Solid-phase extraction column performance verification substance for food vitamin detection and preparation method of solid-phase extraction column performance verification substance

Abstract

The invention discloses a solid-phase extraction column performance verification substance for food vitamin detection and a preparation method of the solid-phase extraction column performance verification substance, and relates to the technical field of food detection. Comprising a water-soluble vitamin composition, a fat-soluble vitamin composition, bovine serum albumin, lactose, maltodextrin, triglyceride and a stabilizer, the bovine serum albumin, the lactose, the maltodextrin, the triglyceride and other components are contained through an elaborately-designed matrix simulation system, and the components can truly simulate the complex environment in food, so that the food quality is improved, and the food quality is improved. Comprise possibly existing interference components such as protein and fat. Due to the design, the solid-phase extraction column can more accurately reflect the performance of the solid-phase extraction column in a real detection environment in a performance verification process, so that the verification accuracy and reliability are improved. The method has the advantages of real matrix simulation, collaborative verification of vitamins, good stability and the like, and a new solution is provided for the field of food vitamin detection.

Description

Technical Field

[0001] This invention relates to the field of food testing technology, specifically to a performance verification material and preparation method for a solid phase extraction column used in the detection of vitamins in food. Background Technology

[0002] In current practices for vitamin testing in food, the performance verification of solid phase extraction columns typically relies on testing the recovery rate using a standard solution of a single vitamin or a simple mixed solution of several vitamins.

[0003] However, current validation methods have several problems. First, the matrices of these validation samples are too homogeneous to accurately simulate the proteins, fats, and other potentially interfering components found in food. This means that the performance of solid-phase extraction columns may not meet expectations in practical applications. Second, water-soluble and fat-soluble vitamins are often processed separately during validation without synergistic validation. This results in an incomplete assessment of the selectivity of the extraction column, failing to ensure good performance across different types of vitamins. Finally, existing validation samples have poor stability and are prone to degradation at room temperature. This necessitates frequent reconstitution of validation solutions, increasing workload and affecting the reproducibility of test results. Therefore, we propose a performance validation sample and preparation method for a solid-phase extraction column for food vitamin detection. Summary of the Invention

[0004] The purpose of this invention is to address the problems of single-matrix validation compounds, which cannot accurately simulate food components, leading to insufficient synergistic validation of vitamins, incomplete selection evaluation of extraction columns, and unreliable performance. Furthermore, existing validation compounds suffer from poor stability, are easily degraded at room temperature, and require frequent re-preparation, increasing workload and affecting the reproducibility of test results. This invention provides a performance validation compound for a solid-phase extraction column used in food vitamin detection, along with its preparation method.

[0005] To achieve the above objectives, the present invention specifically adopts the following technical solution:

[0006] A performance verification compound for a solid-phase extraction column used in the detection of vitamins in food, comprising, by weight: 0.5-1.5 parts of a water-soluble vitamin composition, 0.2-0.8 parts of a fat-soluble vitamin composition, 10-15 parts of bovine serum albumin, 5-10 parts of lactose, 3-8 parts of maltodextrin, 2-5 parts of triglycerides, and 0.05-0.3 parts of a stabilizer.

[0007] Furthermore, the solid-phase extraction column performance verification material comprises, by weight, 0.5 parts of water-soluble vitamin composition, 0.2 parts of fat-soluble vitamin composition, 10 parts of bovine serum albumin, 5 parts of lactose, 3 parts of maltodextrin, 2 parts of triglycerides, and 0.05 parts of stabilizer.

[0008] Furthermore, the solid-phase extraction column performance verification material comprises, by weight, 1 part of water-soluble vitamin composition, 0.5 parts of fat-soluble vitamin composition, 12.5 parts of bovine serum albumin, 7.5 parts of lactose, 5.5 parts of maltodextrin, 3.5 parts of triglycerides, and 0.15 parts of stabilizer.

