Food supplement and production method thereof effective on cognitive functions

Abstract

In particular, the present invention relates to a food supplement that prevents and / or eliminates the effects of cognitive impairment occurring in Alzheimer-like dementia types, prevents toxic effects by obtaining the same from natural materials, and reduces the risk of neurodegenerative diseases occurring and / or progressing with its antioxidant effect by using black carrots and chicory roots, and to the food supplement production method that enables the production of this food supplement.

Description

[0001] DESCRIPTION

[0002] FOOD SUPPLEMENT AND PRODUCTION METHOD THEREOF EFFECTIVE ON COGNITIVE FUNCTIONS

[0003] Field of the Invention

[0004] The present invention relates to a food supplement and production method thereof that prevents and / or eliminates the negative effects of cognitive disorders by affecting the central nervous system.

[0005] In particular, the present invention relates to a food supplement that prevents and / or eliminates the effects of cognitive impairment occurring in Alzheimer-like dementia types, prevents toxic effects by obtaining the same from natural materials, and reduces the risk of neurodegenerative diseases occurring and / or progressing with its antioxidant effect by using black carrots and chicory roots, and to the food supplement production method that enables the production of this food supplement.

[0006] State of the Art

[0007] Cognitive disorders are generally characterized as mental disorders with known or suspected biological causes that negatively affect the patient's cognitive abilities and daily functioning. Disorders that manifest themselves through significant changes in the individual's level of functioning before the onset of the disease generally include impairments in the patient's mental functions such as thinking, learning, memory, attention, decision-making, problem-solving, and perception. These disorders, which can often arise due to neurological, psychiatric, or metabolic diseases, can also manifest themselves due to variable physical or mental conditions, the use of and / or withdrawal from substances that affect brain function, and the patient's advancing age and their negative effects that affect daily life and limit the independence of the individual can increase in a short time. Our ability to perform cognitive functions depends on brain functions such as thinking, reasoning, storing and recalling information, and in progressive deterioration, such as Alzheimer's disease, significant deterioration occurs in these functions, leading to gradual loss of function. In general, the more widespread the damage after the onset of the disease, the greater the dysfunction.These disorders, defined as neurodegenerative diseases, cause brain cells to lose their function and / or malfunction overtime, resulting in various symptoms such as decreased cognitive abilities, movement disorders, loss of balance, and speech difficulties. Neurodegenerative diseases that affect different parts of the brain and cause various symptoms include Alzheimer's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis (ALS), Huntington’s disease, Multiple Sclerosis (MS) and Dementia. Early diagnosis of cognitive disorders, especially those seen in diseases such as Alzheimer's, Lewy body dementia, and Parkinson's, is very important in controlling the progression of the disease.

[0008] Oxidative stress, inflammation, and mitochondrial dysfunction contribute significantly to the pathophysiology of these diseases. The accumulation of all these associated events is thought to be responsible for dopaminergic neurodegeneration by inducing apoptosis. In this context, the biggest problems in the administration of drugs used in the treatment of these diseases are the inability of the drug active ingredients to cross the blood-brain barrier, low bioavailability and toxicity problems. Various nanoformulations have been developed to increase the effectiveness of the drugs used and facilitate their transport to the brain. However, traditional chemical synthesis methods raise concerns regarding their use due to harsh reaction conditions, use of toxic chemicals, and adverse environmental effects. Therefore, more environmentally friendly and sustainable synthesis methods need to be developed, and this issue becomes a critical target for researchers.

[0009] Biosynthesis of nanoparticles using plant extracts is preferred to reduce or prevent the mentioned adverse effects. Phytochemicals used in these applications, as natural compounds, have strong antioxidant and anti-inflammatory properties and offer neuroprotective benefits. However, the low bioavailability of these compounds limits their therapeutic potential and therefore their ability to cross the blood-brain barrier needs to be increased and thus their bioactivity improved. In addition, toxicity problems should be minimized while bioavailability is increased. In addition to all this, due to the fact that the relationship between the gut and the brain has been demonstrated in numerous studies, there is a need to use phytochemicals together with a component that supports gut microbiota in order to support their effect on cognitive functions.

