CN114778723A - Method for detecting aspartame in wine - Google Patents
Method for detecting aspartame in wine Download PDFInfo
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- CN114778723A CN114778723A CN202210402658.XA CN202210402658A CN114778723A CN 114778723 A CN114778723 A CN 114778723A CN 202210402658 A CN202210402658 A CN 202210402658A CN 114778723 A CN114778723 A CN 114778723A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/065—Preparation using different phases to separate parts of sample
Abstract
The method for detecting the aspartame in the wine is provided, after the alcohol, the color and the protein are removed by a sample pretreatment method, the aspartame content in wine products can be quickly and accurately obtained, a potassium dihydrogen phosphate solution buffer salt solution is creatively selected in a mobile phase, the problem that the separation between a main peak and an impurity peak and between adjacent impurity peaks is difficult is thoroughly solved, and the analysis method is quick, high in accuracy, excellent in repeatability and reproducibility, simple to operate and high in sensitivity.
Description
Technical Field
The invention belongs to the technical field of wine analysis, and particularly relates to a method for detecting aspartame in wine.
Background
Aspartame is a substance obtained by condensing two amino acids, namely aspartic acid and phenylalanine, with methanol, has sweetness 180-220 times higher than that of sucrose, and contains less heat than common sucrose; one gram of aspartame has about 4 kcal. However, aspartame is required in such a small amount that it can neglect the calories it contains, and is therefore widely used as a replacement for sucrose. In recent years, liquor products have a high demand in the consumer market of China, but because some small enterprises and small workshops are not operated properly in the brewing process link, liquor bodies easily have bitter taste and foreign taste, some sweeteners can be illegally used for improving the liquor quality, and the intake of foods containing aspartame can cause: cranial nerve abnormality, carcinogenicity, brain tumor, normal bone development, abnormal viscera, etc.; therefore, in order to control and effectively reduce the food safety risk, it is necessary to detect aspartame in wine products, and in order to solve the above problems, an efficient, convenient and high-accuracy aspartame detection method needs to be developed.
Disclosure of Invention
The technical problems solved by the invention are as follows: the method for detecting the aspartame in the wine is provided, after the alcohol, the color and the protein are removed by a sample pretreatment method, the content of the aspartame in the wine product can be quickly and accurately obtained, a potassium dihydrogen phosphate solution buffer salt solution is creatively selected in a mobile phase, the problem of difficult separation between a main peak and an impurity peak and between adjacent impurity peaks is thoroughly solved, and the analysis method is quick, high in accuracy, excellent in repeatability and reproducibility, simple to operate and high in sensitivity.
The technical scheme adopted by the invention is as follows: the detection method of aspartame in wine comprises the following steps:
1) sample pretreatment: carrying out water bath ultrasonic treatment on the liquor product and then removing alcohol; removing impurities from a sample with a dark color in a fermented wine product; removing protein from the fermented wine product with high protein content by using a precipitator;
2) and (3) sample analysis: and acetonitrile and potassium dihydrogen phosphate buffer salt solution are used as mobile phases for analyzing and detecting by a high performance liquid chromatograph.
In the step 1), the liquor product is subjected to water bath ultrasonic treatment at 55-65 ℃ to remove alcohol in the sample pretreatment.
In the step 1), graphitized carbon is adopted to adsorb and remove impurities from the dark-colored products in the fermented wine products in the pretreatment of the sample.
In the step 1), the precipitant is potassium ferrocyanide and zinc acetate, and the fermented alcoholic product with high protein content in the pretreatment of the sample adopts potassium ferrocyanide and zinc acetate to precipitate protein to obtain supernatant.
Further, the water bath ultrasonic treatment time of the liquor product is 8-12 min at the water bath temperature of 55-65 ℃ when the water bath ultrasonic treatment is carried out.
