Preparation method of red amorphous selenium nanoparticle product

Abstract

The invention discloses a red amorphous selenium nanoparticle product preparation method, which comprises: weighing a first mass of purple perilla polysaccharide, and adding a first volume of deionized water to obtain a first solution; adding a second volume of 10mmol / L Na2SeO3 solution into the first solution, and reacting in a dark place to obtain a second solution; adding a third volume of 40mmol / L VC (Vitamin C) solution into the second solution, and reacting in a dark place to obtain a third solution; dialyzing the third solution through a dialysis bag, and removing unreacted reagents and impurities to obtain a fourth solution; the fifth solution is subjected to freeze drying, and the red amorphous selenium nanoparticle product is obtained. According to the purple perilla polysaccharide-selenium nanoparticles (PFPS-SeNPs) prepared by the invention, when the volume of Na2SeO3 and the volume of VC are 4mL, the particle size and PDI reach the minimum value, and the purple perilla polysaccharide-selenium nanoparticles have good dispersity, stability and antioxidant activity, and can be widely applied to the fields of health foods, biological medicines and the like as selenium nanoparticles with relatively good stability or a novel selenium supplementing agent.

Description

Technical Field

[0001] The invention relates to the technical field of selenium particles, and in particular to a method for preparing a red amorphous selenium nanoparticle product. Background Art

[0002] Selenium is one of the essential trace elements for the human body. It plays an important role in reducing the incidence of cancer, preventing cardiovascular disease, diabetes, anti-oxidation, anti-virus, and enhancing immunity. Selenium deficiency in the human body can cause a series of health problems. Studies have shown that the boundary between the minimum acceptable intake of selenium and toxicity is very narrow. Overdose may lead to selenium poisoning. In addition, the form of selenium also affects its toxicity and biological activity. At present, selenium mainly exists in the form of inorganic selenium (such as sodium selenite, etc.), organic selenium (such as selenomethionine, methylselenocysteine, etc.) and nano-selenium. Among them, the low bioavailability and high toxicity of inorganic selenium limit its widespread promotion and application in medicine and food. Organic selenium is comparable to inorganic selenium in terms of subchronic toxic dose, and its complex manufacturing process and high production cost lead to less practical application. In contrast, selenium nanoparticles (SeNPs) formed by nano-sizing are a kind of red elemental selenium. Due to their tiny size and special physicochemical properties, SeNPs have physical and chemical properties such as quantum size effect, surface effect and macroscopic quantum tunneling effect, and thus show broad application prospects in many fields such as physics, chemistry and biology. As a semiconductor material, SeNPs can be used in the manufacture of sensors and exhibit excellent sensing performance; as an adsorbent, SeNPs can efficiently adsorb and remove heavy metal pollutants in sewage; as a selenium supplement, red, zero-valent amorphous SeNPs have lower toxicity and higher biological activity than other forms of selenium, showing potential application value in the prevention and treatment of diseases such as cancer.

[0003] At present, the most commonly used methods for the synthesis of SeNPs include physical methods (pulsed laser ablation, hydrothermal treatment, microwave radiation sonochemical process and physical evaporation, etc.), biological methods (plants, fungi or bacteria) and chemical methods. Physical methods usually require high temperature, high pressure, catalysts, etc. These methods have low efficiency and yield, and expensive equipment makes it impossible to widely produce SeNPs; biological methods are to reduce selenate or selenite to elemental selenium through compounds present in organisms, such as phenols, flavonoids, alcohols, proteins and aldehydes. Since specific microorganisms are used for the production of SeNPs with different characteristics, more research is needed to match microorganisms with corresponding compounds and therapeutic activities; chemical methods mainly reduce the oxidized form of selenium to elemental selenium through reducing agents such as ascorbic acid, traps, etc. Although this method saves time and effort, the formed SeNPs are highly unstable and easily aggregate into gray or black precipitates in aqueous solution, and have low biological activity. Therefore, suitable protective agents or dispersants are usually added to maintain the stability of SeNPs. Summary of the invention

[0004] The invention provides a method for preparing a red amorphous selenium nanoparticle product, aiming to solve the above-mentioned problems existing in the background technology.

