Phytochemical screening, antimicrobial activity, in vitro and in vivo antioxidant activity of Berberis lycium Royle root bark extract

Mughal, T. A.; Ali, S.; Hassan, A.; Kazmi, S. A. R.; Saleem, M. Z.; Shakir, H. A.; Nazer, S.; Farooq, M. A.; Awan, M. Z.; Khan, M. A.; Andleeb, S.; Mumtaz, S.; Mumtaz, S.; Tahir, H. M.; Gulzar, N.

doi:10.1590/1519-6984.249742

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Brazilian Journal of Biology

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Original Article • Braz. J. Biol. 84 • 2024 • https://doi.org/10.1590/1519-6984.249742 copy

Phytochemical screening, antimicrobial activity, in vitro and in vivo antioxidant activity of Berberis lycium Royle root bark extract

Triagem fitoquímica, atividade antimicrobiana, atividade antioxidante in vitro e in vivo do extrato de casca de raiz de Berberis lycium Royle

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Antioxidants are materials that scavenge or remove free radicals from living systems. The oxidation process ends in the production of free radicals. These free radicals are the chief birthplace of cancerous cells. Antioxidizing agents remove free radical intermediates by terminating oxidation processes by being oxidized themselves. On the other hand, infectious diseases affect the world on a large scale. To fight these diseases several synthetic compounds have been used. Plant based medications play important role in this regard. So, the current research aimed to investigate the antibacterial and antioxidant effect of Berberis lycium Royle root bark (BLR) extract. Berberis lycium Royle was used for phytochemical analysis and also as antimicrobial and antioxidant agents. The antimicrobial activity was evaluated by the agar well diffusion method. Current study revealed that BLR was rich in phytochemicals and toxic against tested pathogenic bacteria. BLR showed the highest activity against S. pyogenes (13.3±0.8 mm). The lowest antibacterial activity was reported against E. coli (0±0 mm). In case of minimum inhibitory concentration, it was observed that BLR with 10 μg/mL concentration showed the highest activity while 2.5 μg/mL of BLR showed the least inhibitory activity. The highest In vitro antioxidant activity was recorded as 65% at 100 µg/mL. In case of in vivo antioxidant activity level of CAT, GSH and SOD were decreased while that of MDA was enhanced in groups treated with CCl₄ as compared to the control group. BLR extract treatment reversed all these changes significantly. Current results indicate that BLR is effective against bacterial pathogens and also has antioxidant potential.

Keywords:
Berberis lycium Royle; root bark extract; phytochemicals; antibacterial; antioxidant

S. No.	Phytochemical constituents	Test	Methodology	Positive indications
1	Alkaloids	Wagner	i. Potassium iodide (2 g) + iodine (1.27 g) +Distilled water (5 ml)	Reddish brown and white
			ii. This solution was diluted with distilled water up to 100 ml.	Dull precipitate
			iii. Few drops of this solution + Extract
2	Steroids	Salkowski	Extract (100 mg) + chloroform (2 ml) + con. H₂SO₄ (2 ml)	Formation of brown ring
3	Terpenoids	Salkowski	Extract (100 mg) + chloroform (2 ml) + con. H₂SO₄ (2 ml)	Reddish brown coloration
4	Tannins	Ferric chloride	Extract (0.5 gm) + distilled water (10 mL) +5% ferric chloride (few drops)	Black or blue-green coloration
4	Tannins	Ferric chloride		or precipitate
5	Saponins	Foam	Extract (0.5 g) + distilled water (10 ml)	Formation of frothing
6	Glycosides	Keller-Killiani	Extract (0.5 gm) + distilled water (5 ml) + glacial acetic acid (2 ml containing few drops of ferric chloride) + H₂SO₄ (1 ml)	formation of brown ring
7	Proteins	Ninhydrin	i. Extract (1 ml) + 0.2% ninhydrin reagent (few drops)	Blue color
7	Proteins	Ninhydrin	ii. Heated for 5 minutes	Blue color
8	Free amino acids	Ninhydrin	Extract (1 ml) + ninhydrin reagent (few drops)	Purple, light brown color
9	Carbohydrates	Benedict’s	i. Extract (1 ml) +Benedict’s reagent (2 ml)	Formation of reddish brown colorcolor
9	Carbohydrates	Benedict’s	ii. Heated for few seconds	Formation of reddish brown colorcolor
10	Phenols	Folin ciocalteu	i. Extract (1 ml) + Folin ciocalteu reagent (few drops) + sodium carbonate aquous solution.	Formation of grey or black color
10	Phenols	Folin ciocalteu	ii. Mixture was allowed to stand for 10 minutes	Formation of grey or black color
11	Flavonoids	Ferric	Extract (2 ml) + FeCl₃ (few drops) chloride	Blackish red/ Dark brown color

Sample No.	Phytochemical tested	Results
1	Alkaloids	+
2	Steroids	+
3	Tannins	+
4	Saponins	+
5	Glycosides	+
6	Carbohydrates	+
7	Proteins	+
8	Free Amino acids	+
9	Terpenoids	+
10	Phenols	+
11	Flavonoids	+

Agents	Conc. (µg/ml)	E. coli	K. pneumoniae	S. aureus	S. pyogenes	P. aeruginosa	S. marcesscens	S. epidermidis
Dimethyl sulfoxide (DMSO)	2.5	0	0	0	0	0	0	0
	5	0	0	0	0	0	0	0
	10	0	0	0	0	0	0	0
BLR extract	2.5	0	0	3.3±0.3	5.3±0.3	2±0.5	2.7±0.3	0
	5	7.3±0.3	5.7±0.3	5±0	8±0	5±0.6	5.7±0.3	6±0
	10	9.3±0.7	9.7±0.16	10±1.5	13.3±0.8	8.5±0.2	9.8±0.4	10±0.6

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