A review on various extraction and detection methods of bio-functional components from microgreens: food applications and health properties

ABSTRACT The current review investigates the effects of microgreens on human health and their use in different food items. In recent decades, various extraction methods, such as cold pressing, supercritical CO2 extraction, microwave-assisted extraction, liquid and gas chromatography, hydro-distillation method, Soxhlet extraction, alcohol extraction, steam distillation, and accelerated solvent extraction have been used for extraction under optimal conditions. Microgreens contain essential bioactive compounds which work as functional foods because these microgreens are produced and cut down before ripening. Since these are the tiny cotyledons of different pulses, grains, and vegetable seeds, they possess different nutritional profile than whole one which contains essential fatty acids, in which major one is linoleic acid and functional compounds as important antioxidant enzymes like glucoraphanin which signals the antioxidants, glucosinolates, isothiocyanates, anthocyanins, ascorbate peroxidase, catalase, superoxide dismutase, and peroxidase which work against inflammation and oxidative stress and so work against chronic diseases like hypertension, cardiovascular diseases, diabetes, and cancer. Many clinical and experimental studies have indicated that microgreens of different species fenugreek, brassica, black mustard, mung beans, green peas, red cabbage, and broccoli can be used to reduce the risk of metabolic disorders, goiter, kidney diseases, and other chronic illnesses. This review focuses on different microgreen extraction techniques, their practical applications in the food industry, and their health properties.


Introduction
Microgreens are young seedlings which are tender and edible vegetables and herbs.Unlike other mature longer herbs and vegetables which take months or weeks to grow and ripe, microgreens can be harvested earlier in a few week or 10 days after cultivation at the stage of cotyledon which is a part of seed embryo with small leaves growth. [1,2]These can be grown in small area with less resources and also suitable for hydroponic culture media, an indoor farming practice and for kitchen gardening which make them easily adaptive and suitable as a portable food items and to reduce the scarcity of food in huge urban populations.Microgreens of various species including Fenugreek, Mustard, Red Cabbage, Broccoli, Green Peas, Mung Beans, Black Mustard, and various species of Brassicaceae family are studied. [3,4]oreover, their nutrition-rich profile and high sensory acceptability get attraction of consumers, scientist, researchers as well as nutritionist and health-care professionals. [5]Their readily available macro and micronutrients along with packed bio active compounds make them special food which can not only be used as a daily diet part but also in clinical and dietitian's practice part and recommendations against various metabolic and chronic illnesses and provide valuable information and data to the nutrition policy-makers, health-conscious consumers, nutritionists, and dietitians toward selection of nutrient dense and low caloric foods. [6]Brassicaceae microgreens present a wide array of antioxidant bioactive compounds. [7]While radish, mustards, coriander, and tatsoi possess minerals and functional phenolic compounds which can be readily available to the body due to their high bioavailability and digestibility rate.This content can also be boost up by supplementing some more essential minerals like selenium, which is a strong antioxidant mineral and works as a free radical scavenger and its RDA can be achieved easily through one single food consumption in a day. [8]Supplementing 16 µM of Se in tatsoi and coriander microgreens resulted to increase total phenolic content up to 95% and 21%, respectively. [9]ccording to an epidemiological study, consuming fruits and vegetables help prevent the occurrence of common chronic diseases including diabetes, obesity, and hypertension, which annually cause people's quality of life to decline. [10]In this perspective, microgreens stand out as innovative sources of physiologically active compounds with highly valuable effects. [11]It is vital to take into account the bioavailability of phytochemicals before diving into the bioactivities related to microgreens.It is not enough for these chemicals to be present in food matrices to ensure that their biological capabilities would be confirmed. [12]Although other authors have previously reviewed some aspects of chemical properties, the effects of various pretreatments on its stability, and quality analysis studies, our review provides a more comprehensive analysis of the related studies.

