Pharmacognostic Study of Butea monosperma a versatile medicinal plant

Meenakshi Sharma¹, Raj Kumari¹*, Prasoon Kumar Saxena²

¹ITS College of Pharmacy, Department of Pharmacognosy, Delhi-Meerut Road, Ghaziabad, Uttar Pradesh, India.

²Sunderdeep Pharmacy College, Department of Pharmacognosy,

NH-24, Delhi-Hapur Road, Dasna, Ghaziabad, Uttar Pradesh, India.

*Corresponding Author E-mail: rajkataria80@gmail.com

ABSTRACT:

Throughout human history, plants have been employed as medicine in traditional health care systems and are seen as a source of long, healthy human life. Various plant components, including the roots, leaves, stem, bark, flowers, and seeds, have been utilised to treat a variety of diseases. India has a wide variety of medicinal plants. Out of the many commonly used medical plants, Butea monosperma is a possible medicinal plant widely appreciated for its ability to treat elephantiasis and treat night blindness and other vision problems. Bark is a spicy, acrid, and bitter food and laxative. Its leaves are beneficial for eye conditions and have anti-inflammatory properties. Flowers are helpful for gonorrhoea, scorching urine, and irritation. It is a member of the Fabaceae family, which includes more than 150 genera and 1600 species.The current study was designed to go into greater detail on how macroscopical and microscopical characteristics of leaves and stems were viewed. In the current situation, when there are no regulatory regulations to control the quality of herbal pharmaceuticals, the physicochemical characteristics investigations offered reference information in regard to its identification parameters considered substantially in the path of acceptability of herbal drugs.

KEYWORDS: Anatomy, Herbal medicine, Morphology, Microscopy, Phytochemistry studies, Butea monosperma.

1. INTRODUCTION: 

Mankind had been blessed with lot of intelligence and memory power that has led to the discovery of many remedies and cures for diseases which are major stumbling block for his peaceful and healthy existence. These cures include the drugs of both synthetic and natural origin. The drugs from the natural source have played a vital role in maintaining the health of the kind since the ages¹. B. monosperma in India, the Fabaceae family member(Palash) grows profusely has diverse role in traditional medicine and is known for its oxidative properties. This is a beautiful tree with red-coloured flowers known as forest fire;the plant is useful in treating inflammation, sexually transmitted diseases like Gonorrhoea. Even there are claims in the Unani System of Medicines that the plant is having aphrodisiac, expectorant, tonic property^2,3.

The tree has been widely spread throughout the tropics, and it is a small, dry-season deciduous tree with a modest growth rate that can reach heights of only 15 metres (49 feet) when young. The leaves are tripinnate and have three leaflets that are each 10–20cm long and have an 8–16 cm petiole. Bright orange-red, 2.5cm long flowers are produced in racemes that can reach 15 cm in length^4,5.

The fruit is a pod that is 4-5cm wide and 15-20cm long. According to reports, the stembark has aphrodisiac, astringent, bitter, pungent, alliterative, and anti-obese properties. Roots can treat elephantiasis, as well as night blindness and other vision problems⁶. The qualities of leaves include astringency, tonicity, diuretic, antidiabetic, antidiarrheal, and antibacterial.In order to identify the drug, the current work aims to disclose the essential pharmacognostical and standardisation parameters of B. monosperma leaves^7,8.

2. MATERIAL AND METHOD:

2.1 Collection and Authentication of Plant Material:

Butea monosperma plant parts (leaf and petiole) were collected and authenticated from LVD college Raichur a voucher specimen (No. LVD/05/28) was deposited in the herbarium of the Faculty of Pharmacy. The collected plant parts (leaf and petiole) were dried under shade and powdered with mechanical grinder and stored in air tight containers and used for the further study.

Taxonomical Classification:The gross morphology of plant elements, such as leaf shape, stem type, flower form, fruit form, etc., is used in the ancient science of plant taxonomy to classify plants into comparable groupings^9,10. A list of the plant B. monosperma taxonomical classifications presented in Table 1.

