltwort was collected, washed, dried, crushed, and sieved through a 60-mesh sieve. A culture solution was prepared by dissolving the crushed saltwort in distilled water at a ratio of 1:30. The culture solution was placed in a double-number water bath constant temperature shaker and shaken at a speed of 180 rpm. The temperature was raised to 80°C and maintained for 8 hours. The prepared sample was placed in a 121°C sterilization pot, sterilized for 30 min, and then 8% Bacillus subtilis was added. The sample was cultured in a two-layer shaking incubator at a shaking speed of 18 rpm and a temperature of 40°C for 36 hours to obtain a fermentation broth. The number of bacteria in the fermentation broth was 1.19 x 10 9 CFU / mL. Centrifuged in a centrifuge at 8000 rpm for 15 min, then the centrifugate was filtered under a circulating water vacuum pump, and solid-liquid separation was achieved using a φ50 mm microporous filter membrane. 80 ml of liquid was measured, 6% ZTC 1+1 clarifier was added, and clarification was performed for 70 min. Saltwort biological salt was obtained by crystallization, and the impurity removal rate in this process reached 5.77%. The yield of biological salt obtained according to the above process was 77.75%, and the ion content is shown in Table 1.

[0025] Example 4 Mature saltwort was collected, washed, dried, crushed, and sieved through a 60-mesh sieve. A culture solution was prepared by dissolving the crushed saltwort in distilled water at a ratio of 1:20. The culture solution was placed in a double-number water bath constant temperature shaker and shaken at a speed of 180 rpm. The temperature was raised to 80°C and maintained for 8 hours. The prepared sample was placed in a 121°C sterilization pot, sterilized for 30 min, and then 12% Bacillus subtilis was added. The sample was cultured in a two-layer shaking incubator at a shaking speed of 18 rpm and a temperature of 30°C for 36 hours to obtain a fermentation broth. The number of bacteria in the fermentation broth was 1.35 x 10 9 CFU / mL. Centrifuged in a centrifuge at 8000 rpm for 15 min, then the centrifugate was filtered under a circulating water vacuum pump, and solid-liquid separation was achieved using a φ50 mm microporous filter membrane. 80 ml of liquid was measured, 6% ZTC 1+1 clarifier was added, and clarification was performed for 70 min. Saltwort biological salt was obtained by crystallization, and the impurity removal rate in this process reached 5.77%. The yield of biological salt obtained according to the above process was 77.75%, and the ion content is shown in Table 1.

[0026] Example 5 Matured Salicornia washed, dried, crushed, and passed through a 60-mesh sieve, dissolved in distilled water to make a 1:25 culture solution, and placed in a double number water bath shaker incubator at 180 rpm, heated to 80°C for 8 h. The prepared sample was placed in a 121°C sterilization pot, sterilized for 30 min, and then 10% Bacillus subtilis was added. The mixture was cultured in a two-layer shaking incubator at 35°C for 24 h to obtain a fermentation broth. The number of colonies in the fermentation broth was 1.33 x 10 9 CFU / mL. The broth was centrifuged in an 8000 rpm centrifuge for 15 min, and then the centrifugate was filtered under a circulating water vacuum pump. A φ50 mm microporous filter membrane was used to separate the solid and liquid. 80 ml of the liquid was taken, 10% ZTC 1+1 clarifier was added, and the mixture was clarified for 90 min. Salicornia biological salt was obtained by crystallization. The impurity removal rate in this process was 5.08%. The yield of the biological salt obtained according to the above process was 79.75%, and the ion content is shown in Table 1.

[0027] Example 6 Matured Salicornia washed, dried, crushed, and passed through a 60-mesh sieve, dissolved in distilled water to make a 1:30 culture solution, and placed in a double number water bath shaker incubator at 180 rpm, heated to 80°C for 8 h. The prepared sample was placed in a 121°C sterilization pot, sterilized for 30 min, and then 8% Bacillus subtilis was added. The mixture was cultured in a two-layer shaking incubator at 40°C for 36 h to obtain a fermentation broth. The number of colonies in the fermentation broth was 1.24 x 10 9 CFU / mL. The broth was centrifuged in an 8000 rpm centrifuge for 15 min, and then the centrifugate was filtered under a circulating water vacuum pump. A φ50 mm microporous filter membrane was used to separate the solid and liquid. 80 ml of the liquid was taken, 10% ZTC 1+1 clarifier was added, and the mixture was clarified for 90 min. Salicornia biological salt was obtained by crystallization. The impurity removal rate in this process was 5.08%. The yield of the biological salt obtained according to the above process was 79.75%, and the ion content is shown in Table 1.

