Title:Pharmacognostical Standardization, Isolation of Phytoconstituents
(β-sitosterol), HPTLC Analysis of Extracts of Operculina turpethum (Linn.)
Roots and Evaluation of Cytotoxic, In vitro, and In vivo Anti-inflammatory
Activities
Volume: 19
Issue: 8
Author(s): Akash Ved*, Shweta Gupta, Namrata Singh, Karuna S. Shukla, Om Prakash and Neeru Gaur
Affiliation:
- Department of Pharmacy, Goel Institute of Pharmaceutical Sciences, Lucknow, Uttar Pradesh 226028, India
Keywords:
Convolvulaceae, β-sitosterol, HPTLC, anti-inflammatory, cytotoxic, HepG2.
Abstract:
Operculina turpethum (Linn.) Silva Manso belongs to the family convolvulaceae. It is an
important plant in the Indian conventional system of medicine, which is extensively employed by different
tribes in many countries to cure edema and painful conditions like arthritis, back pain, hyperlipidemia,
diabetes mellitus, liver disorders, skin disorders, and regulation of bowel functions.
The roots of O. turpethum (Linn.) were subjected to physicochemical and phytochemical standardization
and chromatographic separation, which was accomplished by column chromatography, TLC, and
HPTLC. Further, the acute toxicity, cytotoxic and anti-inflammatory activities of Operculina turpethum
roots were estimated by in vivo and in vitro models.
This study includes percentage yield of extraction and organoleptic evaluation, along with the analysis of
its physicochemical investigations and preliminary phytochemical estimation. The isolation of active
phytoconstituents was done by column chromatography, and the isolated compound was then exposed to
TLC and HPTLC analysis. Cytotoxic activity was tested by WST-1 based cell viability assay on HepG2
cells. The anti-inflammatory activity of methanol extract (ME) was evaluated against inflammation occur
by both in vitro and in vivo methods.
The methanolic extract exhibited the presence of most of the phytoconstituents out of all the extracts.
The phytoconstituent phytosterol, i.e., β-sitosterol, was isolated by column chromatography, identified,
and quantified by TLC and HPTLC, which was liable for anti-inflammatory activity. The amount of β-
sitosterol was estimated to be 14.09 μg in a 10.00 mg fraction of MEOT. MEOT was devoid of toxicity
up to 2000 mg/kg in Wistar albino rats. It was analysed that in vitro anti-inflammatory activity of MEOT
by egg albumin denaturation method exhibited an incredible decrement in turbidity and increased the
percentage inhibition of albumin denaturation (60.52%) in MEOT treated group as compared with the
control group. In egg albumin-induced edema in rats, MEOT at the dose of 400 mg/kg reduced the edema
formation (3.03 ± 0.02) induced by egg albumin at 4th h. In cotton pellet-induced granuloma in rats,
MEOT at the dose of 400 mg/kg displayed maximum granuloma inhibition (51.06%), which was similar
to that of indomethacin.
From the obtained findings, it is confirmed that O. turpethum contains β-sitosterol, which is responsible
for potent anti-inflammatory activity without causing cytotoxicity to the plant. The results suggested that
ME of O. turpethum roots had a high potential for application as an anti-inflammatory agent. The recognization
and confirmation of the plant can be obtained from the study and will present data that is aidful
in determining the quality and purity of a crude drug which further helps in preventing its adulteration.