Table 1: B. monosperma taxonomical classifications

1	Kingdom	Plantae
2	Subkingdom	Trachebionta
3	Clade	Tracheophyes
4	Division	Magnoliphyta
5	Class	Magnolipsida
6	Subclass	Rosidae
7	Order	Fabales
8	Family	Fabaceae
9	Genus	Butea
10	Species	monosperma
11	Binomial name	Butea monosperma
12	Synonymus	Butea frondosa Roxb, Erythrina monosperma Lam, Plaso monosperma Kuntze
13	Vernacular names of the plant: English
	English	Flame of forest, Bengal kino tree
	Hindi	Palash, Dhak, Tesu
	Sanskrit	Karaka, Bijasneha, Kamalasana
	Marathi	Palas
	Gujarati	Nim

2.2 Pharmacognostical Study:

2.2.1 Macroscopic and Microscopic Studies:Using accepted techniques, the macro-morphology of the leaves was investigated^11,12 Important physical characteristics were seen to facilitate identification. For documentation, standard microtechniques¹³ and representative diagrams were obtained using an inverted microscope¹⁴. For leaves and petioles, organoleptic characteristics such as colour, texture, flavour, and odour were identified.

2.3 Physico-chemical Evaluation:

The shade dried leaves were subjected tosize reduction to get fine powder (# 40 mesh size) and then evaluated for different physico-chemical parameters such as total ash value, water soluble ash, acid -insoluble ash¹⁵, extractive values, loss on drying¹⁶, moisture content, foaming index, swelling index and nutritive values of leaves were determined as per standard procedures¹⁷.

Analysis of fluorescence: The dried powder sample material from leaf was examined in visible light as well as at two separate ultraviolet light wavelengths, 254nm and 365nm¹⁸.

Making extractand doing a preliminary phytochemical analysis: Hexane, petroleum ether, chloroform, acetone, ethanol, and water were used to extract the powdered leaf material one at a time using the Soxhlet method. The preliminary phytochemical screening was conducted on these extracts.

2.4 HPTLC Fingerprint Profile: HPTLC according to Stahl's TLC approach,profile of the medication was created¹⁹. The dried and coarsely powderedleaf of B. monospermawere extracted with different solvents to obtained four extracts: petroleum ether, chloroform, acetone, and ethanol, to analysed by HPTLC. The extracts were applied in triplicate with band widths of 8mm using CAMAG Linomat V applicators on separate plates on TLC plates (10 ×10cm, precoated silica gel G60 F254, aluminium sheets). Using a previously saturated twin trough chamber, the chromatogram was developed up to a distance of 80mm at room temperature. To identify the optimal wavelength revealing the greatest number of compounds, the generated chromatograms (n = 21) were scanned at various wavelengths using the CAMAG TLC Scanner III with deuterium and tunguston lamps in absorbance/fluorescence mode.

3. RESULTS AND DISCUSSION:

3.1 Morphological study: B. monosperma tree leaves were trifoliate and complex. The lateral leaflets were circular at the apex and obliquely oval. The size ranges from 10cm by 15cm with a border of 10cm to 19cm. It has a rather strong feel; they resemble leather in certain ways, with a glabrescent outside and a silky, hairy interior. Petiolate leaves measure 7.3–18cm in length and have tiny stipules(Figure 1). All leaflets have reticulated venation, they become glabrous at age 15 and are exquisitely silky.

Figure 1: Leaves of Butea Monosperma

Figure 2: Transverse section of B. monosperma leaf midrib enlarged (10 x 5) showing: -parenchyma; Ep: epidermis; vascular bundle (phloem, X: xylem); Tricomes.

3.2: Microscopical characters of leaf:

B. monosperma leaves depicts unicellular, pointed hairs growing from the surface of the upper and lower epidermis. Dorsiventral structure with palisade and spongy tissue is visible in the leaf lamina. Heterogeneous cortex with sclerenchymatous and chlorenchymatous tissue was seen in the midrib area. Vascular bundles were arranged in a ring in the midrib and have bundle caps on the outside. Vascular bundles were lateral and conjoint. Parenchyma with deposits of yellow sticky substance can be seen in the midrib's centre. Phloem rings encircle the one xylem ring that was located toward the centre (Figure 2). The quantitative microscopical data of B. monosperma leaves were presented in the Table 2.

Powder study of leaves: The leaf powder exhibited fibres and vessel elements, fibres were long, thick walled, had narrow lumen and tapering ends. The fibres were 500 - 550 μm long. The vessel elementswere narrow, long and cylindrical; they were 400 – 900 μm long and 30 μm wide.Powder of leaf shows presence of gummy mass, unicellular pointed hairs, vessels with wall thickening and yellow (Figure3A-D).

