Non-Targeted Metabolomic and Chemometric Analysis of Gardeniae Fructus (Zhizi): Linking Processing to Chemical and Bioactive Changes

Ethnopharmacological relevance Gardeniae Fructus (Zhizi, ZZ), the fruit of Gardenia jasminoides Ellis, exhibits anti-inflammatory and hepatoprotective effects, etc. Processing alters its properties, enhances therapeutic effects, and reduces toxicity. Furthermore, The chemical composition of ZZ seed and pericarp is significantly different. Studying the alterations in chemical components of the seed and pericarp under various processing conditions and the resulting variances in biological activities is highly valuable. Aim of the study: This study aimed to investigate the changes in the chemical composition and biological activity of ZZ during processing. Materials and methods This study employed high-performance liquid chromatography-diode array detection (HPLC-DAD) and liquid chromatography coupled to electrostatic orbitrap high-resolution mass spectrometry (LC-Orbitrap-MS), along with chemometric analysis and non-targeted metabolomics, to investigate variances in components of ZZ seed and pericarp and their transformations during processing. Lipopolysaccharide (LPS)-induced RAW 264.7 cells were utilized to evaluate the anti-inflammatory effects of ZZ by measuring nitric oxide (NO) production with the Griess reagent method. Furthermore, the levels of pro-inflammatory cytokines (TNF-α, IL-6) were determined using ELISA technique. Acetaminophen (APAP)-induced HepG2 cells were used to investigate the hepatoprotective of ZZ. Results Using non-targeted metabolomics and chemometrics, the study identified 353 differential features between the ZZ raw seed (RS) and the raw pericarp (RP). During processing, 382 and 386 significant features were found in seed and pericarp, respectively. A total of 210 compounds were identified by reference standards, fragmentation pattern, molecular networking, mzCloud, mzVault, in-house database, and a predictive database. Furthermore, bioassay tests showed reduced anti-inflammatory and hepatoprotective effects during processing. Geniposide and trans -crocin I were identified as primary anti-inflammatory and hepatoprotective compounds. Conclusion The chemical difference between the pericarp and seed of ZZ is obvious, and the chemical changes during processing have a significant impact on the activities. Proper processing is crucial for maximizing the therapeutic effects of ZZ.