[0005] The technical solution provided by the present invention is as follows:

[0006] A method for preparing a red amorphous selenium nanoparticle product comprises the following steps:

[0007] Weighing a first mass of perilla polysaccharide, adding a first volume of deionized water, to obtain a first solution;

[0008] Adding a second volume of 10 mmol / L Na2SeO3 solution to the first solution, and reacting in the dark to obtain a second solution;

[0009] To the second solution was added a third volume of 40 mmol / L C solution, and reacting in the dark to obtain a third solution;

[0010] dialyzing the third solution through a dialysis bag to remove unreacted reagents and impurities to obtain a fourth solution;

[0011] freeze-drying the fifth solution to obtain the red amorphous selenium nanoparticle product;

[0012] Wherein, the first mass: the first volume is 1:500;

[0013] The second volume: the third volume is (1~8): (1~8).

[0014] Furthermore, the obtaining of the second solution after reacting in the dark specifically comprises: obtaining the second solution after reacting in the dark at 40° C. for 30 minutes.

[0015] Furthermore, the obtaining of the third solution after reacting in the dark specifically comprises: obtaining the third solution after reacting in the dark at room temperature for 1 hour.

[0016] Furthermore, the dialysis bag is a 3000da dialysis bag, and the dialysis time is 48h.

[0017] Furthermore, the present invention provides a method for preparing a red amorphous selenium nanoparticle product, comprising the following steps:

[0018] Weigh 0.02 g of perilla polysaccharide in four portions, add 10 mL of deionized water to each portion, and obtain four portions of perilla polysaccharide solutions;

[0019] 0.5 mL, 1 mL, 2 mL and 4 mL of 10 mmol / L Na2SeO3 solution were added to each portion of the perilla polysaccharide solution, respectively, and reacted at 40° C. in the dark for 30 min to obtain four portions of the second solution;

[0020] To the second solution, 0.5 mL, 1 mL, 2 mL and 4 mL of 40 mmol / L L-1 were added respectively. C The solution was reacted in the dark for 1 h to obtain four portions of the third solution;

[0021] After the reaction is completed, the third solution is dialyzed using a 3000da dialysis bag for 48 hours to obtain four fourth solutions;

[0022] The fourth solution was freeze-dried to obtain PFPS-SeNPs. 0.5 , PFPS-SeNPs1, PFPS-SeNPs2, and PFPS-SeNPs4.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The invention discloses a method for preparing a red amorphous selenium nanoparticle product, wherein perilla polysaccharide (PFPS) is used as a stabilizer, Na2SeO3 is used as a selenium source oxidant, and ascorbic acid (V C ) was used as a reducing agent to prepare PFPS-SeNPs. C When the volume was 4 mL, the particle size and PDI reached the minimum. The prepared PFPS-SeNPs have good dispersibility, stability and antioxidant activity, and can be widely used in health food, biomedicine and other fields as a selenium nanoparticle with good stability or a new selenium supplement. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 A flow chart of the method for preparing the red amorphous selenium nanoparticle product provided by the present invention;

[0026] Figure 2 The actual pictures of SeNPs prepared with different stabilizers in the examples of the present invention, wherein (a) is a SeNPs control example; (b) is PFPS-SeNPs; (c) is PFP-SeNPs; (d) is PFPP-SeNPs;

[0027] Figure 3 The particle size distribution diagram of SeNPs prepared by different stabilizers in the examples of the present invention;

[0028] Figure 4 The different Na2SeO3 / V in the embodiments of the present invention CPhysical picture of PFPS-SeNPs prepared under volume, where (a) is SeNPs control example, (b) is PFPS-SeNPs 0.5 ; (c) is PFPS-SeNPs1, (d) is PFPS-SeNPs 2、 (e) PFPS-SeNPs4;

[0029] Figure 5 The particle size distribution diagram of PFPS-SeNPs prepared under different Na2SeO3 / VC volumes in the embodiment of the present invention;

[0030] Figure 6 Schematic diagram of the principle of PFPS-SeNPs in the embodiment of the present invention and the chemical reaction formula of Na2SeO3 and Vc;

[0031] Figure 7 The scanning electron microscopy images of SeNPs and PFPS-SeNPs in the embodiments of the present invention are shown;

[0032] Figure 8 The structural characteristic diagrams of PFPS-SeNPs in the embodiments of the present invention, wherein (A) is a UV-Vis spectrum analysis diagram; (B) is a FTIR spectrum analysis diagram;