[0010] The article, “Blueberry Supplementation in Neuronal Health and Protective Technologies for Efficient Delivery of Blueberry Anthocyanins,” which is a state of the art article, describes the use of blueberries as a daily edible supplement source to minimize the effects of age-related diseases and improve learning and memory in children. Consuming blueberries is described as a healthy fruit that provides various benefits to the nervous system, and it is stated that the most frequentlymentioned compounds among the components in blueberries are anthocyanins. The article's review focuses on neurological research on blueberries, from in vitro cell studies to in vivo studies, including animal and human studies, for their positive results in neuroprotection and intervention in neurodegenerative conditions. In addition, information is provided on the bioavailability of anthocyanins and the key factors affecting them to help consumers make informed decisions about their daily consumption of wild blueberries. In this context, the ways in which blueberries or blueberry supplement forms are consumed and which form is suitable for maximizing the benefits of blueberries are being evaluated. Said document does not mention a dietary supplement and production method aimed at preventing and / or eliminating the effects of cognitive impairment that occur in Alzheimer's-like dementia types using black carrot and chicory roots, while preventing toxic effects and reducing the risk of neurodegenerative diseases developing and / or progressing.

[0011] The patent document numbered CN116035209A, pertaining to the state of the art, describes a chitosan-modified anthocyanin nano liposome based on microfluidic technology and its application. Nano liposome with high encapsulation efficiency for encapsulating anthocyanin is prepared by systematic evaluation taking particle size and encapsulation efficiency as response values through optimized control of microfluidic multiphase channel content ratio, buffer solution condition, flow rate and other coefficients. Chitosan modification improves the storage time, temperature resistance, stability and functionality of the nano liposome under a gastrointestinal tract. Thus, the difficulty of taking anthocyanin as a functional food and using the same by the human body due to environmental sensitivity is overcome. Said document does not mention a dietary supplement and production method aimed at preventing and / or eliminating the effects of cognitive impairment that occur in Alzheimer's-like dementia types using black carrot and chicory roots, while preventing toxic effects and reducing the risk of neurodegenerative diseases developing and / or progressing.

[0012] As a result, there is a need to develop a dietary supplement and the food supplement production method that allows this food supplement to be obtained, particularly using black carrots and chicory roots, that prevents and / or alleviates the effects of cognitive impairment seen in Alzheimer's-like dementia, reduces the risk of developing and / or progressing neurodegenerative diseases by supporting antioxidant effects and gut microbiota, and increases bioavailability while reducing toxic effects.

[0013] Object of the InventionThe present invention relates to food supplement and food supplement production method which fulfils the abovementioned requirements, eliminates all disadvantages and brings some additional advantages.

[0014] The main object of the food supplement and production method which is the subject of the present invention is to obtain a food supplement and production method which prevents and / or eliminates the effects of cognitive disorders by affecting the central nervous system and thus reduces the risk of occurrence and / or progression of neurodegenerative diseases.

[0015] Another object of the present invention is to obtain an effective food supplement and production method that provides neuroprotective benefit by increasing antioxidant and anti-inflammatory activity.

[0016] Another object of the present invention is to obtain a functional food supplement and production method that reduces toxic effects on the patient by reducing harsh reaction conditions, the use of toxic chemicals and environmental adverse effects by using natural ingredients.

[0017] Another object of the present invention is to obtain an effective food supplement and production method that allows increasing the bioavailability effect and therapeutic potential by crossing the blood-brain barrier.

[0018] Another object of the present invention is to obtain an efficient food supplement and production method that acts as a prebiotic and positively affects intestinal microorganisms, thus regulating / supporting the intestinal microbiota and increasing the effect on cognitive functions.

[0019] Another object of the present invention is to obtain a food supplement and production method that ensures the preservation of the effectiveness of the natural ingredients in its structure and the increase of its duration of use.

[0020] Another object of the present invention is to obtain a food supplement and production method that provides ease of use for the user and extends shelf life.

[0021] In order to achieve the above objectives in the most general way, the food supplement developed with the present invention, which prevents and / or eliminates the effects of cognitive disorders by affecting the central nervous system, comprises anthocyanin nanoparticles encapsulated withchitosan and obtained from black carrot; and inulin encapsulated with chitosan and obtained from chicory root.

[0022] The food supplement production method developed with the present invention comprises the process steps of: dissolving chitosan and homogenizing the solution and obtaining chitosan solution; obtaining anthocyanin from black carrot and obtaining homogeneous distribution; adding the homogeneous mixture to sodium citrate solution and precipitating chitosan nanoparticles by stirring; separating the nanoparticles by centrifuging the mixture; freezing the nanoparticle suspension in a deep freezer after washing the nanoparticles; lyophilizing the frozen suspension; washing the chicory roots by rotating them using a heated mixer; removing the water from the roots and purifying them by carbon filtration; completing the inulin extraction by lyophilizing the roots; dissolving chitosan in an acetic acid solution; homogenizing the solution by mixing; obtaining homogeneous distribution by mixing inulin and chitosan solution; observing particle formation by adding sodium tripolyphosphate (TPP) solution to the mixture; rotating the mixture and separating the particles; separating the supernatant from the mixture and removing free inulin by washing; completing inulin encapsulation by freezing and recovering the washed supernatant; obtaining food supplements by adding inulin to chitosan-encapsulated anthocyanins.

