INTRODUCTION:

Traditional medicine is the application of health practises, knowledge, and beliefs through the use of therapies, manual techniques, and exercises derived from plants, animals, and minerals, either separately or in combination, to treat, diagnose, and prevent illnesses or preserve well-being. Any material with qualities that can be employed for therapeutic reasons or as building blocks for the manufacture of different medications is referred to as a medicinal plant¹. Basella rubra L., a perennial vine with rapid growth that can withstand intense heat, is a member of the Basellaceae family. It is also referred to as wine spinach, Malabar spinach, Indian spinach, Ceylon spinach, and other names. It is a perennial, fleshy, glabrous herb with several branches that climbs. Simple, alternating, petiolate, sub-succulent leaves that are cordate at the base and broadly oval in shape.

White or crimson blooms with a spike-like inflorescence and bracteoles longer than the perianth. Red, white, black, and globular fruits². Due to the mineral, protein, oil, carbohydrate, fibre, carotenoid, organic acid, and vitamin content of this plant, it is healthy. Malabar spinach can be used in place of regular spinach throughout the winter because it has comparable nutritional and therapeutic benefits. Since ancient times, anticancer, antiviral, antioxidant, anti-inflammatory, anti-cholesterol, anti-ulcer, anti-microbial, anti-hypoglycemic, wound healing, androgenic diseases have been treated with basella plant species' betacyanin, carotenoids, bioflavonoids, -sitosterol, and lupeol, which are also reported to have these properties. Leaf juice is used to relieve catarrh and is a safe laxative for kids, expecting mothers, and patients with urinary diseases³. A good source of calcium, iron, and vitamins A and C is Basella rubra. Red-stemmed varieties are very nutrient-dense. According to the "Unani" method, basella plant is sour, tonic, antipyretic, improves the voice, and is applied to burns⁴.

MATERIAL AND METHODS:

Collection, identification, and authentication of plant material:

The leaves of Basella rubra (herbarium number-XCH 40489) plant were collected from the surrounding areas of Kerala. It is dried under shade and made into coarse powder.

Identification:

The plant material collected was identified and authenticated by Scientist (Dr) S. Mutheeswaran, M. Sc, M. Phil, PhD, Xavier Research foundation, St Xavier’s college, Tamil Nadu, India.

Morphological and macroscopic features:

The crude medicines' macro- and microscopic characteristics were investigated. The plant matter was carefully pulverised for microscopic characteristics.⁵ Using sensory organs or a magnifying glass, the plant's organoleptic characteristics were investigated.⁶The external borders, apices, texture, external and internal marking, fracture, form, and size of these medicinal plants were among its organoleptic characteristics⁷.

Physiochemical constant:

To assess the quality and purity of the crude medication, ash values are calculated.⁸ The water-soluble extractive value, alcohol soluble extractive value,⁹ total ash content, water soluble ash content, and acid insoluble ash content of powdered leaves of Basella rubra were all determined¹⁰.

Preparation of extracts:

The leaves were cleaned, then dried. Every sample was ground up. Using a soxhlet extractor, accurately weighed 50 g of the relevant plant powder were placed in the thimble and extracted in turn using petroleum ether, chloroform, distilled water, and methanol. All of the extracts were concentrated and stored in an airtight container¹¹.

Preliminary phytochemical screening:

The richest bio-reservoirs of various phytochemicals are found, that are used to treat various diseases and ails. The phytochemical components of plants control their therapeutic qualities. Alkaloids, flavonoids, phenolics, tannins, saponins, steroids, glycosides, terpenes, and others are some of the significant phytochemicals^12,13.

Fluorescent analysis:

On a tiny slide that has not been touched by oil, a small amount of dry plant powder is deposited. The slide is then put inside the UV chamber so that visible light and both short (254nm) and long (365nm) ultra violet radiations can be used to observe the colour. Recorded is the hue seen when various chemicals are applied in various radiations¹⁴.

