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Abstract Marijuana and hemp represent two broad classes ofCannabis sativaplants that are distinguished based on the concentration of the psychoactive cannabinoid delta‐9‐tetrahydrocannabinol (Δ⁹‐THC). In this work, solvent extracts derived from marijuana and hemp were characterized using optical and spectroscopic techniques. The crystalline components of the solvent extracts were first analyzed using polarized light microscopy to determine optical properties, namely, crystal system, optical sign, and principle refractive indices. Crystals from the marijuana‐derived extracts exhibited an orthorhombic crystal system and were optically negative, with n_βbetween 1.6320 and 1.6330 ± 0.0002. In contrast, crystals from hemp‐derived extracts exhibited a monoclinic crystal system and were optically positive, with n_βbetween 1.600 and 1.6040 ± 0.0002. Crystals were further distinguished through infrared spectroscopy, which highlighted structural differences between the two sample types, primarily based on differences in O‐H stretching. Finally, single‐crystal X‐ray diffraction was used to definitively identify the crystalline components, confirming the presence of tetrahydrocannabinolic acid in marijuana‐derived extracts and cannabidiol in hemp‐derived extracts. Given the differences in crystal structure identified between marijuana‐derived and hemp‐derived solvent extracts, optical characterization provides a screening method to differentiate visually similar samples prior to confirmatory analysis.