INTRODUCTION:

In recent years chia seeds have gained a lot of popularity because of their many health benefits. They have a high content of antioxidants that protect our body against the free radicals that damage the cells and lead to chronic diseases. Chia seeds also have a high content of omega-3 fatty acids, which are useful for reducing inflammation and improving heart health and brain health. Additionally, chia seeds make up a good vegetarian diet as it a good source and high content of plant-based proteins.¹The Salvia genus has nearly 900 species. Chia seeds (Salvia hispanica Linn.) belong to the family Lamiaceae grow in tropical and subtropical climates. This herbaceous plant originated from the southern part of Mexico and the northern part of Guatemala; since 1500 BC, food seeds have played a major role in human nutrition.

Eventually, in traditional medicines, chia seeds were used as food and for cosmetics. It is also a part of pre-Columbian civilizations for religious rituals.^2,3It is used in cancer, diabetes, inflammations, and thrombosis. Phytochemical components of this plant have been used for millennia by humans to prevent and treat different diseases. The protein in food has antioxidant and anti-inflammatory activities.²Chia seeds have a diameter of 2 millimeters with oval and grey black and white spots. These edible seeds are a flowering plant from Salvia hispanica of the mint family. Native of the plant is central and southern Mexico. The Aztecs cultivated the crop in pre-Columbian times and used it as a staple food for the Mesoamerican cultures. Chia seeds contain healthy dietary fibre, minerals, proteins, polyunsaturated fatty acids, vitamins, and, importantly, ω-3 fatty acids. They also have a good source of dietary fibre at 10 grams per ounce (which is 2 tablespoons) and contain minerals and proteins, including zinc, iron, magnesium and calcium. These chia seeds are commonly used in hot cereals, salads, cold cereals, and sandwiches. These seeds form a gel-like substance when mixed with juice, water, or milk.^4-10The taxonomical description and vernacular name were shown in Table 1.

Table 1 : taxonomical description and vernacular name of chia seeds

Taxonomical Description

Vernacular Names

Kingdom - Plantae

Division - Magnoliophyta

Class - Magnoliopsida

Order - Lamiales

Family – Lamiaceae

Subfamily - Nepetoideae

Genus - Salvia

Species – hispanica

Mexican – Salba chia

Hindi – Chia beej

Tamil – Sia

Telugu – Chia ginjalu

Kannada – Chia beeja

MACROSCOPICAL CHARACTERISTICS:

The annual herbaceous plant Salvia hispanica, is also called chia. Family- Lamiaceae or Labiate family. The height of the plant is 1m approx. (Fig 1). The leaves are of width 3-5cm and length 4-8cm. The color of the flowers is white or purple, and they are bisexual. The seed is oval with length - 2-2.5mm, width – 1.2-1.5mm, surface - 0.8-1.0, colour - white, brown, grey and blank, taste - bitter, odour - characteristic. The plant is sensitive to daylight. This plant is grown in the region of tropical and subtropical. Salvia hispanica is developed in the sandy and clay loam by itself at a great drainage condition.^11-16

Fig. 1: Single chia seed

MICROSCOPICAL CHARACTERISTICS:

Seed mucilage forms a transparent capsule with an approximate thickness of 414±35µm seen in Fig 2 and 3. When the seed is hydrated for 2hours, it forms a mucilaginous gel that is transparent and reaches its maximum thickness. Then safranin is dyed with it. It has 2 layers; the inner layer is formed due to a branched layer. Outer layer – it is homogeneous and cloudy. Mucilage forms a seed coat in outer cells¹⁷. It is also known as mucilaginous cells or testes. The thickness of the seed coat or testa is 13±0.41µm. It is of 3 layers, the Outer layers – it is mainly structured by rectangular thin-walled cells of size 4.2±1.26µm. Scleroid layer (where the mucilage is present) – These are thin and long cells like the fibers of the endocarp (Fig 4-7). Inner layer – It is not present in S. hispanica. Burt has a similar structure found in Arabidopsis thaliana. Small filaments appear on the surface of the seed when it is in contact with the water, and they slowly begin to stretch unless they are extended fully.