[0033] Fig. 9 The XRD spectrum analysis diagram of PFPS-SeNPs in the embodiment of the present invention;

[0034] Fig.10 The particle size distribution diagram of PFPS-SeNPs at different storage times in the embodiment of the present invention;

[0035] Fig.11 This is a diagram showing the effect of different concentrations of PFPS, SeNPs and PFPS-SeNPs on OH· scavenging ability in the examples of the present invention. DETAILED DESCRIPTION

[0036] In order to make the purpose, technical solution and advantages of the embodiments of the present application clearer, the technical solution in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the embodiments described below are part of the embodiments of the present application, not all of the embodiments.

[0037] In recent years, some studies have used bioactive molecules such as polysaccharides (calcium citrate polysaccharides, chitosan, dextran, etc.), proteins (β-lactoglobulin, bovine serum albumin, etc.), polysaccharide-protein complexes, polyphenols (epigallocatechin-3-gallate, etc.), folic acid and copolymers (polylactic acid / polylactic-co-glycolic acid, etc.) to modify the surface of SeNPs, thereby producing stable surface-covered or nested dispersant nano-selenium forms. Compared with other stabilizers, natural polysaccharides with active functional groups (such as hydroxyl and carboxyl groups formed by monosaccharide molecules connected by glycosidic bonds) have more advantages in enhancing the stability of nanoparticles due to their hydrophilic hydroxyl groups and high specific surface area. In addition, natural polysaccharides also have good water solubility and low toxicity. Therefore, some polysaccharides are considered to be ideal materials for preparing SeNPs with good dispersion and bioactivity in aqueous media. Based on this, the present invention provides a method for preparing a red amorphous selenium nanoparticle product, using perilla polysaccharide (PFPS) as a stabilizer, Na2SeO3 as a selenium source oxidant, ascorbic acid (V C ) was used as a reducing agent to prepare PFPS-SeNPs.

[0038] See also Figure 1 The method for preparing a red amorphous selenium nanoparticle product provided by the present invention comprises the following steps:

[0039] Step S1, weighing a first mass of Perilla polysaccharide (PFPS), adding a first volume of deionized water to obtain a first solution.

[0040] Step S2, adding a second volume of 10 mmol / L Na2SeO3 solution to the first solution, and reacting in a dark place to obtain a second solution.

[0041] Step S3, adding a third volume of 40mmol / LV C solution, and a third solution was obtained after reacting in the dark.

[0042] Step S4, dialyzing the third solution through a dialysis bag to remove unreacted reagents and impurities to obtain a fourth solution.

[0043] Step S5, freeze-drying the fifth solution to obtain the red amorphous selenium nanoparticle product.

[0044] Wherein, the first mass: the first volume is 1:500;

[0045] The second volume: the third volume is (1-8): (1-8). The added Na2SeO3 solution and V C The volume range of the solution is 0.5~6mL.

[0046] Optionally, obtaining the second solution after reacting in the dark specifically comprises: obtaining the second solution after reacting in the dark at 40° C. for 30 minutes.

[0047] Optionally, obtaining the third solution after reacting in the dark specifically includes: obtaining the third solution after reacting in the dark at room temperature for 1 hour.

[0048] Optionally, the dialysis bag is a 3000da dialysis bag and the dialysis time is 48h.

[0049] The present invention illustrates a method for preparing a red amorphous selenium nanoparticle product. The method for preparing a red amorphous selenium nanoparticle product comprises the following steps:

[0050] (1) Weigh 0.02 g of perilla polysaccharide in four portions, add 10 mL of deionized water to each portion, and obtain four portions of perilla polysaccharide solutions;

[0051] (2) adding 0.5 mL, 1 mL, 2 mL and 4 mL of 10 mmol / L Na2SeO3 solution to each portion of the perilla polysaccharide solution, respectively, and reacting at 40° C. in the dark for 30 min to obtain four portions of the second solution;

[0052] (3) Add 0.5 mL, 1 mL, 2 mL and 4 mL of 40 mmol / L L-1 into the second solution respectively. C The solution was reacted in the dark for 1 h to obtain four portions of the third solution;

[0053] (4) After the reaction is completed, the third solution is dialyzed using a 3000da dialysis bag for 48 hours to obtain four fourth solutions;

[0054] (5) freeze-drying the fourth solution to obtain PFPS-SeNPs 0.5 , PFPS-SeNPs1, PFPS-SeNPs2, and PFPS-SeNPs4.

