The leavesof Chenopodium giganteum (herbarium number XCH — 40416) plant was collected from the surrounding areas of Meghalaya. It was dried under shade and made into coarse powder. The plant material collected was identified and authenticated by Scientist (Dr) S. Mutheeswaran, M.Sc., M.Phil., Ph. D., Xavier Research Foundation, St Xavier’s College, Tamil Nadu, India.

The fresh leaves of Chenopodium giganteum were examined for various macroscopic features like colour, odour and taste of leaves. Other external morphological characters like surface, base, margin, size, and shape of leaves were also studied. The microscopic examination of Chenopodium giganteum was done with the help of microscope. The air-dried plant material was then crushed into a coarse powder and used for further research work. The stomatal number and stomatal index of leaves of Chenopodium giganteum was also determined and evaluated by the methods referred from textbook authored by Kokate, Purohit and Gokhale. 19

The physical constants like ash value and extractive value helps in establishing the pharmacopoeial standards of the drug. The leaves of Chenopodium giganteum were examined for Physico-chemical constants like loss on drying, ash value and extractive value as per the methods mentioned in Pharmacopoeias. 20, 21

Successive Solvent extraction: Leaf of Chenopodium giganteum was dried and milled to a coarse powder. One kg of fresh plant material was grounded and defatted using petroleum ether. About 50g of the powdered air-dried defatted plant material was extracted subsequently with chloroform, pet ether, ethanol, and water in a Soxhlet apparatus. Each time before extracting with the next solvent, the marc was air dried below 50°C. The extracts were filtered, and the solvent obtained was evaporated at room temperature and accurate weight of the extracts was taken. The extractive value (%) was calculated with reference to air dried drug.

Preliminary phytochemical investigations of pet ether, chloroform, ethanol and water extracts were done to reveal the presence of different secondary metabolites like Proteins & Amino acids Carbohydrates, Steroids, Phenols, Saponins, Flavonoids, Alkaloids and Tannins. 22, 23

Fluorescent analysis of both extracts and plant powders of Chenopodium giganteum leaves were carried out according to the method mentioned by Chase and Pratt et al. (1949) and Koshiet al. (1958) in day light and in UV light (254 nm and 365 nm). The plant powders and extracts were treated with different solvents and the fluorescence was observed in day light and in near and far UV light. About 10g of drug powder was taken in a petri dish and treated with different reagents and observed under different wavelengths i.e., ultraviolet and visible rays. 24

Using gallic acid as a standard, the total phenol concentration was calculated using the Folin-Ciocalteu test. In this process, 1.5 ml of Folin-reagent Ciocalteu's (FCR) diluted 1:10 v/v and 0.5 ml of plant extracts were combined. After 5 minutes, 1.5 ml of a solution containing 7% sodium carbonate was added. With distilled water, the final volume was made up to 10 ml, and they were then left to remain at room temperature for 90 minutes. A spectrophotometer was used to test the sample's absorbance at 750 nm in comparison to the blank. The entire experiment was done three times for accuracy and data expressed as mean standard deviation in terms of phenol content (gallic acid equivalent, or GAE) per g of dry weight.

Using quercetin as a standard, the total flavonoid content was calculated using the aluminium chloride technique. In this process, 4 ml of water and 1 ml of plant extracts were combined. After 5 minutes, 0.3 ml of 10% Aluminum chloride, 0.3 ml of 5% Sodium nitrite and 1ml of 1 M Sodium hydroxide was added to the reaction mixture after the mixture had been incubated at room temperature for 6 minutes the final volume was made up to 10 ml with distilled water. A spectrophotometer was used to test the sample's absorbance at 510 nm in comparison to the blank. The entire experiment was done three times for accuracy, and data were expressed as mean standard deviation in terms of flavonoid content (Quercetin equivalent, or QE) per g of dry weight. 25

For TLC experiments, one gram of extracts was dissolved in methanol, filtered, and used. On aluminium plates that had already been coated with silica gel G, about 6µg of Chenopodium giganteum extract were applied. In a TLC chamber, the plate was produced using several solvent systems. Utilizing a photo documentation equipment, produced plates were seen and documented in both short and long UV. 26

The morphological data of Chenopodium giganteum showed the colour of young leaves was pink, magenta colour and older leaves were green with smooth under surface as shown in Figure 1. It has odour and bitter characteristic taste. The shape of leaves was extremely variable as simple, deltoid, ovate to lanceolate, upper entire, rhomboid, lower toothed or irregularly lobed. The size of leaf was about 1-3 cm; petioles were of 1-2 cm in length and as long as thick blade. Its length varied from 9 to 4.5 cm broad having dentate margin. It has acute apex and upto 5 cm base.

