Extraction and Industrial Applications of Macro Molecules: A Review

Abstract

Macromolecules, large molecules composed of repeating subunits called monomers, play a crucial role in living organisms, performing diverse functions such as energy storage, structural support, information storage and transfer, and catalysis of chemical reactions. Carbohydrates, lipids, proteins, and nucleic acids are the four major classes of biological macromolecules. Extracting macromolecules from natural resources is critical in developing analytical processes and subsequent fortified products. Public awareness has grown due to using natural resources for environmental preservation and sustainable development. Extraction might be the first step in developing analytical methods and give room to product development. However, conventional techniques use organic solvents like acetone, ethanol, methanol, and ethyl acetate along with heated or boiling water. As a result, high temperatures and lengthy extraction times are produced when procedures like maceration, percolation, and solvent extraction are utilized. Due to these drawbacks, other extraction techniques have recently started to replace these conventional methods. These conventional procedures frequently entail using an energy source to hasten the transfer of the macromolecules compounds for further processing. This paper explores emerging techniques, such as pulse electric field-assisted extraction, Ionic liquid-based extraction, Subcritical water extraction, pressurized liquid extraction, Enzyme-assisted extraction, supercritical fluid extraction, ultrasonication-assisted extraction, and microwave-assisted extraction. The extraction of macromolecules for fortification purposes offers significant health and commercial benefits, addressing nutritional deficits and malnutrition. By understanding each macronutrient's specific benefits and purposes, effective fortification strategies can be developed to maintain a healthy body.

Introduction

Macromolecules

A macromolecule is a large molecule composed of repeating subunits known as monomers. These subunits are typically covalently bonded to form a polymer, a long chain of repeating units. It can be natural materials like sugars and natural fibers (like cotton) or synthetic materials like plastics, synthetic fibers, and adhesives ¹. Large molecules called macromolecules fill a cell with essential life-sustaining tasks. For instance, macromolecules provide structural support, act as a storage space for electrical energy, enable the storage and retrieval of genetic information, and hasten biological processes. Giant molecules that are biologically necessary for life can be found in key nutrients. These organic macromolecules (polymers) are combinations of monomers, which are smaller organic molecules. In Table 1 below, everyone of the four major biological macromolecules—carbohydrates, lipids, nucleic acids, and proteins,—is briefly reviewed about their sources, structure and composition, and their commercial applications. The human diet's primary caloric components are macromolecules present in food systems. The main source of carbohydrates in our daily diets is starch, which accounts for about 50% of our daily energy needs. Among other useful properties of food, starch may affect the texture, mouthfeel, moisture, viscosity, and shelf life of food products. Proteins are also necessary for the human body to operate. The amino acid chain that makes up proteins can be found in both plant and animal sources. The three kinds of amino acids are essential, nonessential, and conditional ². Protein has the desirable abilities to thicken, gelatinize, emulsify, foam, and have a high capacity to hold water. Our everyday diet must contain lipids, such as fats and oils because they are crucial to nutrition and food preparation. While vital fatty acids and fat-soluble elements are essential in generating the particular flavour of cooked foods, the lipid is ingested in daily meals as a crucial energy source ³. It is crucial to remember that while lipids are advantageous in moderation, consuming too many bad fats can lead to health issues like obesity and cardiovascular illnesses. For maintaining life, nucleic acids are the most crucial macromolecules. Nucleic acids are vital biopolymers that are found in all living organisms and are utilized to transmit, encode, and express genes. Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are the two major forms of nucleic acids¹. This study aims to show macromolecule extraction and present some cutting-edge technologies for food fortification, health, and commercial applications.

