# Comparative Analysis of Morphological, Molecular, and Physicochemical Markers to Evaluate Trollius ledebouri Rchb. as a Potential Alternative Source to Trollius chinensis Bunge for High-Quality Flos Trollii Supplements

## TL;DR

This study compares Trollius ledebouri and Trollius chinensis to determine if the former can serve as a high-quality alternative for Flos Trollii supplements.

## Contribution

The study provides a systematic comparison of morphological, genetic, and chemical markers between two Trollius species for potential medicinal use.

## Key findings

- Trollius ledebouri has shorter sepals than petals and brown secretory structures not found in Trollius chinensis.
- Trollius ledebouri shows higher levels of orientin and vitexin compared to Trollius chinensis.
- Genetic analysis reveals significant divergence in psbA-trnH sequences between the two species.

## Abstract

Trollius chinensis Bunge is a medicinal herb whose dried flowers (Flos Trollii) exert heat-clearing and detoxifying effects. Trollius ledebouri Rchb., a morphologically similar species in the Great Xing’an Mountains with inconsistent regulatory status, lacks a systematic comparison with it. This study used morphological assessment, microscopy, DNA barcoding, and physicochemical analysis to explore its potential as an alternative. Results showed distinct morphological and genetic differences between the two species, but Trollius ledebouri Rchb. had slightly higher orientin and vitexin levels. It holds potential as a Flos Trollii alternative, requiring further metabolomic studies.

Trollius chinensis Bunge (TCB), a perennial Ranunculaceae herb, produces Flos Trollii-dried flowers with medicinal properties including heat clearing, detoxification, and relieving oral/throat discomfort, eye pain, and cold-induced fever. TCB is mainly cultivated in northern China, while Trollius ledebouri Rchb. (TLR), distributed in Heilongjiang’s Great Xing’an Mountains, is morphologically similar to TCB. However, their regulatory statuses are inconsistent, and comprehensive comparative studies are lacking. This study adopted morphological assessment, microscopy, DNA barcoding, and physicochemical analysis to explore whether TLR could be a potential alternative source of Flos Trollii. Key differences were identified: TLR’s sepals are shorter than petals, whereas TCB’s sepals and petals are nearly equal in length; TLR has brown secretory structures absent in TCB. Genetic distance analysis showed high conservation in ITS2 and trnL-trnF sequences between the two species, but psbA-trnH sequence divergence exceeded the 0.05 threshold. HPLC quantification revealed that TLR contained slightly higher levels of orientin and vitexin than TCB. HPLC quantification revealed that TLR contained slightly higher levels of orientin (5.370–5.377 mg/g) and vitexin (1.954–2.053 mg/g) compared to TCB (orientin: 4.493–4.620 mg/g; vitexin: 1.361–1.451 mg/g). Collectively, TLR exhibits comparable flavonoid content and holds potential as an alternative Flos Trollii source. Given the limited bioactive compounds analyzed, future research should conduct comprehensive metabolomic profiling to fully evaluate its phytochemical composition and medicinal value. These data establish chemotaxonomic markers for Trollius authentication in herbal medicine.