High-Throughput Chromatographic and Spectroscopic Profiling for Reliable Differentiation of Hyoscyamus niger and Cleome viscosa L.

Abstract

Quality evaluation of herbal medicines is essential to ensure their safety, efficacy, and consistency. The present study focused on the quality control and authentication of Hyoscyamus niger (henbane) and its adulterant Cleome viscosa L. using modern analytical techniques. Both plants possess significant medicinal value but are often adulterated, affecting therapeutic reliability. The study aimed to differentiate and authenticate H.niger and C. viscosa through macro- and microscopic characterization, followed bychemical profiling using High-Performance Thin-Layer Chromatography (HPTLC) andHigh-Performance Liquid Chromatography coupled with Mass Spectroscopy (HPLC–MS). These advanced tools enabled rapid, simultaneous analysis of multiple phytoconstituents, facilitating efficient quality assessment. The results revealed distinct differencesin macro- and microscopic features between the two species. Chemical profiling identified characteristic marker compounds specific to each plant, allowing clear discrimination between authentic and adulterated samples. Quantitative estimation of total alkaloidcontent showed a notable variation, with H. niger containing 0.1737 ± 0.0023% andC.viscosa 0.0441 ± 0.0018% (w/w). Overall, the findings highlight the effectivenessof combining high-throughput screening with advanced analytical approaches for theauthentication of medicinal plants. This integrated strategy strengthens quality controlmeasures, ensuring the authenticity, potency, and safety of herbal formulations, and supports the development of robust quality assurance protocols in the herbal industry.