[0009] Furthermore, the solid-phase extraction column performance verification material comprises, by weight, 1.5 parts of water-soluble vitamin composition, 0.8 parts of fat-soluble vitamin composition, 15 parts of bovine serum albumin, 10 parts of lactose, 8 parts of maltodextrin, 5 parts of triglycerides, and 0.3 parts of stabilizer.

[0010] Further, the water-soluble vitamin composition comprises 0.1-0.2 parts of VB1, 0.15-0.25 parts of VB2, 0.1-0.15 parts of VB6, 0.01-0.03 parts of VB12, and 1.0-1.5 parts of VC.

[0011] Furthermore, the stabilizer is a mixture of ascorbate palmitate and α-tocopherol in a mass ratio of 3:1.

[0012] A method for preparing a performance verification compound for a solid-phase extraction column used in food vitamin detection includes the following steps:

[0013] S1. Dissolve bovine serum albumin, lactose, and maltodextrin in phosphate buffer solution at pH 7.2-7.6 at 40-45℃;

[0014] S2. Add triglycerides and emulsifier, and homogenize at 18-25 MPa pressure 3-5 times;

[0015] S3. Dissolve water-soluble vitamins in deionized water and fat-soluble vitamins in ethanol, then gradually mix them into the base solution.

[0016] S4. After repackaging, a three-step freeze-drying method is adopted: pre-freezing stage -40℃ / 4h → first drying -20℃ / 10Pa / 20-24h → second drying 25℃ / 5Pa / 5-8h.

[0017] Furthermore, the homogenizer parameters in S2 are 20000-25000 rpm, with a 2-minute interval between each cycle.

[0018] Furthermore, in the S4 three-step freeze-drying process, the moisture content is detected using near-infrared online monitoring, and the drying process is terminated when the moisture signal drops to 3%.

[0019] The beneficial effects of this invention are as follows:

[0020] 1. This invention utilizes a meticulously designed matrix simulation system containing bovine serum albumin, lactose, maltodextrin, and triglycerides. These components realistically simulate the complex environment in food, including potential interfering components such as proteins and fats. This design allows the solid-phase extraction column to more accurately reflect its performance in a real detection environment during performance validation, thereby improving the accuracy and reliability of the validation. Water-soluble and fat-soluble vitamins were synergistically processed and validated during the validation process. This approach not only considers the interactions between vitamins but also comprehensively evaluates the selectivity of the solid-phase extraction column. Through synergistic validation, the solid-phase extraction column can maintain good performance when faced with different types of vitamins, thus improving its applicability in the detection of complex samples. The stability of the validated samples was improved by adding stabilizers. This combination of stabilizers effectively prevents the degradation of vitamins under room temperature conditions, thereby extending the shelf life of the validated samples, reducing the frequency of reconstitution, reducing workload, and improving the reproducibility of detection results. With advantages such as realistic matrix simulation, synergistic vitamin validation, and good stability, this invention provides a new solution for the field of food vitamin detection.

[0021] 2. By precisely controlling the proportions of each component and the preparation process, this invention also enables a comprehensive and accurate evaluation of the performance of solid-phase extraction columns, including their extraction efficiency, selectivity, and enrichment capacity for trace substances. This not only improves the accuracy and reliability of vitamin detection in food but also provides strong support for the research and optimization of solid-phase extraction columns. This invention has broad application prospects and significant practical importance in the field of vitamin detection in food. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below.

[0023] This invention provides a solid-phase extraction column performance verification material for food vitamin detection, wherein the solid-phase extraction column performance verification material comprises, by weight: 0.5-1.5 parts of water-soluble vitamin composition, 0.2-0.8 parts of fat-soluble vitamin composition, 10-15 parts of bovine serum albumin, 5-10 parts of lactose, 3-8 parts of maltodextrin, 2-5 parts of triglycerides, and 0.05-0.3 parts of stabilizer.

