Method for detecting content of betaine in beet molasses

Abstract

The invention relates to the technical field of food detection, in particular to a method for detecting the content of betaine in beet molasses, which is characterized in that anion resin is used for pretreatment, and then high performance liquid chromatography is used for detection. The method comprises the following steps: mixing resin, diatomite, activated carbon and a molasses diluted solution according to a ratio of (1-7): (1-2): 1: 10 (v / v), mixing for 4-5 hours by using a shaking table at 30-35 DEG C and 150-200rpm, centrifuging at 8000-10000 rpm, and taking supernate to obtain a test sample, and the technical scheme provides a good detection environment for later detection of betaine, so that the detection interference is small, and the accuracy is high.

Description

[Technical field]

[0001] The invention relates to the technical field of food detection, and in particular to a method for detecting the content of betaine in beet molasses. [Background technology]

[0002] Beet molasses is a byproduct of sugar production using beets as raw materials. It is a dark black or brown viscous substance with a slightly alkaline nature. Beet molasses contains about 80% dry matter, of which the total sugar content can reach 50-60%, mainly sucrose, and other sugar substances such as raffinose, glucose, fructose, and inositol galactoside. In addition, beet molasses also contains nutrients such as betaine, lipids, pectin, and amino acids. Betaine is a kind of R-(CH3)3N + A general term for CH2COO structured substances. Betaine is widely found in animals, plants and microorganisms. Due to its good biological activity, it is used in feed, daily chemicals and food additives. Betaine is most abundant in beets. The content of betaine in fresh beets can reach 0.3-0.7%, and the content of betaine in beet molasses is as high as 8%. At present, regarding the detection method of betaine content, titration is a method of determining the content of the substance to be tested by consuming a standard solution through a chemical reaction. In the detection of betaine, a perchloric acid standard solution is usually used for titration. Spectrophotometry is a method of quantitative analysis using the absorption characteristics of a substance to light. In the detection of betaine, spectrophotometry has the advantages of simple operation and high sensitivity. High performance liquid chromatography is a highly sensitive analytical method suitable for the separation and determination of components in complex mixtures. In the detection of betaine, HPLC has the advantages of good separation effect and high sensitivity. It also mainly includes: alkaloid precipitation method, alkaloid colorimetry, colorimetry, non-aqueous titration method, capillary electrophoresis, gas chromatography and liquid chromatography. These methods have their own characteristics and are suitable for different sample types and analysis requirements. Liquid chromatography is a commonly used method for detecting betaine content. However, the complex components in beet molasses seriously affect the detection of betaine content, which is not conducive to the process of separating betaine from beet molasses and limits the high-value utilization of beet molasses.

[0003] Therefore, it is necessary to develop a betaine detection method with simple operation, low interference and high accuracy. [Summary of the invention]

[0004] In view of the above, it is necessary to develop a method for detecting the betaine content in beet molasses. This technical solution is a betaine detection method with simple operation, low interference and high accuracy.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] A method for detecting betaine content in beet molasses comprises: performing detection by high performance liquid chromatography after pre-treatment with anion resin.

[0007] Further explanation, the method of pre-treatment using anionic resin is:

[0008] Using anionic resin, resin: diatomaceous earth: activated carbon: molasses diluted solution = 1-7:1-2:1:10 (v / v) were mixed, and then mixed on a shaker at 30-35°C, 150-200 rpm for 4-5 hours, and centrifuged at 8000-10000 rpm to obtain the supernatant to obtain the test sample.

[0009] To further illustrate, the molasses dilution solution is prepared by mixing molasses: deionized water at a ratio of 1:1-10 (w / w), and then oscillating and mixing to prepare the molasses dilution solution.

[0010] Further description, the specific conditions of the high performance liquid chromatography detection are:

[0011] Chromatographic column: hydrophilic interaction (Hilic) column;

[0012] Detector: UV detector / diode array detector;

[0013] Mobile phase: 0.1% acetic acid deionized water: acetonitrile = 12-16: 88-84 (v / v);

[0014] Flow rate: 0.2~0.5mL / min;

[0015] Detection wavelength: 195nm;

[0016] Column temperature: 30-35°C;

[0017] Injection volume: 5-30μL.

