Comprehensive analysis of commercial willow bark extracts by new technology platform: Combined use of metabolomics, high-performance liquid chromatography–solid-phase extraction–nuclear magnetic resonance spectroscopy and high-resolution radical scavengin

Here, proof-of-concept of a new analytical platform used for the comprehensive analysis of a small set of commercial willow bark products is presented, and compared with a traditional standardization solely based on analysis of salicin and salicin derivatives. The platform combines principal component analysis (PCA) of two chemical fingerprints, i.e., HPLC and 1H NMR data, and a pharmacological fingerprint, i.e., high-resolution 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS+) reduction profile, with targeted identification of constituents of interest by hyphenated HPLC–solid-phase extraction–tube transfer NMR, i.e., HPLC–SPE–ttNMR. Score plots from PCA of HPLC and 1H NMR fingerprints showed the same distinct grouping of preparations formulated as capsules of Salix alba bark and separation of S. alba cortex. Loading plots revealed this to be due to high amount of salicin in capsules and ampelopsin, taxifolin, 7-O-methyltaxifolin-3′-O-glucoside, and 7-O-methyltaxifolin in S. alba cortex, respectively. PCA of high-resolution radical scavenging profiles revealed clear separation of preparations along principal component 1 due to the major radical scavengers (+)-catechin and ampelopsin. The new analytical platform allowed identification of 16 compounds in commercial willow bark extracts, and identification of ampelopsin, taxifolin, 7-O-methyltaxifolin-3′-O-glucoside, and 7-O-methyltaxifolin in S. alba bark extract is reported for the first time. The detection of the novel compound, ethyl 1-hydroxy-6-oxocyclohex-2-enecarboxylate, is also described.

► A new platform for comprehensive analysis of herbal preparations is presented.
► Proof-of-concept was performed with a small set of Salix preparations.
► HPLC and NMR provided chemical fingerprints for multivariate data analysis.
► A high-resolution ABTS reduction assay provided radical scavenging profiles for metabolomics.
► Structural identification was performed by the hyphenated HPLC–SPE–NMR technique.