Table 1: Known Macromolecules as Recommended by Food and Drug Administration

Macro molecule	Carbohydrates	Proteins	Fats	Nucleic Acids	Reference
Sources	Fruits and Vegetables; Apples, Oranges, Carrots, and Bananas. Legumes; Kidney beans, Chickpeas, and Lentils. Cereals and Grains; Rice, Corn, Wheat, and Barley. Tubers and Root Vegetables; Cassava, Potatoes, and sweet potatoes. Milk and Dairy products; Yoghurt, Lactose, and Cheese. Shellfish and Seafood; Chitin and Glycogen.	Meat; Poultry, Pork, Beef, and Lamb. Seafood; Fish and Shellfish. Dairy products; Cheese, Milk, Yoghurt, and Eggs. Legumes; Chickpeas, lentils, Beans, and Peas. Nuts and Seeds; Walnuts, Hemp seeds, Almond, and Chia seeds. Grains; Quinoa, Barley, Rice, and Wheat. Soy Products; Tempeh, Soy milk, and Tofu.	Animal sources: Red meat; Beef, Pork, and Lamb. Poultry; Turkey and Chicken. Dairy products; Cream, Butter, Whole milk, and Cheese. Plant-Based fat Sources: Nuts and Seeds; Flaxseeds, Chia seeds, Walnuts, and Almonds. Oils; Olive oil, coconut oil, Avocado oil, and Flaxseed oil.	Nucleic acids are found in cells, particularly in the nucleus; sources include animal tissues and organs; Kidney, Liver, and Muscle. Plant tissues and organs; Seeds, Roots, and Leaves. Microbial sources; Bacteria and Yeast.	1,2,4–9
Structure & Composition	Carbohydrates are classified into four types: Monosaccharides; Glucose, Fructose, and Galactose. Disaccharides; Sucrose, Lactose, and Maltose. Oligosaccharides; Raffinose and Stachyose. Polysaccharides; Starch, Cellulose, and Glycogen.	Classification of Proteins: Complete Protein; Animal-based, and Plant-based sources. Incomplete Protein; Plant-based sources. There are four levels of protein structure; primary, secondary, tertiary, and quaternary. Proteins are water-soluble polymers of small building blocks known as amino acids. Twenty distinct amino acids; Nine are essential, five are nonessential while six are conditional.	Classification of Dietary fats: Saturated Fatty Acids; Plant-based, and Animal-based sources. Monounsaturated fatty acids; Plant-based sources. Polyunsaturated Fatty Acids; Omega-3 fatty acids, and Omega-6 fatty acids. Trans Fats; Natural sources, and Artificial sources.	Classification of Nucleic acids: Deoxyribonucleic Acid (DNA); Animal-based sources and Plant-based sources. Ribonucleic Acid (RNA): Messenger RNA (mRNA), Ribosomal RNA (rRNA), and Transfer RNA (tRNA).	1,7,10–15
Commercial Applications	Food and Beverage Industry; Sweeteners, Fiber, Starch, Bakery and Confectionery. Healthcare and Pharmaceutical Industry; Stabilizers and Fillers, Injectable Carbohydrates, Wound healing and Tissue Engineering, and Excipients. Textile Industry; Starch and Dextrins. Biofuel Production; Ethanol. Personal Care and Cosmetics Industry; Emulsifiers and Thickeners, and Humectants. Bioplastics and Biomaterials; Polysaccharides like cellulose, starch, and chitosan.	Food and Beverage Industry; Dairy and Meat Alternatives, Protein Fortification, Bakery and Confectionery. Healthcare and Pharmaceutical Industry; Vaccines, Therapeutic Proteins, Diagnostic Assays. Personal Care and Cosmetics Industry; Anti-aging and Anti-wrinkle, Hair and Skin Care. Animal Feed; Animal Protein Supplement. Sports Nutrition and Supplements; Protein drinks, Protein Powders and Bars. Bioplastics and Biomaterials; Protein-based materials such as silk and collagen.	Food Industry; Cooking and Frying, Shelf-Life Extension, Flavour and Texture Enhancers, Emulsifiers and Stabilizers, Fat-based Spreads and margarines. Personal Care and Cosmetics Industry; Lipsticks and Lip balms, Moisturizers and Emollients. Healthcare and Pharmaceutical Industry; Excipients, Controlled-Release System. Industrial Application; Biofuels and Lubricants. Animal Feed; Energy Source. Industrial Chemicals; Surfactants.	Agriculture and Food Industry; Genetically Modified Organisms (GMOs), Food Safety Testing. Genetic Testing and Personalized Medicine; DNA Sequencing, Pharmacogenomics. Disease Detection and Diagnostics; Nucleic Acid Probes, Polymerase Chain Reaction, Gene Expression Analysis. Forensic Science; DNA Profiling. Biotechnology and Biopharmaceuticals; Gene Therapy, Recombinant DNA Technology, RNA Interference (RNAi). Environmental Monitoring; Biodiversity Assessment.	16–25