[0018] The solution preparation before the test includes the preparation of molasses dilution solution and the preparation of standard solution. The preparation of standard solution is as follows: accurately weigh 2.00g of betaine standard, add appropriate amount of deionized water to dissolve, place in a 100mL volumetric flask, add deionized water to the scale line of the volumetric flask, and prepare 20.00mg / mL betaine standard solution. Accurately measure 1.00, 2.00, 3.00, 4.00, 5.00mL of 20.00mg / mL betaine standard solution and place in a 10mL volumetric flask, add deionized water to the scale line of the volumetric flask, and prepare 2.00, 4.00, 6.00, 8.00, 10.00mg / mL betaine standard solution.

[0019] The present invention has the following beneficial effects:

[0020] Generally, the process of preparing betaine uses large-scale impurity removal equipment, which requires anion exchange resin and cation exchange resin to remove impurities from the supernatant. This large-scale equipment mainly passes the treated separated liquid through a column and is mainly used in large-scale filtration plants. However, this application is used in a laboratory.

[0021] The invention uses anion resin pretreatment and then uses high performance liquid chromatography for detection. The method of using anion resin pretreatment is as follows: using anion resin, mixing resin: diatomaceous earth: activated carbon: molasses dilution solution = 1-7: 1-2: 1: 10 (v / v), using a shaker at 30-35 DEG C, 150-200 rpm to mix for 4-5 hours, centrifuging at 8000-10000 rpm to take supernatant, and obtaining a test sample, thereby providing a good detection environment for the later detection of betaine, so that the interference of the detection is small and the accuracy is high. [Specific implementation method]

[0022] In order to make the above-mentioned purpose, features and advantages of the present invention more obvious and easy to understand, a detailed description is given below in conjunction with the specific embodiments of the present invention. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific implementation disclosed below.

[0023] Embodiment 1:

[0024] A method for pre-treating and determining betaine in beet molasses comprises the following steps:

[0025] S1. Solution configuration:

[0026] (1) Preparation of standard solution: Accurately weigh 2.00 g of betaine standard, add appropriate amount of deionized water to dissolve, place in a 100 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare a 20.00 mg / mL betaine standard solution. Accurately weigh 1.00, 2.00, 3.00, 4.00, 5.00 mL of 20.00 mg / mL betaine standard solution and place in a 10 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare 2.00, 4.00, 6.00, 8.00, 10.00 mg / mL betaine standard solutions.

[0027] (2) Preparation of molasses dilution solution: molasses: deionized water = 1:10 (w / w) and oscillate to prepare molasses dilution solution.

[0028] S2. Pretreatment: Use anionic resin, mix resin: diatomaceous earth: activated carbon: molasses diluted solution = 1:1:1:10 (v / v), use a shaker at 30°C, 150rpm for 4h, ensure that the resin, diatomaceous earth and activated carbon are fully adsorbed, then centrifuge at 8000rpm to obtain the supernatant to obtain the test sample.

[0029] S3. Chromatographic analysis: The HPLC system was an Agilent 1200 series liquid chromatograph equipped with a UV detector, ZORBAX Hilic Plus (2.1×150 mm, 1.8 μm), the mobile phase was deionized water-acetonitrile (12:88, V / V) containing 0.1% acetic acid, the column temperature was 35°C, the flow rate was 0.3 mL / min, the injection volume was 10 μL, and the detection wavelength was 195 nm.

[0030] Methodological investigation of the detection method:

[0031] (1) Precision test

[0032] Take 20.00 mg / mL betaine standard solution, and repeat the experiment 5 times according to the method of Example 1. The peak area is shown in Table 1. The calculated peak area RSD is 0.04%. The results show that this method has high precision in detecting betaine content.