[0023] The structural and characteristic features of the present invention will be understood clearly by the following drawings and the detailed description made with reference to these drawings and therefore the evaluation shall be made by taking these figures and the detailed description into consideration.

[0024] Detailed Description of the Invention

[0025] In this detailed description, the preferred embodiments of food supplement and food supplement production methods of the present invention are described by means of examples only for clarifying the subject matter.

[0026] Food supplement developed with the present invention, which prevents and / or eliminates the effects of cognitive disorders by acting on the central nervous system, increases antioxidant, antiinflammatory and prebiotic effects and reduces toxic effects, it comprises 200-400mg of anthocyanin nanoparticles per dose, preferably obtained from black carrot and encapsulated with chitosan, to obtain the highest neuroprotective effect; 40-60gr of inulin per dose, preferably obtained from chicory root and encapsulated with chitosan, to obtain the highest probiotic effect; and preferably 400-600mg of vitamin C per dose, to obtain the highest effect.In an illustrative embodiment of the food supplement developed with the present invention, anthocyanin, a phytochemical, exhibits a neuroprotective effect in the patient with the help of its strong antioxidant and anti-inflammatory effects and increases bioavailability by crossing the blood-brain barrier thanks to its encapsulated structure with chitosan. In this way, it supports short-term and long-term memory by increasing communication between brain cells while reducing the risk of neurodegenerative diseases. Chitosan-coated inulin contributes to the bioavailability of anthocyanin by supporting the intestinal microbiota, and with the joint work of these two components, the effect on the central nervous system is increased, resulting in a strong therapeutic effect in preventing and / or eliminating the effects of cognitive impairment. In addition, since both components are natural materials, toxic effects on the patient are prevented during or after use.

[0027] In this sense, Anthocyanins are naturally occurring antioxidants and are well known for their preventive effects on neurodegenerative disorders. Black carrots [Daucus carota L. ssp. sativus var. atrorubens Alef.] are a rich source of anthocyanins and contain mostly acylated cyanidin- based derivatives, making them more stable. However, in terms of the potential neuroprotective effect of black carrot anthocyanins (BCA), especially at high concentrations (50, 100 Ig / ml), they show a cytoprotective effect by increasing metabolic activity and reducing membrane damage. BCA shows antioxidant activity by scavenging MPP+-derived reactive oxygen species (ROS) and protecting dopaminergic neurons from ROS-mediated apoptosis. This suggests that BCA has a neuroprotective effect due to its high antioxidant and antiapoptotic activity and lack of cytotoxicity. In conclusion, BCA functions as a neuroprotective agent with high stability and potential neuroprotective effects.

[0028] The relationship between antioxidant intake (vitamins C and E, flavonoids, carotenoids) and cognitive function or risk of dementia is included. Vitamin C deficiency causes metabolism to deteriorate, leading to symptoms of folic acid deficiency. Another important example of this is the dependence of iron absorption from dietary sources on ascorbic acid. In addition, studies have shown that high levels of vitamin C reduce oxidative stress markers, mitochondrial fragmentation, and apoptosis in cortical peri-infarct neurons [Morris-Blanco KC, Chokkalla AK, Kim T, Bhatula S, Bertogliat MJ, Gaillard AB, Vemuganti R. High-Dose Vitamin C Prevents Secondary Brain Damage After Stroke via Epigenetic Reprogramming of Neuroprotective Genes. Transl Stroke Res. 2022 Dec;13(6):1017-1036. doi: 10.1007 / s12975-022-01007-6. Epub 2022 Mar 20. PMID: 35306630; PMCID: PMC9485293. Ramakrishna T. Vitamins and brain development. Physiol Res.