Benefits of Macromolecules

Macromolecules, as essential components of cells, perform a variety of crucial functions necessary for sustaining life. These macromolecules, which include proteins, carbohydrates, lipids, and nucleic acids, offer several advantages and benefits. Carbohydrates, a type of biomolecule, are divided as four categories depending on their structure and chemistry: Monosaccharides: Monosaccharides are the simplest form of carbohydrates and cannot be further hydrolyzed into smaller sugar units. Common monosaccharides include glucose, fructose, and galactose. Glucose is the primary source of energy for many organisms, while fructose is found in fruits and is often used as a sweetener. Galactose is a component of lactose, a disaccharide found in milk ⁴. Disaccharides: Disaccharides are formed by the condensation of two monosaccharide units. They can be hydrolyzed into their constituent monosaccharides by enzymatic or chemical reactions. Common disaccharides include sucrose, lactose, and maltose. Sucrose, popularly known as white sugar, is made up of two sugars: fructose and glucose. Lactose, found in milk, consists of glucose and galactose. Maltose is a product of starch digestion and consists of two glucose units ¹. Oligosaccharides: Oligosaccharides are composed of a small number of monosaccharide units, typically between 3 to 10 sugar residues. They are often found in plant-based foods. Examples of oligosaccharides include raffinose and stachyose. These compounds are not readily hydrolyzed by human digestive enzymes but can be fermented by gut bacteria, providing prebiotic benefits. Polysaccharides: Polysaccharides are large, complex carbohydrates composed of many monosaccharide units joined together by glycosidic linkages. They serve as storage forms of carbohydrates in plants and animals and can be classified as either structural or storage polysaccharides. Starch, found in plants, is a storage polysaccharide composed of glucose units. Cellulose, also found in plants, is a structural polysaccharide that forms the cell wall. Glycogen is the storage polysaccharide in animals, including humans, and is structurally similar to starch but has more extensive branching. These different types of carbohydrates play vital roles as energy sources, structural components, and functional molecules in various biological processes ⁴. According to Kaur ²⁶, rye is a good source of carbohydrates, proteins, fiber, minerals and antioxidants. Starch takes up a sizable amount of the carbohydrates in rye and other cereal grains (55-70%). Starch is a major human diet's energy source. It also improves the utility of food products. Dietary fibers are just as essential as carbohydrates because they support healthy weight management and digestive health. Rye's grain nutritional fiber composition indicates that lignins, arabinoxylan, beta-glucan, and fructan are all present in varying degrees. Cereal grains are a rich source of carbohydrates in general. By far the most common constituent group, carbohydrates comprise 66 to 80% of rye grains. Starch (56-75%) makes up the majority of the carbohydrates found in rye grains, followed by fiber (15-22%) and sugars (0.61%, 0.18%, and 2.58%) at 3%.

The majority of organic molecules in biological systems are proteins, which also play the greatest range of functions of any macromolecule. In general, proteins are enzymes that serve as structural, regulatory, contractile, or protective components for membranes, transport, and storage ²⁷. Each cell in a living organism may have thousands of proteins, each with a distinct function. Protein activities and structures are very different from one another. However, they are all amino acid polymers that are linearly organized. Proteins are made up of amino acid polymers. Each amino acid has a core carbon that is joined to a R group, also referred to as a side chain, an amino group, a carboxyl group, and a hydrogen atom. There are 20 commonly occurring amino acids, each with a unique R group. Each amino acid is joined to its immediate neighbours by a peptide bond. A polypeptide is an extended chain of amino acids. Proteins are categorized at the primary, secondary, tertiary, and (optionally) quaternary levels of the organization ¹. The specific arrangement of amino acids determines the primary structure. The secondary structure is produced when the polypeptide locally folds to produce structures like the helix and pleated sheet. The total three-dimensional structure is known as the tertiary structure. When two or more polypeptides combine to form the full protein structure, the configuration is known as the quaternary structure of a protein. Protein form and function are closely connected; any alteration brought on by variations in temperature or pH may cause denaturation of the protein and a loss of function ²⁸. The protein that is most supported among all other cereal proteins in terms of amino acid makeup is rice protein. The most popular type of rice is refined, which has only the endosperm of the grain left after being milled to remove the husk and bran. According to Li ¹⁴, it has been found that the husk and bran layer contains a sizable amount of rice protein—between 11.3% and 14.9%. The limiting amino acid in grains, lysine, is also present in second-highest concentration. Compared to other plant proteins, rice protein is considered the best source of protein for functional foods since it has a protein efficiency ratio similar to milk casein and is hypoallergenic. Whole grains are far healthier to eat than pearled rice because they contain endosperm, germ, and bran. Not only does eating whole grains improve basic nutrition, but it also reduces the risk of various chronic diseases.

