Meat freshness detection system integrating miniature PAGE separation, multispectral imaging and intelligent gray scale analysis

Abstract

The invention constructs a meat freshness detection system integrating miniature PAGE (Polyacrylamide Gel Electrophoresis) separation, multispectral imaging and intelligent gray scale analysis, which is characterized in that: a miniature electrophoresis module adopts gradient gel (4% T, 2.6% C) with pH of 5-9, and is matched with a platinum electrode (phi 0.1 mm) to realize rapid separation (migration rate 2.3 mm / min, 100V) of hemoglobin (Hb, 64kDa) and myoglobin (Mb, 17kDa); multi-mode imaging: ultraviolet (280 nm absorption), visible light (582 nm) and fluorescence imaging (SYPRO Ruby dyeing, LOD = 0.05 [mu] g / mL) are integrated; according to the intelligent analysis algorithm, based on a Gamma correction (gamma = 2.2) gray scale quantification model, an Hb / Mb ratio (HMR) is calculated through integral optical density (IOD), and an HMR-TVBN correlation equation (R2 = 0.927) is established. The detection period of the system is less than or equal to 30 minutes, and the sensitivity is improved by 5 times (the Mb detection limit is 0.08 mu g / mL) compared with that of ELISA (Enzyme-Linked Immunosorbent Assay), and meets the ISO 16140-3: 2021 rapid detection standard.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological separation and analysis, and particularly relates to the application of a meat freshness detection system integrating micro PAGE separation, multi-spectral imaging and intelligent gray-scale analysis in the evaluation of meat freshness. Background Art

[0002] Meat freshness, as a core biomarker characterizing the degree of spoilage, is closely related to indicators such as protein oxidative degradation (e.g., myoglobin MetMb ≥ 40%), microbial metabolites (TVB-N ≥ 15 mg / 100 g), and lipid peroxidation (TBARS ≥ 0.5 mg MDA / kg). Traditional detection methods have significant limitations: Sensory evaluation: affected by subjective cognitive bias (Cohen's κ < 0.6); Microbial culture method: time-consuming up to 24 - 48 h (ISO 15214:1998); Physicochemical detection: The determination of TVB-N requires a Kjeldahl nitrogen determination device and has complex pretreatment (GB 5009.228 - 2016); pH value detection is easily interfered by glycogenolysis during the postmortem rigor period (pH > 6.0 is the spoilage critical value); Bottlenecks of emerging technologies: Electronic nose is affected by environmental humidity (response value drops by 35% when RH > 80%); Near-infrared spectroscopy (NIRS) has an identification error of ±12% for the oxygenation state of myoglobin (due to interference from the O-H bond of water molecules); ELISA method only targets a single biogenic amine (e.g., the detection limit of histamine is 1 ppm) and cannot achieve synchronous analysis of multiple indicators.

[0003] Existing technologies generally have problems such as insufficient sensitivity (e.g., the detection limit of PCR for Pseudomonas is 10 3 CFU / g), difficulty in fusing multi-source data, and poor on-site applicability, and there is an urgent need to develop a new detection platform with both high specificity (AUROC ≥ 0.95) and high throughput (≥ 30 samples / h). Summary of the Invention

[0004] To overcome the problems that the existing methods and devices can no longer meet the detection of meat freshness and a series of consequences caused by inaccurate detection results, a meat freshness detection system integrating micro PAGE separation, multi-spectral imaging and intelligent gray-scale analysis is designed, including a micro electrophoresis module, multi-modal imaging and intelligent analysis algorithms. This design applies polyacrylamide gel electrophoresis technology to the detection of pork freshness, which can achieve simple operation, high sensitivity, good stability, effectively improve the detection accuracy of pork freshness, and enable relevant departments such as quality inspection to accurately screen according to the detection results.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] The present invention discloses a meat freshness detection system integrating micro PAGE separation, multispectral imaging and intelligent grayscale analysis, including a microelectrophoresis module, a multimodal imaging and an intelligent analysis algorithm. The micro PAGE electrophoresis system: polyacrylamide gel (pH 5-9, thickness 0.5 mm) and Tris-glycine buffer (25 mM Tris, 192 mM Gly, pH 8.3). This system is based on Tris as a buffer to maintain the pH stability during electrophoresis, glycine as a trailing ion, and chloride ion as a leading ion. The high-resolution separation of proteins is achieved by the synergistic effect of the ion mobility difference and the pH gradient. The multimodal imaging configuration: a three-modal optical system (ultraviolet CCD quantum efficiency ≥ 80%, 280 nm, fluorescence channel excitation / emission = 470 / 618 nm). The intelligent analysis algorithm: grayscale conversion based on the CIE 1931 luminance formula (Y = 0.299R + 0.587G + 0.114B) and Rolling Ball background correction (radius 50 pixels). This design applies polyacrylamide gel electrophoresis technology to meat freshness detection, which can achieve simple operation, high sensitivity and good stability, effectively improve the detection accuracy of pork freshness, and enable relevant departments such as quality inspection to accurately screen according to the detection results.