Total Phenolic Content Determination:

The assessment of crude extracts' overall phenolic content. The resultant solutions were allowed to stand for 15 minutes at room temperature after being combined with the extracts (0.1mL), 0.25mL of Folin-Ciocalteu Reagent (FCR), and 1.5mL of 10% Na2CO3. The absorbance was then measured right away at 725nm after they had been diluted to 5mL. Gallic acid standard solutions (0.1, 0.2, 0.3, 0.4, and 0.5mg/mL) were used to produce calibration curves. Using the same procedures and materials as for the creation of the standard and sample solutions, a blank solution was also created¹⁵.

Total Flavonoid Content Determination:

Using quercetin as the reference substance, TFC was calculated using the absorbance method. All extracts were combined into a test tube containing 0.8ml, 0.1ml of 10% Aluminium trichloride, 0.4ml of 1M potassium acetate, and 5 ml of distilled water. Use Quercetin as the standard and incubate the reaction mixture for 25 minutes at room temperature before measuring the absorbance at 415nm. Per gramme of extract, the amount of flavonoid was represented as mg Quercetin equivalent (QE)¹⁵.

RESULT AND DISCUSSIONS:

Morphology of Basella rubra leaves:

Basella rubra leaves are morphologically identified. It is a few metres long succulent, branching, silky, twining herbaceous vine. Green or purple stems are common. The leaves are 5 to 12cm long, stalked, fleshy, oblong or heart-shaped, tapering to a point with a cordate base. Axillary, solitary, 5-29cm long spikes are present. When fully grown, the fruit is juicy, stalkless, ovoid or spherical, 5–6mm long, purple, and contains just one seed. About 4 millimetres long and pink, the flowers are. (Figure 1)

Whole plant Leaf

Fig 1: Morphological characters of leaves of Basella rubra

Microscopy of Basella rubra leaves:

Using a microscope, the microscopic examination of Basella rubra leaves was carried out. Transversal views of the leaves reveal that the Basella rubra leaf is present of parenchymatous cells. The middle, which has a helical form, is where the vascular bundles are organised. There are a lot of clustered calcium oxalate crystals in the leaf's ground tissue. Few crystals are also present near to the meristele region; however, the majority of the calcium oxalate crystals are found in the narrow-winged extensions in the laminar region. Parenchyma cells appeared to be spongy. The figure 2 and figure 3 show a transversal segment of basella rubra leaves under 10X magnification.

(A) T.S of Basella rubra

(B) Paranchyma cells (C) stomata

(D) Vascular bundles (E) Helical vessels

Fig 2 (A – E): Transverse section of leaves of Basella rubra

(A) Oxalate crystals (B) Helical xylem vessels

(C) Fibres

Fig. 3: (A-C): Powder microscopy of leaves of Basella rubra (magnification 10X)

Determination of physiochemical constants:

The findings of the evaluation of the total ash, acid insoluble ash, water soluble ash value, and loss of drying of leaves of Basella rubra are shown in table 1.

Table 1: Physiochemical constant and leaf constant of leaves of Basella rubra

Loss of drying	Total ash	Acid insoluble ash	Water soluble ash
12.34%W/W	27.192%W/W	1.46%W/W	15.46%W/W

Extraction of plant materials:

The yield and appearance of various Basella rubra extracts were assessed, and the results are shown in the table.2.

Table 2: Yield of extract obtained from extraction of leaves of Basella rubra

Plant name	Type of Extract	Appearance/ State	Yield (% w/w)
*Basella rubra leaves*	Pet ether	Pale green / Semisolid	3.2% w/w
	Chloroform	Dark green / Semisolid	4.4% w/w
	Ethanol	Green / Semisolid	7.6% w/w
	Water	Dark Brown / Semisolid	12.4% w/w

The results of several research' observed values have allowed for the preliminary identification of species and the regular measurement of quality in the herbal sector. ¹⁶

Preliminary phytochemical screening of leaf extracts of Basella rubra:

Scientists have been interested in phytochemical as a result of the creation of new and advanced methodologies. The plant synthesises a wide range of chemical compounds that can be categorised by chemical class, biosynthetic origin on functional groups into primary and secondary metabolites. These techniques are important in providing solutions to systematic problems on the one hand and the search for additional resources of raw materials for pharmaceutical industries on the other.¹⁷

Steroids were found in petroleum ether and chloroform extracts, while flavonoids, carbohydrates, saponins, proteins, alkaloids, phenols, steroids, and tannins were found in ethanol and aqueous extracts, respectively (Table 3).

