Background: Lawsonia inermis Linn popularly known as Henna, plays an important
role in ayurvedic or natural herbal medicines. The presence of phytoconstituents in henna, that
may affect the animal or human health adversely, needs to be elucidated for L. inermis Linn species
grown in India.
Introduction: The aim of this research was to perform phytochemical screening, and study cytotoxicity
and anti-inflammatory activities to understand the potential of leaves of Lawsonia inermis
of Indian origin to provide a way forward for therapeutic use in medicine.
Methods: We assessed the phytochemical profile for the presence of phytoconstituents (alkaloids,
carbohydrates, glycosides, steroids, flavonoids, saponins, tannins, proteins/amino acids and
gums/mucilage) in various extracts of the plant leaves. The extracts were further purified by column
chromatography for the isolation of plant constituents and monitored by TLC, analyzed by
Fourier transform infrared FT-IR spectroscopy, H1NMR, and GC-MS analysis. Fractions were assessed
for cytotoxicity and anti-inflammatory properties at various concentrations. We assessed the
anti-inflammatory activity by nitric oxide production in various leaf extracts determined by Griess
Results: All the spectral results suggest that the compounds from the extract contain an aromatic
nucleus and OH group along with the methoxy group, allyl as well as vinyl group. Fractions of
chloroform/methanolic (7:3) leaf extract of Lawsonia inermis confirmed the presence of the two
constituents i.e. fraxetin and 1(3H)-isobenzofuranone. We observed a significant difference in
cytotoxicity at higher concentrations in methanol and chloroform: methanol (8:2) leaf extracts
(p>0.05), we could not find any significant differences amongst other leaf extracts at different
concentrations. Some leaf extracts have potential cytotoxic activity on Vero cells. Reducing the
chloroform concentration during extraction decreases the cytotoxic effect on cells. Nitric oxide
levels decreased from 1000 μg/ml concentration to lower concentrations with varying degrees.
Overall the highest nitric oxide production by CHCl3 (70%)/ MeOH (30%) was observed amongst
various fractions at different concentrations.
Conclusion: Phytochemical screening and the study of cytotoxicity and anti-inflammatory activities
highlight the potential of leaves of the plant to provide a way further for their use in medicine.
Fraxetin 1(3H) and isobenzofuranone structures were confirmed in fractions of CHCl3 (70%)/
MeOH (30%) extract as potent constituents. Some leaf extracts have potential cytotoxic activity on
Vero cells. Reducing the chloroform concentration during extraction, it decreases the cytotoxic effect
on cells. The cytotoxicity studies indicate the presence of cytotoxic compounds in some of
these extracts, warranting research for the fabrication of suitable formulations comprising these
constituents to reduce dose/toxicity for beneficial effects of the plant components.