Peucedanum ostruthium (L.) Koch has a longstanding history as herbal remedy in the Alpine region of Austria,
where the rhizomes (= Radix Imperatoriae) are traditionally used to treat disorders
of the gastro-intestinal and respiratory tract as well as the cardiovascular system
[1]. A recent ethnopharmacological study on 71 Austrian traditional herbal drugs revealed
a distinct anti-inflammatory activity by Radix Imperatoriae [2]. In the present work, the aim was to unravel the constituents responsible for the
observed anti-inflammatory activity of a dichloromethane-methanol extract prior to
isolation. Therefore, a recently developed biochemometric approach named ELINA (Eliciting Nature’s Activities) [3] was applied. Here, the objective was to simplify the crude extract by generating
micro-fractions with quantitative variances of constituents over several consecutive
fractions. This was achieved via an optimized high-performance counter-current chromatographic
(HPCCC) fractionation. After this single fractionation step, 1H-NMR data and bioactivity data from an in vitro assay on the cell adhesion molecules E-selectin and VCAM-1 were collected for all
31 micro-fractions. In parallel to a quantitative variance of 1H-NMR signals over consecutive fractions, bioactivity patterns relating to this variation
were obtained. To reveal chemical features crucial for the observed activities, statistical
heterocovariance analyses (HetCA) [4] were performed. In addition, LC-MS-CAD data were used to facilitate the identification
of the bioactive constituents. As a result, we identified two coumarins (Imperatorin
and Ostruthol) and one chromone (Peucenin) responsible for the anti-inflammatory activities.
The applied HetCA approach enables an early identification and dereplication of even
minor bioactives prior to any isolation without wasting resources.