Table 3: Preliminary phytochemical screening of leaf extract of Basella rubra

Chemical tests	*Basella rubra* leaf extracts
Chemical tests	Pet ether	Chloroform	Ethanol	Water
Proteins and Amino acids	-	-	+	+
Carbohydrates	-	-	+	+
Steroids	-	-	-	-
Phenols	+	+	+	+
Saponins	+	+	+	+
Flavonoids	-	-	+	+
Alkaloids	-	-	-	-
Glycosides	+	+	-	-
Tannins	+	+	-	-

+ indicates presence and – indicates absence of phytoconstituents

Fluorescent analysis:

The chosen plant is ground into a coarse powder, treated with the necessary chemical reagents, and then examined with visible and ultraviolet light. (18) Tables 4 and 5 present the findings.

Table 4: Fluorescence analysis of powder leaves of Basella rubra

S. No	Treatment	DAY LIGHT	SHORT UV (254 nm)	Long UV (366 nm)
1.	Powder	Green	Green	Dark green
2.	Powder+ Water	Pale green	Green	Green
3.	Powder+1N HCl	Pale green	Pale green	Pale green
4.	Powder+1N H2SO4	Pale green	Green	Pale green
5.	Powder+1N HNO3	Green	Green	Pale green
6.	Powder+ Acetic acid	Green	Green	Red
7.	Powder+1N NaOH	Green	Green	Green
8.	Powder+1N Alc. NaOH	Green	Green	Green
9.	Powder+1N KOH	Green	Green	Pale green
10.	Powder+1N Alc. KOH	Green	Green	Pale Green
11.	Powder+ Ammonia	Pale green	Green	Pale green
12.	Powder+ Iodine	Reddish brown	Dark green	Dark green
13.	Powder+Fecl3	Brownish green	Green	Brownish green
14.	Powder+ Ethanol	Pale green	Green	Red

Table 5: Fluorescence analysis of various extracts of Basella rubra

SL No	Extracts	Daylight	UV Light
SL No	Extracts	Daylight	Short 254 nm	Long 365 nm
1	Pet ether	Green	Green	Red
2	Chloroform	Green	Green	Red
3	Ethanol	Pale green	Green	Red
4	Water	Yellowish brown	Green	Green

Total Phenolic and Flavonoid content:

Table 6 lists the findings of estimating the total phenolic and flavonoid Basella rubra aqueous, ethanol, chloroform, and petroleum ether extracts.

*Basella rubra* plant extract	Total phenolic content	Total flavonoid content
Ethanol	497.57±0.25	380.78±0.013
Water	89.76±0.98	8.76±0.76

CONCLUSION:

In the current study, the leaves of the Basella rubra plant were analysed for phytochemical screening, physiochemical parameter measurement, and pharmacognostic characterization. The chosen plants were confirmed, and macroscopic investigations were done to show their veracity and cleanliness. A microscopic analysis was performed. Physical-chemical tests such ash value, acid insoluble ash value, and extractive value were performed according to protocol. Additionally, fluorescence analysis was performed using various solvents. The significant phyto-constituents, which are well-known for their therapeutic potentials, were present as shown by phytochemical screening. The screening of leaves reveals a high phenolic content and a high flavonoid content, which may be caused by the presence of antioxidant-rich phenol, flavonoid, and tannin. Consequently, it may be said that our study gives the data for future research on the same plant.

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Received on 28.06.2023 Modified on 18.12.2023

Res. J. Pharmacognosy and Phytochem. 2024; 16(2):73-77.

DOI: 10.52711/0975-4385.2024.00014