Characterization of White Frost on Exocarpium Citri Grandis: Flavonoid Crystallization Enhances Anti-Inflammatory Activities

(ECG) is a distinctive medicinal and edible product originating from southern China and is often covered with a layer of characteristic "white frost" (WF). This study investigated the composition, formation mechanism, microbial safety, and anti-inflammatory activity of the WF. Multi-technique analyses revealed that WF mainly consisted of crystalline naringin (~80% of total mass). Drying-induced shrinkage and rupture of oil glands on ECG suggested metabolite migration and surface crystallization as the key mechanisms for WF formation. Microbial profiling revealed no significant differences in fungal and bacterial communities between WF and non-frost (NF) samples, and none of eight common mycotoxins was detected, confirming its microbial safety. Brewing tests demonstrated that water boiling for 30 min achieved efficient extraction of naringin, with higher yields in WF samples than in NF samples. In RAW264.7 cells, both WF and NF extract significantly inhibited lipopolysaccharide-induced NO production as well as the secretion and transcription of TNF-α, IL-6, IL-1β, iNOS, and NF-κB, with WF extract showing a stronger effect. Overall, these findings indicate that WF originates from endogenous flavonoid crystallization rather than microbial contamination and enhances the anti-inflammatory activity. This study provides a scientific basis for quality evaluation, processing optimization, and standardization of ECG products.