[0024] In this embodiment, preferably, the water-soluble vitamin composition comprises 0.1-0.2 parts of VB1, 0.15-0.25 parts of VB2, 0.1-0.15 parts of VB6, 0.01-0.03 parts of VB12, and 1.0-1.5 parts of VC. The content of these water-soluble and fat-soluble vitamins in the composition is carefully designed to simulate the vitamin distribution in real food, thereby ensuring the accuracy and reliability of the solid-phase extraction column during the detection process.

[0025] In this embodiment, preferably, the stabilizer is a mixture of ascorbyl palmitate and α-tocopherol in a mass ratio of 3:1.

[0026] The synergistic effects of the components in the formula are as follows:

[0027] (1) Matrix simulation system

[0028] Bovine serum albumin (10-15 parts): This simulates protein interference components in food. Its isoelectric point (pI≈4.7) is similar to that of common food matrices. It can bind to vitamins through hydrogen bonds. This test examines the ability of the extraction column to release bound vitamins.

[0029] Triglycerides (2-5 parts): As a lipid-soluble carrier, they dissolve vitamins A / D / E while simulating lipid interference. Their melting point (-20℃~40℃) can verify the lipid adsorption efficiency of the extraction column under low temperature / normal temperature conditions.

[0030] Lactose + maltodextrin (8-18 parts): Construct a carbohydrate matrix and test the selectivity of the extraction column in the presence of sugars by competitive adsorption of hydroxyl groups against silica filler.

[0031] (2) Vitamin synergistic protection mechanism

[0032] Ascorbyl palmitate (0.05-0.3 parts): It has both water-soluble and fat-soluble antioxidant capabilities and is preferentially oxidized to protect vitamins B and C (oxidation potential: ascorbyl palmitate 0.28V < vitamin C 0.39V).

[0033] α-Tocopherol (combined with ascorbic acid): forms an oxidation-reduction cycle system (α-tocopherol → tocopherol quinone → ascorbic acid regeneration), making the degradation rate of vitamins A / D / E during the freeze-drying process <5%.

[0034] (3) Gradient concentration design

[0035] Water-soluble vitamins: fat-soluble vitamins = (0.5-1.5): (0.2-0.8): This ratio is designed based on 10 times the detection limit of vitamins in infant formula, such as GB 10765-2010, to ensure that low concentrations of vitamins, such as B12, can be accurately traced and to verify the trace enrichment capacity of the extraction column.

[0036] By precisely proportioning proteins, fats, and carbohydrates, the physicochemical properties (logP = -0.3 to 5.2) of the solid-phase extraction (SPE) column are validated across more than 90% of food samples. Simultaneously, a gradient concentration design simulates the actual vitamin content range in different foods, thereby comprehensively evaluating the extraction efficiency and selectivity of the SPE column at different concentrations. This design not only improves the practicality of the validation samples but also enhances the accuracy and reliability of SPE column performance validation. In practical applications, operators simply pass the prepared validation samples through the SPE column and then measure the recovery rate to quickly and accurately evaluate the SPE column's performance.

[0037] This invention utilizes a meticulously designed matrix simulation system containing bovine serum albumin, lactose, maltodextrin, and triglycerides. These components realistically simulate the complex environment in food, including potential interfering components such as proteins and fats. This design allows the solid-phase extraction column to more accurately reflect its performance in a real detection environment during performance validation, thereby improving the accuracy and reliability of the validation. Water-soluble and fat-soluble vitamins were synergistically processed and validated during the validation process. This approach not only considers the interactions between vitamins but also comprehensively evaluates the selectivity of the solid-phase extraction column. Through synergistic validation, the solid-phase extraction column can maintain good performance when faced with different types of vitamins, thus improving its applicability in the detection of complex samples. The stability of the validated samples was improved by adding stabilizers. This combination of stabilizers effectively prevents the degradation of vitamins under room temperature conditions, thereby extending the shelf life of the validated samples, reducing the frequency of reconstitution, reducing workload, and improving the reproducibility of detection results. With advantages such as realistic matrix simulation, synergistic vitamin validation, and good stability, this invention provides a new solution for the field of food vitamin detection.