[0029] 1999;48(3): 175-87. PMID: 10523053],The food supplement production method developed with the present invention, which enables the prevention and / or elimination of cognitive impairment effects by affecting the central nervous system, increasing antioxidant, anti-inflammatory and prebiotic effects and reducing toxic effects, comprising the process steps of; dissolving chitosan in 2% by weight glacial acetic acid solution and homogenizing the solution by stirring the same using a magnetic stirrer, preferably for 14 hours and obtaining a chitosan solution with a pH value in the range of 4.5-5 to obtain optimum solubility by preventing protonation, viscosity increase, loss of homogenization and the risk of precipitation; obtaining anthocyanin from black carrots and adding the obtained anthocyanin to the chitosan solution and mixing it in a homogenizer, preferably at 18000rpm for 6-10 minutes to obtain a homogeneous distribution; adding the homogeneously dispersed mixture to a sodium citrate solution of preferably 100ml and 2% by weight and stirring the mixture using a mechanical mixer for preferably 1 hour to precipitate the chitosan nanoparticles; centrifuging the mixture using a refrigerated centrifuge device after precipitation, preferably at 11000 rpm for 8-10 minutes, to separate the nanoparticles from the supernatant and preferably analysing the supernatant for free drug content. Here, while precipitation provides nanoparticle formation, the nanoparticles are separated from the supernatant by centrifugation, and by using the centrifugation processes together, homogeneous, pure and desired sized chitosan nanoparticles are obtained. Subsequently, it comprises the process steps of; freezing the nanoparticle suspension in a deep freezer at (-) 70-90 ° C, preferably for 24 hours after washing the separated nanoparticles with distilled water at least 3 times; lyophilizing the frozen suspension using a lyophilizer, preferably at 10-30 ° C, for 24 hours; keeping the chicory roots in distilled water for inulin extraction, preferably at 85-95 ° C for 5 minutes, then washed by rotating for preferably 2 hours using a heated mixer; removing water from washed roots using vacuum evaporator and purifying by carbon filtration; completing the inulin extraction by lyophilizing the purified roots for 12 hours, preferably at (-) 70- 90 ° C; dissolving chitosan in 1% by weight acetic acid solution for inulin encapsulation; homogenizing the solution by mixing the same with a magnetic stirrer for preferably 12 minutes and adjusting the pH value between 4, 5-5, 5; mixing the obtained inulin and chitosan solution and mixing the same using a homogenizer, preferably for 10 minutes, at 10000-15000rpm to obtain a homogeneous distribution; observing particle formation by adding 0.1 -0.5% w / v sodium tripolyphosphate (TPP) solution to the homogeneous mixture of inulin and chitosan; rotating the mixture at 15000 rpm, preferably for 30 minutes, using a refrigerated centrifuge device to separate the particles; separating the supernatant from the mixture and removing free inulin by washing the same with distilled water at least 4 times; completing the inulin encapsulation by freezing the same preferably at (-) 70-90 ° C and completing the same using a lyophilizer; obtaining the mentioned food supplement by adding chitosan encapsulated inulin and preferably vitamin C to the obtained chitosan encapsulated anthocyanins.In an example analysis application, EEG recordings were taken twice from patients diagnosed with dementia, before and after the use of the developed food supplement. EEG recordings are monopolar and in the 10-20 placement system. M electrodes placed on the mandibular bone were used as reference electrodes and recordings were taken in the resting state with eyes closed. Recordings were taken with the AntNeuro eego sports model EEG device in suitable environments in the building where the patients were located, and electrical noise that may occur due to environmental changes was controlled by performing noise analysis before and after recording. The ability of the system to operate without being connected to the network line significantly reduces network noise (50 Hz). In addition, after the EEG recordings were taken, noise extraction was performed using the Independent Component Analysis (ICA) method. Although most of the analysis processes were performed in Matlab and eeglab, the Python program was also used.

[0030] The method for comparison is Spectral Power Density (PSD) analysis and the resulting spatial map of frequency bands on the head. In addition, an entropy measure, Quadratic Sample Entropy (QSE), was applied to examine the increase of disorder. Decreased complexity and disorganization (signal disorganization not physiological) are reported in the literature in patients exhibiting cognitive impairment.

[0031] The pwelch method was used in the implementation of PSD. After noise reduction, all signal processing was performed and the average value was used for each electrode. A temporal analysis was not performed in the applied method. After calculating the individual differences and the PSDs of individual electrodes for each individual, the total average was taken for both individual and electrode basis for general comparison and the following graph was obtained (Graph 1).Group Comparison

[0032] ower Spect i Density

[0033]

[0034] Graph 1. Spectral Power Density (PSD) analysis graph

[0035] The “first” line in Graph 1 shows before use, and the “second” line shows after use. When the general average obtained here is examined, changes are observed in the delta (1-4 Hz), beta (12-30 Hz) and low gamma (30-45 Hz) bands. As a result of the statistical analysis (t-test) performed for PSD analysis, it was observed that a significant change was observed in the beta3 (25-30 Hz) and gamma (30-45 Hz) bands. In this context the overall average shows that there is a power increase in these frequency bands after use. Spatial analysis was performed to see where this difference occurs topologically. Topological head maps are given below (Graph 2).