[0033] Table 1 Example 1 Precision test results

[0034]

[0035] (2) Linear relationship investigation

[0036] 2.00, 4.00, 6.00, 8.00, 10.00, and 20.00 mg / ml betaine standard solutions were prepared and analyzed according to the method of Example 1. Data regression analysis was performed based on concentration (X) and peak area (Y). The linear regression equation was: Y = 4251.2X + 4593.1, R 2 =0.9973. The results showed that within the concentration range of 2 mg / mL-20.0 mg / mL, the peak area and concentration of betaine showed a good linear relationship.

[0037] (3) Repeatability test

[0038] Nine test sample solutions were taken and the experiment was carried out according to the method of Example 1. The peak areas are shown in Table 2. The peak area RSD was calculated to be 0.9%. The results showed that the method had good reproducibility in detecting betaine content.

[0039] Table 2 Repeatability test results of Example 1

[0040]

[0041] (4) Sample recovery investigation

[0042] Take 2.00mL of the test sample solution, add 6.8, 7.6, 7.9, 9.7, 9.6, 9.9, 12.9, 11.0, 13.7mg of betaine standard, respectively, analyze the betaine content in different samples by the method of Example 1, and calculate the recovery rate. After calculation, the average recovery rate is 100.3%, and the RSD is 1.5%, indicating that the method has a good recovery rate.

[0043] Embodiment 2:

[0044] A method for pre-treating and determining betaine in beet molasses comprises the following steps:

[0045] S1. Solution configuration:

[0046] (1) Preparation of standard solution: Accurately weigh 2.00 g of betaine standard, add appropriate amount of deionized water to dissolve, place in a 100 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare a 20.00 mg / mL betaine standard solution. Accurately weigh 1.00, 2.00, 3.00, 4.00, 5.00 mL of 20.00 mg / mL betaine standard solution and place in a 10 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare 2.00, 4.00, 6.00, 8.00, 10.00 mg / mL betaine standard solutions.

[0047] (2) Preparation of molasses dilution solution: molasses: deionized water = 1:1 (w / w) and oscillate to prepare molasses dilution solution.

[0048] S2. Pretreatment: Use anionic resin, mix resin: diatomaceous earth: activated carbon: molasses diluted solution = 2:1:1:10 (v / v), use a shaker at 35°C, 200 rpm for 5 hours, ensure that the resin, diatomaceous earth and activated carbon are fully adsorbed, then centrifuge at 10000 rpm to obtain the supernatant to obtain the test sample.

[0049] S3. Chromatographic analysis: The HPLC system was an Agilent 1200 series liquid chromatograph equipped with a UV detector, ZORBAX Hilic Plus (2.1×150 mm, 1.8 μm), the mobile phase was deionized water-acetonitrile containing 0.1% acetic acid (15:85, V / V), the column temperature was 35°C, the flow rate was 0.3 mL / min, the injection volume was 20 μL, and the detection wavelength was 195 nm.

[0050] Methodological investigation of the detection method:

[0051] (1) Precision test

[0052] Take 20.00 mg / mL betaine standard solution, and repeat the experiment 5 times according to the method of Example 2. The peak area is shown in Table 3. The calculated peak area RSD is 0.03%. The results show that this method has high precision in detecting betaine content.

[0053] Table 3 Example 2 Precision Experiment Results

[0054]

[0055] (2) Linear relationship investigation

[0056] 2.00, 4.00, 6.00, 8.00, 10.00, and 20.00 mg / ml betaine standard solutions were prepared and analyzed according to the method of Example 2. Data regression analysis was performed using concentration (X) and peak area (Y). The linear regression equation was: Y = 4250.2X + 4594.1, R 2 =0.9991. The results showed that within the concentration range of 2 mg / mL-20.0 mg / mL, the peak area and concentration of betaine showed a good linear relationship.

[0057] (3) Repeatability test

[0058] Nine test sample solutions were taken and the experiment was carried out according to the method of Example 2. The peak areas are shown in Table 4. The peak area RSD was calculated to be 0.9%. The results showed that the method had good reproducibility in detecting betaine content.

