Antimicrobial Screening and HPLC-DAD-MS Characterization of the Flavonoid–Rich Fractions of the Methanol Leaf-Extract of Lawsonia inermis Linn doi.org/10.26538/tjnpr/v5i8.28

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Vivian O. Okeke
Nkeoma N. Okoye
Kenneth G. Ngwoke
Festus B. Chiedu Okoye

Abstract

Flavonoids are known for their wide biological activities, including antimicrobial activity. This study was aimed at investigating the antimicrobial potential of the flavonoid-rich fractions of Lawsonia inermis leaf extract. The ethylacetate fraction from the methanol extract of L. inermis leaves was subjected to Vacuum Liquid Chromatography (VLC) using binary combinations of Hexane: Ethylacetate and Dichloromethane:Methanol to obtain fractions F1-F14. Fractions F11 and F12 which contain flavonoids were further subjected to High Performance Liquid Chromatography–Diode Array Detector (HPLC-DAD) and Liquid Chromatography-Mass Spectrometry (LC-MS) analysis, and subsequently subjected to in vitro test for antimicrobial activity against laboratory strains of Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, Candida albicans using Agar well diffusion method at concentrations of 0.0625-1 mg/mL. Erythromycin (50 µg/mL) and Miconazole (50 µg/mL) were used as positive controls for bacteria and fungi, respectively. HPLC-DAD and LC-MS analysis of the flavonoid-rich fraction (F12) led to the detection and identification of four flavone pigments namely; vitexin, luteolin 4-glucoside, apigenin monoglycoside and isoorientin (luteolin 6C-β-glucoside). Fraction F11 (constituents not identified) and fraction F12 (the flavone-rich fraction) showed moderate antibacterial activity, with E. coli being the most susceptible bacteria with Minimum Inhibition Concentrations (MIC) of 62.5 μg/mL for F11 and 500 μg/mL for F12. Very mild anti-fungal activity was only observed against C. albicans but not against A. niger. The unidentified polar compounds (the major components of F11) and luteolin 6C-β- glucoside (the major flavones detected in F12) may contribute to the observed antimicrobial activity of L. inermis leaf extract.

Article Details

How to Cite
O. Okeke, V., Okoye, N. N., Ngwoke, K. G., & Okoye, F. B. C. (2021). Antimicrobial Screening and HPLC-DAD-MS Characterization of the Flavonoid–Rich Fractions of the Methanol Leaf-Extract of Lawsonia inermis Linn: doi.org/10.26538/tjnpr/v5i8.28. Tropical Journal of Natural Product Research (TJNPR), 5(8), 1500–1505. Retrieved from https://tjnpr.org/index.php/home/article/view/473
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References

Okoye FBC, Odimegwu DC, Anyasor CN, Ajaghaku DL, Gugu TH, Osadebe PO, Proksch P. A new antioxidant and antimicrobial compound isolated from Alchornea cordifolia leaves. Afr J Pharm Res Dev. 2015; 7(2):87-94

Tong Y, Deng Z. An aurora of natural products-based drug discovery is coming. Synth Syst Biotechnol. 2020; 5(2): 92–96

Wolfender JL, Litaudon M, Touboul D, Queiroz EF. Innovative omics-based approaches for prioritisation and

targeted isolation of natural products – new strategies for drug discovery. Nat Prod Rep. 2019; 36(6): 855–868

Al-Snafi AE. A review on lawsonia inermis: a potential medicinal plant. Int J Curr Pharm Res. 2019; 11 (5):1-13

Orwa C, Mutua A, Kindt R, Jamnadass RH, Simons A. Agro forest tree Data base: a tree reference and selection guide version 4.0 [Online] 2009. Available from http://www.worldagroforestry.org/sites/treedbs/treedatabases.asp(http://www.worldagroforestry.org/sites/treedbs/treedatabases.asp)

Sharma RK, Goel A, Bhatia AK Lawsonia Inermis Linn: A Plant with Cosmetic and Medical Benefits. Int J Appl Sci Biotech. 2016; 4(1): 15-20.

Chetty KM, Sivaji K, Rao KT. Flowering Plants of Chittoor District, Andhra Pradesh, (26th ed.) Tirupati Students Offset 2008: 132 p.

Babu PD, Subhasree RS. Antimicrobial activities of Lawsonia inermis-A review. Acad J Plant Sci. 2009; 2(4):

-232.

Habbal O, Hasson SS, El-Hag, AH, Al-Mhrooqi Z, AlHshmi, N, Al-Binami Z, Al-Balushi, MS, Al-Jabri AA Antibacterial activity of Lawsonia inermis Linn (Henna) against Pseudomonas aeruginosa. Asian Pac J Trop

Biomed. 2011; 1(3):173-176

Okoye FBC, Agbo MO, Nworu CS, Nwodo NJ, Esimone CO, Osadebe PO, Proksch P. New neolignan glycoside and an unusual benzoyl maleic acid derivative from Maytenus senegalensis leaves. Nat Prod Res. 2015; 29(2):109-115.

Okoye NN, Okoye COB Antimicrobial and antioxidant flavonoid glycosides from the leaves of Alstonia boonei De Wild. J Pharm Res Int. 2016; 10(6):1-9.

Basile A, Giordano S, Lopez-Saez JA, Cobianchi RC. Antibacterial activity of pure flavonoids isolated from

mosses. Phytochem. 1999; 52(8):1479-1482.

Chung JG, Hsia TC, Kuo HM, Li YC, Lee YM, Lin SS, Hung CF. Inhibitory actions of luteolin on the growth and

arylamine N-acttyl transferase activity in strains of Helicobacter pylori form ulcer patients. Toxicol In vitro.

; 15(3):191-198

De Campos MP, Cechinel FV, Da Silva RZ, Yunes RA, Zacchino S, Juarez S, Bella Cruz CA Evaluation of

antifungal activity of Piper solmsianum.C. DC. Var. solmisianum (Piperacaea). Biol Pharm Bull. 2005; 28(8):

-1530.

Tshikalange TE, Meyer JJM, Hussein AA Antimicrobial activity, toxicity and the isolation of a bioactive compound from plants used to treat sexually transmitted diseases. J Ethnopharmacol. 2015; 96(3):515-519

Akroum S, Bendjeddou D, Satta D, Lalaoui K Antibacterial activity and acute toxicity effect of flavonoids extracted from Mentha longifolia. Am-Euras. J Sci Res. 2009; 4(2): 93-96.

Cushnie TPT, Lamb AJ. Antimicrobial activity of flavonoids. Int J Antimicrob Agents. 2005; 26(5):343-356.

Al-Kakei SNH. Antibacterial Activity of Henna Plnt Lawsonia inermis on Gram Negative Bacteria. Al-Mustansiriyaj J Sci. 2011; 22(7):15-20.

Karpiński TM, Adamczak A, Ozarowski M. Antibacterial activity of Apegenin, Luteolin and their C-glycosides. 5th International Electronic Conference on Medicinal Chemistry [online] 2019 [cited 2021 Mar 15] Available from: http://www.sciforum.net/paper/view/6321(http://www.sciforum.net/paper/view/6321)