The present study was intended to evaluate the pharmacognostic attributes including macroscopy, microscopy, physico‐chemical and phytochemical screening of the aerial part of Xanthium spinosum, belonging to family Asteraceae. Morphologically, the leaf was covered with sparsely scattered non‐glandular, multicellular hair on adaxial surface, while abaxial surface was reported with dense silvery furs. Leaf showed lamphystomatic nature bearing anomocytic stomata. Various other leaf constant values like vein islets number, vein termination number and palisade cell ratio were also observed and recorded as 4.36 ± 0.5, 3.36 ± 0.5, and 11.82 ± 2.04, respectively. Cross section of the stem showed typical dicot anatomy differentiated into a single epidermal layer followed by few rows of collenchyma cells, thin‐walled parenchyma cells followed by closed and collateral vascular bundles having prephloemic sheaths and a central pith. Light and scanning electron microscopy of the powdered drug revealed a variety of tissues and cells fragments having different measurements. Fluorescence analysis of the fine powder treated with different reagents revealed a variety of colors implying as a sign for the presence of various biochemical constituents. Highest and lowest percent extractive value was found in methanol and distilled water which were 29.10% and 9.80%, respectively. Qualitative phytochemical screening revealed the presence of various secondary metabolites like flavonoids, phenols, tannins, glycosides, phytosterols, glycosides, saponins, and coumarins, while quantitative phytochemical screening detected maximum quantity of flavonoids in chloroform followed by methanol and ethyl acetate. The aforementioned studies have been reported first time for the said species, which may be helpful for the standardization of the current taxon.
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