[0059] Table 4 Repeatability test results of Example 2

[0060]

[0061] (4) Sample recovery investigation

[0062] Take 2.00mL of the test sample solution, add 7.1, 7.8, 8.0, 10.0, 10.1, 11.2, 11.9, 12.5, 14.1mg of betaine standard, analyze the betaine content in different samples by the method of Example 2, and calculate the recovery rate. After calculation, the average recovery rate is 99.8%, and the RSD is 1.2%, indicating that the method has a good recovery rate.

[0063] Embodiment 3:

[0064] A method for pre-treating and determining betaine in beet molasses comprises the following steps:

[0065] S1. Solution configuration:

[0066] (1) Preparation of standard solution: Accurately weigh 2.00 g of betaine standard, add appropriate amount of deionized water to dissolve, place in a 100 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare a 20.00 mg / mL betaine standard solution. Accurately weigh 1.00, 2.00, 3.00, 4.00, 5.00 mL of 20.00 mg / mL betaine standard solution and place in a 10 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare 2.00, 4.00, 6.00, 8.00, 10.00 mg / mL betaine standard solutions.

[0067] (2) Preparation of molasses dilution solution: molasses: deionized water = 1:5 (w / w) and oscillate to prepare molasses dilution solution.

[0068] S2. Pretreatment: Use anionic resin, mix resin: diatomaceous earth: activated carbon: molasses diluted solution = 5:2:1:10 (v / v), use a shaker at 32°C, 160rpm for 4.5h, ensure that the resin, diatomaceous earth and activated carbon are fully adsorbed, then centrifuge at 8500rpm to obtain the supernatant to obtain the test sample.

[0069] S3. Chromatographic analysis: The HPLC system was an Agilent 1200 series liquid chromatograph equipped with a UV detector, ZORBAX Hilic Plus (2.1×150 mm, 1.8 μm), the mobile phase was deionized water-acetonitrile containing 0.1% acetic acid (15:85, V / V), the column temperature was 35°C, the flow rate was 0.3 mL / min, the injection volume was 20 μL, and the detection wavelength was 195 nm.

[0070] Methodological investigation of the detection method:

[0071] (1) Precision test

[0072] Take 20.00 mg / mL betaine standard solution, and repeat the experiment 5 times according to the method of Example 3. The peak area is shown in Table 5. The calculated peak area RSD is 0.03%. The results show that this method has high precision in detecting betaine content.

[0073] Table 5 Example 3 Precision Experiment Results

[0074]

[0075] (2) Linear relationship investigation

[0076] 2.00, 4.00, 6.00, 8.00, 10.00, and 20.00 mg / ml betaine standard solutions were prepared and analyzed using the method of Example 3. Data regression analysis was performed using concentration (X) and peak area (Y). The linear regression equation was: Y = 4250.2X + 4594.1, R 2 =0.9991. The results showed that within the concentration range of 2 mg / mL-20.0 mg / mL, the peak area and concentration of betaine showed a good linear relationship.

[0077] (3) Repeatability test

[0078] Nine test sample solutions were taken and the experiment was carried out according to the method of Example 3. The peak areas are shown in Table 6. The calculated peak area RSD was 0.7%. The results showed that the method had good reproducibility in detecting betaine content.

[0079] Table 6 Repeatability test results of Example 3

[0080]

[0081]

[0082] (4) Sample recovery investigation

[0083] Take 2.00mL of the test sample solution, add 5.2, 5.5, 6.0, 7.5, 12.1, 11.9, 11.8, 14.5, 14.1mg of betaine standard, analyze the betaine content in different samples by the method of Example 3, and calculate the recovery rate. After calculation, the average recovery rate is 99.1%, and the RSD is 1.4%, indicating that the method has a good recovery rate.