Functional compounds in microgreen
Due to higher bioactive components and functional compounds, multivitamins, and more mineral bioavailability compared to mature greens, microgreens consumption increased day by day to improve human health.The most and utmost benefit of the microgreen is that they are being consumed in raw form which reduces the side effects of overcooking, food processing, and destruction of delicate B complex vitamins which means more bioavailability of the bioactive compounds and nutrients, especially of phenolic compounds and water-soluble micronutrients which are sensitive toward heat and processing. [8]A study conducted by Fuente et al. [7] in which they observed the bioactive content of different species of microgreen which includes kale, broccoli, red mustard, and radish.They found that all the microgreens consist of a handsome amount of total carotenoids and vitamin C, along with isothiocyanates and total polyphenols as major anti-oxidants with 43-70% oxidative capacity in 31-63% bio accessibility rate.The availability of minerals was also high with up to 90% bioavailability.However, a low potassium level was observed which would be suitable for renal patients for avoiding salt load. [6]he analysis of different species of Brassicaceae and Asteraceae family showed that the microgreens of these contain good amounts of phenolic compounds carotenoids, fat-soluble vitamin E with calcium and potassium minerals.Among these, the Asteraceae microgreens possess more carotenoids while of Broccoli contains more amount of tocopherol. [13]While purple and green basil species microgreen found rosmarinic acid and cichoric acid as major antioxidants.kaempferol-3-O (caffeoyl) sophoroside-7-O-glucoside and caffeic acid are abundant in tatsoi with rutin and chlorogenic acid as a major phenols of coriander. [9]Many conclusion compositions of 13 different species of microgreens and he came to a conclusion that under controlled environment microgreen species shows genotypic variation which affects carotenoid content which was significantly increased in Brassicaceae.With high phenolic concentration as flavones, flavonol glycosides and hydroxycinnamic acids in Lamiaceae and Apiaceae microgreens. [14]inerals are the important micro nutrients with functional properties of anti-oxidants, and among these potassium pays its role in balancing sodium level and is good for hypertension.In this regard, hemp microgreens are a good contributor of calcium and potassium with iron, selenium, and zinc mineral as well.Zinc works in synergy with calcium and helps its absorption. [8]A study conducted on microgreens in Thailand reported that total chlorophyll content, ascorbic acid and carotenoids content are highest in lentil microgreens while of buck wheat also possess phenolic compounds with highest scavenging capacity against reactive oxygen species and other oxides.Red cabbage and purple radish contain anthocyanin as functional compounds. [6,15]he proximal analysis of microgreens of Fabaceae, Brassicaceae, Convolvulaceae, Malvaceae, Polygonaceae, and Pedaliaceae found that along with healthy functional compounds these microgreens possess good macro-nutrient ratio with low-calorie carbohydrates comprising 7.16% and fats of only 0.15% to 0.66% of sample but high protein content with 6.5% with low calories.About 20-54% kcal among these varieties. [15]With these micro and macro nutrients, the glucoraphanin, an important bioactive compound of microgreens might play a major role against metabolic diseases because it activates nuclear factor erythroid 2-related factor 2 Nrf 2 by working as a precursor of sulforaphane.This Nrf 2 provides signaling service to the antioxidants, against the cytotoxic effect of oxidative stress, as cellular defense system.This Nrf 2 is not available in clinics due to biosafety concerns.These anti-oxidants positively reduce the obesity and obesity induced inflammation which further leads to diabetes by browning of adiposities, enhancing expenditure and polarizing M2 macrophages and decreases endotoxemia.In this way, glucoraphanin acts strongly against insulin resistance, low inflammation rate, and ameliorating obesity (Figure 1). [16]

Microgreen extraction and detection techniques
Extraction of microgreens and detection of their phenolic compounds and active substance are described in Table 1.

Water extraction
Water Extraction Micro greens are small, nutrient-dense plants that can be extracted using a variety of techniques depending on the compounds that are desired.Here are some common extraction techniques for micro greens.Water extraction is a gentle and effective method for extracting bioactive compounds from micro greens without the use of harsh solvents.It is particularly useful for extracting water-soluble compounds such as polyphenols, flavonoids, and vitamins.However, water extraction may not be suitable for extracting lipophilic compounds or compounds that are sensitive to heat or oxidation.The micro greens are harvested, washed, and dried to remove any moisture.The micro greens are blended with distilled water in a blender or food processor to create a smooth paste.The paste is mixed with additional distilled water to create slurry with a desired concentration.The slurry is then strained through a fine mesh strainer or cheesecloth to separate the liquid extract from the solid residue.The liquid extract is then filtered using filter paper or a coffee filter to remove any remaining solid particles.
Storage: The resulting extract can be stored in a sealed container in the refrigerator for future use. [29]l extraction Oil extraction is a commonly used method for extracting lipophilic compounds such as essential oils and other fat-soluble compounds from microgreens.Here is an overview of the steps involved in oil extraction of microgreens.Oil extraction is a gentle and effective method for extracting lipophilic compounds from microgreens. [4]The use of oil as a solvent can also help to protect and stabilize sensitive compounds from degradation.However, oil extraction may not be suitable for extracting water-soluble compounds or for use in certain applications where the presence of oil is not desirable. [30]To get rid of any moisture, the microgreens are collected, cleaned, and dried.The microgreens are blended with a suitable oil, such as olive oil, coconut oil, or sunflower oil, in a blender or food processor to create a smooth paste.Pat the microgreens dry with a clean kitchen towel to remove any excess water. [31]Place the microgreens in a clean glass jar.Pour the oil over the microgreens until they are completely covered.Seal the jar with a lid and let it sit at room temperature for 24-48 h.Shake the jar gently once or twice a day to ensure that the oil is evenly distributed.After 24-48 h, strain the oil using a fine mesh strainer or cheesecloth to remove the microgreens.Transfer the oil to a clean glass jar or bottle and store in a cool, dark place.