[0055] In this embodiment, according to the ratio of perilla polysaccharide mass to deionized water volume of 1:500, 0.02g perilla polysaccharide was weighed in four portions, and 10mL deionized water was added to each portion. In other embodiments, the mass of perilla polysaccharide of other components and deionized water can also be selected to prepare the first solution according to the principle of perilla polysaccharide mass to deionized water volume of 1:500, such as weighing 0.04g perilla polysaccharide and adding 20mL deionized water. Correspondingly, 10mmol / L Na2SeO3 and 40mmol / L C The volume of the solution was doubled accordingly.

[0056] At the same time, the present invention also provides a comparative example, using perilla protein (PFP) and perilla polysaccharide-protein complex (PFPP) as raw materials, Na2SeO3 as a selenium source oxidant, ascorbic acid (V C ) as a reducing agent, PFP-SeNPs and PFPP-SeNPs were prepared by using the same conditions as described above in the present invention. Figure 2 As shown in the figure, (a) is SeNPs prepared by conventional methods; (b) is PFPS-SeNPs prepared by the present invention using perilla polysaccharide (PFPS); (c) is PFP-SeNPs; (d) is PFPP-SeNPs. Figure 3 The results in Table 1 show that under the same preparation conditions, the SeNPs prepared using PFPS as a template have smaller particle size and more uniform distribution.

[0057] Table 1 Effects of different stabilizers on SeNPs particle size and PDI

[0058]

[0059] After analysis, PFPS was used as a stabilizer, and by changing the Na2SeO3 and V C volume ratio, prepared size-controlled PFPS-SeNPs (e.g. Figure 4 shown), Figure 4 Medium, (a) SeNPs; (b) PFPS-SeNPs 0.5 ; (c) PFPS-SeNPs1; (d) PFPS-SeNPs2; (e) PFPS-SeNPs4. The results show that when Na2SeO3 and V C When the volume was 4 mL, the particle size of PFPS-SeNPs reached the minimum value of 105.93 ± 12.12 nm (see Figure 5 and Table 2 ).

[0060] Table 2 Different volumes of Na2SeO3 / V C Effects of PFPS-SeNPs particle size and PDI

[0061]

[0062] The principle of preparing PFPS-SeNPs in the present invention is as follows: First, the precursor SeO3 2-It is adsorbed on the PFPS chain through electrostatic interactions and then reduced to elemental selenium in situ. These selenium atoms aggregate into selenium cores, which further aggregate into SeNPs as the reaction proceeds. Since the selenium atoms on the surface of SeNPs bind to the OH groups (OH…Se) and -CONH- groups (NH…Se) of PFPS, a PFPS-SeNPs nanocomposite is produced to protect the SeNPs from further aggregation. Finally, the selenium surface is strongly adsorbed and passivated by the PFPS molecules, resulting in the SeNPs being well dispersed and stable in the PFPS molecular microenvironment. Based on the formation mechanism of PFPS-SeNPs, the size of SeNPs can be controlled by adjusting the ratio of selenium to PFPS (see Figure 6 ).

[0063] Perilla polysaccharides (PFPS) are a class of complex soluble bioactive macromolecules. Many studies have shown that PFPS has potential biological efficacy. After research, it was found that four single components of perilla leaf polysaccharides were obtained by separation and purification with DEAE-cellulose-52 and Sephadex G-100, all of which showed different degrees of in vitro antioxidant activity; perilla cake polysaccharides are medium molecular weight polysaccharides with β-glycosidic bonds, mainly composed of rhamnose, arabinose, xylose, mannose, glucose and galactose, and have good antioxidant activity. Because natural polysaccharides contain functional groups such as -OH, -COOH, -NH2, and -C=O, these functional groups can combine with nano-selenium to form intermolecular and intramolecular hydrogen bonds along the polymer chain, thereby enhancing the stability of nano-selenium.