[0036]

[0037] Graph 2. Topological head maps graph

[0038] Graph 2 shows electrode-based topological density maps for Betal (12-15 Hz), Beta2 (15-22 Hz), Beta3 (22-30 Hz) and Low Gamma (30-45 Hz) frequency bands, respectively. Accordingly, an increase in concentrated power is observed in the F7, F3, T5 and C3 electrodes.

[0039]

[0040] Graph 3. QSE analysis graph

[0041] Additionally, in the QSE analysis performed in Graph 3, significant changes were observed in the delta and gamma frequency bands. How the QSE values changed before and after in the graph can be seen. The color map determines the direction and amount of change. Green color (-3) indicates an increase after use, while red color (+3) indicates a decrease. Electrodes highlightedin blue (thick peripheral line) are where statistically significant changes (wilcoxon test) were observed. Accordingly, it is observed that there is an increase in the QSE value in the delta and gamma bands, that is, the complexity and disorder increase.

[0042] F7 and T5 electrodes are particularly focal points in EEG studies in Alzheimer's patients due to their proximity to the hippocampal region. The change in high beta and low gamma frequency in this region is significant, especially considering the role of gamma frequency in information processing in the brain.

[0043] In this context, 10-second sections from the EEG traces of patients in whom the detected differences were clearly observed are included (Graph 4):

[0044]

[0045] Graph 4. First subject 10-second resting state EEG trace graphs

[0046] In Graph 4, the 10-second resting state EEG trace (Left) of an 88-year-old male patient diagnosed with dementia before using the developed food supplement is shown, with the horizontal axis set at 10 seconds and the vertical axis set at 100 μV, and the 10-second resting state EEG trace (Right) of an 88-year-old male patient diagnosed with dementia after using the developed food supplement is shown, with the horizontal axis set at 10 seconds and the vertical axis set at 100 μV.

[0047]

[0048] Graph 5. Second subject 10-second resting state EEG trace graphs

[0049] In Graph 5, the 10-second resting state EEG trace (Left) of an 81-year-old female patient diagnosed with dementia before using the developed food supplement is shown, with the horizontal axis set at 10 seconds and the vertical axis set at 100 μV, and the 10-second resting state EEG trace (Right) of an 81-year-old female patient diagnosed with dementia after using the developed food supplement is shown, with the horizontal axis set at 10 seconds and the vertical axis set at 100 μV.

[0050] In the trace images of two patients, it can be understood from the rapid changes that there is an increase in high frequencies in the recordings after use. In the first recording of the male patient the prominence of slow wave activity is noticed. Additionally, after use, an improvement can be noticed in the form of alpha fluctuations between recordings. The decrease in energy in the delta band and the increase in high frequency (gamma) energy are also seen on this trace by people familiar with EEG examination.

[0051] With the help of the food supplement effective on cognitive functions developed with the present invention and the production method of this supplement, especially by using black carrot and chicory roots, the effects of cognitive impairment occurring in Alzheimer-like dementia types are prevented and / or eliminated. In addition, the natural ingredients in its content provide antioxidant effects and support intestinal microbiota, thus reducing the risk of neurodegenerative diseases occurring and / or progressing, thus this results in an effective, efficient and functional food supplement with increased bioavailability and reduced toxic effects, as well as a food supplement production method that enables the production of this food supplement.

Claims

CLAIMS1. A food supplement that prevents and / or eliminates the effects of cognitive disorders by acting on the central nervous system and increases antioxidant, anti-inflammatory and prebiotic effects and reduces toxic effects, comprising:anthocyanin nanoparticles encapsulated with chitosan, wherein the anthocyanin nanoparticles are obtained from black carrots;inulin encapsulated with chitosan, wherein the inulin is obtained from chicory root.

2. A food supplement according to claim 1, wherein the amount of anthocyanin nanoparticles is 200-400 mg per dose to obtain the highest neuroprotective effect.

3. A food supplement according to claim 1, wherein the amount of inulin is 40-60g per dose to obtain the highest probiotic effect.

4. A food supplement according to claim 1, wherein the supplement also comprises 400-600mg of vitamin C per dose to obtain the highest effect.