Lipids, a class of macromolecules, are non-polar and hydrophobic by nature. This is because most of the bonds in hydrocarbons are non-polar carbon-carbon or carbon-hydrogen bonds. They are hydrophobic, or “hating water,” non-polar chemicals that do not dissolve in water ¹. Lipids have several functions within a cell. Cell membranes, which preserve the consistency and order of cells, primarily consist of lipids. Particularly significant for the cell membrane's structural integrity are phospholipids. Lipids are a more effective energy source than carbohydrates since they may be stored as energy reserves in the body. Cell signaling pathways involve lipids like phospholipids and sphingolipids.

They can serve as signal relayers between cells and are also important in controlling cellular reactions to environmental cues. All biological membranes depend on lipids, which are also the building elements of many hormones. The generation of hormones that control numerous physiological processes, including growth and development, metabolism, and immunological function, is aided by certain lipids, notably cholesterol and steroid hormones. Additionally, lipids can serve as a barrier around some organs, such as the kidneys, preventing harm from physical trauma or injury. Animals and plants are protected from their surroundings by lipids. For example, they help keep aquatic birds and mammals dry by producing a protective layer over fur or feathers due to their water-repellent hydrophobic nature ^15,29.

Triglycerides, phospholipids, and steroids are the three main types of lipids. Although waxes are a less popular category, they are nonetheless regarded as lipids. Triglycerides: The most prevalent lipids are triglycerides, also referred to as fats and oils. They comprise three fatty acids and glycerol, which the body uses to store energy and provide insulation and cushioning ³⁰. Phospholipids: The lipids that make up the cell membrane are called phospholipids. They include two fatty acids, a phosphate group, and a glycerol molecule. While the fatty acid tails are hydrophobic (fearful of water), the phosphate group makes one end of the molecule hydrophilic (loving water). Because of their special structure, phospholipids can form a bilayer in cell membranes, helping to regulate what enters and leaves the cell ¹³. Steroids: A four-ring structure is a characteristic of steroids, a category of lipid. They are crucial for a number of physiological activities, such as the control of the immune system, metabolism, and reproductive procedures. In addition to being a component of cell membranes, cholesterol is a sort of steroid that serves as a precursor for the creation of other steroids, including hormones like testosterone ³¹. A long-chain fatty acid and a long-chain alcohol make up the lipid known as waxes. They frequently serve as a protective layer in both plants and animals. Although waxes are a form of lipid, their discussion is less frequent than that of the other three groups indicated above. Lipids are important and serve a purpose in food because they improve flavour, satiety, and nutrition. As a result, lipid quality is a crucial consumer concern and may be connected to a variety of medical issues ³⁰. Each of these categories has a certain purpose. Olives, avocados, almonds, pecans, pumpkin, and sesame seeds are high in monounsaturated fats. Canola oil, corn, fish, walnuts, linseed oils, and flaxseed are some foods containing polyunsaturated fats. Saturated fats can be present in some plant-based foods, such as coconut and palm oils, although they are mostly found in animal products, such as beef or cheese. In terms of overall structure, all macromolecules—aside from lipids—are thought of as polymers. A polymer is a chain of covalently connected homologous monomers, or subunits. Proteins, carbohydrates, and nucleic acids all have monomers that are called sugars, amino acids, and nucleotides, respectively. This diverse collection of molecules is composed of a large variety of nonpolymeric compounds known as lipids.³².

