INTRODUCTION:

The wild edible plants are essentially crucial for biochemical and nutritional aspects. They produce a wide range of compounds to protect themselves against pathogens. The Antimicrobials derived from plants are a vast untapped source of medicines that has widespread therapeutic promise and effectiveness in the treatment of infectious diseases; further research on plant antimicrobials is required¹.

They contain rich sources in amino acids, carbohydrates, fibers, vitamins, and active compounds like antioxidant and antimicrobial compounds. These plants are valuable sources to fulfill the daily requirements of rural people. Upon consumption of these vegetables and plants helps in fighting against multiple diseases^2-5. Among such plants, Natsiatum herpeticum Buch.-Ham. ex Arn. and Sphenoclea zeylanica Gaertn. are two commonly known plants for their medicinal properties. N. herpeticum belongs to the family Icacinaceae. It has twining climber, young branches yellow-brown strigose-hairy, mature stems conspicuously lenticellate. These plants are a rich source of Camptothecin. Camptothecin (CPT) has become this century's third most crucial anti-cancer chemotherapeutic. However, there are other medicinal properties that this plant retains. The N. herpeticum also has shown other medicinal properties like anti-malarial and anti-bacterial properties. It is found in the Northeastern part of Baksa district, Assam. This plant's young shoots are essential for constipation and other gastrointestinal problems. The whole plant is used for various other ailments.

The S. zeylanica is an herbaceous annual plant. It belongs to Sphenocleaceae. It grows up to 6-10 inches in height and grows in marshy land, paddy field, or water banks. It is a seasonal herb. It is an erect annual herb, and the inflorescence is a dense, terminal spike with small, greenish-yellow flowers. It thrives in damp ground and prefers stagnant water. It is locally known as Sibung in Bodo in Assam (India). This plant is readily available in the months from July to October. The stems and leaves are fragile, with a smooth leaf margin. S. zeylanica showed allelopathic properties in plant growth inhibitor compounds isolated from it. It was reported that these inhibitory compounds restrict the growth of roots of rice saplings at a 3.0mM concentration⁶. This plant showed antimicrobial properties in methanol extract against some pathogenic bacteria⁷. The S. zeylanica has shown antimicrobial activity due to different secondary metabolites. So, this plant can be used to identify the specific bioactive natural products that may lead to the development of new antimicrobial agents. The bioethanol production from the dry powder of S. zeylanica was also reported with an ethanol concentration of 11.84g/L⁸.

1. Phytochemical and chemistry screening:

a. Phytochemistry:

The preparatory study of phytochemical components existing in N. herpeticum and S. zeylanica plants was performed using methanol extract⁹. The phytochemical constituents investigation shows the presence of alkaloids, saponins, cardiac glycosides, steroids, coumarins, phenols, tannins, flavonoids, anthocyanins, phlorotannins, carbohydrates, proteins, and lignin¹⁰ though proteins absent in S. zeylanica. The proximate analysis results of N. herpeticum show that the moisture, ash, fat, fiber, protein, carbohydrate content, and calorific value are 83.72g, 3.42g, 0.666g, 1.99g, 5.37g, 6.80g, 54.72 kcal per 100g of fresh weight respectively. For S. zeylanica, these values are 92.81g, 1.14g, 0.124g, 1.25g, 3.08g, 2.55g, and 23.70kcal per 100g of fresh weight^10-12. Foods with high content of fibers are essential for digestion and the effective elimination of wastes. Fibers can lower serum cholesterol, risk of coronary heart disease, hypertension, constipation, diabetes, and colon and breast cancer¹². Carbohydrates, fats, and proteins are the essential nutrients of life. High carbohydrate content indicates high energy content in food. The primary function of carbohydrates in the body is to supply energy, and it is responsible for doing activities in our daily life^9,12. Quantitative phenolic, flavonoid, and vitamin C contents of N. herpeticum and S. zeylanica are estimated. The total phenolic content in the methanol extract of N. herpeticum and S. zeylanica plants are 10.45mg and 36.39mg GAE (gallic acid equivalents)/g DE (dried extract), respectively. For flavonoid, these values are 0.29mg and 0.23mg QE (quercetin equivalents)/g DE, and for vitamin C, these values are 85.71mg and 40.37mg/100g FW (fresh weight), respectively¹⁰. The chemical constituents present in the plant S. zeylanica are (1S,3R,4R)-(+)-4-hydroxy-3-hydroxy-methyl-1,2-dithiolane-1-oxide, (1R,3R,4R)-(y)-4-hydroxy-3-hydroxy-methyl-1,2-dithiolane-1-oxide, (2R,3R,4R)-(y)-4-hydroxy-3-hydroxy-methyl-1,2-dithiolane-2-oxide, and (2S,3R,4R)-(+)-4-hydroxy-3-hydroxy-methyl-1,2-dithiolane-2-oxide¹³.