Ethanol extraction
Ethanol extraction is a common method of extracting bioactive compounds from microgreens.Here are the steps for extracting microgreens using ethanol. [13]Rinse the microgreens under cold running water to remove any large debris or soil.Pat the microgreens dry with a clean kitchen towel to remove any excess water.Place the microgreens in a clean glass jar.Pour enough ethanol (such as ethanol or 95% ethanol) over the microgreens to completely cover them. [32]Seal the jar with a lid and let it sit for 24-48 h.After 24-48 h, strain the ethanol using a fine mesh strainer or cheesecloth to remove the microgreens.Transfer the ethanol extract to a clean glass jar or bottle and store in a cool, dark place.If desired, you can evaporate the ethanol to concentrate the extract.This can be done by placing the extract in a shallow dish and allowing the ethanol to evaporate naturally, or by using a rotary evaporator or other specialized equipment. [33]

Alcohol extraction
Alcohol extraction is a popular method for extracting bioactive compounds from microgreens. [13]The microgreens are ground into a fine powder using a mortar and pestle or a blender.Due to its capacity to extract a variety of chemicals, ethanol is frequently utilized as the solvent for alcohol extraction of microgreens. [34]The mixture is then agitated or stirred for a certain amount of time, usually several hours, to facilitate the extraction of the desired compounds from the microgreens.The mixture is then filtered to separate the liquid extract from the solid residue.The liquid extract is then evaporated to remove the ethanol and concentrate the desired compounds.The resulting extract can be stored in a sealed container in a cool, dry place for future use. [35]

Enzyme extraction
Enzyme extraction is a gentle and effective method for extracting specific compounds from microgreens.It can be used to extract compounds that are difficult to extract using other methods, such as polysaccharides and glycosides. [36]However, enzyme extraction can be time-consuming and expensive, as specific enzymes can be costly.Additionally, the conditions for enzyme extraction need to be carefully optimized to ensure optimal yield and quality of the extract.Enzyme extraction is a method of extracting bioactive compounds from microgreens using specific enzymes. [37]Here is an overview of the steps involved in enzyme extraction of microgreens.An appropriate enzyme is selected based on the target compounds to be extracted.For example, cellulase can be used to extract cellulose, while pectinase can be used to extract pectin. [38]The microgreens are mixed with the enzyme in a container, such as a beaker or flask.The mixture is then incubated at an appropriate temperature and pH for a certain amount of time to allow the enzyme to break down the cell walls of the microgreens and release the desired compounds. [39]he mixture is then filtered to separate the liquid extract from the solid residue.Concentration: The liquid extract is then concentrated, usually by evaporation, to increase the concentration of the desired compounds.The resulting extract can store in a sealed container in a cool, dry place for future use. [3]percritical CO 2 Supercritical CO 2 extraction is a clean and efficient method for extracting bioactive compounds from microgreens. [25,40]It can be used to extract a wide range of compounds, including lipids, terpenes, and phenolics.The use of CO 2 as a solvent is advantageous because it is nontoxic, nonflammable, and leaves no residue. [41]However, the equipment required for supercritical CO 2 extraction can be expensive, and the process can be complex and time-consuming.It is a method of extracting bioactive compounds from microgreens using carbon dioxide in its supercritical state.The dried microgreens are loaded into an extraction vessel, such as a column or extractor.CO 2 is compressed and heated to become supercritical, a state in which it exhibits both liquid and gas properties. [42]The supercritical CO 2 is then passed through the extraction vessel containing the microgreens.The supercritical CO 2 acts as a solvent and extracts the desired compounds from the microgreens.The pressure, temperature, and flow rate of the CO 2 can be adjusted to optimize the extraction efficiency.The mixture of supercritical CO 2 and extracted compounds is then passed through a separator, which separates the CO 2 from the extract. [43]

Cold pressing extraction
It is important to note that cold pressing is a slow and gentle process that helps to preserve the nutrients and enzymes present in the microgreens. [44]This is in contrast to other extraction methods that use heat or chemicals, which can damage the delicate nutrients and flavors of the microgreens. [45]he resulting juice extracted from microgreens using the cold pressing technique is a highly concentrated source of vitamins, minerals, and antioxidants.It is also believed to have anti-inflammatory and detoxifying properties, making it a popular health drink.Overall, the cold pressing extraction technique of microgreens is a preferred method of extracting the valuable nutrients and health benefits of these young vegetable greens. [46,47]The cold pressing extraction technique of microgreens involves using a hydraulic press to extract the juice from the young vegetable greens without using heat.The microgreens are first washed thoroughly to remove any dirt or debris.The microgreens are then cut into small pieces to make them easier to press.The cut microgreens are loaded into a hydraulic press, which applies pressure to extract the juice.The extracted juice is then filtered to remove any pulp or solids.The filtered juice is then bottled and sold as a nutritional supplement. [48]