Nucleic acids are molecules made up of nucleotides that regulate biological processes including cell division and protein production. Each nucleotide consists of a pentose sugar, a nitrogenous base, and a phosphate group. The two different types of nucleic acids are DNA and RNA. Parents pass DNA on to their children (in the form of chromosomes), contains the genetic code for every cell. It has a double-helical structure, in which the two strands run counterclockwise, complement one another, and are connected by hydrogen bonds. Single-stranded RNA is made up of the pentose sugar ribose, a nitrogenous base, and a phosphate group. RNA is involved in both the regulation and production of proteins. The messenger RNA (mRNA), which is exported from the nucleus to the cytoplasm, contains the instructions for constructing proteins. Ribosomal RNA (rRNA), on the other hand, is a component of the ribosmes at the site of protein synthesis, whereas transfer RNA (tRNA) delivers the amino acid there. How mRNA is used to produce proteins is regulated by microRNA ³³.

Commercial Applications of Macromolecules

Essential macromolecules in the food industry include proteins, carbohydrates, and lipids, which are added to and supplement foods to enhance their diverse nutritional qualities. They are necessary parts of human nutrition. They are also employed in manufacturing a number of culinary items, including cheese, yoghurt, and bread. Proteins are frequently employed in foods as texturizing agents to enhance their consistency and texture. While gelatin is used to create desserts and gummy candies, casein enhances the texture of cheese and yoghurt. In products like salad dressings and mayonnaise, proteins can also be employed as emulsifiers to facilitate mixing water and oil ¹⁶. In processed foods, carbohydrates like cellulose, pectin, and carrageenan are frequently employed as thickeners, stabilizers, and gelling agents. Foods like sauces, soups, and desserts can benefit from them by having a better mouthfeel and texture. Gums are one of the few food additives with the proper structural and functional characteristics, and they can be obtained from various sources. Polysaccharide gums, additionally referred to as hydrocolloids, have been used in the food, medicinal, fabric, power, water, the field of biotechnology, surroundings, personal care products, and pharmaceutical sectors for a range of applications. This is due to its desirable attributes like affordability, sustainability, recycling potential, accessibility, compositional variety, high potential for binding to water, emulsion formation stabilization, distinctive rheological features, non-toxicity, and gel-making capability ¹⁷. Lipids, such as vegetable oils and animal fats, are frequently added to processed meals as ingredients to improve flavour and texture. They can also be employed as preservatives to increase the shelf life of food products and as emulsifiers to aid in blending ingredients. Proteins and nucleic acids are used extensively in producing various drugs and vaccines in the pharmaceutical industry. The capacity of proteins to interact with specific targets in the body makes them a common component of therapeutic medicines used in medicine. One such protein hormone is insulin, which is used to treat diabetes and control blood glucose levels. Enzymes, growth factors, and antibodies are examples of additional protein medications ¹⁸. Gene therapy, a possible method of treating genetic diseases, uses nucleic acids like DNA and RNA. Gene therapy includes adding new or altered genes to the body to supplement or replace damaged genes. In vaccines, genetic material instructing cells to generate particular proteins that stimulate an immune response is delivered through nucleic acids. Macromolecules are essential to the production of many drugs and vaccines, and their diverse properties and functions make them valuable tools in the field of medicine ²⁰. Macromolecules—particularly nucleic acids and proteins—are widely exploited in the study and development of biotechnology. Recombinant DNA technology is one such example where nucleic acids are utilized to manufacture desired proteins. Examples include recombinant DNA technology and protein expression systems like yeast, bacteria, and mammalian cells. Proteins are necessary molecules that provide various bodily activities, including signalling, catalysis, and structure ^19,20.

Genetic engineering is changing an organism's genetic makeup to add new features or characteristics. This can be accomplished by modifying the organism's DNA or RNA using methods like gene cloning, PCR, and CRISPR-Cas9. The procedure of DNA sequencing is used to establish the nucleotide sequence within a DNA molecule. It is a crucial tool in biotechnology research to examine organisms' genetic makeup, spot genetic diseases, and create novel medications and treatments. Using computational tools and algorithms to analyse and understand biological data is the focus of the interdisciplinary field of bioinformatics. The analysis of DNA and protein sequences, the prediction of the structure and function of proteins, and the development of novel medications and treatments all make substantial use of this technology in the biotechnology research ^20,34.

