Micro-encapsulation as a noble technique for application of Bio-active compounds in the food industry.

In recent years, in functional foods, plant-derived nutraceuticals and dietary supplements have embodied health-promoting ingredients able to enhance human well being. Functional foods, dietary supplements and nutraceuticals contain, beyond the nutritional components, bioactive compounds with health benefits.
Bioactive compounds have gained lot of importance due to their role in prevention of several chronic diseases. These compounds are found in edible plants and foods. These natural bioactives, as part of whole diets, ingredients, or supplements, can be extremely beneficial for human health and wellness. Microencapsulation (ME) is a useful tool to improve the delivery of bioactive compounds into foods and
thus promote the successful delivery of bioactive ingredients to the gastrointestinal tract.
More research needs to be done to investigate the additional benefits of ME on the stability of bioactive ingredients in the gastric environment and on the release of bioactive ingredients into the GI tract.

1. Master’s Seminar on Microencapsulation as a noble technique for application of Bioactive compounds in the Food Industry Presented by Ms. Rozee Behera Roll no.- 27132203023 Seminar In charge Dr. Bhavnita Dhillon Assistant professor Department of Food Science and Technology Guru Nanak Dev University, Amritsar Presented by Ms. Rozee Behera Roll no.- 27132203023 Seminar In charge Dr. Bhavnita Dhillon Assistant professor Department of Food Science and Technology Guru Nanak Dev University, Amritsar Master’s Seminar on Microencapsulation as a noble technique for application of Bioactive compounds in the Food Industry

2. CONTENTS 1. 2. 5 . 3. 4. 6. 7. 8. 9. 10.

3. In recent years, in functional foods, plant-derived nutraceuticals and dietary supplements have embodied health-promoting ingredients able to enhance human well-being. Functional foods, dietary supplements and nutraceuticals contain, beyond the nutritional components, bioactive compounds with health benefits. Bioactive compounds have gained lot of importance due to their role in prevention of several chronic diseases.  These compounds are found in edible plants and foods. These natural bioactives , as part of whole diets, ingredients, or supplements, can be extremely beneficial for human health and wellness. Microencapsulation (ME) is a useful tool to improve the delivery of bioactive compounds into foods and thus promote the successful delivery of bioactive ingredients to the gastrointestinal tract. M ore research needs to be done to investigate the additional benefits of ME on the stability of bioactive ingredients in the gastric environment and on the release of bioactive ingredients into the GI tract. Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9916361/ https://www.sciencedirect.com/science/article/pii/S0958166907000328 INTRODUCTION

4. B ioactive compounds   are extra-nutritional constituents that typically occur in small quantities in foods providing health benefits beyond the basic nutritional value of the product. Bioactive compounds are secondary metabolites extracted from plants, fungi, microbes, or animals They are being intensively studied to assess their effects on the human health. B ioactive compounds have shown to have valuable physiological, immunological and behavioural effects. Natural bioactive compounds have significant contributions in the development of functional food products with better quality attributes. Examples of plant bioactive compounds are  carotenoids , polyphenols, or phytosterols. Some animal Bioactive compounds are fatty acids found in milk and fish etc. Other examples include  flavonoids ,  caffeine , phytoestrogens , and  prebiotics and many more. Several bioactive compounds have been discovered which vary widely in their chemical structure and function and are grouped accordingly. BIOACTIVE COMPOUNDS Source: https://tinyurl.com/2dwftb6n

5. Metabolites are the intermediate products produced during metabolism, catalyzed by various enzymes that occur naturally within cells. Metabolites produced by each living cell can be generally divided into two groups: Primary metabolites (PMs) and Secondary metabolites (SMs). PMs aim at growth and development of plant. They include the components of processes such as glycolysis, the Krebs or citric acid cycle, photosynthesis. T he SMs help plant to survive and overcome local challenges. Secondary plant metabolites are numerous chemical compounds produced by the plant cell through metabolic pathways derived from the primary metabolic pathways.  TYPES OF BIOACTIVE COMPOUNDS Source: https://rb.gy/1en39l https://www.intechopen.com/chapters/61866

6. CLASSIFICATION OF BIOACTIVE COMPOUNDS Source: https://rb.gy/fbolsq

7. Fatty acids present in milk and fish, peptides from milk protein, polysaccharides and amino polysaccharides (chitin, chitosan, chondroitin), ammonium compounds (L-carnitine, choline, glucosamine), as well as quinones (coenzyme Q 10 ) are examples of bioactive compounds in animal products. Bioactive peptides can be derived from meat proteins. M eat also has bioactive substances including L-carnitine, taurine, creatine and choline . BIOACTIVE COMPOUNDS OF ANIMAL ORIGIN Source: https://link.springer.com/referenceworkentry/10.1007/978-3-642-36605-5_14