Microwave-Assisted Extraction (MAE)
Microwave-assisted extraction is a rapid and efficient method for extracting bioactive compounds from microgreens.It can be used to extract a wide range of compounds, including phenolics, flavonoids, and carotenoids. [49]Additionally, MAE can be a more environmentally friendly method compared to traditional solvent extraction methods, as it reduces the amount of solvent required and can shorten the extraction time.However, the process needs to be carefully optimized to avoid overheating the sample, which can result in degradation of the target compounds. [4]Microwaveassisted extraction is a method of extracting bioactive compounds from microgreens using microwave energy.Here is an overview of the steps involved in microwave-assisted extraction of microgreens.After being gathered, the microgreens are cleaned and dried to remove any moisture.The microgreens are ground into a fine powder using a mortar and pestle or a blender.An appropriate solvent is selected based on the target compounds to be extracted.Water or ethanol is commonly used solvents for MAE.The ground microgreens are mixed with the solvent in a container, such as a microwave-safe beaker or flask.The mixture is then subjected to microwave irradiation at an appropriate power and time.The microwave energy causes the solvent to boil, creating pressure inside the container, which helps to facilitate the extraction of the desired compounds from the microgreens. [50]The mixture is then filtered to separate the liquid extract from the solid residue.The liquid extract is then concentrated, usually by evaporation, to increase the concentration of the desired compounds.The resulting extract can be stored in a sealed container in a cool, dry place for future use. [51]

Ultrasound assisted extraction
Ultrasound assisted extraction involves ultrasound energy sound waves and elastic medium includes liquid solvents, ultrasound uses high-frequency sound waves which are beyond the human hearing capacity.The sound waves are in the range of 2 MHz to 20 kHz. [52]The ultrasound mechanism involves four steps.The first one is the sound waves pass through the food matrix and the medium causes the generation of cavitation bubbles.The bubble then collapsed increasing the temperature and releases a microjet with pressure toward the surface.This pressure ruptures the surface of food matrix, then active ingredients inside the cell surface and solvent are in direct contact with the solution.All the bioactive compounds are then released from the matrix into the solvent. [53]The ultrasound assisted extraction words significantly only in liquid medium and increases the mass transfer.It is not resisted with any solvent.Ultrasound sound waves are used in the extraction of essential oils, bioactive compounds, polyphenols, flavonoids, anthocyanins and plant extracts. [54]

