An efficacy-linked strategy for the quality evaluation of traditional medicine: Chaenomeles speciosa as an example

Chaenomeles speciosa (Sweet) Nakai, a traditional remedy for "Bi-syndrome", possesses potent anti-inflammatory and analgesic activities. It is widely utilized in Traditional Chinese Medicine (TCM) and ethnic medical systems of Mongolian, Tibetan, and Miao nationalities.

However, in practical application, Chaenomeles speciosa is frequently confused with its related species Chaenomeles sinensis. This study aims to establish a holistic strategy to screen and validate Quality Markers (Q-markers) for Chaenomeles speciosa by integrating metabolomics, GNPS-guided isolation, and in vivo anti-inflammatory assay.

Initially, a plant metabolomics strategy was employed to screen differential markers. Then, GNPS molecular networking was utilized for the targeted isolation of differential markers from the core active fractions. Next, the anti-inflammatory effects of candidates from Chaenomeles speciosa on LPS-induced Tg(fli1:EGFP) zebrafish were evaluated by confocal microscopy and assessment of inflammatory factor levels. Finally, the selected Q-markers were quantified across 26 batches using a validated HPLC method.

A total of 134 components were characterized, with 15 differential markers prioritized to distinguish Chaenomeles speciosa from Chaenomeles sinensis. GNPS-guided targeted isolation successfully yielded 9 of the 15 differential markers, along with 8 additional constituents. Zebrafish screening identified oleanolic acid, chlorogenic acid, procyanidin B1, (-)-epicatechin, naringenin chalcone, apigenin, and diosmetin as potential Q-markers. The established HPLC method proved rapid and robust for routine quality control across 26 batches of Chaenomeles speciosa.