[0038] A method for preparing a performance verification compound for a solid-phase extraction column used in food vitamin detection includes the following steps:

[0039] S1. Dissolve bovine serum albumin, lactose, and maltodextrin in phosphate buffer solution at pH 7.2-7.6 at 40-45℃;

[0040] S2. Add triglycerides and emulsifier, and homogenize at 18-25 MPa pressure 3-5 times;

[0041] S3. Dissolve water-soluble vitamins in deionized water and fat-soluble vitamins in ethanol, then gradually mix them into the base solution.

[0042] S4. After repackaging, a three-step freeze-drying method is adopted: pre-freezing stage -40℃ / 4h → first drying -20℃ / 10Pa / 20-24h → second drying 25℃ / 5Pa / 5-8h.

[0043] In this embodiment, preferably, the homogenizer parameters in S2 are 20,000-25,000 rpm, with a 2-minute interval between each cycle. This precise homogenization process ensures that triglycerides and emulsifiers are uniformly dispersed in the base liquid, thereby further improving the stability and homogeneity of the test sample.

[0044] In this embodiment, preferably, in the S4 three-step freeze-drying process, the moisture content is detected using near-infrared online monitoring, and drying is terminated when the moisture signal drops to 3%. This near-infrared online monitoring technology not only improves the accuracy of the drying process but also ensures that the moisture content of the test sample after freeze-drying meets the standard, further enhancing the stability and long-term preservation capability of the test sample.

[0045] Working principle and usage process of this invention:

[0046] In step S1, bovine serum albumin, lactose, and maltodextrin are dissolved under suitable pH and temperature conditions to form a stable matrix solution. Subsequently, in step S2, triglycerides and an emulsifier are added, and homogenization ensures thorough mixing of all components, forming a stable emulsion. The homogenizer parameters and circulation interval settings in this step ensure the stability and homogeneity of the emulsion. In step S3, water-soluble and fat-soluble vitamins are dissolved separately and then gradient-mixed into the matrix solution. This gradient mixing method simulates the actual vitamin content range in different foods, thus comprehensively evaluating the extraction efficiency and selectivity of the solid-phase extraction column at different concentrations. Finally, in step S4, the prepared validation sample is dried using a three-step freeze-drying method. The parameter settings for the pre-freezing stage, primary drying, and secondary drying ensure the drying effect and stability of the validation sample. Simultaneously, near-infrared online monitoring of moisture content accurately controls the drying process, preventing over-drying or under-drying.

[0047] By employing a three-step freeze-drying process (pre-freezing → primary drying → secondary drying) combined with a stabilizer, vitamin retention is >95% after 12 months of storage at 25°C, ensuring long-term stability and reproducibility of the validation samples. This significantly reduces the frequency of re-formulation and improves work efficiency. Furthermore, through precise control of the component ratios and preparation processes, this invention enables a comprehensive and accurate evaluation of the performance of solid-phase extraction columns, including their extraction efficiency, selectivity, and enrichment capacity for trace substances. This not only improves the accuracy and reliability of vitamin detection in food but also provides strong support for the research and optimization of solid-phase extraction columns. This invention has broad application prospects and significant practical value in the field of vitamin detection in food.

[0048] Example 1:

[0049] In this embodiment, preferably, the solid-phase extraction column performance verification material comprises, by weight, 0.5 parts of water-soluble vitamin composition, 0.2 parts of fat-soluble vitamin composition, 10 parts of bovine serum albumin, 5 parts of lactose, 3 parts of maltodextrin, 2 parts of triglycerides, and 0.05 parts of stabilizer.

[0050] Example 2:

[0051] In this embodiment, preferably, the solid-phase extraction column performance verification material comprises, by weight, 1 part of water-soluble vitamin composition, 0.5 parts of fat-soluble vitamin composition, 12.5 parts of bovine serum albumin, 7.5 parts of lactose, 5.5 parts of maltodextrin, 3.5 parts of triglycerides, and 0.15 parts of stabilizer.

