Objective Given that the liquid formulation of the medicine-food homology formula Wuzhiyin is susceptible to non-enzymatic browning during storage, this study systematically investigates the underlying browning mechanism and evaluates the gastroprotective effect of Wuzhiyin prepared using an anti-browning processing technology against alcoholic gastric injury. Methods The differential metabolites and browning intermediates before and after browning of Wuzhiyin were analyzed by non-targeted ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and Spe-Pak microcolumn separation technology. The degree of browning was characterized by color changes, browning degree, and alterations in nutritional components. An anhydrous ethanol-induced acute gastric injury mouse model was established to evaluate the gastroprotective function of anti-browning prepared Wuzhiyin. Results Total sugars, total polyphenols, and total amino acids were significantly decreased by browning (P < 0.05). Nineteen key differential compounds were screened, among which fructose, sucrose, valine, isoleucine, tyrosine, and p-coumaric acid were identified as major driving factors. 2-Hydroxy-3-methyl-2-cyclopenten-1-one was identified as the core browning intermediate, and the browning mechanism was elucidated as a synergistic process involving the Maillard reaction, acid-promoted sugar degradation, and oxidative cross-linking of phenolic compounds. High-dose anti-browning prepared Wuzhiyin (43.72 g/kg) significantly reduced the gastric ulcer index (inhibition rate 72.42%) and alleviated pathological damage to the gastric mucosa. The underlying mechanisms include inhibiting nuclear factor-κB (NF-κB) pathway, reducing inflammatory cytokines [interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6], regulating oxidative stress [elevation of superoxide dismutase (SOD)/malondialdehyde ratio], and protecting mucosal barrier [modulation of NO, prostaglandin E₂ (PGE₂), trefoil factor 2 (TFF2)]. Conclusion Fructose, sucrose, valine, isoleucine, tyrosine, and p-coumaric acid are the key drivers of browning in Wuzhiyin; the Maillard reaction, acid-catalyzed sugar degradation, and phenolic oxidative cross-linking constitute the major browning pathways; and Wuzhiyin prepared via the anti-browning processing technology exhibits a significant gastroprotective effect against alcoholic gastric injury.

Wuzhiyin  /  non-enzymatic browning  /  browning mechanism  /  alcoholic gastric injury  /  metabonomics  /  fructose  /  sucrose  /  valine  /  isoleucine  /  tyrosine  /  p-coumaric acid  /  2-hydroxy-3-methyl-2-cyclopenten-1-one  /  Maillard reaction