[0028] Example 7 Matured Salicornia washed, dried, crushed, and passed through a 60-mesh sieve, dissolved in distilled water to make a 1:20 culture solution, and placed in a double number water bath shaker incubator at 180 rpm, heated to 80°C for 8 h. The prepared sample was placed in a 121°C sterilization pot, sterilized for 30 min, and then 10% Bacillus subtilis was added. The mixture was cultured in a two-layer shaking incubator at 30°C for 12 h to obtain a fermentation broth. The number of colonies in the fermentation broth was 1.42 x 10 9CFU / mL. Centrifugation in 8000 rpm centrifuge for 15 min, then the centrifugal liquid in the circulating water vacuum pump filtration, using φ50 mm microporous filter membrane to realize the solid-liquid separation, take 80 ml of liquid, add 10% ZTC 1+1 clarifier clarification 80 min, crystallization to get the Salicornia bigelovii biological salt, the process of impurity removal rate reached 6.63%. The yield of biological salt obtained according to the above process is 77.82%, and the ion content is shown in Table 1.

[0029] Example 8 The mature Salicornia bigelovii was collected, washed, dried, crushed, and passed through a 60-mesh sieve. The culture solution was prepared by dissolving the crushed Salicornia bigelovii in distilled water at a ratio of 1:30. The culture solution was placed in a double number water bath constant temperature oscillator and shaken at a shaking speed of 180 rpm. The temperature was raised to 80°C and maintained for 8 hours. The prepared sample was placed in a 121°C sterilization pot and sterilized for 30 min. Then 8% Bacillus subtilis was added, and the sample was cultured in a two-layer shaking incubator at a shaking speed of 18 rpm at 40°C for 24 hours to obtain a fermentation broth. The number of bacterial colonies in the fermentation broth was 1.32×10 9 CFU / mL. Centrifugation in 8000 rpm centrifuge for 15 min, then the centrifugal liquid in the circulating water vacuum pump filtration, using φ50 mm microporous filter membrane to realize the solid-liquid separation, take 80 ml of liquid, add 10% ZTC 1+1 clarifier clarification 80 min, crystallization to get the Salicornia bigelovii biological salt, the process of impurity removal rate reached 6.63%. The yield of biological salt obtained according to the above process is 77.82%, and the ion content is shown in Table 1.

[0030] Example 9 The mature Salicornia bigelovii was collected, washed, dried, crushed, and passed through a 60-mesh sieve. The culture solution was prepared by dissolving the crushed Salicornia bigelovii in distilled water at a ratio of 1:30. The culture solution was placed in a double number water bath constant temperature oscillator and shaken at a shaking speed of 180 rpm. The temperature was raised to 80°C and maintained for 8 hours. The prepared sample was placed in a 121°C sterilization pot and sterilized for 30 min. Then 8% Bacillus subtilis was added, and the sample was cultured in a two-layer shaking incubator at a shaking speed of 18 rpm at 40°C for 24 hours to obtain a fermentation broth. The number of bacterial colonies in the fermentation broth was 1.32×10 9 CFU / mL. Centrifugation in 8000 rpm centrifuge for 15 min, then the centrifugal liquid in the circulating water vacuum pump filtration, using φ50 mm microporous filter membrane to realize the solid-liquid separation, take 80 ml of liquid, add 10% ZTC 1+1 clarifier clarification 80 min, crystallization to get the Salicornia bigelovii biological salt, the process of impurity removal rate reached 6.63%. The yield of biological salt obtained according to the above process is 77.82%, and the ion content is shown in Table 1.

[0031] Table 1 Mineral content and biological salt yield of Salicornia bigelovii biological salt in each example and comparative example

[0032] In combination with the data in Table 1, the above example results show that in the core process of extracting biological salt by coupling Bacillus subtilis and ZTC 1+1 natural clarifier, the solid-liquid ratio is the main control factor in the fermentation stage, and the clarifier addition amount is the key parameter in the clarification stage. The process realizes the dual effects of targeted purification and nutrient enrichment type preparation of biological salt through fermentation of Bacillus subtilis coupled with ZTC 1+1 natural clarifier. The process can effectively remove impurities and clarify, and provides a new solution to solve the technical bottlenecks of nutrient competition, incomplete impurity removal, and insufficient quality and other problems in the traditional biological salt extraction method.