8. Encapsulation  involves the incorporation of food ingredients, enzymes, cells or other materials in small capsules. It is a process of trapping Active components into a secondary material called Encapsulant. These capsules can then be able to release the active components at specific conditions. For ex- Encapsulated Vitamins. Microencapsulation is the process of enclosing tiny particles or droplets within a protective coating, often referred to as a microcapsule. This coating acts as a barrier, shielding the encapsulated material from external factors such as moisture, heat, and light. On the other hand, encapsulation involves encapsulating a substance within a larger structure or container, providing physical protection and controlled release . INTRODUCTION TO MICROENCAPSULATION Source: https://rb.gy/y76v0k

9. The natural Bioactive compounds can impart functional effects in Food systems (Flavour enhancers, colorants etc.) as well as in consumers (Anti-oxidants, anti-microbials etc.) Encapsulation and Microencapsulation effect on Human health : Increased Bio accessibility, Bioavailability and absorption efficiency of the Bioactive compounds. Example- Phenolic compounds have lesser stability, poor solubility and slow intestinal permeability which causes their poor bio availability when used in their free form. The beneficial properties of Bioactive compounds can be lost because of the gastro-intestinal environment. Their stability and bio-activity can be enhanced by encapsulating them in colloidal matrices. MICROENCAPSULATION OF BIOACTIVE COMPOUNDS But Why?? Source: https://rb.gy/vx6z9f https://www.mdpi.com/2076-3417/12/3/1424

10. In case of Food systems: Their bioactivity is affected by processing and storage conditions because they are sensitive to high temperature, pressure, extreme pH, light, etc. So, their direct incorporation into food product formulations may result in their- Degradation and Transformation at faster rates. Even a slight increase or decrease in their concentration may disturb the flavour and overall acceptability of the food products. So, to overcome these challenges, there was a need for a delivery system for these Bioactive compounds into the food products. One of them is Microencapsulation. - Example- A stud y on encapsulation of vitamin B 12  in water-in-oil-in-water double emulsions to produce functional cream for cheese milk standardization. It lead to lower loss of vit in vitro gastric digestion and their reduced losses in whey portion. Casein and inulin were used for the microencapsulation of citric acid powder, and the microcapsules were incorporated in the formulation of chewing gums. The samples containing microcapsules had higher sensory characteristics than control samples.  MICROENCAPSULATION OF BIOACTIVE COMPOUNDS But Why?? Source: https://www.mdpi.com/2076-3417/12/3/1424

11. The shell or wall material should preferably be insoluble, non-reactive to active ingredients, have excellent film-forming, and have the desired protective properties against various environmental conditions. The encapsulation method, properties of core and encapsulant materials, and load quantity result in the different microstructures of encapsulates . The microstructure of capsules has an impact on the efficiency and retention of the core material and the encapsulation efficiency and regulates the release of active ingredients . The microstructure of encapsulates varies in the external structure such as rough, porous, hollow, cracked, shrunken, etc., or the internal structures such as the arrangement and configuration of the core material. Encapsulates with lower core load- higher encapsulation efficiency, larger shell thickness, more compact shell, better protection , and slower release of ingredients STRUCTURE AND PROPERTIES OF ENCAPSULATES Source: https://www.mdpi.com/2076-3417/12/3/1424

12. F ood biopolymers are used in the encapsulation process. Many polymers are used as a coating material for encapsulating bioactive compounds, such as β- glucans, dextran, starch, alginate, cellulose, chitin, chitosan, pectin, collagen, gums, zein, hyaluronic acid, and gelatin , among others.  Finally, since encapsulation is accomplished by the continual formation of a film on each particle, all coating materials must be able to spread over the particle surface. These materials are suitable for microencapsulation, because of their ability to form films, viscosity and resistance to the gastrointestinal tract, solid content, biodegradability, safety, and low price. POLYMERS USED FOR ENCAPSULATION OF BIOACTIVE COMPOUNDS Source : https://www.thepharmajournal.com/archives/2020/vol9issue7/PartA/9-8-16-293.pdf https://www.mdpi.com/2073-4360/14/19/4194