[0084] Embodiment 4:

[0085] A method for pre-treating and determining betaine in beet molasses comprises the following steps:

[0086] S1. Solution configuration:

[0087] (1) Preparation of standard solution: Accurately weigh 2.00 g of betaine standard, add appropriate amount of deionized water to dissolve, place in a 100 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare a 20.00 mg / mL betaine standard solution. Accurately weigh 1.00, 2.00, 3.00, 4.00, 5.00 mL of 20.00 mg / mL betaine standard solution and place in a 10 mL volumetric flask, add deionized water to the mark on the volumetric flask to prepare 2.00, 4.00, 6.00, 8.00, 10.00 mg / mL betaine standard solutions.

[0088] (2) Preparation of molasses dilution solution: molasses: deionized water = 1:8 (w / w) and oscillate to prepare molasses dilution solution.

[0089] S2. Pretreatment: Use anionic resin, mix resin: diatomaceous earth: activated carbon: molasses diluted solution = 7:2:1:10 (v / v), use a shaker at 35°C, 180 rpm for 5 hours, ensure that the resin, diatomaceous earth and activated carbon are fully adsorbed, then centrifuge at 8000-10000 rpm to obtain the supernatant to obtain the test sample.

[0090] S3. Chromatographic analysis: The HPLC system was an Agilent 1200 series liquid chromatograph equipped with a UV detector, ZORBAX Hilic Plus (2.1×150 mm, 1.8 μm), the mobile phase was deionized water-acetonitrile (16:84, V / V) containing 0.1% acetic acid, the column temperature was 35°C, the flow rate was 0.3 mL / min, the injection volume was 20 μL, and the detection wavelength was 195 nm.

[0091] Methodological investigation of the detection method:

[0092] (1) Precision test

[0093] Take 20.00 mg / mL betaine standard solution, and repeat the experiment 5 times according to the method of Example 4. The peak area is shown in Table 7. The calculated peak area RSD is 0.03%. The results show that this method has high precision in detecting betaine content.

[0094] Table 7 Example 4 Precision Experiment Results

[0095]

[0096] (2) Linear relationship investigation

[0097] 2.00, 4.00, 6.00, 8.00, 10.00, 20.00 mg / ml betaine standard solutions were prepared and analyzed by the method of Example 4. Data regression analysis was performed using concentration (X) and peak area (Y), and the linear regression equation was: Y = 4248.5X + 4589.1, R2 = 0.9981. The results showed that within the concentration range of 2 mg / mL-20.0 mg / mL of betaine, the peak area and concentration showed a good linear relationship.

[0098] (3) Repeatability test

[0099] Nine test sample solutions were taken and the experiment was carried out according to the method of Example 4. The peak areas are shown in Table 8. The calculated peak area RSD was 0.8%. The results showed that the method had good reproducibility in detecting betaine content.

[0100] Table 8 Repeatability test results of Example 3

[0101]

[0102] (4) Sample recovery investigation

[0103] Take 2.00mL of the test sample solution, add 6.0, 5.5, 6.5, 7.8, 11.4, 11.9, 11.5, 14.1, 14.6mg of betaine standard, analyze the betaine content in different samples by the method of Example 1, and calculate the recovery rate. After calculation, the average recovery rate is 98.8%, and the RSD is 1.5%, indicating that the method has a good recovery rate.

[0104] Embodiment 5:

[0105] A method for pre-treating and determining betaine in beet molasses comprises the following steps:

[0106] S1. Solution configuration: same as in Example 1.

[0107] S2. Pretreatment: Use anionic resin, mix the resin and molasses dilution solution at a ratio of 1:10 (v / v), use a shaker at 30°C and 150 rpm for 4 hours, ensure that the resin is fully adsorbed, and then centrifuge at 8000 rpm to obtain the supernatant to obtain the test sample.

[0108] S3. Chromatographic analysis: same as in Example 1.