Food applications of microgreens
It is common knowledge that food and health are related.Plant-based diet is essential for a healthy diet.According to study, consuming fruits and vegetables help prevent the occurrence of common chronic diseases including diabetes, obesity, and hypertension, which annually cause people's quality of life to decline. [10]In this perspective, microgreens are innovative sources of active compounds with valuable effects. [11]It is vital to take into account the bioavailability of phytochemicals before diving into the bioactivities related to microgreens.It is not enough for these chemicals to be present in food matrices to ensure that their biological capabilities were confirmed. [12]t is known that colon epithelial cells absorb isothiocyanates in free state along with conjugates, which are absorbed by peripheral organs and accumulate in cell bodies by interacting with thiols of glutathione and proteins. [55]While phenolic substances do not pass through intestinal barrier as quickly.According to a number of publications, polyphenols' due to chemical instability, poor absorption, gut microbial modification, and excessive metabolism cause low bioavailability.Nonetheless, many phenolic substances exhibit biological effects even when plasma concentrations were low.The impact of the dietary matrix should be evaluated during this process. [56]n a rapidly growing world population, environmental sustainability and crop production to meet human nutrition become a most focusing challenge of the era as malnutrition increases day by day.Scientists and developers are working to develop new agricultural techniques for growing.Repeatedly mass production of staple crops reduces the nutrient density of the food day by day. [57]The application potential of microgreens varies from normal food sources to the enriched foods because of its high protein content and can be used as functional food-packed with essential antioxidants, mineral, and vitamins.Due to easily cultivated in hydroponic medium, makes them best food application as space foods for astronauts. [58]icrogreens are the cotyledons of the various vegetables, aromatic herbs, herbaceous plants, pulses, and seeds which are young and tender edible seedlings which are harvested mostly in 7-21 days after germination.In food engineering, human sensory analysis is typically used as the standard method to evaluate food properties.Different types of microgreen species were sensory evaluated to a panel whose results showed that self -produced microgreens are highly accepted by the consumers which ensures the good acceptance. [59]eanwhile, microgreens provide promising edible safety as well.A study conducted to check the oral acute and sub-acute toxicity level by consuming Brassica carinata A. Braun microgreens ethanoic extract in Wistar rats resulted no any sort of toxicity in rats. [60]Arugula, broccoli, bull's blood beet, red cabbage, red garnet amaranth, and tendril pea (Six microgreen species) were checked in a study for consumer acceptability.The resulted data showed high acceptability score for sensory properties, flavor and acceptance for all microgreen species with few negatives with food neophobia. [5]Microgreens are used in salads, shakes, in sandwiches, in sweets and savory dishes as a garnish which complement the color, texture, flavor, flatbreads, pizza, soups due to nutritional properties and long shelf life (Figure 2).
Microgreens come in a diverse array of flavors, vibrant colors (including red, purple, green, yellow, and more), varied appearances, and textures (ranging from tender and juicy to crisp).Microgreens are extensively used in beverage industry such as juices [61] and fermented drinks. [62]Broccoli sprouts are categorized as functional foods due to their natural high concentration of glucoraphanin (GR), which serves as the precursor to the anticancer compound known as sulforaphane (SFN).Because of their potential health benefits, broccoli sprout juice is produced through mechanical pressing.The juice is then collected in chilled tubes, clarified through centrifugation (at 4°C for 30 min), and subsequently blended with various other juices. [61,63]Broccoli sprouts are known for their potential protective effects against a range of health conditions, including cardiovascular disease, neurodegenerative disorders, diabetes, and various inflammatory conditions. [64,65]icrogreens and the leaves of Brassica species like pak choi and kale have the potential to offer consumers a range of health-promoting bioactive substances, including breakdown products of glucosinolates (GLS), carotenoids, chlorophyll metabolites, and phenolic compounds. [66,67]icrogreens and fresh leafy plant material from Brassica and kale can serve as valuable additions to enhance the levels of bioactive, health-promoting substances in a typical Western diet, especially when incorporated into commonly consumed food products like bread. [68]These characteristics render them versatile ingredients suitable for incorporation into salads, soups, sandwiches, and as decorative elements to enhance the sensory appeal of various dishes. [63]Their versatility positions them as a valuable resource for the development of nutritious food products.Incorporating fresh kale and pak choi microgreens into blends, it is possible to enhance the levels of secondary metabolites, which offer health-promoting benefits. [69]Microgreens can also be utilized to create innovative food items such as beverages, seasonings, weaning foods [70] probiotic-infused products, yogurt, [66] flour enriched with powdered microgreens, [71] bakery items, and teas, [72] in addition to being enjoyed fresh. [73,74]n addition to being consumed fresh, sprouts and microgreens offer diverse possibilities for creating innovative food products.These include juices, fermented drinks, breakfast or snack foods, seasonings like vinegar or sauce, weaning foods, probiotic-infused beverages or foods, yogurt, flours enriched with powdered sprouts and microgreens, bakery items, and even teas.Recently, there has been a growing interest in incorporating sprouted wheat pomace powder, a by-product from juice production, into flour mixtures.This addition was found to enhance the nutritional value of the wheat flour significantly. [69,75]read made with sprouted flour emerges as a notable source of antiradical and chelating compounds, along with phytochemicals capable of safeguarding lipids against oxidation.Both the elicitation process and sprouting have an impact on the profile of phenolic acids and the antioxidant capabilities of wholemeal wheat flour. [67]The assessment of the antioxidant potential of functional products takes into account the potential for bioactive compounds to be accessed and absorbed by the body.Additionally, these bioactive compounds exhibit resistance to high temperatures and possess the potential to be effectively accessed by the body, making them valuable additions to functional food products. [73,74]icrogreens are also involved the production of vegetable juice and fermented vegetable juice, which contained a mixture of vegetables, seeds, and germinated lentils and cowpeas (Lens culinaris and Vigna sinensis).The seeds were initially soaked in three times their volume of water at room temperature for 8 h.Subsequently, they were allowed to germinate in a refrigerated incubator at a temperature of 30°C for a period of 5 days for lentils and 3 days for cowpeas. [61]These vegetable and sprout-based products have demonstrated numerous beneficial effects, including antidiabetic activity, the ability to lower cholesterol levels, ACE (Angiotensin-Converting Enzyme) inhibitory activity, hemagglutinating activity, and antioxidant properties. [76,77]

Health properties of microgreens
Microgreens are abundant in nutritional and functional compounds which provide protection against chronic and metabolic diseases.Due to this, these are considered as health promoting diets. [58]These get promotion based on their high nutritional, bioactive compound content which promote good human health by reducing chronic illnesses like CVDs, hypercholesterolemia, and other metabolic disease risk. [5]These can be add in daily diet with less cultivation cost, recycle able, easy to grow without any weather dependency and also transportable medium and provided to the malnourished communities with less than 1 week as a good food. [78]Many researcher conducted a study to check antihypertensive, anti-diabetic, and hypolipidemic effect, in the perspective of cardiovascular disease burden, of five different kinds of sprouts in which he found that out of five species (mung beans, broccoli, radish, buckwheat, and soybeans) mung beans, buckwheat, and broccoli possess strong antihyperlipidemic and hypertensive effects (Figure 3 and Table 2). [79]umerous leafy vegetables are rich sources of bioactive compounds.These bioactive compounds, when highly bioaccessible following digestion, have the potential to exert antiinflammatory, anticancer, antimicrobial, and anti-diabetic effects. [80]A study has provided evidence demonstrating the impact of microgreens from kale, radish, mustard, and broccoli on tumoral colon cells, underscoring the antiproliferative properties of microgreens against colon cancer cell proliferation.Mustard and radish microgreens exhibited more pronounced antiproliferative effects than kale and broccoli, aligning with their higher content of vitamin C, total carotenoids, and total isothiocyanates. [81]icrogreens are frequently regarded as preferable substitutes for sprouts owing to their elevated nutritional profile and heightened gustatory attributes.Furthermore, microgreens have the propensity to harbor augmented quantities of phytochemicals, minerals, and vitamins relative to their mature counterparts.Consequently, the integration of microgreens into dietary regimens holds promise for amplifying overall nutritional adequacy and fostering favorable health outcomes among consumers. [82]The substantial presence of bioactive compounds in microgreens, encompassing vitamins, minerals, and phytochemicals are capable of counteracting free radicals and mitigating oxidative stress-related damage.These antioxidants include vitamin C (VC), various phytochemicals such as carotenoids and phenolics, as well as specific minerals like copper (Cu), zinc (Zn), and selenium (Se). [83]Promoting digestive wellness.[109,110]   (Continued)