Macromolecules, particularly polymers, are used to produce various materials such as plastics, adhesives, and coatings. Polymer-based materials, known as plastics, are employed in various industries, including electronics, building, and packaging. Polystyrene, PVC, polyethylene, and polypropylene are examples of common plastic kinds. Depending on their use, these polymers can be generated as films, fibres, or moulded items ³⁵. Materials are bound together by chemicals called adhesives. Polymers, such as cyanoacrylate, epoxy, and polyurethane, are used to make a variety of adhesives. Metals, plastics, and wood can all be joined together with the help of these adhesives. Materials are put to a surface as coatings to improve or protect it. Acrylic, polyurethane, and epoxy are just a few of the polymers that are used to make many coatings. These coatings protect against wear and tear, UV rays, and corrosion. Macromolecules are important in producing many different materials, and their unique properties and functions make them valuable in a wide range of applications ²¹.

Macromolecules like proteins and lipids are used to make different cosmetic items like creams and lotions. The appearance and texture of the skin can be improved by using proteins like collagen and elastin, which are frequently utilized in cosmetic goods. These proteins may contribute to fewer wrinkles, firmer skin, and better skin moisture ¹⁶. Ceramides and fatty acids, two lipids that can aid in increasing the skin's moisture content, are also frequently found in cosmetic products. The skin can be kept soft and supple by using these lipids to help reinforce the skin barrier and stop moisture loss. Polymers like polyvinylpyrrolidone (PVP) and polyethylene glycol (PEG) are frequently employed in cosmetic products as thickeners, emulsifiers, and stabilisers. These polymers can enhance cosmetic items' stability and consistency while making them simpler. Macromolecules are crucial in creating cosmetics due to their wide range of attributes and use as ingredients in creating creams, lotions, and other skincare products ²².

To increase agricultural productivity and quality, fertilizers, insecticides, and herbicides are made from macromolecules like proteins and carbohydrates. For plants to grow and develop, fertilizers are compounds that supply them with vital nutrients like nitrogen, phosphorus, and potassium. A lot of fertilizers are composed of macromolecules, such as proteins and carbohydrates, which soil microbes can break down to release nutrients for plant absorption. Insects, weeds, and fungi that can harm crops and lower production are examples of pests that can be controlled or eliminated by applying pesticides ²³. The metabolic or nervous system of pests can be disturbed by several pesticides, which are often manufactured from macromolecules like proteins and lipids. Herbicides are chemicals that are applied to weeds to prevent or reduce their ability to compete with crops for nutrients, water, and sunlight. Numerous herbicides are composed of macromolecules that might obstruct weed growth or metabolism, such as lipids and carbohydrates ²⁴. The creation of fertilizers, insecticides, and herbicides depends on macromolecules, and their distinctive characteristics and uses make them valuable for enhancing crop productivity and quality.

In bioremediation procedures, contaminants are broken down, and contaminated places are cleaned using macromolecules like enzymes and bacteria. Using natural biological processes to remove or degrade contaminants is known as bioremediation, and it is a cost- and environmentally-friendly method of cleaning up contaminated locations. In bioremediation, proteins called enzymes are frequently utilized to break down contaminants into less harmful chemicals and accelerate chemical reactions ²⁵. For instance, certain enzymes can degrade chlorinated solvents like trichloroethylene (TCE), which is frequently discovered in contaminated areas. In bioremediation, microorganisms like bacteria and fungi are frequently utilized because they can break down contaminants and turn them into less toxic compounds. For instance, certain bacteria may break down petroleum hydrocarbons, which are frequent contaminants in soil and groundwater and include gasoline and diesel 36. Macromolecules like enzymes and microorganisms play an important role in bioremediation processes, and their unique properties and functions make them valuable tools in cleaning up contaminated sites and reducing environmental pollution. Macromolecules can be extracted using various cutting edge technologies as discussed below.