5. A method of producing a food supplement that prevents and / or eliminates the effects of cognitive disorders by acting on the central nervous system and increases antioxidant, anti¬ inflammatory and prebiotic effects and reduces toxic effects, comprising the steps of:dissolving chitosan in 2% by weight glacial acetic acid solution and mixing the same by using a magnetic stirrer to make the solution homogeneous and obtaining a chitosan solution with a pH value in the range of 4.5-5;obtaining anthocyanin from black carrots and adding the obtained anthocyanin to the chitosan solution and mixing the same in the homogenizer to obtain a homogeneous distribution;adding the homogeneously dispersed mixture to 2% by weight sodium citrate solution and precipitating the chitosan nanoparticles by mixing the mixture by using a mechanical mixer;separating nanoparticles from the supernatant by centrifuging the mixture by using a refrigerated centrifuge after the precipitation;washing the separated nanoparticles at least three times with distilled water and then freezing the nanoparticle suspension by storing the same in a deep freezer;lyophilizing the frozen suspension by using a lyophilizer;soaking chicory roots in distilled water for inulin extraction and then washing them by stirring with a heated mixer;removing water from washed roots by using a vacuum evaporator and purifying the same with carbon filtration;completing inulin extraction by lyophilizing the purified roots;dissolving chitosan in an acetic acid solution at a concentration of 1% by weight for inulin encapsulation;homogenizing the solution by stirring in a magnetic stirrer and adjusting the pH value to between 4,5 and 5,5;achieving a homogeneous distribution by mixing the obtained inulin and chitosan solution by using a homogenizer;observing particle formation by adding a 0.1 -0.5% w / v sodium tripolyphosphate (TPP) solution to a homogeneous inulin and chitosan mixture;spinning the mixture by using a refrigerated centrifuge and separating the particles;separating the supernatant from the mixture and removing the free inulin by washing at least 4 times with distilled water;completing inulin encapsulation by freezing the washed supernatant and lyophilizing the same;obtaining the food supplement by adding the chitosan encapsulated inulin to the chitosan encapsulated anthocyanins obtained.

6. A food supplement production method according to claim 5, comprising the step of dissolving the chitosan and mixing the same in a magnetic stirrer for 14 hours.

7. A food supplement production method according to claim 5, comprising the step of mixing anthocyanin in the homogenizer at 18000 rpm for 6-10 minutes after adding the same to the chitosan solution.

8. A food supplement production method according to claim 5, wherein the amount of the sodium citrate solution is 100ml.

9. A food supplement production method according to claim 5, comprising the step of mixing for 1 hour in a mechanical mixer to precipitate chitosan nanoparticles.

10. A food supplement production method according to claim 5, comprising the step of analyzing the supernatant for free drug content after separation of the nanoparticle-containing supernatant.

11. A food supplement production method according to claim 5, comprising the step of centrifugation at 11000 rpm for 8-10 minutes to separate the supernatant containing nanoparticles.

12. A food supplement production method according to claim 5, comprising the step of freezing the nanoparticle suspension by keeping the same in a deep freezer at (-) 70-90 ° C for 24 hours.

13. A food supplement production method according to claim 5, comprising the step of lyophilizing the frozen suspension at 10-30 °C for 24 hours.

14. A food supplement production method according to claim 5, comprising the step of keeping the chicory roots at 85-95 °C for 5 minutes and then washing them by rotating them for 2 hours using a heated mixer.

5. A food supplement production method according to claim 5, comprising the step of lyophilizing the roots at (-) 70-90 °C for 12 hours.

16. A food supplement production method according to claim 5, comprising the step of dissolving the chitosan in an acetic acid solution and then stirring the same for 12 minutes in a magnetic stirrer to homogenize the solution.

17. A food supplement production method according to claim 5, comprising the step of obtaining a homogeneous distribution by mixing the inulin and chitosan solution using a homogenizer for 10 minutes at 10000-15000 rpm.

18. A food supplement production method according to claim 5, comprising the step of separating the particles by rotating the homogeneous inulin and chitosan mixture at 15000 rpm for 30 minutes using a refrigerated centrifuge.

19. A food supplement production method according to claim 5, comprising the step of freezing the washed inulin and chitosan supernatant at (-) 70-90 °C.

20. A food supplement production method according to claim 5, comprising the step of obtaining the food supplement by adding chitosan-encapsulated inulin and vitamin C to the chitosan- encapsulated anthocyanins obtained.

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