Hypercholesterolemia
According to some studies, intake of red cabbage microgreens transferred cholesterol synthesis in the body and positively affects its metabolism by reducing the cholesterol rate, meanwhile it provides protection against weight gain due to hypercholesterolemia. Hypercholesterolemia is a major cause of atherosclerosis and plaque in coronary arteries which resulted in cardiovascular diseases.The chances would be reduced by lowering the plasma concentration of low-density triglycerides-TGs in liver about 23%, cholesterol ester formation with 65.5% and lipoproteins-LDL (up to 34%) with red cabbage microgreens in animal model fed with high-fat diet.The underline mechanism was the inhibition of TGs synthetic enzymes glycerol-3-phosphate acyltransferase enzyme (GPAM), diacylglycerol O-acyltransferase 1 known as DGAT1, lecithin-cholesterol acyltransferase (LACT), and cholesterol ester synthetic enzyme acetyl COA acetyl transferase 3-ACAT3 in animal model study.[84]    Contains strong anticancer properties, antiinflammatory properties.[116,117]  Microgreens have potentials by comparing these with normal growing plant.According to them, it is only the potential of red cabbage microgreens, not of red cabbage, to lower the effect of cholesterol ester synthetic enzyme SOAT 1, down regulation of triacylglycerol synthesis enzyme DGAT1 and fatty acid coenzyme substrate molecules in liver, the potential benefit of microgreens which would be otherwise not achieved by whole vegetable plant, the potential benefit of microgreens which would be otherwise not achieved by whole vegetable plant. [84]Therefore, microgreens supplementation significantly keeps in control the plasma level of LDL, TGs concentration, and hepatic cholesterol ester formation by reducing the expressions of inflammatory cytokines in the liver as the over production of cholesterol also shares its mechanism with inflammation rate.In an animal research model explains that the broccoli microgreen juice was introduced in the mice to check the hypolipidemic effects.
Effectively broccoli microgreen juice intake showed good results by significantly lowering serum lipid profile with decreased concentration of total cholesterol-TC and LDL levels also. [85]

Anti-oxidative capacity and inflammation
The antioxidant potential of the cell which is very important for the normal biological system.All of the microgreens had estimated levels of four important antioxidant enzymes ascorbate peroxidase, catalase, superoxide dismutase, and peroxidase.A raised activity suggests that reactive oxygen species (ROS) are accumulating less.In order to prevent oxidative damage in extremely adverse circumstances [86] plants had evolved an antioxidant defense system that comprises antioxidants. [87]ethionine and homocysteine are the amino acid thiols which work against stress and reduce the oxidative damage resulted by it.In this way, these amino acids provide protection to the cells against inflammation by working as anti-oxidants.Along with these, microgreens are also packed with other essential amino acids in different species with vary concentrations.Like L-asparagine is the abundant amino acid of radish and green bean microgreens, L-glutamine is of amaranth and kale microgreens. [3]anthine oxidase enzyme takes part in the metabolism of hypoxanthine and conversion of xanthine into uric acid and cause uric acid elevation in gout patients.In 2020, Le and his colleagues found that broccoli is a major source of functional compounds which are xanthine oxidase inhibitors.So by incorporating these bioactive compounds into the treatment therapy will work for gout and other XO-based diseases. [80]ealthy intestinal microflora is very important for a healthy gut to keep the body immune from outer pathogens and harmful bacteria and also provide anti-inflammatory effect by producing shortchain fatty acids (SCFAs).When broccoli microgreen juice was given to the animals under study, along with other positive aspects, the microgreens intake also resulted to boost up the healthy bacteria of the animal intestine.As compared to control group which was fed with high-fat diet, the treatment groups had abundance of Bacteroidetes and Bacteroids acidifaciens with reduced burden of Firmicutes.This modification of microflora will then lead to increase production of SCFAs with a reduction in lipoproteins parameters (Figure 4). [85]ti-diabetic properties Diabetes, a metabolic syndrome that is the development of insulin resistance in the cells or by the pancreas' inability to generate enough insulin.High blood glucose levels are a defining feature of diabetes, which is often managed or treated by reducing blood glucose levels by strict dietary restrictions, insulin injections, supplemental insulin production, and improved insulin sensitivity. [88,89]Diabetes is one of the major chronic diseases nowadays which are mainly due to poor lifestyle and dietary habits.Environmental factors along with genetics and lifestyle add for it. [90,91]To reduce the burden of diabetes, many scientists work on dietary modifications and new techniques in food technology.
However, extended pharmaceutical use may have negative side effects, as a result, eating fruits and vegetables with little to no sugar is typically advised.Microgreens are becoming more and more popular, and they are expected to be more effective and have a great potential to lower diabetes.They are also very nutrient-dense.In HepG 2 cells, fenugreek microgreen extract reduced α-amylase activity about 70%, and in L6 cells, glucose absorption was increased by 44% (2 mg/mL) in the presence of insulin. [92]icrogreens were also tested against it and showed positive results.α-amylase inhibition and αglucosidase inhibition mechanism splay vital role against disease prevention.These inhibitors are found in black medic, beetroot, mung beans, and broccoli microgreens.Radish microgreens also showed significant potential against high inhibitory activity toward pancreatic lipase. [3]Downregulation of genes related to lipid accumulation and increase in Nrf2 activity is also important in this regard.The glucoraphanin which can be extracted from broccoli sprouts/microgreens helps to do reduce this burden also down regulating the other gene expressions which are major involved in lipogenesis and gluconeogenesis. [16]In another study, the anti-diabetic effect of lyophilized broccoli microgreens in type-2 diabetic mice in 18 weeks experiment was surprisingly effective with the significant results of reduced weight and improved glucose level along with other factors which were plasma lipid level, antioxidant capacity, and bio-inflammatory markers. [57]The histological study of the organs showed improved gut microbiota with increase concentration of propionic acid and pathological changes against insulin resistance, which concluded that broccoli microgreens possess the ability to reduce T2D symptoms which make this new class vegetable a strong functional food. [93]he fenugreek microgreens' high phenolic content, flavonoids, and antioxidant levels are likely to regulate anti-diabetic efficacy in vitro.Moreover, the non-enzymatic glycation of protein was prevented by this extract.Additionally, rats on high-fat diet showed a hypoglycemic response to lyophilized broccoli microgreen powder used with dose rate of 2 g per BW. [56]Wadhawan and his cofellows examined the impact of microgreens on enzymatic glucose release.He observed that, in comparison to the control groups of, curry leaves, fennel seeds, and asafetida, 23.3 mg extract of fenugreek microgreen reduced the activity of porcine -amylase -an enzyme that breaks down the α-1,4-glycosidic linkages in starch for producing glucose.The extract of 2 mg also dramatically reduced non-enzymatic glycation up to 70% compared to control, revealing that it has a positive result on blood glucose levels. [94]

