Antimicrobial activity by bee bread and their production method

Abstract

Disclosed is a biocomposite product with high microbial activity obtained by using bee bread, the method of obtaining this product and using of this biocomposite material for the coating of products like artificial materials, packaging, etc. to be used in areas requiring hygiene or their use as an intermediate raw material.

Claims

1. A method of producing a biocomposite nanofiber, the method comprising: extracting bee bread so as to obtain an extract; lyophilizing of the extract; turning a thermoplastic polymer into a polymer solution by using a magnetic stirrer; adding the lyophilized extract into the polymer solution; stirring the added lyophilized extract and the polymer solution with the magnetic stirrer so as to obtain a homogenous solution; and electrospinning the homogenous solution so as to form the biocomposite nanofiber.

2. The method of claim 1, wherein the bee bread has a mixture of 20% to 50% bee bread with methanol, wherein the step of extracting comprises: extracting the bee bread with water.

3. The method of claim 1, wherein the step of extracting in carried out at 50 C. for ten hours in a shaken water bath.

4. The method of claim 1, wherein the step of extracting the bee bread comprises: filtering the bee bread with a filter at least two times for one hour so as to obtain the extract.

5. The method of claim 1, further comprising: freezing the extract at a temperature of 80 C., wherein the step of lyophilizing is carried out in lyophilizer.

6. The method of claim 1, wherein the thermoplastic polymer is selected from the group consisting of polyurethane, polylactic acid, polyglycolic acid, poly (epsilon-caprolactone), and polyethylene.

7. The method of claim 1, wherein the step of turning comprises: preparing the polymer solution in a homogenous structure with the magnetic stirrer for twenty-four hours at room temperature.

8. The method of claim 1, wherein the step of adding the lyophilized extract into the polymer solution comprises: adding 5% to 50% lyophilized extract to the polymer solution.

9. The method of claim 1, wherein the step of stirring comprises: stirring the added lyophilized extract and the polymer solution for between one hour and two hours at room temperature.

10. The method of claim 1, further comprising: filling a plastic syringe with ten milliliters to twenty milliliters of solution.

11. The method of claim 10, wherein the step of electrospinning comprises: passing the homogenous solution from the plastic syringe through a stainless steel needle into an electrospinning device.

12. The method of claim 11, wherein the step of passing is at a rate of between one milliliter per minute and twenty milliliters per minute.

13. The method of claim 12, wherein a tip of the stainless steel needle is spaced by five centimeters to twenty centimeters from a collector surface of the electrospinning device.

14. The method of claim 1, further comprising: winding the biocomposite nanofiber onto a collector surface by rotating the collector surface, the collector surface being covered with an aluminum film.

15. A biocomposite nanofiber produced by the method of claim 1.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) The invention is a polymer biocomposite product with high microbial activity obtained using bee bread, the method of obtaining this product and its use for coating material or as an intermediate raw material for artificial materials, packaging, etc. to be used in areas requiring hygiene.

(2) The basic steps of the proposed method are as follows: 1obtaining an extract by extracting bee bread, 2lyophilization of (freeze-drying) extract, 3bringing the thermoplastic polymer (preferably polyurethane, polylactic acid, polyglycolic acid, poly(epsilon-caprolactone), polyethylene) into solution with magnetic stirrer, 4adding the polymer solution into lyophilized bee bread extract and obtaining a homogeneous solution with magnetic stirrer, 5converting the solution consisting of polymer and bee bread extract into nanofibers (biocomposite) by electrospinning method, 6using of the obtained biocomposite material as a coating or intermediate raw material for artificial materials, packaging, and etc. products that wanted to gain hygienic properties.

(3) In the extraction process to be carried out in the first step, 20-50% bee bread is left to extraction with methanol: water (80:20) at 50 C. for 10 hours in a shaking water bath. At the end of the period, it is filtered through Whatman 1 filter. The bee bread remaining on the filter is left to be extracted with the same amount of solvent (methanol: water) for 1 hour under similar conditions, then the extract is removed by filtering through the filter paper. In the last stage, bee bread is left for 1 more hour extraction under similar conditions. The extracts obtained at the end of the three stages are combined. Then the methanol is removed by keeping it in the rotary evaporator under vacuum.