In the step 1), accurately weighing 4 g-6 g of sample in a beaker, carrying out ultrasonic treatment in water bath at 55-65 ℃ for 8 min-12 min to remove alcohol smell, transferring the sample liquid to a 22 mL-28 mL volumetric flask, continuously adding 4 mL-6 mL of distilled water into the beaker, carrying out ultrasonic treatment for 4 min-6 min, transferring the liquid to the same volumetric flask, repeating the operation, and carrying out constant volume treatment on the white spirit product with water to scale lines.
In the step 1), accurately weighing 4g to 6g of sample in a beaker, carrying out ultrasonic treatment in a water bath at 55 ℃ to 65 ℃ for 8min to 12min without alcohol smell, adding 8mL to 12mL of 50% methanol water and 0.8g to 1.2g of graphitized carbon, shaking up, continuing to carry out ultrasonic treatment for 8min to 12min, transferring the sample solution to a 22mL to 28mL volumetric flask, continuing to add 4mL to 6mL of 50% methanol water in the beaker, carrying out ultrasonic treatment for 4min to 6min, transferring the liquid to the same volumetric flask, repeating the above operations, fixing the volume to a scale line by using water, carrying out vortex mixing, pouring into a centrifuge tube, and centrifuging at 3000r/min for 2min to 4 min.
In the step 1), accurately weighing 4-6 g of sample of the fermented wine product with high protein content, putting the sample into a beaker, carrying out water bath ultrasonic treatment at 55-65 ℃ for 8-12 min without alcohol smell, transferring the sample liquid into a 50mL colorimetric tube, continuously adding 4-6 mL of distilled water into the beaker, carrying out ultrasonic treatment for 4-6 min, transferring the liquid into the same volumetric flask, repeating the operation, adding a potassium ferrocyanide solution and a zinc acetate solution into the colorimetric tube, then carrying out constant volume treatment on the colorimetric tube to a scale line by using water, carrying out vortex mixing, pouring into a centrifuge tube, and centrifuging at 3000r/min for 2-4 min.
Further, when the protein of the fermented alcoholic beverage product with high protein content is removed, 0.8-1.2 mL of potassium ferrocyanide solution is added, 0.8-1.2 mL of zinc acetate solution is added, vortex is carried out, and the supernatant is obtained by centrifugation after constant volume.
Compared with the prior art, the invention has the advantages that:
1. according to the technical scheme, after the alcohol, the color and the protein are removed by the sample pretreatment method, the content of the aspartame in the wine product can be quickly and accurately obtained;
2. the technical scheme creatively selects the potassium dihydrogen phosphate solution buffer salt solution in the mobile phase, and thoroughly solves the problem of difficult separation between a main peak and impurity peaks and between adjacent impurity peaks;
3. the technical scheme has the advantages of high speed, high accuracy, excellent repeatability and reproducibility, simple operation and high sensitivity.
Drawings
FIG. 1 is a HPLC analysis chart of aspartame in white spirit in the first embodiment of the present invention;
FIG. 2 is a HPLC analysis chart of aspartame from kiwi fruit wine according to example two of the present invention;
FIG. 3 is a HPLC analysis chart of aspartame in fermented glutinous rice in example three of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to fig. 1 to 3 in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
It should be noted that, in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation. The use of the verb "to comprise a" does not exclude the presence of other, identical elements in a process, method, article or apparatus that comprises a stated element.
