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CN-122025020-A - Method for determining quality requirements of intestinal canal improvement type functional food and application thereof

CN122025020ACN 122025020 ACN122025020 ACN 122025020ACN-122025020-A

Abstract

The application relates to the technical field of functional food quality control and research and development, and provides a method for establishing intestinal canal improvement type functional food quality requirements and application thereof. The method is based on obtaining digestion products by in-vitro simulated digestion, remarkably up-regulating metabolites after non-targeted metabonomics locking digestion, screening potential core metabolites by combining multiple evidence chains such as disease target point association, network topology analysis, molecular docking and the like, further reversely pushing and determining corresponding prototype compounds by in-vitro transformation verification, determining the core function prototype compounds as quality control markers by multiple linear regression analysis, finally establishing a quantitative effect relationship by using multimode indexes such as anti-inflammatory/barrier, mucous membrane permeability, microecological metabolism and the like, determining the lowest effective concentration, and forming quality qualification standards which are driven by functional threshold values, quantifiable, traceable and suitable for compound complex systems according to the minimum effective concentration.

Inventors

  • FENG JIANYING
  • CHEN SHUANGSHUANG
  • DU YING
  • ZHONG ZHIGANG
  • YING JIAN
  • LI TING
  • WU JING
  • LIU YANCHEN
  • WANG ZEYU
  • WANG JIALE
  • YU SHIYI
  • Wan Huaizhou

Assignees

  • 武汉轻工大学

Dates

Publication Date
20260512
Application Date
20260123

Claims (9)

  1. 1. A method for establishing the quality requirement of an intestinal canal improvement type functional food is characterized by comprising the following steps: S1, in-vitro simulated digestion, namely taking food to be researched, and selecting stomach-intestine, intestine-colon or stomach-intestine-colon full-stage for digestion simulation according to the action part of the food to be researched to obtain digestion products; s2, potential core metabolite primary screening S21, carrying out non-targeted metabonomics detection on the food raw materials before digestion and the digestion products in the step S1, screening to obtain a differential metabolite which is obviously up-regulated after digestion, and constructing a differential metabolite set; s22, predicting potential action targets based on the differential metabolite set in the step S21, taking intersection with related intestinal tract disease targets, constructing a metabolite-target-disease interaction network, and screening potential active metabolites and core targets; S23, carrying out molecular docking screening on the potential active metabolites and the core targets in the step S22 to obtain potential core metabolites; s3, reversely pushing the prototype compound by the active ingredient, namely performing metabolic transformation path analysis on the potential core metabolite in the step S23 and reversely pushing the prototype candidate, and then determining the prototype compound corresponding to the potential core metabolite through in vitro transformation verification; s4, establishing a standard, namely taking food to be researched in the step S1, constructing a sample library with different raw material ratios, carrying out chemical component spectrum measurement on the prototype compound in the step S3, carrying out in-vitro digestion, acting on an anti-inflammatory barrier protection model, setting an effective threshold, analyzing the contribution rate and the optimal ratio of each component to activity by using linear regression, and reversely deducing the minimum content requirement of each prototype component for ensuring the product to have the intestinal canal improvement function according to the output result of the model, thereby preparing the quality standard.
  2. 2. The method according to claim 1, wherein in step S21, the first 20 differential metabolites that are significantly up-regulated and most abundant after digestion are selected based on VIP not less than 1 and P <0.05 as a standard, and a differential metabolite set is constructed.
  3. 3. A method for establishing intestinal tract improving functional food quality requirements according to claim 1 wherein in step S22 network moderate, mid-centrality and near-centrality are screened for potential active metabolites and core targets of the first 20%.
  4. 4. A method for establishing a quality requirement of an intestinal tract improving functional food according to claim 1, wherein in step S23, molecular docking is performed using AutoDock 4.2 software.
  5. 5. The method for establishing a quality requirement of a functional food with improved intestinal tract as claimed in claim 4, wherein in step S23, potential core metabolites with strong binding capacity to the target are screened out based on the binding capacity of less than or equal to-5 kcal/mol.
  6. 6. The method according to claim 1, wherein in step S4, the effective threshold is a barrier repair rate 20% improvement and an anti-inflammatory inhibition rate not lower than 30%, or the minimum concentration of SCFA production 20% improvement is used as a quality criterion.
  7. 7. A method of establishing a gut-modified functional food quality claim according to any one of claims 1 to 6, further comprising the following: S5, multi-dimensional function verification, namely, respectively acting the core function prototype compound in the step S3 on an in-vitro anti-inflammatory barrier protection model, an intestinal mucosa permeability model and an artificial colon microecological model according to the quality standard concentration in the step S4, detecting TNF-alpha, IL-6 inflammatory factors, NO, tight junction protein expression and TEER values, MCU2 expression and fluorescein sodium permeability, and detecting intestinal gas and short chain fatty acid content to verify the intestinal improvement function.
  8. 8. The use of a method for establishing the quality requirements of an improved intestinal functional food according to claim 7 in the development and quality control techniques of improved intestinal functional foods.
  9. 9. The use according to claim 8, wherein the gut-modified functional food comprises one or more of a food of the group consisting of a food homolog, a probiotic and a dietary fiber.