The microscopic study of Chenopodium giganteum leaves was done with the help of microscope. The transverse section of the leaves reveals that the Chenopodium giganteum leaf is dorsiventral with palisade cells pointing upward toward the top epidermal layer. Nearly half of the leaflet is covered by the lengthy palisade cells. In the upper epidermis, there were spherical, thin-walled collenchymas in place of the thick-walled parenchymatous cells that were present there. The palisade layer is broken in the midrib area underneath the top epidermis by collenchymatous cells. Near the vascular bundles, thick-walled parenchymatous cells revealed the existence of spiral and annular vessels. While the cells in the lower epidermis are spherical, those in the upper epidermis are flattened. In the midrib area, behind the collenchyma cells, the vascular bundles are organized like a bunch of grapes. The transverse section and powder microscopy (magnification 10X) of leaves of Chenopodium giganteum is given below in Figure 2 and Figure 3.

Figure 3

The total stomatal number and stomatal index of upper epidermis of leaves was found to be 12 and 22.2% respectively and lower epidermis was found to be 11 and 12.5% respectively.

The total ash, acid-insoluble ash and water-soluble ash value and loss on drying of leaves of Chenopodium giganteum were evaluated and the results are mentioned in Table 1.

The appearance and yield of different extracts of Chenopodium giganteum were evaluated and the results are mentioned in Table 2.

Petroleum ether and chloroform extracts revealed the presence of steroids whereas ethanol and aqueous extracts indicated the presence of flavonoids, carbohydrates, saponins, proteins, alkaloids, phenols, steroids, and tannins respectively (Table 3).

The selected plant is made into course powder and treated with required chemical reagents and observed under visible and ultraviolet rays. The results are given in Table 4 and Table 5.

The total phenolic and flavonoidal content for aqueous, ethanol, chloroform, and petroleum ether extracts of Chenopodium giganteum were estimated and results are given in Table 6.

Table 6

Chenopodium giganteum plant extract	Total phenolic content	Total flavonoid content
Pet ether	14.477±0.0226 µg/ml	17.2227±0.0729 µg/ml
Chloroform	17.764±0.0216 µg/ml	23.7224±0.0878 µg/ml
Ethanol	19.518±0.0173 µg/ml	49.8601±0.0303 µg/ml
Water	27.686±0.0233 µg/ml	64.7705±0.0375 µg/ml

The extracts were undertaken for TLC profiling to assess the nature of phytochemicals present in it. A number of developing solvent systems were tried for all the extract and fractions. The solvent system, which gave the best resolution, was considered optimized, valid, and useful. The satisfactory resolution was obtained in the mobile phase mentioned in Table 7 and photo documentation is shown in Figure 4. The Rf values of different extracts was also calculated and are mentioned in Table 8. Blue and orange spots were observed which indicated the presence of phenolic compounds. 27

Figure 4

In the current study, Chenopodium giganteum plant’s leaves were examined for pharmacognostic characterisation, physiochemical parameter measurement, phytochemical screening, and TLC examinations of the crude extracts. The chosen plants were verified, and to prove their authenticity and purity, macroscopic examinations were carried out. To ascertain the fundamental cellular makeup of the leaf petiole, stem, kind of stomata, etc., a microscopic investigation was conducted. Physicochemical studies were carried out as per standard procedure such as ash value, acid insoluble ash values and extractive values. Fluorescence analysis was also carried out using different solvents. The important phyto-constituents were present as depicted in phytochemical screening which are well-known for their medicinal potentials. The screening of leaves indicates the presence of high phenolic content and high flavonoid content which may be due to presence of phenol, flavonoid and tannin which possess antioxidant properties. Identification of the numerous phytochemical ingredients found in the raw medicine is aided by the TLC fingerprint profile. The TLC fingerprint profile aids in locating significant phyto-constituents as well. Thus, it is concluded that our study provides the data which helps to isolate, identify, and characterize the various medicinal potential of Chenopodium giganteum leaves.