INTRODUCTION:

Acacia catechu Willd. or Senegalia catechu (Fabaceae family; Mimosoideae subfamily) is also known as Catechu, Cachou and Black Cutch. It is a deciduous, thorny tree with height up to 15m and widely distributed throughout India (Sub-Himalayan tract of Punjab to Assam, and dried parts like Madhya Pradesh, Maharashtra, Gujarat, Bihar, Rajasthan and Tamil Nadu and eastern slopes of Western Ghats), Ganjam, Burma, Konkam and Deccan¹. Plant is well known source for katha extracted from heartwood.

Plant is rich in tannins such as catechin and catechutannic acid, acacatechin, epicatechin, catechin tetramer, dicatechin, gallocatechin, gossypin, kaempferol and dihydro derivative, taxifolin, procyanidine, isorharnnetin, (+) afzelchin and flavonoids like quercetin^2,3. The seeds of the plant yield oil (3.5%) mainly consisting of neutral lipids (55%) and polar lipids (44.5%). The major fatty acids are oleic and linoleic acid⁴. The leaves contain trace elements such as Cu (8.90ppm), Fe (126.08ppm), Mn(25.31ppm) and Zn (24.26ppm) on dry matter basis⁵. Plant is used for antipyretic, hepatoprotective, hypoglycaemic, antidiarrhoeal^6-8, antimicrobial activity⁹, antileprotic¹⁰, anti-mycotic¹¹, hypotensive¹², antifertility¹³, and immunomodulatory activity¹⁴. Keeping in view the shortcomings related to synthetic drugs more researches have been conducted in the field of herbal drugs over the past few years which constitute a large part of the current pharmacopoeia. Till now, no scientific eye has been opened for establishing its pharmacognostical and phytochemical parameters using standard procedures. The present study was done for establishing pharmacognostic and physicochemical standards of the plant stem using pharmacognostical, chemical and analytical means.

MATERIALS AND METHODS:

Plant material and extract preparation: Stem of A. catechu Willd. was collected from the Sadhora forest region, Yamunanagar, Haryana. The plant material was identified by Dr. Sunita Garg (Emeritus Scientist, CSIR-NISCAIR, NewDelhi) under a voucher specimen number- NISCAIR/RHMD/Consult/2018/3295-96-1dated Nov. 28, 2018. Further, specimen was submitted in Department of Pharmacognosy, Hindu College of Pharmacy, Sonepat, Haryana for reference (AC 01). The stem was minced and dried under shade at room temperature. The dried stem was crushed into coarse powder for further studies and packed in vacuum container.

The plant material so obtained was undergone through soxhlation by using various solvents according to their polarity viz. petroleum ether (60-80˚C), chloroform, ethanol (95%) and water. Extracts were dried under rota-evaporator and stored in a desiccator for further use. All the chemicals and reagents were procured from RFCL, Mumbai, India.

Organoleptic Studies: Colour of the sample drug was observed under day light. Size and shape were also noticed. Texture of the drug was observed by touching the surface. Powdered drug was rubbed between two fingers for observation of odour^15,16

Microscopic Studies: Microscopic evaluation of thin, free hand transverse section and fine powder (passed through #60) of stem was done using various mounting medium and stains like phloroglucinol, hydrochloric acid, safranin, ruthenium red and glycerine. Microscopic photographs were taken using Labomed ATC-2000 microscope attached with Sony camera.^15-19

Physicochemical Studies: Water-soluble extractive value, alcohol-soluble extractive value and successive extractive values were determined according to standard procedures. Moisture content, swelling index, total ash content, water soluble ash value, acid-insoluble ash value has been determined as per procedure given in IP^15-22. Preliminary phytochemical screening was carried out by using standard methods to identify the presence of various phytoconstituents^23-27. Fluorescence study was performed as per standard procedure^28,29. Heavy metal analysis, aflatoxin content, microbial contamination and pesticide residue were assessed for plant as per WHO guidelines^18,30.

RESULTS AND DISCUSSION:

A. Pharmacognostic Studies:

a) Morphological studies: Heartwood of the plant stem was brownish red which was surrounded by brownish white sapwood. Fracture was hard and fibrous. Trunk was cylindrical in shape with fibrous texture and rough to touch. Size and colour of the trunk varies as tree grows. External portion of the bark is brownish grey while internal portion is brownish red in colour. Powdered drug is brown in colour. Stem powder has no specific taste and odour. (Figure 1 and 2)

Figure (1-2):- 1: A. catechu plant showing leaves and pods; 2: Plant stem with heartwood, sapwood and bark

b) Microscopical studies: Transverse section of stems of A. catechu has a spherical transaction which showed the presence of medullary rays, vessels are isolated or clustered as two to four, xylem fibres, secondary xylem cells forming a sheath around vessel, parenchyma, epidermis, some of the cells were filled with brown content and few cells observed with calcium oxalate crystals (Figure 3 and 4).

Figure (3-4) Microscopy of transverse section:- 3:Medullary rays, xylem, vessels, parenchyma, cells filled with brown fluid and epidermis at 10x; 4: Vessels and cells with calcium oxalate crystals at 45x

Powder microscopy showed the presence of xylem fibres, xylem tracheid, parenchyma and pitted cells (Figure 5 and 6).

