Nanoencapsulation of the black carrot extraction with complex coacervation method: Optimization with response surface method

Foods contains many compounds as well as traditional nutrients. Many of these compounds have the capacity to change enzymatic and chemical reactions and thus can affect human health both positively and negatively. Phenolic compounds, also called polyphenols, are one of the most abundant and most common groups of plant metabolites. They have a high biological activity spectrum such as antioxidant, antiinflammatory, antibacterial and antiviral functions. When these compounds are consumed as part of a balanced diet, they are beneficially beneficial to human beings and have positive effects on various chronic diseases such as cancer and cardiovascular disease related to oxidative stress. Today, one of the main areas of research in food science and technology is the characterization of new natural additives with biological activity that can contribute to consumer welfare, which is part of new functional additives, as well as extraction. Phenolic compounds can be found in leaves, fruites, flowers, woody parts, and in many plant parts of various plants. Phenolic compounds, phenolic acids and flavonoids are separated into two groups. Flavanoids are polyphenolic antioxidants found in the natural structures of herbal teas, fruits and vegetables. Some of the phenolic compounds are effective in the formation of the flavors of fruits and vegetables, especially in the formation of two important flavors such as bitterness and bitterness in the mouth. Some of them provide the colors of fruits and vegetables in yellow, yellow-brown, red-blue tones. It has been shown that phenolic compounds, secondary metabolites of plants, show antioxidant properties, cleans radicals, extinguishes oxygen atoms and acts as an ion or hydrogen donor. Phenolic compounds, and especially anthocyanins, can be quickly deteriorated by being affected by environmental conditions. Alkaline environment, temperature, oxygen, etc. Many factors can affect the distortion. In addition, phenolic compounds are affected by ambient pH during gastrointestinal conditions during the digestion period, and the presence of enzymes and other nutrients also inhibits the potential health effects and activities of these compounds. In this context, it is possible to benefit from encapsulation technology by aiming to protect these health-worthy components. In encapsulation technology, various bioactive substances are encapsulated in capsules, thus being protected from environmental factors. The use of encapsulated polyphenols in place of the free compounds can come from overcoming the disadvantages of instabilities, compensating for unpleasant tastes, as well as increasing the bioavailability and half-life of the compound in vivo and in vitro. In the encapsulation method, the size of the capsules can be very wide. Nano-, micro-, macro-encapsulation studies are available. However, nano-sized encapsules seem to be more and more active in order to increase the bioavailability and biocompatibility of foods, ie to increase digestion in the intestines. Currently there are many methods used for the encapsulation of phenolic compounds and anthocyanins: spray drying, freeze drying, emulsification, etc. However, according to the literature review, there is no study on the encapsulation of phenolic rich black carrot extracts by the coevaporation method. Black carrots are traditionally grown in temperate regions and are regarded as one of the most important of natural food colorants. The high content of phenolic constituents and, most importantly, the amount of anthocyanin provides many benefits for black carrot and health as well as its availability as a coloring agent. The main component of the black carrot is an acylated and unsubstituted cyanide derivative. The anthocyanins commonly found in black carrots can be defined as cyanide-3-xylosyl-galactose and cyanide-3-xylosyl-glucosyl-galactose, which are more monoacylated with ferulic, sinapic and p-gumaric acids. Besides the anthocyanins, p-coumaric acid, caffeic acid, ferulic acid, 3-hydroxybenzoic acid derivatives and 1quercetin glycoside, chlorogenic acid, mono- and dihydroxycinnamoylquinic acid are other phenolics determined in black carrot. The reaction surface method (RSM) was used to determine the optimum conditions for the encapsulation of phenolic rich black carrot extracts by coacervation method. RSM represents a combination of mathematical and statistical techniques aimed at optimizing the final response. When the encapsulation conditions are determined, it is decided by taking into consideration the conditions and the raw materials used for coaching. The pH parameter, which is one of the main criteria for coacervation. The trial design center was prepared with 3 factors and 3 levels with composite design. In study, 5 different features/effects of 3 independent variables were examined. The selected value range for Ph is 3.0-5.0, the selected range for the% extraction ratio is 0.2-1.0, and the ratio of whey protein used in the different formulations selected as coating