Extraction Methods for Macro Molecules

The extraction process is a crucial step in the food industry for obtaining macromolecules and bioactive compounds, such as flavourings, pigments, and antioxidants, from natural sources. The elements that make up our food, whether they come from plants or animals, are all combinations of these macromolecules. Over the past few decades, there has been a lot of research on the most effective ways to extract valuable molecules from natural resources. The use of natural resources to promote sustainable development and environmental protection has raised public awareness. Extraction can be thought of as the initial stage in the development of analytical methods and ensuing products. The use of organic solvents or high temperatures in conventional extraction techniques, such as solvent extraction, maceration, percolation, and steam distillation, harms the environment and causes the extracted molecules to degrade. However, conventional techniques use organic solvents like acetone, ethanol, methanol, and ethyl acetate along with heated or boiling water. High temperatures and extended extraction times are the results of this. Due to these drawbacks, other extraction techniques have recently started to replace these conventional methods. These procedures frequently entail using an energy source to hasten the transfer of the macromolecule compounds to the solvent. Additionally, techniques that are more eco-friendly and require less solvents have been sought for. Over the past 10 years, new methods and emerging techniques have been developed and used to decrease the amount of sample that needs to be handled.

Green extraction technologies (GETs) have emerged as a sustainable alternative to conventional extraction methods. GETs are characterized by their eco-friendliness, high efficiency, and selectivity. Supercritical fluid extraction, microwave-assisted extraction, ultrasonication-assisted extraction, pressurized liquid extraction, pulse electric field-assisted extraction, subcritical water extraction, ionic liquid-based extraction, and enzyme-assisted extraction are some of the most cutting-edge extraction methods. This paper reviews the recent advances in GETs and their applications in the food industry.

Methods of Green Extraction Technologies

Green extraction technologies prioritize efficiency, sustainability, and safety when extracting macromolecules from various sources. Here are a few of the most popular ways to extract green materials that would be discussed in this article: Supercritical fluid extraction (SFE), Microwave-assisted extraction (MAE), Ultrasound-assisted extraction (UAE), Pressurized liquid extraction (PLE), Pulse-Electric-Field Assisted Extraction (PEF), Subcritical water extraction (SWE), Ionic liquid-based extraction (ILE), Enzyme-assisted extraction (EAE) ^79,80,89 for better green extraction of bioactive compounds have developed over the past few decades. The methods, applications, advantages, and disadvantages of these techniques are presented in Table 2.

Supercritical fluid extraction (SFE)

SFE uses supercritical carbon dioxide as a solvent to extract compounds at high pressure and low temperatures. Supercritical fluid (SF) serves as the extraction solvent in supercritical fluid extraction. Due to its similar liquid solubility and gas diffusivity properties, SF can dissolve various natural substances. Near their critical points, slight variations in pressure and temperature resulted in substantial modifications in their solvating properties. Supercritical fluids having lower surface tension, lower viscosity, and higher diffusion coefficients than normal solvents. In addition, they have characteristics with both gases and liquids. Supercritical fluids can be used to selectively extract from both liquid and solid matrices, and the ability to select of the extraction can be modified merely adjusting the degree of heat. Because supercritical fluids cannot dissolve the extracted matrices, the extraction pressure also depends on the density of the extracted matrices. Highly widely utilized solvent in SFE is carbon dioxide (CO2) since that it's accessible, safe, non-toxic, recyclable, and can reach supercritical conditions quickly. Additionally, CO2 supercritical extraction produces cleaner results than traditional extraction techniques ⁹⁰. To execute SFE, the fluid is pressurized and heated to the necessary temperature and pressure before entering the extractor, bringing it to its supercritical condition (Figure 1). The solvent is then injected into the extractor and uniformly disseminated throughout the fixed bed by solid matter. During the extraction, the solvent passes across the fixed bed and dissolves the soluble components. In the separator, the solutes are separated from the supercritical fluid by rapidly lowering the pressure, increasing the temperature, or both. As a result, the supercritical fluid will become gaseous; the solutes will no longer be solubilized in the supercritical fluid and will be separated by gravity. Extracts are gathered at the separator's bottom. Depending on the equipment, the solvent is cooled and compressed before being returned to the extractor or discharged into the atmosphere ⁸⁰.

Table 2: Methods, principles, applications, advantages, and disadvantages of GETs

Method

Principle

Applications

Advantages

Disadvantages

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