[0109] Methodological investigation of the detection method:

[0110] (5) Precision test

[0111] Take 20.00 mg / mL betaine standard solution, and repeat the experiment 5 times according to the method of Example 5. The peak areas are shown in Table 9. The peak area RSD is calculated to be 1.47%. The results show that this method has high precision in detecting betaine content.

[0112] Table 9 Example 5 Precision Experiment Results

[0113]

[0114] (5) Linear relationship investigation

[0115] 2.00, 4.00, 6.00, 8.00, 10.00, and 20.00 mg / ml betaine standard solutions were prepared and analyzed using the method of Example 5. Data regression analysis was performed using concentration (X) and peak area (Y). The linear regression equation was: Y = 4012.4X + 4101.3, R 2=0.8256. The results showed that within the concentration range of 2 mg / mL-20.0 mg / mL, the peak area and concentration of betaine showed a good linear relationship.

[0116] (6) Repeatability test

[0117] Nine test sample solutions were taken and the experiment was carried out according to the method of Example 5. The peak areas are shown in Table 10. The calculated peak area RSD was 2.83%. The results showed that the method had good reproducibility in detecting betaine content.

[0118] Table 10 Repeatability test results of Example 5

[0119]

[0120]

[0121] (7) Sample recovery investigation

[0122] Take 2.00mL of the test sample solution, add 5.9, 6.3, 7.8, 7.9, 9.5, 10.5, 11.3, 12.8, 13.4mg of betaine standard, respectively, analyze the betaine content in different samples by the method of Example 1, and calculate the recovery rate. After calculation, the average recovery rate is 94.9%, and the RSD is 2.7%, indicating that the method has a good recovery rate.

[0123] Embodiment 6:

[0124] A method for pre-treating and determining betaine in beet molasses comprises the following steps:

[0125] S1. Solution configuration: same as in Example 1.

[0126] S2. Pretreatment: using anionic resin, mix resin: diatomaceous earth: molasses dilution solution = 1:1:10 (v / v), use a shaker at 30°C, 150rpm for 4h, ensure that the resin and diatomaceous earth are fully adsorbed, then centrifuge at 8000rpm to obtain the supernatant to obtain the test sample.

[0127] S3. Chromatographic analysis: same as in Example 1.

[0128] Methodological investigation of the detection method:

[0129] (6) Precision test

[0130] Take 20.00 mg / mL betaine standard solution, and repeat the experiment 5 times according to the method of Example 6. The peak area is shown in Table 11. The calculated peak area RSD is 0.59%. The results show that this method has high precision in detecting betaine content.

[0131] Table 11 Example 6 Precision Experiment Results

[0132]

[0133]

[0134] (8) Linear relationship investigation

[0135] 2.00, 4.00, 6.00, 8.00, 10.00, and 20.00 mg / ml betaine standard solutions were prepared and analyzed using the method of Example 6. Data regression analysis was performed using concentration (X) and peak area (Y). The linear regression equation was: Y = 4152.1X + 4015.1, R 2 =0.9256. The results showed that within the concentration range of 2 mg / mL-20.0 mg / mL, the peak area and concentration of betaine showed a good linear relationship.

[0136] (9) Repeatability test

[0137] Nine test sample solutions were taken and the experiment was carried out according to the method of Example 6. The peak areas are shown in Table 12. The peak area RSD was calculated to be 1.17%. The results showed that the method had good reproducibility in detecting betaine content.

[0138] Table 12 Repeatability test results of Example 6

[0139]

[0140] (10) Sample recovery investigation

[0141] Take 2.00mL of the test sample solution, add 7.5, 7.8, 8.3, 9.1, 10.2, 11.2, 12.0, 13.6mg of betaine standard, analyze the betaine content in different samples by the method of Example 1, and calculate the recovery rate. After calculation, the average recovery rate is 96.3%, and the RSD is 2.2%, indicating that the method has a good recovery rate.

[0142] Embodiment 7:

[0143] A method for pre-treating and determining betaine in beet molasses comprises the following steps:

[0144] S1. Solution configuration: same as in Example 1.