13. Spray drying Freeze drying Coacervation Co-crystallisation Emulsification Ionic gelation Supercritical fluid-based techniques Extrusion based techniques TECHNIQUES OF MICROENCAPSULATION OF BIOACTIVE COMPOUNDS

14. TECHNIQUES OF MICROENCAPSULATION OF BIOACTIVE COMPOUNDS Source: https://www.mdpi.com/2073-4360/14/19/4194

15. Most widely employed technique. A chief advantage is that this technique can be used for heat-labile materials. SPRAY DRYING Hydration of the Encapsulant Homogenisation of the core material and the encapsulant (1:4) The mixture fed into the Spray dryer and atomised with a nozzle Water evaporated by hot air contacting the atomised material Capsules are collected as they fall to the bottom of the dryer Source : https://www.thepharmajournal.com/archives/2020/vol9issue7/PartA/9-8-16-293.pdf

16. Freeze-drying is a suitable microencapsulation technique for sensitive bioactive compounds. B ased on the dehydration by sublimation of a frozen sample . FREEZE DRYING Hydration of the Encapsulant Mixing of the core material and the encapsulant Homogenisation Freeze drying of the samples Collection of the capsules/Freeze dried powder and packaging Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7248874/ https://www.sciencedirect.com/science/article/pii/S0268005X20313643

17. COACERVATION Addition of Core material and oppositely charged Biopolymer Addition of electrolyte or non-solvent compound to allow precipitation to occur Precipitation takes place Collection of precipitated complex of Biopolymer and core material Drying and packaging P roduction of complexes between oppositely charged biopolymers upon mixing core materials with charged biopolymer solutions. In this, an oppositely charged biopolymer is added, which forms a complex entrapping the core material and precipitates. Source : https://www.mdpi.com/2076-3417/12/3/1424

18. It is a relatively recent development. It is is a process whereby a second (active) ingredient is embedded inside the conglomerate of crystals. Sucrose is used as a primary ingredient. Advantages- Improved solubility, wettability, homogeneity, dispersibility, hydration, anticaking, stability and flowability of the encapsulated materials CO-CRYSTALLISATION Mixing of the Sucrose and Bio-active compound Heating of the mixture with continuous agitation Removal of heat while maintaining agitation Drying of the co-crystallised product Milling and packaging of co-crystallised product Source: http://www.ijpab.com/form/2018%20Volume%206,%20issue%202/IJPAB-2018-6-2-1366-1371.pdf

19. The release mechanism of encapsulated bioactives is controlled by the chemical composition and characteristics of the carrier wall and the quality of the material used to make bioactives . The physical properties, such as size, shape, and morphological characteristics of the carriers, must be considered. Generally, the stability of these bioactive compounds is low, and encapsulation results in higher stability against variations in temperature, light, pH, or oxygen, increasing the release rate of these active ingredients. The controlled release strategy is recently being applied in food delivery systems and is considered a characteristic that assists the release mechanism of the encapsulated bioactives to the target site while maintaining the biological and functional characteristics. Many of the bioactives cannot permeate into the small intestine in a sufficient concentration for efficacy without an efficient oral delivery system. T h e encapsulation technique aims to protect and deliver the bioactive compounds to the target tissue of the human body . MECHANISM OF RELEASE OF BIOACTIVE COMPOUNDS Source: https://www.mdpi.com/2504-5377/7/2/25#:~:text=5.2.-,Controlled%20Release,the%20biological%20and%20functional%20characteristics .

20. The Releasing action is triggered by several internal (such as diffusion, degradation, and swelling) and external stimuli (such as changes in temperature, pH, light, ultrasound, ionic strength, and magnetic field). The release process can be time-specific, site-specific, rate specific, and stimulus-specific which can be achieved by one or a combination of different release mechanisms. Bioactive components are released from encapsulants in the following steps: 1. Surface Release 4. Degradation 2. Diffusion 5. Swelling 3. Matrix erosion 6. Dissolution MECHANISM OF RELEASE OF BIOACTIVE COMPOUNDS Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9571964 /