[0033] The Bacillus subtilis coupled ZTC 1+1 natural clarifier extraction process for biological salt provided by the application successfully realizes the perfect unity of high yield (73.51%~87.13%), nutrient enrichment and efficient targeted impurity removal (6.77%~5.08%) of biological salt by precisely controlling the key variables such as fermentation solid-liquid ratio (1:20~1:30), fermentation temperature of 30 ℃~40 ℃ and clarifier addition amount (6%~10%). The detailed experimental data undoubtedly prove that in the high-salt biological salt system, ZTC 1+1 natural clarifier still shows excellent impurity removal and clarification efficiency; at the same time, Bacillus subtilis shows excellent survival rate in high-salt environment, ensuring the fermentation efficiency. The core innovation of the process lies in the synergistic effect of "Bacillus subtilis fermentation coupled with ZTC 1+1 clarification" in high-salt environment. This combination significantly improves the retention rate of active substances and the removal efficiency of impurities under high-salt conditions, and the deep coupling of the two links finally produces the effect of "1+1>2". The process has significant advantages and innovation in solving the core bottleneck problems of traditional biological salt extraction methods such as nutrient content, impurities and quality, and provides a new efficient, stable and reliable technical solution for the industrialized production of high-quality biological salt.

[0034] The protection scope of the application is not limited to the above-mentioned embodiments. Obviously, those skilled in the art can make various modifications and changes to the application without departing from the scope and spirit of the application. If these modifications and changes belong to the scope of the claims of the application and equivalent technologies, the intention of the application also includes these modifications and changes.

Claims

1. A method for preparing Salicornia herba biosalt, characterized in that: The steps include: Step 1, raw material pretreatment: collecting mature Salicornia herba raw materials, washing them, then drying them under natural conditions, crushing them, and sieving them to obtain Salicornia herba powder samples, and dissolving the powder in distilled water to prepare a culture solution; Step 2, mixed extraction: weigh the culture medium in step 1 and place it in an even-numbered water bath constant temperature oscillator for mixed extraction reaction; Step 3, sterilization: sterilize the culture solution obtained in step 2 in an autoclave; Step 4, inoculating Bacillus subtilis: inoculating Bacillus subtilis into the culture solution sterilized in step 3, controlling the inoculation amount; Step 5, fermentation: ferment the sample solution obtained in step 4 in a two-layer shaking incubator, controlling the fermentation time and temperature; Step 6, centrifugation: placing the fermentation broth obtained in step 5 into a centrifuge for centrifugation to obtain a centrifuge; Step 7, solid-liquid separation: the impurity-containing centrifuge liquid obtained in step 6 is filtered in a vacuum pump to achieve solid-liquid separation; Step 8, clarification: using ZTC 1+1 clarifier to remove impurities from the liquid obtained after solid-liquid separation in step 7 to make the solution clear and bright; Step 9, crystallization: the clarified solution obtained in step 8 is placed under vacuum conditions. As the solvent in the solution gradually evaporates, the crystallization water in the sample is gradually removed, and the solute in the sample reaches a saturated state, thereby precipitating crystals to obtain Salicornia herba biosalt.

2. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: The sieving conditions in step 1 are: 60 mesh sieve, and the solid-liquid ratio of Salicornia herba powder to distilled water is 1:20 to 1:

30.

3. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: The shaking speed of the shaker during the mixing and extraction process in step 2 is 180 rpm.

4. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: The sterilization time in step 3 is 30 minutes and the sterilization temperature is 121°C.

5. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: In step 4, the inoculum volume is controlled at 8% to 12% of the sample volume.

6. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: The fermentation time in step 5 is 12 hours to 36 hours, and the fermentation temperature is 30° C. to 40° C.

7. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: In step 6, the centrifuge speed is 8000 rpm and the centrifugation is performed for 15 minutes.

8. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: The specific clarification process using the ZTC 1+1 clarifier in step 8 is as follows: 80 ml of the solution obtained in step 7 is measured, component A is dissolved in distilled water at a ratio of 1:99, and the mixture is swelled for 12 hours with stirring; component B is dissolved in 1% acetic acid at a ratio of 1:99, and the mixture is swelled for 12 hours with stirring. Component B is added to the solution first, followed by component A, with the addition ratio being B:A = 2:

1. After the addition of component B, fine suspended matter is observed to aggregate after about 5 minutes of treatment, and then component A is added for auxiliary treatment for 30 minutes. The temperature is maintained at 60°C throughout the process, and the total amount of clarifier added to the A+B solution is 6% to 10% of the volume of the measured solution.

9. The method for preparing a Salicornia herb biosalt according to claim 1, wherein: In step 9, the clarification time is 70 min to 90 min.

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