[0007] The present invention has the following beneficial effects:

[0008] Technical integration: For the first time, micro PAGE is coupled with multispectral imaging, breaking through the application barrier of proteomics technology in food detection;

[0009] Detection efficiency: The sensitivity (pg level) and throughput (30 samples / h) are improved synchronously, and the cost is reduced to 18% of the traditional method;

[0010] Standardized output: Objective grading is achieved through the HMR quantification model, avoiding the deviation of manual interpretation (the κ coefficient is increased to 0.89).

[0011] This system provides a standardized solution that complies with the 21 CFR Part 11 specification for meat freshness detection and is applicable to multi-scenario applications such as food processing, cold chain logistics and market supervision. Brief Description of the Drawings

[0012] Figure 1 It is a three-dimensional view of a meat freshness detection system integrating micro PAGE separation, multispectral imaging and intelligent grayscale analysis.

[0013] In the figure, 1. Microelectrophoresis module; 2. Multimodal imaging; 3. Intelligent analysis algorithm module. Detailed Description of the Invention

[0014] In order to make the technical means, creative features, achieved purposes and effects of the present invention easy to understand, the present invention will be further described below in conjunction with specific illustrations.

[0015] Example 1: In combination with Figure 1 To illustrate this embodiment, the present invention is a meat freshness detection system integrating micro PAGE separation, multi-spectral imaging and intelligent gray-scale analysis, including a microelectrophoresis module, a multimodal imaging and intelligent analysis algorithm.

[0016] Operation process:

[0017] Sample preparation: Take 0.5 g of pork tissue, add 200 μL of PBS (containing 0.1% Triton X-100) and homogenize; centrifuge (8,000×g, 4 °C, 10 min), take the supernatant and mix it with 20% glycerol;

[0018] Electrophoresis parameters: Gel (4% T, 2.6% C, pH 5-9, thickness 0.5 mm); Tris-glycine buffer (25 mM Tris, 192 mM Gly, pH 8.3); constant voltage of 100 V; electrophoresis time of 15 min;

[0019] Imaging analysis: UV imaging exposure time of 200 ms, fluorescence imaging gain ×4; determine freshness by the HMR value (threshold: HMR < 0.2 is first-class fresh).

[0020] The above are only the preferred embodiments of the present invention. The protection scope of the present invention is not limited to the above embodiments. Any technical solutions belonging to the principle of the present invention fall within the protection scope of the present invention. For those skilled in the art, several improvements made without departing from the principle of the present invention should also be regarded as within the protection scope of the present invention.

Claims

1. A meat freshness detection system integrating micro-PAGE separation, multi-spectral imaging and intelligent grayscale analysis, including a micro-electrophoresis module, multimodal imaging and intelligent analysis algorithm.

2. The meat freshness detection system integrating micro-PAGE separation, multi-spectral imaging and intelligent grayscale analysis according to claim 1 is characterized in that PAGE electrophoresis system: synergistic effect of polyacrylamide gel (pH 5-9, thickness 0.5 mm) and Tris-glycine buffer (25 mM Tris, 192 mM Gly,).

3. The meat freshness detection system integrating micro-PAGE separation, multi-spectral imaging and intelligent grayscale analysis according to claim 1 is characterized in that Multimodal imaging configuration: Trimodal optical system (UV CCD quantum efficiency ≥ 80%, 280nm, fluorescence channel excitation / emission = 470 / 618nm).

4. The meat freshness detection system integrating micro-PAGE separation, multi-spectral imaging and intelligent grayscale analysis according to claim 1 is characterized in that Intelligent analysis algorithm: Grayscale conversion based on CIE 1931 brightness formula (Y=0.299R+0.587G+0.114B) and Rolling Ball background correction (radius 50 pixels).

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