(4) In the lyophilization stage of the method, the remaining dense extract is frozen at 80 C. and then freeze-dried in a lyophilizer. Lyophilization process is carried out in 3 main stages; 1. Freezing the Product: The process of completely freezing the product after passing through the appropriate pre-preparation processes, turning it into crystal form. 2. Sublimation (Primary Drying): Creating the energy required for sublimation to take place in the frozen product and performing sublimation. The stage where the water content is 90-95% removed. 3. Desorption (Secondary Drying): The process of removing water that cannot be removed in primary drying and is bound to the substance.

(5) In the third step, the polymer is brought into solution using magnetic stirrer to prepare the polymer. Thermoplastic polymer (preferably polyurethane, polylactic acid, polyglycolic acid, poly (epsilon-caprolactone), polyethylene) is preferred as the polymer here.

(6) The reason why the mentioned thermoplastic polyurethane material preferred is to have the properties of softening when heated, hardening when cooled, and being easily processed in heat-treatment processes such as extrusion and injection.

(7) Furthermore, it shows a good performance during processing with its unique molecular structure and it can have different chemical properties and strengths by regulating the chemical structures in its formulation. This structure makes it a good packaging material.

(8) In the fourth step, lyophilized bee bread extract is added to the polymer solution and a homogeneous solution is obtained with magnetic stirrer.

(9) In the fifth step, this homogeneous solution (polymer, bee bread extract, solvent used to form the polymer solution) is transformed into nanofibers (biocomposite) by electrospinning method.

(10) The above thermoplastic polymer solutions can be selected from solvents such as dimethyl sulfoxide or dimethyl formamide, as well as solvents such as tetrahydrofuran, acetone, methyl ethyl ketone, chloromethane, dichloromethane can be used to form the polymer solution.

(11) In the preferred embodiment of the invention, lyophilized bee bread extract was prepared into a total polymer solution up to 50% by weight, by the following method: In the first step, a homogeneous polymer solution (in the range of 6-15% by weight) is prepared with magnetic stirrer for 24 hours at room temperature. In the second stage, 5-50% by weight of lyophilized bee bread extract is added into the total polymer solution and mixing continues with magnetic stirrer at room temperature for 1-2 hours until a homogeneous structure is obtained.

(12) Bee bread extract added thermoplastic polymer solutions prepared at room temperature, are filled into a plastic syringe in the range of 10 mL-20 mL and this syringe is attached to a stainless-steel needle as a nozzle. The polymer solution in the syringe is produced in the form of nanofibers by using an electrospinning device (Inovenso-Nanospinner24). Here, the emitter electrode of the high-voltage power supply, which will allow the polymer solution to be drawn into fibers, is charged by connecting it to a conductive jet. The other or grounding electrode of the high-voltage power supply is connected to the conductor collecting device to complete the cycle. To adjust the optimum voltage for each polymer solution, different voltages were applied to the polymer solutions obtained as nanofibers by electrospinning. The feeding rate was set as 1 mL/min-20 mL/min and the distance between the needle tip and the collector surface was set as 5 cm-20 cm. The samples (Nanofibers) are wrapped in a collector surface covered with an aluminum film and in rotation.

(13) The recommended packaging material is the use of pollen, propolis, honey, brood homogenate, as well as biologically active products including vitamins, food coloring, mineral and medicinal substances in the form of wax. The product also allows for effective oral or sublingual use. The product can be used as food, packaging, medical products (all implant materials remaining in the body) by covering them with appropriate coating techniques. For example, it can be applied locally in artificial vessels, prostheses, tissue support materials, hemorrhage stopping materials, wound infections and wound care products, open wounds, as well as it is used systemically for balancing the immune system in immunosuppressive or transplant patients, textile, automotive, furniture, chemistry fields.