Development of an HPTLC Method for Differentiating the Botanical Origins of Cuscutae Semen

Cuscutae Semen has a long-standing history as a traditional herbal medicine widely utilized in East Asian countries, including Korea, China, Hong Kong, and Taiwan. The Korean Herbal Pharmacopoeia (KHP) exclusively recognizes Cuscuta chinensis Lam. as the authentic source of Cuscutae Semen, whereas other countries also accept Cuscuta australis R.Br. This discrepancy has resulted in imports containing both species, which contravenes local regulations. Furthermore, a significant portion of domestically produced Cuscutae Semen is actually Cuscuta japonica Chois., regarded as an adulterant, which further complicates the market. This situation underscores the urgent need for clear differentiation of the botanical origins of Cuscutae Semen. In this study, a high-performance thin-layer chromatography (HPTLC) method was developed to simultaneously differentiate the three origins of Cuscutae Semen: C. chinensis, C. australis, and C. japonica. By utilizing a TLC scanner and TLC-MS interface, the chemical fingerprints of the samples were analyzed in detail. The results revealed that while C. chinensis and C. australis share similar profiles, they can be distinguished by the presence or absence of astragalin and kaempferol. In contrast, C. japonica showed distinct differences, characterized by the presence of chlorogenic acid derivatives. This method demonstrated the ability to rapidly and accurately differentiate between Cuscutae Semen origins, making it a cost-effective and reliable tool for ensuring quality control. It is expected to contribute to improving the standardization and reliability of herbal medicine quality management in the domestic market.