[0064] The PFPS-SeNPs prepared by the present invention are C When the volume was 4 mL, the particle size and PDI reached the minimum values, which were 105.93 ± 12.12 nm and 0.136 ± 0.008, respectively. SEM observations showed that SeNPs had a larger specific surface area and particle size due to their low water solubility in the absence of PFPS, while PFPS-SeNPs were surrounded by some substances and showed a uniform spherical shape and low size distribution, with a size smaller than SeNPs (see Figure 7 ); ICP-MS analysis showed that the selenium contents of SeNPs and PFPS-SeNPs were (3.96±0.001)% and (28.56±0.05)%, respectively; UV and FTIR spectroscopy analysis showed that the formation of PFPS-SeNPs may be achieved by the hydrogen bond-like interactions (OH…Se) and (NH…Se) between the selenium atoms of SeNPs and the OH groups and -CONH- groups of PFPS (see Figure 8 ), Figure 8Figure 1 shows the structural characteristics of PFPS-SeNPs, including (A) UV-Vis spectrum analysis; (B) FTIR spectrum analysis. XRD analysis shows that PFPS-SeNPs are amorphous (see Fig. 9 ); Stability analysis showed that PFPS-SeNPs had good stability when placed in the dark at 4°C for 21 days (see Fig.10 and Table 3); the antioxidant activity analysis showed that the scavenging ability of PFPS-SeNPs on hydroxyl radicals was significantly higher than that of PFPS and SeNPs (see Fig.11 ). All these results indicate that SeNPs constructed by PFPS have good dispersibility, stability and antioxidant activity, and can be widely used in health food, biomedicine and other fields as a selenium nanoparticle with good stability or a new selenium supplement.

[0065] Table 3 Changes in absorbance ratio (A410 / A490), particle size and PDI of PFPS-SeNPs at different storage times

[0066]

[0067] The above is only the best specific implementation of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions within the technical scope disclosed in the present application shall be included in the protection scope of the present application. Therefore, the protection scope of the present application shall be based on the protection scope of the claims.

Claims

1. A method for preparing a red amorphous selenium nanoparticle product, characterized in that: The following steps are involved: Weighing a first mass of perilla polysaccharide, adding a first volume of deionized water, to obtain a first solution; Adding a second volume of 10 mmol / L Na2SeO3 solution to the first solution, and reacting in the dark to obtain a second solution; Add a third volume of 40 mmol / L L C solution, and reacting in the dark to obtain a third solution; dialyzing the third solution through a dialysis bag to remove unreacted reagents and impurities to obtain a fourth solution; freeze-drying the fifth solution to obtain the red amorphous selenium nanoparticle product; Wherein, the first mass: the first volume is 1:500; The second volume: the third volume is (1~8): (1~8).

2. The method for preparing the red amorphous selenium nanoparticle product according to claim 1, characterized in that: The step of obtaining the second solution after reacting in a dark place specifically comprises: obtaining the second solution after reacting in a dark place at 40° C. for 30 minutes.

3. The method for preparing the red amorphous selenium nanoparticle product according to claim 1, characterized in that: The step of obtaining the third solution after reacting in the dark specifically includes obtaining the third solution after reacting in the dark at room temperature for 1 hour.

4. The method for preparing a red amorphous selenium nanoparticle product according to any one of claims 1 to 3, characterized in that: The dialysis bag is a 3000da dialysis bag, and the dialysis time is 48h.

5. The method for preparing a red amorphous selenium nanoparticle product according to claim 1, characterized in that: The following steps are involved: Weigh 0.02 g of perilla polysaccharide in four portions, add 10 mL of deionized water to each portion, and obtain four portions of perilla polysaccharide solutions; 0.5 mL, 1 mL, 2 mL and 4 mL of 10 mmol / L Na2SeO3 solution were added to each portion of the perilla polysaccharide solution, respectively, and reacted at 40° C. in the dark for 30 min to obtain four portions of the second solution; To the second solution, 0.5 mL, 1 mL, 2 mL and 4 mL of 40 mmol / L L-1 were added respectively. C The solution was reacted in the dark for 1 h to obtain four portions of the third solution; After the reaction is completed, the third solution is dialyzed using a 3000da dialysis bag for 48 hours to obtain four fourth solutions; The fourth solution was freeze-dried to obtain PFPS-SeNPs. 0.5 , PFPS-SeNPs1, PFPS-SeNPs2, and PFPS-SeNPs4.

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