material is 0-20%. For this purpose, encapsulation efficiency, total flavonoid amount, total antioxidant amount, particle size and zeta potential analysis were applied. In total antioxidant assays, a single assay was not considered sufficient because of the antioxidant properties produced by the different components and was determined by two different methods. Antioxidant capacity The ability to capture free radicals by the DPPH (Diphenyl-1pictylhydrazyl) method and the ability to reduce copper ions by the Cupric Reducing Antioxidant Capacity (CUPRAC) method were analyzed. In this study, black carrot was extracted with chemical methods and then dried in freezer dryer. Black carrot samples were divided into small pieces in liquid nitrogen and in the grinder to avoid reaching high temperatures. Thus, the efficiency is expected to increase when extractions are applied. Black carrots were extracted with 70% methanol. After extraction, the solution was left in the freeze dryer for one night and the water was removed. Samples taken from the freeze dryer were stored at -20 ° C for subsequent analysis The extracts ready for encapsulation were mixed with maltodextrin, gum arabic and whey protein solutions (10%) suitable for formulation and then treated in Ultratrox and Ultrasound devices for homogenization and particle size reduction. After adjusting the pH with HCl and NaOH solutions, particle sizes and zeta potentials were measured immediately and then dried in spray drier. The same procedure was applied to 15 different formulations according to RSM design. Subsequent analyzes were repeated for each sample. According to the analysis results, the lowest encapsulation yield was 60% while the highest encapsulation yield was 89,36%. The results of DPPH analysis for the total antioxidant activity were 21,09 mg Trolox / 100 mg to 68,09 mg Trolox / 100 mg, CUPRAC analysis results were 27,50 mg Trolox / 100 mg and 141,58 mg Trolox / 100 mg. The total flavanoid amount analysis results ranged from 2.51 mg Trolox / 100 mg to 9.54 mg Trolox / 100 mg. The encapsulation efficiency was found to be the effect of coating material and coating material * pH interaction (p <0,05). In the DPPH analysis, which is the first analysis to identify antioxidant activity, the effect of concentration was determined to have a significant effect of p <0.05 and for CUPRAC, the second antioxidant determination analysis, it was determined that the effect of coating material was p <0.01. In the analysis to determine the amount of flavanoid, the pH effect is significant at p <0.05. When the effects of the independent variables on particle size and zeta potential were examined, the importance of coating material in p <0.005 degree was determined. As the ratio of whey protein to coating material increases, the zeta potential shrinks, the particle size grows. Optimization of the RSM design result of 0.84 is obtained. This value indicates that the analyzes can be repeated at the desired rate and the desired optimum conditions can be achieved. The results from the optimization RSM design were repeated with the same conditions as the previous analyzes. Analyzes of the capsules prepared according to the optimization values were carried out. In this context, analyzes of particle size, zeta potential, encapsulation efficiency, antioxidant activity determination and total flavanoid amount were carried out. The results of each analysis are compared to the values required to reach the optimization endpoint. It is important that this comparison is smaller than 5% when compared. In our study, estimated values are compared to actual values and it is determined that the error margin is less than 5%. Coacervates produced under optimal conditions have been examined for gastric and intestinal digestion in various products for the purpose of examining the effect of different food matrices. Phenolic rich capsules were added to samples of skim milk and apple juice to determine protein richness and the effect of carbohydrate rich matrices. In addition, capsules were added to the chocolate beverage powder to examine the effect of temperature on the stability of the capsules and were prepared warm (85°C) and cold (4°C) with semi-oily milk according to the preparation instructions. The different foods enriched with capsules were then analyzed in vitro for bioavailability studies. Studies were carried out in stomach and intestinal environment and the total phenolic and antioxidant activities of phenolic substances released in liquid (SGF) mimicing stomach environment and liquid (SIF) mimicking intestinal environment were analyzed by spectrophotometric methods. Phenolic extraction was performed by breaking down the capsules in an amount of the samples before performing bioavailability analysis so that the SGF and SIF emissions can be compared to the actual total amount of phenolic material and the antioxidant activity, for example. In addition, in vitro bioavailability analysis has been carried out on non-capsular products, and thus it has been found that products enriched with capsules have more antioxidant properties as a result of digestion.