Fig. 2: Hydrated form

Fig. 3: Inside mucilage and encase

Fig. 4: Inside mucilage and encase

Fig. 5: Three layer of seeds

Fig. 6: Columella

Fig. 7: tiny clusters

The filaments of seeds are fully developed when they are completely hydrated. A volcano columella structure has become apparent. Around the seed surface, it is uniformly distributed. The chia seed has testa epidermal cells where the mucilage is present. It immediately expands when it comes in contact with the water and the primary cell layer is ruptured which is produced by the epidermal cell that is around the seed(Fig 8-9).^18-19

Fig 8 and 9 Depicts the SEM of the chia seed

The chia seeds are in different colors, and seeds are different in length and width when viewed laterally. The transverse section and longitudinal section of chia seeds correspond to each other. The chia seeds contain 2 cotyledons. They are visible; when the seeds are hydrated, we can observe hexagonal thickened cell walls and epidermal cells. The columella is present in the centre.²⁰

POWDER MICROSCOPY:

S. hispanica seed in the hypodermis showed parenchyma cells and rounded collenchyma cells. The mucilage was filled with polygonal epidermis cells, pigmented layer, lignified sclereids, yellow-colored fibres, pitted wall, aleurone grains, and oil globules (Fig 10).²¹

Fig 10(a) Parenchyma cells Fig 10(b) Oil globules

Fig 10(c) Lignified fibres.

NUTRITIONAL INFORMATION:

Chia seeds contain energy, carbohydrates, vitamins, minerals, dietary fibre(18-30%), ash (4-5%), lipids(40%), protein(20.70), fat(30.47), fibre (34.4) amino acids such as cysteine (0.41), leucine (1.37), serine (1.05), proline(0.77), aspartic acid (1.67), tyrosine (0.56), threonine (0.71), isoleucine (0.80), histidine (0.53), glutamic acid (3.50), phenylalanine (1.01), methionine (0.59), arginine (2.14), lysine (0.97), valine (0.95), tryptophane (0.44), alanine (1.05) and minerals like calcium, phosphorous and potassium content are present.^8,22,23

CHEMICAL CONSTITUENTS:

The chemical constituents which are present are Omega 6 fatty acid, omega 3 fatty acid, omega 6 LA, and omega 3 ALA. Flavonoids - Mycertin, quercetin, kaempferol, caffeic acid, rosmarinic acid, chlorogenic acid, vitamins. Fibre, protein, amino acids and minerals.⁷

EXTRACTION PROCESS:

The study used Muñoz et al.'s (2012) method for mucilage extraction. In a beaker, place a whole seed weighing 20g in distilled water, then stir it and for 2 hours hydrate it. In an air convection heat oven, the drying tray was exposed to 50˚C and 10 hours after the suspension was filled. By using a 40-mesh screen, the dried mucilage was separated from the seed. An analysis was performed using AOAC methods, while the Anthrone method was used for total carbohydrate content determination.¹¹Chia seeds were extracted mucilage using the freeze-drying method, adding distilled water, and stirring at 45°C for about 30 mins. Then, the mixture was separated with the help of centrifugation at 11600 xg for about 30 mins. The leachate was then stored in the Lyophilizer-76 M for future use.²⁴The method involves grinding and drying chia seeds with a 1:20 dilution of distilled water. The mixture was stirred on a magnetic stirrer, separated by centrifuge, and was dried in the oven at 60°C for nearly 4-5 hours. The same process is applied to unmilled seeds.²⁴

PHYTOCHEMICAL SCREENING:

The phytochemical screening of different extracts of chia seeds was performed with the conventional method of screening, and results are given in Table 2.