b. Metal contents:

Quantitative metal contents of N. herpeticum and S. zeylanica were detected. Potassium is one of the essential nutrients which plays various biophysical and biochemical roles. Potassium-riched plant foods are generally used to treat rheumatoid arthritis and heart disease^14,15.Magnesium is vital in the ionic balance and enzyme cofactors, and calcium is required to build skeletal structures and muscle functioning¹⁶. Iron is needed for the normal functioning of the central nervous system and for synthesizing hemoglobin in RBC, which is required for oxygen transportation to all body parts. Iron deficiency is linked to anemia and causes immune system dysfunction, associated with an increased risk of infection^9,17. When dietary iron consumption is sufficient, about 15% of the body’s iron stored for future supplies is mobilized¹⁸. Copper plays a vital role in biological electron transport and is essential for the production of enzymes in the body. The deficiency of copper leads to reduced energy production, abnormal glucose, and cholesterol metabolism, and increased oxidative damage^9,19. The suggested dietary allowance for Zinc is 8 mg and 11 mg per day for adult women and men, respectively²⁰. Zinc is a necessity element for human growth, which increases resistance to infection, and excessive zinc intake has been reported to be toxic ^9,21. Zinc is an essential element for the antioxidant enzyme superoxide dismutase. It is required to function over 300 enzymes and many enzymatic reactions involved in carbohydrate and protein metabolism¹⁸. Manganese is an important element of metalloenzymes and plays an essential role in several physiological processes as a constituent or activator of some enzymes, which are necessary for the metabolism of carbohydrates, cholesterol, and amino acid¹⁹. The metal contents per 100g dried of N. herpeticum and S. zeylanica are Na (92.15mg and 48.73mg), K (8436.10 mg and 7684.29mg), Ca (5.47mg and 6.22mg), Mg (4.58mg and 4.90mg), Fe (2.48mg and 1.83mg), Cu (2.10mg and 2.58mg), Zn (0.70mg and 0.70mg ), Mn ( 0.70mg and 0.54mg), Ni ( 4.70mg and 2.60mg), Cr ( 0.74mg and 0.58mg), Co ( 0.59mg and 0.39mg), Se (0.78mg and 0.81mg), Pb (0.75mg and 0.59mg), Cd ( Below detectable level ), As ( Below detectable level ) respectively¹⁰.

c. Amino acid analysis:

It was observed that a total of sixteen non-essential and essential amino acids were detected in S. zeylanica. The total amino acid present in S. zeylanica was 42.87mg/g. The complete essential and non-essential amino acids found in S. zeylanica are 13.17mg/g DW). The total non-essential amino acids (NEAA) were higher in S. zeylanica at 6.40mg/g DW. The asparagine range was 2.86mg/g, which was the highest, followed by alanine (1.53mg/g) and aspartic acid at 0.90mg/g²². The glutamic acid was 0.39mg/g in S. zeylanica²³. The serine was found at 0.18 mg/g by S. zeylanica²⁴. The Alanine content was detected as 1.53 mg/g in S. zeylanica, and the proline concentration was found to be 0.42mg/g in S. zeylanica²⁵. Arginine was observed only in S. zeylanica as 0.06mg/g DW²⁶. Essential amino acids (EAA) content was detected in S. zeylanica at 6.77mg/g DW, the valine at 2.32mg/g, followed by leucine at 1.65mg/g and isoleucine at 1.57mg/g, and lysine was detected the lowest in the plant with the range of 0.01mg/g²⁷. The aromatic amino acid viz. phenylalanine was higher in S. zeylanica at 0.60mg/g²⁸. Threonine was increased in S. zeylanica by 0.39mg/g²⁹. Norvaline was obtained at 6.88 mg/g in S. zeylanica Choi et al.³⁰. Ethanolamine, β-amino butyric acid, and α-amino adipic acid were also found in S. zeylanica Choi et al.³⁰.