[0145] S2. Pretreatment: Use anionic resin, mix resin: activated carbon: molasses dilution solution = 1:1:10 (v / v), use a shaker at 30°C, 150rpm for 4h, ensure that the resin and activated carbon are fully adsorbed, then centrifuge at 8000rpm to obtain the supernatant to obtain the test sample.

[0146] S3. Chromatographic analysis: same as in Example 1.

[0147] Methodological investigation of the detection method:

[0148] (7) Precision test

[0149] Take 20.00 mg / mL betaine standard solution, and repeat the experiment 5 times according to the method of Example 7. The peak areas are shown in Table 13. The calculated peak area RSD is 0.61%. The results show that this method has high precision in detecting betaine content.

[0150] Table 13 Example 7 Precision Experiment Results

[0151]

[0152] (11) Linear relationship investigation

[0153] 2.00, 4.00, 6.00, 8.00, 10.00, and 20.00 mg / ml betaine standard solutions were prepared and analyzed using the method of Example 7. Data regression analysis was performed using concentration (X) and peak area (Y). The linear regression equation was: Y = 4152.7X + 4610.1, R 2 =0.9364. The results showed that within the concentration range of 2 mg / mL-20.0 mg / mL, the peak area and concentration of betaine showed a good linear relationship.

[0154] (12) Repeatability test

[0155] Nine test sample solutions were taken and the experiment was carried out according to the method of Example 7. The peak areas are shown in Table 14. The peak area RSD was calculated to be 1.27%. The results showed that the method had good reproducibility in detecting betaine content.

[0156] Table 14 Example 7 Repeatability Experiment Results

[0157]

[0158]

[0159] (13) Sample recovery investigation

[0160] Take 2.00mL of the test sample solution, add 6.5, 7.1, 8.6, 9.7, 10.8, 11.4, 11.8, 12.9mg of betaine standard, analyze the betaine content in different samples by the method of Example 1, and calculate the recovery rate. After calculation, the average recovery rate is 95.3%, and the RSD is 2.3%, indicating that the method has a good recovery rate.

[0161] From the above experimental comparison, it can be seen that the technical solution of the present application will use anion resin pretreatment and then use high performance liquid chromatography for detection. The method of using anion resin pretreatment is: using anion resin, resin: diatomaceous earth: activated carbon: molasses dilution solution = 1-7: 1-2: 1: 10 (v / v) is mixed, and a shaker is used at 30-35 ° C, 150-200 rpm for 4-5 hours, and the supernatant is centrifuged at 8000-10000 rpm to obtain a test sample, which provides a good detection environment for the later detection of betaine, so that the interference of the detection is small and the accuracy is high.

[0162] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for detecting betaine content in beet molasses, characterized in that: The specific method is: use anion resin pretreatment and then use high performance liquid chromatography for detection.

2. The method for detecting betaine content in beet molasses according to claim 1, characterized in that: The method of using anion resin pretreatment is: Using anionic resin, resin: diatomaceous earth: activated carbon: molasses diluted solution = 1-7:1-2:1:10 (v / v) were mixed, and then mixed on a shaker at 30-35°C, 150-200 rpm for 4-5 hours, and centrifuged at 8000-10000 rpm to obtain the supernatant to obtain the test sample.

3. The method for detecting betaine content in beet molasses according to claim 2, characterized in that: The molasses dilution solution is specifically prepared by mixing molasses: deionized water at a ratio of 1:1-10 (w / w), and then oscillating and mixing to prepare the molasses dilution solution.

4. The method for detecting betaine content in beet molasses according to claim 1, characterized in that: The specific conditions for the high performance liquid chromatography detection are: Chromatographic column: hydrophilic interaction (Hilic) column; Detector: UV detector / diode array detector; Mobile phase: 0.1% acetic acid deionized water: acetonitrile = 12-16: 88-84 (v / v); Flow rate: 0.2~0.5mL / min; Detection wavelength: 195nm; Column temperature: 30-35°C; Injection volume: 5-30μL.

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