Figure (5-6) Powder microscopy at 45x:- 5: (a) parencyma, (b)xylem tracheids; 6: (a)xylem fibre, (b) unstained pitted cells, (c) stained pitted cells

B. Physicochemical parameters:

All the parameters were performed as per standard procedures given in IP. Values for total ash content, water-soluble ash content, acid-insoluble ash content, ethanol-soluble extractive value, water-soluble extractive values, loss on drying and swelling index are given in Table 1. Successive extraction was carried out according to increasing polarity of the solvents i.e. petroleum ether, chloroform, ethanol and water respectively. Percentage extractive values and colour of extracts observed are given in Table 2. Preliminary phytochemical screening of stems revealed the presence of flavonoids, tannins, and phenolic compounds, saponin glycosides, carbohydrate and lipids. Results for preliminary phytochemical screening are given in Table 3. Observations for fluorescence behaviour are presented in Table 4. Heavy metal content, aflatoxins, microbial content and pesticide residues were evaluated according to WHO guidelines and results are shown in Table 5.

Table 1: Result for physicochemical parameters

Sr. No.	Parameter	*A. catechu*
1.	Total Ash Content	1.8% w/w
2.	Acid-insoluble ash value	0.6% w/w
3.	Water soluble ash value	0.9% w/w
4.	Loss on drying	12% w/w
5.	Alcohol soluble extractive value	5.6% w/w
6.	Water soluble extractive value	3.4% w/w
7.	Swelling Index	NIL

Table 2: Result for Successive extracts

Sr. No.	Extract	Colour	%age Extractive Value
1.	Petroleum ether	Brown	0.16%
2.	Chloroform	Brown	0.94%
3.	Ethanol	Dark brown	7.19%
4.	Aqueous	Brownish black	5.07%

Table 3: Phytochemical Screening of A. catechu Stem

Sr. No.	Compounds	PE	CH	Et.	Aq.
1.	Alkaloid	-ve	-ve	-ve	-ve
2.	Carbohydrate	-ve	-ve	+ve	-ve
3.	Steroid	-ve	-ve	-ve	-ve
4.	Saponin	-ve	-ve	+ve	+ve
5.	Protein	-ve	-ve	-ve	-ve
6.	Fixed Oil/ Fat	+ve	-ve	-ve	-ve
7.	Flavonoid	-ve	-ve	+ve	+ve
8.	Tannin and Phenol	-ve	-ve	+ve	+ve
9.	Gum and Mucilage	-ve	-ve	-ve	-ve
10.	Glycoside	-ve	-ve	-ve	-ve

# PE- Petroleum ether extract; CH-Chloroform extract; Et- Ethanol extract; Aq- Aqueous extract

Table4: Fluorescence behavior of stem powder of A. catechu with different reagents.

Sr. no.	Reagent	Day light	Short UV (254 nm)	Long UV (365nm)
1.	Powder as such	Light Brown	Light Green	Brown
2.	1N NaOH in Methanol	Brown	Dark Green	Greenish Brown
3.	1N NaOH in Water	Brown	Yellowish Brown	Brown
4.	1N HCl	Brown	Yellowish Brown	Dark Brown
5.	50% HNO₃	Dark Brown	Dark Green	Black
6.	50% HCl	Brown	Dark Green	Dark Brown
7.	50% H₂SO₄	Brown	Light Green	Blackish Brown

Table 5: Physicochemical parameters

Parameter	Value	Specified limit
Microbial contamination test
Total Bacterial Count	140	1 X 10⁵c.f.u./g
Total yeast mould count	Nil	1 X 10³c.f.u/g
E. coli	Nil	Nil
Salmonella sp.	Nil	Nil
S. aureus	Nil	Nil
P. aeruginosa	Nil	Nil
Aflatoxin content
Aflatoxin B1	Nil	0.5
Aflatoxin B2	Nil	0.1
Aflatoxin G1	Nil	0.5
Aflatoxin G2	Nil	0.1
Heavy metal analysis
Arsenic	ND	5
Cadmium	0.06	0.3
Lead	0.04	10
Mercury	ND^*	0.2
Pesticides
Alachlor	ND	0.02
Atrazine	ND	-
BHC (sum of all isomers)	ND	0.3
Bifenthrin	ND	-
Butachlor	ND	-
Carbofuran	ND	-
Carbofuran, 3-Hydroxy	ND	-
Chlordane (sum of cis-, alpha-)	ND	0.05
Cypermethrin peak 1	ND	1.0
DDD(sum of all isomers)	ND	1.0
DDE (sum of all isomers)	ND	1.0
Dieldrin	ND	0.05
Dimethoate	ND	0.5
Edifenphos	ND	-
Endosulfan peak 1	ND	3.0
Endosulfan peak 2	ND	3.0
Endosulfan sulphate	ND	3.0
Endrin	ND	0.05
Ethion	ND	2.0
Fenthion	ND	0.5
Fenvalerate	ND	1.5
Heptachlor	ND	0.05
Heptachlor epoxide	ND	0.05
Malathion	ND	1.0
Methoxychlor	ND	-
Parathion-methyl	ND	0.2
Phorate	ND	-
Phoratesulfone	ND	-
Phosalone	ND	0.1

ND = Not Detected

CONCLUSION:

Pharmacognostical and physicochemical evaluation of plant is essential step for plant standardization. These parameters reveal about identification, quality and purity of the plant. For this purpose, A. catechu stem was screened for various parameters. Preliminary phytochemical screening showed the presence of tannins, phenolic compounds and flavonoids in its stem. Physicochemical parameters were found within limits. Thus, in the present research work, plant has been standardized and can be used further for pharmacological evaluation.

CONFLICT OF INTEREST:

Article has no conflict of interest.

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Received on 27.04.2020 Modified on 30.06.2021

Research J. Pharm. and Tech 2022; 15(10):4485-4489.

DOI: 10.52711/0974-360X.2022.00752