d. Anti-nutritional content:

High oxalate content was found in S. zeylanica 11.02 mg/g DW. . In the human body, a high concentration of oxalate forms insoluble calcium oxalate complex which hinders the absorption of soluble calcium ions and as a result, the kidney stone is formed³¹. The oxalate due to its water solubility can be removed by washing, boiling, and soaking methods³². Tannin content was found in S.zeylanica 2.74mg/g DW. The complex nature of structures of saponins present in the plants, they impart several chemical and biological properties such as bitterness, foaming and emulsifying, hemolytic, insecticidal and antimicrobial properties³³. The saponin content was found as 11.00mg/g in S. zeylanica. Alkaloids are organic molecules containing nitrogen of plant origin. They have antiphysiological and pharmacological effects on humans and animals. Some plant alkaloids have also been reported to cause infertility. Alkaloids when consumed in excess cause gastrointestinal upset and neurological disorders^34,35. The alkaloid content was found as 1.68mg/g in S. zeylanica.

2. Pharmacological properties:

a. Antioxidant property:

DPPH free radical scavenging activity was investigated in the methanolic extract of N. herpeticum and S. zeylanica. The DPPH method is widely used to see the antioxidant property of plant extracts. A stable free radical DPPH has a characteristic UV-Visible absorption at 517nm. The reaction of antioxidants molecule present in the plant extracts with DPPH converts 1,1-diphenyl-2- picrylhydrazyl (DPPH, deep violet color) to 1, 1-diphenyl-2-picrylhydrazine which is a stable molecule (yellow color or bleached product) by gaining an electron or hydrogen radical at a very rapid rate which leads to decrease in absorbance at 517nm⁹. The IC₅₀ value is defined as the inhibitory concentration of the crude extract that scavenges 50% of reactive oxygen species or inhibits the oxidation process by 50%. It is oppositely related to antioxidant capacity, and a lower IC₅₀ value signals better antioxidant activity. The methanol extract of N. herpeticum and S. zeylanica showed DPPH free radical scavenging activity, and the antioxidant activity increased with increasing concentration of sample extract. The result indicates that N. herpeticum has 24.85% DPPH radical scavenging activity with an IC₅₀ value of 1658.47μg/mL whereas S. zeylanica has 46.25% DPPH radical scavenging activity with an IC₅₀ value of 519.90μg/mL. In contrast, the standard ascorbic acid showed 98.84% inhibition with an IC50 value of 25.01μg/mL¹⁰.

ABTS free radical scavenging activity of methanolic extract of N. herpeticum and S. zeylanica was also performed. ABTS IC₅₀ value is the plant's actual concentration of methanol extract in µg/mL, inhibiting the ABTS activity by 50%. The ABTS+ radical is a stable free radical that accepts an electron or hydrogen radical from antioxidant compounds and becomes a stable molecule. This stable ABTS or ABTSH molecule inhibits the initiation or propagation of the free radical chain of oxidation. It has been reported that if the molecular weight of phenolic (tannins) is very high, they have more potential to quench free radicals (ABTS+), and their efficacy depends on the molecular weight, several aromatic rings, and character of the hydroxyl group's substitution than the specific functional groups¹⁶. The ABTS radical scavenging activity of N. herpeticum was 54.27%, with an IC₅₀ value of 283.23 μg/mL and these value for S. zeylanica was 91.71%, with an IC₅₀ value of 115.99 μg/mL. Trolox was utilized as standard in the ABTS assay and displayed an IC₅₀ value of 73.67 μg/mL¹⁰.

Hydrogen peroxide, a non-radical oxidant molecule, is present in living organisms. However, H₂O₂ is not a very reactive molecule; sometimes, it may cause poison to the body because it has the potential to penetrate cell membranes which may give rise to hydroxyl radicals and singlet oxygen which initiate oxidation in the cells ³⁶. Therefore, natural antioxidant sources pull out H₂O₂ from the body and protect the biological system. Its scavenging activity of methanolic extract of N. herpeticum and S. zeylanica plants was determined with the standard ascorbic acid. The percentage of scavenging activity of methanolic extract of N. herpeticum was found at 37.12 %, with an IC₅₀ value of 38.54 µg/ml, whereas for S. zeylanica, it was found at 8.51 %, with an IC₅₀ value of 123.19 µg/ml while standard ascorbic acid exhibited an IC₅₀ value of 19.02 μg/ml¹⁰. Food polyphenols have been displayed to defend mammalian and bacterial cells from cytotoxicity induced by hydrogen peroxide, significantly compounds with the orthodihydroxy phenolic structure, catechin, quercetin, caffeic acid ester, and gallic acid ester³⁷.

