Untargeted Metabolomics and anti-Helicobacter pylori Activity of Amphipterygium adstringens Bark: Influence of Tree Sex and Seasonal Variation.

Cuachalalate bark (Amphipterygium spp.), relevant in Mexican herbal medicine for its gastroprotective, and anti-Helicobacter pylori effects, as well as its reported antiproliferative/cytotoxic and antitumor effects, lacks systematic studies on bioactive metabolites influenced by biotic and abiotic factors. This study used untargeted 1H NMR metabolomics to characterize chemical profiles, quantify anacardic acids and 3α-hydroxymasticadienoic acid (3α-HMDA) according to climatic season, tree sex, and bark type (stem or branch), and evaluate anti-H. pylori activity. Three hundred sixty samples from 40 juvenile-adult A. adstringens trees were collected in Chiautla de Tapia, Puebla, Mexico. Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) of chemical profiles discriminated variations by collection month, tree sex, and bark type. Only minor variations in biomarker concentrations were recorded in specific cases (e.g., July > September/December [3α-HMDA: 28.9 vs. ∼19 mM; anacardic acids: 18.7 vs. ∼12 mM]; males > females in July [19.6 vs. 17.8 mM; 30.3 vs. 23.6 mM]; stem > branch in December [15.7 vs. 9.6 mM; 20.8 vs. 15.1 mM] and September [14.3 vs. 10.4 mM; 20.6 vs. 17.1 mM]), with no significant overall differences by seasonality, sex, or bark type. Despite some differences in biomarker concentrations in specific cases, anti-H. pylori activity remained stable (59.4 ± 14.6% at 0.125 mg/mL; 82.7 ± 11.8% at 0.25 mg/mL; and 99.5 ± 6.5% at 0.5 mg/mL). In conclusion, PCA/PLS-DA discriminated chemical variations by month, sex, and bark type, but biomarkers showed overall stability and consistent anti-H. pylori activity, supporting phytotherapeutic standardization.