21. 1. Surface release: It may be caused by inadequate entrapment of bioactives inside the matrix or by a polar bioactive compound’s tendency to move towards the hydrophilic surface of an emulsion. 2. Diffusion: T he Diffusion of a bioactive compound from the interior to the exterior of an encapsulation system depends on the solubility in the encapsulated system and its permeability through the capsule material. T he total transport of molecules from a higher concentration to a lower concentration field.  3. Erosion: It happens when the encapsulated system faces a specific environmental condition, the chemical degradation of the particle matrix, leading to release of the bioactive compound. It may be caused by various factors, such as high temperatures, strong acids or bases or enzymatic reasons. 4. Degradation: It is the disruption of bio-polymers microorganisms . The core compounds dispersed in the polymer matrix are released after the biodegradation of the polymer. 5. Swelling: T he release of bioactive compounds happens when the capsule swells because of the solvent absorption; this release mechanism may be controlled by the selection of the polymeric matrix and the environmental conditions, such as temperature and pH. 6. Dissolution: In the case of encapsulated system, bioactives are dissolved in the release media. The theory behind this process is that the ions or molecules of bioactives are transferred to the surrounding environment. MECHANISM OF RELEASE OF BIOACTIVE COMPOUNDS Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9818261/ https://www.mdpi.com/2504-5377/7/2/25 https://rb.gy/j59b6v

22. APPLICATIONS IN FOOD INDUSTRY Encapsulation allows for better stability and bioavailability of the Bioactive compounds after food digestion. Essential oils have also been microencapsulated. Commercial Food flavours can be encapsulated or microencapsulated to protect them against oxygen light and heat. In case of fortification of food with Iron , Bioavailability of Iron is affected by its interaction with food ingredients. This can be prevented by microencapsulation. Omega 3 fatty acids are unsaturated in nature, so they are susceptible to oxidation. Microencapsulation can play a significant role here in their protection. The ME of probiotics for their addition into foods and beverages. Source: https://www.thepharmajournal.com/archives/2020/vol9issue7/PartA/9-8-16-293.pdf

23. In a very recent study, the authors proposed the use of different encapsulated fish oils in the formulation of chicken nuggets. Moreover, these authors also highlighted that the addition of unencapsulated fish oil in chicken nuggets led to a decrease in sensory acceptance, while the use of encapsulated oil did not affect sensory quality, which demonstrated the high potential to use encapsulated fish oil to produce healthy meat products. Development of a functional bread using encapsulated garlic and flaxseed oils and reported that these functional breads were stable against oxidation and simultaneously received a high sensory rating and acceptability. Vitamin D 3   was encapsulated in two flaxseed oil emulsion formulations to fortify the cheese with omega-3 and other polyunsaturated fatty acids. Emulsions were stabilized with calcium caseinate in the presence or absence of lecithin; these were used to standardize cheese milk. Another important application of encapsulated bioactive molecules is their use for extending meat products’ shelf-lives. In a very recent study, the authors proposed the encapsulation of garlic essential oil using the spray drying technique and maltodextrin and gum Arabic as wall materials to prevent minced meat deterioration. Finally, other authors proposed the use of encapsulated (ionic gelation) betalain -rich extract of  Opuntia- ficus -indica  fruit as a colorant for the development of gummy candies. STUDIES ON THE APPLICATIONS OF MICROENCAPSULATED BIOACTIVE COMPOUNDS Source: https://www.mdpi.com/2076-3417/12/3/1424

24. BENEFITS Protection of active ingredients Allows longer release time Controlled release in the body Increase stability of Bioactive compounds Masking of undesirable flavours Prevents unwanted reactions and interactions LIMITATIONS A single microencapsulation process not adaptable to all core materials Incomplete or discontinuous coating Requirement of high temp., and water- soluble carriers in spray-based techniques. High cost involved in some techniques. Complex and lengthy process Source : https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9571964/ https://www.researchgate.net/figure/Importance-of-microencapsulation-of-bioactive-compounds_fig1_366123434 https://www.mdpi.com/2076-3417/12/3/1424

25. Microencapsulation has proved to be a very efficient method for the controlled and the targeted delivery of functional ingredients through the food to the gastrointestinal tract of human being. It not only provides a controlled release of active ingredients but also protects the sensitive compounds by acting as a protective barrier, masks the undesirable flavour of the compounds in food and increases their bioavailability in food. More techniques are being researched to find their suitability in ME of Bioactive compounds. At present and in the future, the availability of suitable wall materials that fit within the requirement of personal beliefs, food habits, and clean label requirements poses a challenge in the food industry. N eed for in-depth research at microscopic and nanoscopic levels to identify various health and safety issues and alleviate them to make this method more acceptable and compatible with green food processing. With increasing education and awareness leading to an increasing interest by consumers in how foods are prepared, and ingredients added, this technology will be immensely helpful in the success of the food industry, and microencapsulated food with natural active ingredients will play important role in the future. SCOPE, CHALLENGES AND THE WAY FORWARD Source: https://www.mdpi.com/2076-3417/12/3/1424