Another method, the Ferric reducing antioxidant power (FRAP) assay, was used to investigate the antioxidant activity of the wild plants in this study. The Ferric reducing antioxidant power (FRAP) assay is a simple, low-cost, and widely utilized method mainly employed for the evaluation of antioxidant activity and is based on the capability of antioxidants for reduction of ferric (III) ions to ferrous (II) ions³⁶. It is another antioxidant activity used to study wild plants. A higher FRAP value indicates higher antioxidant activity. The FRAP value for N. herpeticum and S. zeylanica was 60.01 µM TE/g DE and 38.57 µM TE/g DE¹⁰. The increased activity may be due to the antioxidant compounds in plants' methanol extracts, which could react with free radicals to stabilize and stop radical chain reactions. Free radical scavenging is an essential function of antioxidants as free radicals encourage oxidation of proteins, DNA, and lipids, resulting in disturbances and functional loss of biological membranes and enzymes and ultimately producing toxins³⁸.

b. Antimicrobial activity:

The antimicrobial activity of the methanol extracts of the leaf, stem, and flower of S. zeylanica at different concentrations was studied using the organisms B. subtilis, P. Vulgaris, S. aureus, E. coli, C. Albicans, A. niger, S. Typhi and B. cereus^39-41. It was seen that the inhibition displayed by the extracts of concentrations 50, 60, and 70 was higher than the inhibition shown by control in some cases. No inhibition was seen at the lowest concentration except for the inhibition of B. cereus by the leaf extract. The leaf extract showed maximum antimicrobial activity against the organisms at the highest concentration. The most susceptible organisms to the leaf extract were found to be P. Vulgaris, C. Albicans, A. niger, and B. cereus. The least vulnerable organisms were S. aureus and E. coli. The stem extract of S. zeylanica had the lowest antimicrobial activity compared to the other two extracts. Thus, it can be concluded that the leaves of S. zeylanica have significant antimicrobial activity⁴².

3. CONCLUSION:

In this study, two plants were used for the phytochemistry and pharmacological review, S. zylenica and Natsiatum herperticum. The phytochemistry studied shows the presence of alkaloids, saponins, cardiac glycosides, steroids, coumarins, phenols, tannins, flavonoids, anthocyanins, phlorotannins, carbohydrates, proteins, and lignin in both the plant. The total phenolic content in the methanol extract of N. herpeticum and S. zeylanica plants are 10.45 mg and 36.39mg GAE (gallic acid equivalents)/g DE (dried extract), respectively. For flavonoids, these values are 0.29mg and 0.23mg QE (quercetin equivalents)/g DE respectively. The alkaloid content was found as 1.68mg/g in S. zeylanica. Both plants show high content of potassium, 8436.10mg, and 7684.29mg, respectively, per 100g of dried samples. The total amino acid present in S. zeylanica was 42.87mg/g. The complete essential and non-essential amino acids found in S. zeylanica are 13.17mg/g DW. The antioxidant property of N. herperticum shows 24.85% DPPH radical scavenging activity with an IC50 value of 1658.47μg/mL, whereas S. zeylanica has 46.25% DPPH radical scavenging activity with an IC₅₀ value of 519.90 μg/mL. The ABTS radical scavenging activity of N. herpeticum was 54.27%, with an IC₅₀ value of 283.23 μg/mL, and the value for S. zeylanica was 91.71%, with an IC₅₀ value of 115.99μg/mL. The S. zeylancia shows maximum inhibition in B cereus at the lowest concentration of leaf extract.

4. ACKNOWLEDGEMENT:

The authors are thankful to Director General and Deputy Director General (Technical), Central Council for Research in Ayurvedic Sciences, New Delhi under Ministry of Ayush, Govt. of India for providing necessary support in preparation of this article.

5. CONFLICT OF INTEREST:

Author has no conflict of interest.

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Received on 08.09.2022 Modified on 09.02.2023

Research J. Pharm. and Tech. 2024; 17(3):1403-1407.

DOI: 10.52711/0974-360X.2024.00222