Table 2: Phytochemical screening of different extracts of chia seeds

Test	Aqueous extract	n-hexane extract	Ethanol Extract	Chloroform extract	Ethyl acetate Extract
Alkaloids
Wagner’s test	+	-	+	+	+
Meyer’s test	+	-	+	+	+
Hager’s test	+	-	+	+	-
Dragondroff’s test	+	-	+	+	-
Carbohydrates
Barfoed’s test	+	-	+	-	-
Test for hexose sugar	+	-	-	-	-
Molish’s test	+	-	+	-	-
Fehling’s test	+	-	+	-	-
Test for pentose sugar	+	-	+	-	-
Benedict’s test	+	-	+	-	-
Saponins
Foam test	+	-	+	-	-
Phenols
Lead acetate test	+	-	+	-	+
Ferric chloride test	+	-	+	-	+
Glycosides
Fluorescence test	+	-	+	-	-
Keller killian test	+	-	+	-	-
Legal test	+	-	+	-	-
Modified brontrager’s test	+	-	+	-	-
Sodium picrate test	+	-	+	-	-
Fats and fixed oils
Saponification test	-	+	-	+	-
Spot test	-	+	-	+	-
Flavonoids
Shinoda test	+	-	+	-	+
Vanillin test	+	-	+	-	+
Ammonia test	+	-	+	-	-
Steroids and triterpenoids
Liebermann bur chard’s test	-	+	+	-	+
Salkovaski test	-	+	+	-	+

Chia seeds include phytochemicals such as phenolic elements and essential fatty acids. Quercetin, omega 3, myricetin, caffeic acid, omega 6 and kaempferol are among the active ingredients. The composition of phytochemicals is influenced by nutrient availability, soil conditions, environmental variations, cropping year or climatic changes. Dietary fibre (18-30%), antioxidants, high fat (30-33%), vitamins, proteins (15-25%), minerals and carbs (26-41%) are all present in chia seeds.⁸These are also known as a family of natural products that are crucial; these secondary metabolites frequently have a polyphenolic structure. The polyphenol group is made up of 2 benzene rings and 15 carbon atoms. The biochemical capabilities such as hepatoprotective, anti-cancer, antiviral, anti-inflammatory, antioxidant and antibacterial activities are present in flavonoids. These substances are already present in chia seeds, their synthesis is boosted by microbial infection. The flavonoids such as kaempferol, myricetin and quercetin are most frequently found in chia seeds.⁸This is an essential flavonoid. Antidiabetic, anti-cancer, antioxidant, cardioprotective, anti-inflammatory, neuroprotective and antibacterial actions are only a few of the pharmacological traits that Kaempferol exhibits. Nowadays researchers are paying a lot of attention to foods rich in kaempferol such as chia seeds have a lot of uses in the treatment of cancer. Studies show that to increase Kaempferol's low bioavailability and increase its bio-efficacy, it should be coupled with quercetin shown in (Fig 11).⁸It contains antioxidant capabilities that can help prevent several diseases by scavenging free radicals and LDL oxidation is reduced. Quercetin's (Fig 12) antioxidant properties enable the inhibition of lipid peroxidation and prevent the transformation of unsaturated fatty acids into free radicals. Furthermore, it also reduces inflammation by scavenging free radicals.⁸Myricetin (Fig 13) goes via autoxidation; it contains myricetin compounds, which makes it a pro-oxidant. Superoxide, a byproduct of autoxidation that happens in the presence of cyanide and is known to cause cellular damage, is produced. Since it can increase the hydroxyl radicalization by the interaction of iron ions and hydrogen peroxide. It is also a prooxidant. In DNA deterioration, radicals are produced and linked.⁸Hydroxycinnamic acid is present in caffeic acid (Fig 14) and, found in the human diet, is created through the secondary metabolism of food. Caffeine acid is essential from the biological and chemical perspectives on chia seeds. Chia seed caffeic acid has anti-inflammatory, anti-cancer, anti-hypertensive, antioxidant, and neuron-protective properties.⁸Omega 3α fatty acid (Fig 15) reduces triglyceride and cholesterol levels, promoting low blood pressure and cardioprotective, heart-related diseases, hepatoprotective, anti-diabetic, anti-inflammatory action, and protection against cancer, autoimmune diseases, and arthritis.^25-29. Chia seeds contain omega-6 fatty acids (Fig 16), which are very useful fatty acids that the body cannot synthesize and must be obtained through food. Chia seeds have about 20% of this fatty acid content. ^29-31

Fig. 10: Kaempferol Fig. 11: Quercetins Fig. 12: Myricetins

Fig. 13: Caffeic Acid Fig. 14: Omega 3 fatty acids

Fig. 15: Omega 6 fatty acids

Furthermore, chia seeds have a good content of alpha-linolenic acid (ALA), and omega-3 fatty acid which accounts for approximately 75% of all fatty acids in chia seeds.³²

