Integrated chemical and molecular authentication of Pinellia tuber: A multinational collaboration within the FHH ATLAS project with government regulatory agencies from seven countries and regions.

Pinellia ternata, a widely used herbal medicine in the Western Pacific region, is frequently adulterated with morphologically similar species, posing challenges to quality control and safety. As part of the Forum for the Harmonization of Herbal Medicines (FHH) ATLAS project, this multi-national collaborative study aimed to evaluate and validate the applicability of practical and reliable methods to distinguish P. ternata from its adulterants, thereby ensuring standardized quality control across member countries. Multiple analytical approaches applicable to member countries were evaluated. Morphological analysis and high-performance thin-layer chromatography proved ineffective in distinguishing adulterants, whereas high-performance liquid chromatography (HPLC) and polymerase chain reaction (PCR) successfully identified them. HPLC, using triglochinic acid as a marker compound, differentiated P. ternata from P. pedatisecta but could not detect Typhonium flagelliforme. In contrast, PCR demonstrated higher specificity by amplifying distinct DNA fragments for P. ternata, P. pedatisecta, and P. tripartita, as well as a specific fragment for T. flagelliforme. Cross-validation among countries confirmed the reproducibility of these findings. By integrating chemical and molecular methods, this collaborative effort provides an effective framework for authenticating P. ternata, contributing to safer and more reliable herbal medicines in the Western Pacific and beyond.