Anti-cancerous properties
Microgreens as a functional food are incredibly high in bioactive substances like tocopherols, carotenoids, chlorophyll, polyphenols, glucosinolates, and ascorbic acids.Traditional recommendations against cancer, obesity, and people with heart disease include Brassicaceae microgreens.The most common malignancies in different persons are liver, gall bladder, gallbladder, colon, and lung cancers.It is advised that these cancer patients consume vegetables on a regular basis and in high quantities. [95]According to an in vitro study, radish, kale, broccoli, mustard, and microgreens possess anti-proliferative activity against colon cancer cell growth because they significantly reduced the viability of tumor colon cells compared to the control group.In accordance with their greater levels of total isothiocyanates and total carotenoids, radish and mustard microgreens which showed excellent anti-proliferative effects compared with kale and broccoli. [4]any studies on the connection between nutrition and cancer have recently been conducted, and cruciferous vegetable consumption is strongly advised for cancer prevention.The effects of broccoli, radish, kale, and mustard, grown in hydroponic systems, on colon Caco-2 cells as opposed to healthy colon CCD18-Co cells. [7]The bioactivity of the four microgreens' bio-accessible fractions was compared to that of the colon cancer medication 5-fluorouracil after the cells had been exposed to it for 24 h.Increased ROS-reactive oxygen species with general cell cycle arrest in G2/M, a decreased glutathione intracellular concentration, and apoptotic cell death were all seen in the cells treated with microgreens. [48]n addition, human hepatocellular carcinoma (HepG2) cells and human breast adenocarcinoma (MCF-7) demonstrated an anti-migratory effect in response to microgreen extract of Thai red-tailed radish (from cold-plasma-treated seeds), which also inhibited cancer cell proliferation and induced apoptosis through regulating Caspase-3 and Bax expressions.This extract also effectively inhibits MMP-9 and MMP-2 genes linked to metastasis and tumor invasion. [96]Kaimuangpak found that extracellular vesicles of Raphanus sativus L. var.caudatus Alef microgreens possessed functional macromolecules which played role in HCT116 cells proliferation inhibition.These proteins passed βpleated sheet secondary structures which showed anti-cancerous properties against colon cancer. [97]