SCREENING METHODS:

Thermal analysis and Fourier transform infrared spectroscopy (FT-IR):

On Bruker vector 22 the FTIR spectrum of xanthan gum and chia seed mucilage is recorded using KBr pellets. To get the thermograms differential scanning calorimetry (DSC) and thermogravimetric analysis (TG) were done under the nitrogen atmosphere which has a flow rate of 150 and 50mL/min. In every experiment, a sample of 3- 4±0.1mg was used. At a heating rate of 10˚C min^-1 using the TG the thermal stability study was performed. To determine the lower solution critical temperature with the help of Mettler Toledo star system 822e DSC measurements were carried out with a heating rate of 10˚C min^-L.

HPLC determination of monosaccharides:

Ribotta et al. using HPLC determined the uronic acids and monosaccharides content in S.hispanica mucilage. From Sigma-Aldrich, the Standard D-glucose, D-arabinose, L-rhamnose, glucuronic acid, D-xylose, galacturonic acid, D-mannose and D-galactose were purchased. The hydrolysis of mucilage was done to determine the uronic acids and neutral sugar composition. Firstly the sample was made to dissolve in 0.5M sulphuric acid and was heated for about 12 to 24 hours approx, at 95˚C and then it was neutralized in 0.2M NaOH. Now the volume was graduated to 50ml and through a nylon membrane, the supernatant was filtered. Now a known amount of neutral sugars and uronic acids was doped with a hydrolyzed mucilage sample for the purpose of quantification of sugars. In HPLC the hydrolyzates of 20µL were injected with the help of Shimadzu HPLC system with an autosampler and then a spectrophotometric detector was used for uronic acids and a refractive index detector for sugars. A flow rate of 0.5mL/min and a mobile phase of sulphuric acid 5mM was used to perform the separation of monosaccharide using BioRad HPX 87-H column at the temperature of 65˚C. Three times every sample was injected.³³

PHARMACOLOGICAL ACTION:

Anti-hyperlipidemia and anti-hypercholesterolemia:

Chia seeds and chia oil have shown impressive health benefits in studies with obese rats. They reduced oxidative stress, improved lipid metabolism, and prevented dyslipidemia and insulin resistance. Chia oil also had positive effects on fat cells. When given to pigs and rabbits, chia seeds increase healthy fatty acids in meat and enhance flavour. Mixtures of different seeds increase healthy fats in animals' bodies, providing additional immune and heart benefits due to their antioxidant properties. Chia seeds and oil offer promising ways to improve health and fight metabolic issues.³⁶

Anti-inflammatory property:

The inflammatory mediator’s control is done by n3 PUFA in Chia seed oil, suggesting it to compete with arachidonic acid for the incorporation into the membrane. Hence, generate slightly modified prostaglandins and eicosanoids viz., LTE5, LTB5, and PGE3, which induce a lesser extent of inflammation via reduced induction of COX-2.³⁷

Hypoglycemic Effects:

The author evaluated the protein quality of chia on glucose levels, lipid profile, and liver and intestine function in Wistar rats. They divided the chia into two portions: untreated and heated. After 14 days, various protein ratios were calculated, and the liver and intestine morphologies were determined. Results showed that chia-fed animals had lower glucose levels, triacylglycerides, and lower VLDL, LDL, and HDL concentrations. The liver weight was lower, and the thickness of intestinal muscle layers was high.^{38, 39}Ayerza et al. (2005) studied the effect of chia seed, a rich source of omega-3-ALA, on cholesterol, triacylglycerol, and lipid profile. They found that rats fed with chia seeds had reduced triacylglycerol content and increased HDL levels with increased x-3 fatty acid content. The study indicates that chia oil rich in ALA may be a suitable alternative for individuals who are allergic to fish or are vegetarians.⁴⁰

Antihypertensive and Antioxidant Potential:

Chia protein is beneficial for hypertension patients due to its antioxidant and ion-chelating properties, while chia seeds have anti-hypertensive and antioxidant properties, with globulin and albumin providing the highest antiradical activity against ABTS and DPPH. Chia seed exhibits the inhibitory action for angiotensin-converting enzymes. Studies have shown that chia has strong blocking abilities, ranging from 53.84% to 69.31%, indicating its potential as an antihypertensive agent.⁴¹

Anticancer Activities:

In a study by Espada et al. (2007), S. hispanica SaO and Carthamus tinctorius CO which are vegetable oil sources of ω-3 and -6 PUFAs, and a commercial diet as control were used. They analyzed fatty acids in neoplastic cells and found lower levels of arachidonic acid and eicosanoids compared to SaO and CO. SaO inhibited growth and metastasis in the tumor model.⁴²

Post-menopausal Effects:

FuxiaJin et al.'s 2012 experiment on 10 postmenopausal women showed an increase in alpha-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid levels after a seven-week supplementation period.⁴³

Hepatoprotection activity:

The effects of S hispanica seeds in an animal model of antitubercular drug-induced hepatic damage were studied at doses of 250, 500 and 1000mg/day for 21 days. Hepatotoxicity was induced by administering isoniazid with rifampicin at a dose of 100mg/kg each intraperitoneal. There was a significant decrease in liver enzymes and total bilirubin in the drug-treated and silymarin-treated groups when compared to the control. The hepatoprotective activity was further evidenced by the histopathological examination, which was like that of normal control.⁴⁴

Anti - Obesity effects:

Poudyala et al. (2012) found that chia seed treatment improved insulin sensitivity, glucose tolerance, and decreased visceral adiposity, hepatic steatosis, and inflammation without affecting plasma lipids or B.P. in rats.²⁹In a study, Luciana Tavares Toscano et al. (2015) found that chia supplementation significantly reduced body weight and waist circumference in overweight or obese individuals. They also observed a reduction in total cholesterol, VLDL-c, and HDL-c, but only in groups with abnormal initial values.⁴⁶The study concluded that chia consumption for 12 weeks promoted a discrete reduction in weight and waist circumference while enhancing lipid profile dependent on initial values. In obese rats, adding chia oil to their diet had positive effects on their muscles. It increases the levels of protective proteins and antioxidants, which help fight stress. Also, eating chia seeds, whether for a long or short time, improved energy metabolism and muscle function. Chia oil and seeds show promise for promoting muscle health in obesity.³⁶

Effect on the immune system:

Fernandez et al. (2008) noted the immune system effects of the chia seeds on 23-day-old Weanling male Wistar rats and found that body weight had no difference and even IgE when the chia was given in oil or seed form. The inclusion of chia in any form did not induce abnormal behaviour, diarrhoea, dermatitis, or other symptoms.⁴⁶

Relieve Dry Skin:

A small, randomized study by Jeong SK et al. and Purnamawati S et al found that applying chia seed oil 4% to dry skin for almost 8 weeks showed significant improvement in the conditions of normal individuals and those who have their skin itchy due to diabetes or kidney disease. The study suggests that applying chia seed oil immediately after bathing or showering can maximize its benefits.^{26, 47}

CONCLUSION:

Salvia hispanica is very useful and has a lot of medicinal properties. Scientists have proven with various experiments that it possesses activities like anti-hyperlipidemia, anti-hypercholesterolemia, anti-inflammatory properties, hypoglycaemic effects, antihypertensive, antioxidant potential, anticancer activities, post-menopausal effects, hepatoprotection, effects on obesity, anti-diabetic effects, effect on the immune system and relieve dry skin. Still further research is needed to explore the other pharmacological activities.

AUTHORS CONTRIBUTION:

All authors contributed equally.

CONFLICT OF INTERESTS:

None to be declared.

ACKNOWLEDGMENT:

The authors would like to thank Dr S.N.V.L Narasimha Raju, Chairman, The Oxford Group of Institutions, and The Oxford College of Pharmacy, for their kind support for the project work.

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Received on 17.10.2023 Revised on 12.06.2024

Accepted on 05.11.2024 Published on 28.01.2025

Available online from February 27, 2025

Research J. Pharmacy and Technology. 2025;18(2):912-918.

DOI: 10.52711/0974-360X.2025.00134

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