Description

Method for determining quality requirements of intestinal canal improvement type functional food and application thereof Technical Field The invention relates to the technical field of functional food quality control and research and development, in particular to a method for establishing intestinal canal improvement type functional food quality requirements and application thereof. Background The intestinal health is closely related to the physiological functions of human bodies, the intestinal improvement type functional food becomes an industry hotspot, the quality of the functional food directly determines the effect realization, and establishment of scientific quality requirements is the core of the standard market. At present, related methods in the industry are mainly divided into two types, and have obvious limitations, so that the requirements of accurate control and efficacy guarantee cannot be met. The first type is a single component guiding method, which takes the content of single active components (such as total flavonoids and probiotics) in raw materials as a core index, and refers to national standard set content threshold value to judge quality. The method only focuses on the initial content of the components, and does not consider the digestion and transformation process and bioavailability of the components in the gastrointestinal tract, so that quality evaluation is disjointed from actual efficacy. The second type is a single model validation method, which relies on a single cell model (such as Caco-2 cell permeability detection) or only focuses on flora changes, lacks full chain analysis of "metabolite-receptor-function", and cannot systematically validate the relevance of components to intestinal tract improvement efficacy. The prior art has obvious defects that firstly, the degradation and transformation path of the component gastrointestinal tract cannot be tracked, the raw materials and the efficacy are unhooked, secondly, metabolites are primarily screened only through non-targeted metabonomics, multi-technology fine screening is not combined, false positive components are easy to mix, thirdly, a verification system is imperfect, an anti-inflammatory-barrier repair-microecology regulation full chain is not covered, core microecology index detection is omitted, and fourthly, a quantitative threshold based on biological activity is lacking, so that superior and inferior products are difficult to define. Aiming at the defects, an optimization method is needed in the field, but multiple technical problems are faced, namely, the gastrointestinal digestion process is complex, the data matching difficulty of each technical link is high, a coherent system is difficult to form in the prior art, the reverse thrust of metabolites to raw material prototypes is easy to be subjected to multiple interference and lacks an accurate verification means, the accurate correspondence of function quantification indexes, component concentration thresholds and quality standards cannot be realized, and the traditional method is more aimed at single raw materials, is difficult to be suitable for preparing and eating homologous compound raw materials and has insufficient universality. In conclusion, the existing method has the problems of weak efficacy relevance, incomplete verification, missing quantitative standards, poor universality and the like, and is difficult to form a scientific evaluation system due to the restriction of technical problems. Therefore, a combined method which covers the whole process, precisely locks the core components and gives consideration to universality and quantitative standards is developed, and the combined method has great significance to industry standardization development. Disclosure of Invention In view of the above, the invention provides a method for establishing the quality requirements of intestinal canal improvement type functional foods and application thereof, and aims to overcome the defects of disjointed indexes and functions, lack of prototype tracing, low screening accuracy, single verification dimension, unscientific standard setting, and the technical difficulties of establishing a closed-loop system, verifying transformation association, associating functions and standards and the like in the existing establishment method of the quality requirements of the intestinal canal improvement type functional foods. The technical scheme of the invention is realized in such a way that on one hand, the invention provides a method for establishing the quality requirement of the intestinal canal improvement type functional food, which comprises the following steps: S1, in-vitro simulated digestion, namely taking food to be researched, and selecting stomach-intestine, intestine-colon or stomach-intestine-colon full-stage for digestion simulation according to the action part of the food to be researched to obtain digestion products. S2, potential core metabolite primary