Microscopical characters of Petiole: T.S of petiole of B.monospermashowed roundabout diagram. Inside it was separated into epidermis, cortex and stele. Hairs were unicellular and pointed emerging from surface of epidermis. External cortex was 1-3 layered and sclerenchymatous while internal cortex was parenchymatous. Stele was horse shoe or wedge molded in layout. Vascular packswith sclerenchymatous cap on the external side and were conjoint. Xylem vessels were organized in outspread groups and were of end arch type (Figure 4).

                              A                                                 B                                                        C                                                         D

Figure 3: Powder microscopy of leaves of B. monospermaA. yellow matter (40 x); B.Tricomes (40 x); C. Spiral vessel and fragment of cortex (10 x 2.5); D. fibers (Fi)(40 x);

Figure 4: Transverse section through petiole of B.monosperma leaf (10 x 5) showing: - Ep: epidermis; vascular bundle (P: phloem, X: xylem) Bundle Cap; yellow gummy mass.

Table 2: Quantitative Microscopical Data of B. monosperma Leaves

S. No.	Leaf Constant Parameters	Range	S. No.	Leaf Constant Parameters	Range
1	Stomatal number in upper epidermis	10-14	4	Vein islet number	5-8
2	Stomatal number in lower epidermis	27-32	5	Vein termination number	10-13
3	Stomatal index in upper epidermis /lower epidermis	15-20/21-29	6	Palisade ratio	2-4

Table 3: Physicochemical observation of B. monosperma leaves

S. No.	Parameters	Values	S. No.	Parameters	Values
1	Ash Value (%w/w): Total ash value	2.45±0.01	8	WaterExtractive values	12.72 ± 0.03
2	Acid insoluble ash value	1.89±0.02	9	Moisture content	1.99 ± 0.02
3	Water soluble ash value	0.67±0.01	10	Foaming index	<100
4	Extractive values (% w/w): Petroleum ether	0.87±0.01	11	Swelling index (%)	23
5	Chloroform Extractive values	0.61±0.02	12	pH value	6.1
6	Acetone Extractive values	2.21 ± 0.01	13	Loss on drying	2.00 ± 0.02
7	AlcoholExtractive values	8.92 ± 0.12

Table 3: Physicochemical observation of B. monosperma leaves

S. No.	Parameters	Values	S. No.	Parameters	Values
1	Ash Value (%w/w): Total ash value	2.45±0.01	8	WaterExtractive values	12.72 ± 0.03
2	Acid insoluble ash value	1.89±0.02	9	Moisture content	1.99 ± 0.02
3	Water soluble ash value	0.67±0.01	10	Foaming index	<100
4	Extractive values (% w/w): Petroleum ether	0.87±0.01	11	Swelling index (%)	23
5	Chloroform Extractive values	0.61±0.02	12	pH value	6.1
6	Acetone Extractive values	2.21 ± 0.01	13	Loss on drying	2.00 ± 0.02
7	AlcoholExtractive values	8.92 ± 0.12

Table 4: Fluorescence analysis of B. monosperma leaf powder

S. No.	Chemical Treatment	Observations
		254 nm	366 nm
1	1N NaOH in methanol	Light Green	Brownish Green
2	1N NaOH in water	Light Yellow	Dark Green
3	50% HCl	Light Green	Blackish Green
4	50% HNO3	Yellowish Green	Brown
5	50% H2SO4	Brownish Black	Dark Black
6	Hexane	Clear	Clear
7	Petroleum Ether	Green	Brown
8	Chloroform	White	Creamish White
9	Ethanol	Clear	Buff White
10	Drug as such	Creamish Yellow	Greenish Black

3.3 Physicochemical parameters: The physico-chemical characters of powdered drug of leaves of B.monosperma such as successive extractive values with petroleum ether, chloroform, alcohol, and water-soluble extractive, ash value, acid insoluble ash, water-soluble ash, loss on drying, and foreign matter were presented in Table3.

Fluorescence analysis: Color and fluorescence activity of powdered drug observed at 254, 365nm and visible light were given in Table 4.

2.3 Phytochemical Analysis: For the purpose of detecting different chemicals, consecutive extracts were taken in petroleum ether, chloroform, ethanol, and water, and the extractive value was estimated using the accepted practises outlined by Harborne¹⁷ and Khandelwal¹⁸Result shown in (Table 5).

