Phytochemical Profiling, Physicochemical Evaluation, and In Vitro Biological Activities of Blepharis integrifolia Extracts

Abstract

Blepharis integrifolia (Acanthaceae) is a traditionally valued medicinal herb utilized for a wide
spectrum of ailments, yet its systematic phytochemical and pharmacological validation remain
limited. This study aimed to comprehensively evaluate the qualitative and quantitative
phytochemical profiles, physicochemical parameters, and in vitro biological activities of B.
integrifolia extracts. Shade-dried plant material was subjected to parallel cold maceration using
70% ethanol (BIE) and chloroform (BIC). Baseline physicochemical analysis established a
moisture content of 2.41% and a total ash value of 8.31%. Preliminary qualitative screening
confirmed the presence of phenols, flavonoids, proteins, and carbohydrates in both extracts.
Notably, solvent polarity influenced the extraction profile: terpenoids were exclusive to the
hydroethanolic extract, while alkaloids were specifically extracted by chloroform. Quantitative
spectrophotometric analysis revealed that the chloroform extract yielded a significantly higher
total phenolic content (342.24 ± 16.13 μg GAE/mg), whereas the 70% ethanol extract exhibited
a superior total flavonoid concentration (197.51 ± 8.41 μg QE/mg). The extracts were
subsequently evaluated for their in vitro biological efficacies. Both extracts demonstrated
potent, broad-spectrum antibacterial activity against S. aureus and E. coli (zones of inhibition
up to 15.66 mm at 3 mg). Furthermore, the ethanol extract exhibited superior antioxidant
capacity (DPPH IC₅₀ = 88.84 μg/mL), whereas the chloroform extract demonstrated stronger
antidiabetic potential (α-glucosidase IC₅₀ = 60.16 μg/mL). These findings validate the rich
secondary metabolite profile of B. integrifolia, substantiating its ethnomedicinal applications
and providing a concrete scientific rationale for its potential therapeutic use in managing
oxidative stress, microbial infections, and metabolic conditions.