Metabolic disorders and inflammations
The metabolic disorders are commonly linked to diets that are high in calories, fat, and fruits.The bioactive substances found in microgreens may affect a variety of inflammatory pathways.Hence, by following the swelling guideline, microgreens can help prevent diabetes, CVD, and weight issues.Red cabbage microgreens, as previously mentioned, tended to control weight brought by high-fat diet intake.
Using red cabbage microgreens has been shown to reduce the levels of C-reactive protein (CRP) and lump death aspect alpha (TNF-a) in hepatocytes which are brought on due to high-fat diet intake. [98]This result explained that the microgreen's capacity to control hepatic lipids, the exceed amount of which is associated with inflammation. [84]Red cabbage microgreen study shows that desulfo-glucosinolate concentrations are significantly higher which was about 17.15 mol/g on dry weight basis compared with mature equivalents (8.30 mol/g).
This might be due to microgreen retention with its capacity to mimic adipogenesis, even though the system is unclear.Several phytochemicals of the microgreens and of vegetables including I3C and retinoic acid (RA), a carotene metabolite, have been demonstrated to inhibit adipogenesis.According to estimates, one-third of all cancer cells can be stopped by eating fruits and vegetables along with diet which should be low in, fat, calories, and fattening foods.Many bioactive generated from diet have been shown anti-cancer properties, those against important cancer cells including prostate, breast, and colon cancer.Nevertheless, the mechanisms are still unexplained. [19]esity and weight management C57BL/6J mice were used to examine the preventative effect of obesity using broccoli microgreen juice.The study model comprised of induced obesity fed a high-fat diet were gavaged broccoli microgreen juice.Melbin was administered as a positive control at a dose of 300 mg/kg-bw/d, which markedly reduced body weight, adipocyte size, and the mass of white adipose tissues while dramatically increasing water consumption.Moreover, it improved glucose tolerance, decreased insulin levels, and reduced insulin resistance.According to our results, the short-chain fatty acid -lipopolysaccharide -inflammatory axis in the gut microbiota may be involved in the protective benefits of juice of broccoli microgreens against obesity. [85]rdiovascular disorders Among other chronic conditions, cardiovascular disorders rank among the leading killers.Hypercholesterolemia is a chronic metabolic illness that contributes to cardiovascular disease. [99]Increased consumption of fruits and vegetables was found to lower the rate of cardiovascular illnesses according to several epidemiological data.The consumption of microgreens is strongly advised as a part of the diet to enhance metabolic health and reduce fatal CVDs because of their rich nutrients and phyto-chemicals. [58]In vivo investigation, Huang, looked for the efficacy of microgreens against cholesterol and lipid.They supplemented their meals with microgreens and employed a high fat diet to induced obesity model. [84]In addition to losing weight, the animals also significantly decreased their hepatic levels of triacylglycerol, cholesterol ester, inflammatory cytokines and LDL production. [3]Also, it was found that inhibition of the triglyceride-synthesizing enzymes acetyl-CoA acetyltransferase-3, glycerol-3-phosphate acyltransferase, and lecithin-cholesterol acyltransferase was associated with a decrease in cholesterol and inflammatory cytokines present in the liver.

Kidney disorder
A kidney ailment is a new, worldwide problem that falls under the metabolic illnesses because of its connections to many other abnormalities that are caused by a sedentary lifestyle. [100]Foods having a high potassium level are banned for patients with chronic renal diseases.They are therefore advised to eat fruits and vegetables with little to no potassium content.While potassium-rich foods put high risk for kidney-impaired patients, eating enough dietary potassium is necessary to either keep blood pressure lower or normal it.The patients also susceptible to diseases associated with a sedentary lifestyle, including hypertension, diabetes, hyperlipidemia, and several other heart-related conditions. [101]ue to the high potassium content in vegetables, research has focused on the production of microgreens, whose nutritional value may be changed by adjusting the cultivation conditions, resulting in microgreens with low potassium level.Researcher investigated lettuce and chicory microgreen in a hydroponic culture media with significantly less potassium.Its low content of potassium was made possible by the nutrient solution with lack of potassium salts during the microgreen growth.They came to the conclusion that growing microgreens without potassium in the nutrient solution did not affect the nutritional value, form, texture, or color of the microgreens, but it did result in a minor decrease in biomass yield. [59]

Iron deficiency
Many studied bioavailability for the iron content in different microgreen kinds as fenugreek, broccoli and rocket while compared them with corresponding whole plant food equivalents in light of the growing culinary trend of these microgreens.Using an inductively coupled plasma optical emission spectrometer (ICP-OES) and microwave digestion, they assessed the bioavailability and mineral content of the microgreens.The findings showed that fenugreek microgreens had increased iron concentrations, indicating a high degree of bioavailability to the cells.They concluded that microgreens from fenugreek could be employed to boost the body's ability to absorb iron and its enhancer mechanism. [102]

Conclusion
There is a lot of evidence showing that microgreen and its active compounds have effective hypocholesterolemia, anti-oxidant, anti-inflammatory as well as anti-diabetic properties against many chronic diseases in humans and animals which make them a safe food to use and consume.These can be used directly as salad, in dressings and also in the form of juices and drinks.The new goal of the researchers and scientists is to apply the extractions in nutraceutical industry as concentrated active ingredients so that they can be supplement to the boy in less time with more effective way despite any food-food interactions.Clinical and experimental research work is needed to further assess the efficiency of microgreen on body.

Figure 4 .
Figure 4. Working of different functional compounds of microgreens against metabolic diseases.