关键词: 松茸, 制备工艺, 响应面法, 抗氧化活性, 食物过敏, 超高效液相色谱-串联质谱联用技术

Abstract: To explore the antioxidant and anti⁃food allergy activities of Tricholoma matsutake extract and enhance its high value⁃added application potential，using ethanol as the extraction solvent and total antioxidant capacity（T⁃AOC）as the evaluation index，single⁃factor tests combined with response surface methodology were adopted to systematically optimize the extraction process parameters.Food allergy model of BALB/c mice sensitized by ovalbumin（OVA）was constructed，and the levels of serum allergy⁃and immune⁃related factors were detected by ELISA to evaluate the potential intervention effect of the extract on food allergies. Furthermore，untargeted metabolomics analysis was performed using ultra⁃high performance liquid chromatography⁃tandem mass spectrometry（UPLC⁃MS/MS） to fully clarify the material basis underlying its anti⁃allergic effect.The results showed that after optimization by response surface methodology，the optimal preparation conditions for the Tricholoma matsutake extract were ethanol volume fraction of 88%，a liquid⁃to⁃solid ratio of 31∶1，extraction temperature of 31 ℃，and extraction time of 60 min.Under these conditions，the T⁃AOC of the extract reached 357.00 μmol/g.In vivo animal experiments confirmed that intervention with the Tricholoma matsutake extract effectively alleviated the clinical symptoms of allergic mice；significantly down⁃regulated the levels of Th2⁃type allergic mediators such as immunoglobulin E，mouse mast cell protease⁃1，and histamine（p<0. 01）；significantly up⁃regulated the level of the Th1⁃type cytokine IFN⁃γ（p<0.01）；and exhibited significant protective and repair effects against intestinal and splenic tissue damage caused by food allergies in mice.UPLC⁃MS/MS analysis revealed that the extract was rich in key substances involved in allergic immune regulation，such as L⁃phenylalanine and L⁃tryptophan，as well as a variety of bioactive components including brassinolide and α⁃eleostearic acid.In summary，the extract of Tricholoma matsutake prepared by this process has good antioxidant activity and has a significant intervention effect on food allergic reaction.The study clarifies that the active material basis of Tricholoma matsutake extract is L⁃phenylalanine and L⁃tryptophan which provides theoretical basis and data support for the development of high value⁃added Tricholoma matsutake products and new natural anti⁃allergic preparations.

Key words: Tricholoma matsutake, Preparation process, Response surface methodology, Antioxidant activity, Food allergy, Ultra?high performance liquid chromatography?tandem mass spectrometry