3.4 HPTLC fingerprints of Butea monosperma extracts: Petroleum ether, chloroform, acetone and methanol successive extracts investigated by HPTLC for development of fingerprints. The chromatograms obtained after development in different solvent system followed by scanning at 254nm in absorbance mode depicted presence of number of substances in the extracts. chloroform, and ethanol showed presence of 2,13,16,17 spots respectively, with different R_f values as given in Table 6.

Table 5: Phytochemical screening of leaf extracts of B. monosperma

S. No.	Constituents	Hexaneext.	Pet.ether ext.	Chloroform ext.	Acetone ext.	Ethanol ext.	Aqueous ext.
1.	Alkaloids	-	-	-	-	+	+
2.	Carbohydrates	+	-	+	-	++	+++
3.	Glycosides	-	-	-	-	++	+++
4.	Phenolic compounds and Tannins	+	+	+	+	+	++
5.	Flavonoids	-	-	-	-	++	+++
6.	Proteins and free amino acids	+	-	-	-	-	-
7.	Saponins	-	-	++	-	++	-
8.	Sterols	+ +	++	++	++	+	-
9.	Amino acid	-	-	-	-	+	++

[+ Present ; – Absent]

The phytochemical screening of various extracts of leaf indicated the presence of carbohydrates, phenolic compounds and tannins, flavonoids, proteins and free amino acids, saponins, sterols etc.

Table 6: HPTLC fingerprint profile of Butea monosperma extracts

Extract	Solvent System	No. of Spots	Rf Values	Visualizing Agents
Petroleum ether extract of leaves	Toluene: Ethyl acetate: Glacial Acetic acid (9.5: 0.5: 0.2)	2	0.01, 0.26	Anisaldehyde in Sulphuric acid
Chloroform extract of leaves	Toluene: Ethyl acetate: Chloroform: Glacial Acetic acid (5.0:2.0:3.0:0.2)	13	0.00,0.06,0.09,0.20,0.22,0.35,0.45, 0.49, 0.54,0.61,0.68,0.78,0.85	Anisaldehyde in Sulphuric acid
Acetone extract of leaves	Toluene: Hexane: Glacial Acetic acid (6.0:4.0:0.1)	16	0.01,0.04,0.19,0.21,0.34,0.40,0.46,0.49,0.58,0.62,0.64,0.73,0.76,0.78,0.86,0.88	Anisaldehyde in Sulphuric acid
Ethanol extract of leaves	Toluene: Ethyl acetate: Chloroform: Glacial Acetic acid (4.0:3.0:3.0:0.5)	17	0.04,0.08,0.16,0.20,0.25,0.27,0.30,0.33,0.35,0.38,0.43,0.44,0.48,0.60,0.72,0.78,0.80	Anisaldehyde in Sulphuric acid

Chloroform 366nm

Chloroform 254 nm

Ethanol 366nm

Ethanol 254nm

CONCLUSION:

The bulk of crude medications used in the Ayurvedic medical system are derived from plants. Before using a plant, it is important to verify its authenticity and quality. Therefore, a thorough pharmacognostic examination is a crucial prerequisite. The petiole of B. monosperma has a half-moon-shaped or wedge-shaped stele, which is a distinctive characteristic (Lamk.) Taub. These characteristics could be viewed as anatomical traits that help identify the plant. The precise anatomy, physico-chemical parameters, and phytochemical ingredients of the drug, all of which were covered in this work, are some of the basic characteristics that the current pharmacognostic investigation of B. monosperma leaves helps us to understand.In preliminary and quantitative tests, it was shown that B. monosperma leaves include flavonoids, sterol, saponin, and phenols. The favourable results of these phytoconstituents point to the necessity for additional research on the isolation and characterization of active principle ingredients. The research of the separation of flavonoids and saponins was determined to be appropriate for the mobile phase. The makers will be assisted in identifying adulterants and standardising herbal formulations by the new phytochemical analysis.

The goal of the current work is to provide insight into the anatomical characteristics and phytochemical analyses of several plant sections belonging to B. monosperma (Lamk) Taub. These checklists will aid in species identification, and it may also be used to establish the botanical identity of herbal medicines. It can also be helpful for quantitative and qualitative standardisation of real drugs in herbal preparations. Positive results for flavonoids, phenols and saponins is indicative of scope for future analysis.

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Received on 10.09.2022            Modified on 07.05.2023

Accepted on 21.11.2023           © RJPT All right reserved

Research J. Pharm. and Tech 2023; 16(12):5828-5833.

DOI: 10.52711/0974-360X.2023.00944