[0052] Example 3:

[0053] In this embodiment, preferably, the solid-phase extraction column performance verification material comprises, by weight, 1.5 parts of water-soluble vitamin composition, 0.8 parts of fat-soluble vitamin composition, 15 parts of bovine serum albumin, 10 parts of lactose, 8 parts of maltodextrin, 5 parts of triglycerides, and 0.3 parts of stabilizer.

[0054] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A performance verification compound for a solid-phase extraction column used in the detection of vitamins in food, characterized in that, The solid-phase extraction column performance verification material comprises, by weight, 0.5-1.5 parts of water-soluble vitamin composition, 0.2-0.8 parts of fat-soluble vitamin composition, 10-15 parts of bovine serum albumin, 5-10 parts of lactose, 3-8 parts of maltodextrin, 2-5 parts of triglycerides, and 0.05-0.3 parts of stabilizer.

2. The performance verification material for a solid-phase extraction column for vitamin detection in food according to claim 1, characterized in that, The solid-phase extraction column performance verification material comprises, by weight, 0.5 parts of water-soluble vitamin composition, 0.2 parts of fat-soluble vitamin composition, 10 parts of bovine serum albumin, 5 parts of lactose, 3 parts of maltodextrin, 2 parts of triglycerides, and 0.05 parts of stabilizer.

3. The performance verification material for a solid-phase extraction column for vitamin detection in food according to claim 1, characterized in that, The solid-phase extraction column performance verification material comprises, by weight, 1 part of water-soluble vitamin composition, 0.5 parts of fat-soluble vitamin composition, 12.5 parts of bovine serum albumin, 7.5 parts of lactose, 5.5 parts of maltodextrin, 3.5 parts of triglycerides, and 0.15 parts of stabilizer.

4. The performance verification material for a solid-phase extraction column for vitamin detection in food according to claim 1, characterized in that, The solid-phase extraction column performance verification material comprises, by weight, 1.5 parts of water-soluble vitamin composition, 0.8 parts of fat-soluble vitamin composition, 15 parts of bovine serum albumin, 10 parts of lactose, 8 parts of maltodextrin, 5 parts of triglycerides, and 0.3 parts of stabilizer.

5. The performance verification material for a solid-phase extraction column for vitamin detection in food according to claim 1, characterized in that: The water-soluble vitamin composition comprises 0.1-0.2 parts of VB1, 0.15-0.25 parts of VB2, 0.1-0.15 parts of VB6, 0.01-0.03 parts of VB12, and 1.0-1.5 parts of VC.

6. The performance verification compound for a solid-phase extraction column for vitamin detection in food according to claim 1, characterized in that: The stabilizer is a mixture of ascorbyl palmitate and α-tocopherol in a mass ratio of 3:

1.

7. A method for preparing a performance verification compound for a solid-phase extraction column used in the detection of vitamins in food, characterized in that, It includes the following steps: S1. Dissolve bovine serum albumin, lactose, and maltodextrin in phosphate buffer solution at pH 7.2-7.6 at 40-45℃; S2. Add triglycerides and emulsifier, and homogenize at 18-25 MPa pressure 3-5 times; S3. Dissolve water-soluble vitamins in deionized water and fat-soluble vitamins in ethanol, then gradually mix them into the base solution. S4. After repackaging, a three-step freeze-drying method is adopted: pre-freezing stage -40℃ / 4h → first drying -20℃ / 10Pa / 20-24h → second drying 25℃ / 5Pa / 5-8h.

8. The method for preparing a performance verification compound for a solid-phase extraction column for vitamin detection in food according to claim 7, characterized in that: The homogenizer parameters in S2 are 20000-25000 rpm, with a 2-minute interval between each cycle.

9. The method for preparing a performance verification compound for a solid-phase extraction column for vitamin detection in food according to claim 7, characterized in that: In the S4 three-step freeze-drying process, the moisture content is detected using near-infrared online monitoring, and the drying process is terminated when the moisture signal drops to 3%.