Instrumentation used in the examples:
25mL volumetric flask, 50mL volumetric flask, electronic balance (accurate to 0.001g), constant temperature water bath ultrasound and high performance liquid chromatograph;
reagents used in the examples:
acetonitrile (chromatographically pure), potassium dihydrogen phosphate, potassium ferrocyanide, zinc acetate, glacial acetic acid, methanol;
standard substance aspartame standard solution (concentration is 1000 mug/mL), diluted gradually with distilled water to prepare standard working solution 10 mug/mL, and the concentration of standard curve points is drawn as follows: 0.5. mu.g/mL, 1.0. mu.g/mL, 2.0. mu.g/mL, 5.0. mu.g/mL, 10.0. mu.g/mL;
the conditions of the HPLC apparatus used in the examples were as follows:
and (3) chromatographic column: c18A column; specification: 250mm multiplied by 4.6mm, 5 um; mobile phase: 20% acetonitrile: 80% of 0.01mol/L potassium dihydrogen phosphate; the wavelength of the ultraviolet detector is 200 nm; column temperature: 35 ℃; sample injection amount: 20 uL; flow rate of mobile phase: 1.0 mL/min;
example one
Accurately weighing about 5g of Chinese liquor in a beaker, and performing ultrasonic treatment in water bath at 60 ℃ for 10min without alcohol smell. And transferring the sample liquid into a 25mL volumetric flask, continuously adding 5mL of distilled water into the beaker, performing ultrasonic treatment for 5min, transferring the liquid into the same volumetric flask, repeating the operation, and performing constant volume to a scale mark by using water. And analyzing the constant volume liquid according to the instrument conditions of the high performance liquid chromatograph.
Example two
Accurately weighing about 5g of kiwi fruit wine in a beaker, carrying out ultrasonic treatment in a water bath at 60 ℃ for 10min without alcohol smell, adding 10mL of 50% methanol water and 1g of graphitized carbon, shaking up, continuing ultrasonic treatment for 10min, transferring the sample liquid to a 25mL volumetric flask, continuing adding 5mL of 50% methanol water into the beaker, carrying out ultrasonic treatment for 5min, transferring the liquid to the same volumetric flask, repeating the operation, and carrying out constant volume treatment with water to a scale mark. And (4) after vortex mixing, pouring into a centrifuge tube, centrifuging for 3min at 3000r/min, and taking supernate to analyze according to the conditions of a high performance liquid chromatograph.
EXAMPLE III
Accurately weighing about 5g of fermented glutinous rice (or rice wine) in a beaker, carrying out ultrasonic treatment in a water bath at 60 ℃ for 10min for no alcoholic smell, transferring the sample liquid to a 50mL volumetric flask, continuously adding 5mL of distilled water into the beaker, carrying out ultrasonic treatment for 5min, transferring the liquid to the same volumetric flask, and repeating the operation. Adding 1.0mL of potassium ferrocyanide solution and 1.0mL of zinc acetate solution into a volumetric flask, then adding water to a constant volume to a scale mark, uniformly mixing by vortex, pouring into a centrifuge tube, and centrifuging for 3min at 3000 r/min. And taking the supernatant to analyze according to the conditions of a high performance liquid chromatograph instrument.
And (4) performing analysis by a standard adding recovery quality control means on the three samples. The aspartame content (μ g/g) in the sample and its precision measured in the same laboratory are shown in table 1.
Precision: the absolute difference between two independent measurements obtained under repeated conditions must not exceed 10% of the arithmetic mean.
TABLE 1
As can be seen from Table 1, the reproducibility and reproducibility of the present invention is very good. Meanwhile, the recovery rate of the standard addition recovery rate experiment is 93.6-107.6%, which shows that the method has higher accuracy.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (9)
1. The method for detecting aspartame in wine is characterized by comprising the following steps:
1) sample pretreatment: carrying out water bath ultrasonic treatment on the liquor product and then removing alcohol; removing impurities from a sample with a darker color in a fermented wine product; removing protein from the fermented wine product with high protein content by using a precipitator;
2) and (3) sample analysis: acetonitrile and potassium dihydrogen phosphate buffer salt solution are used as mobile phases for analyzing and detecting by a high performance liquid chromatograph.
2. The method for detecting aspartame in wine according to claim 1, characterized in that: in the step 1), the liquor product is subjected to water bath ultrasonic treatment at 55-65 ℃ to remove alcohol in the sample pretreatment.
3. The method for detecting aspartame in wine according to claim 1, characterized in that: in the step 1), graphitized carbon is adopted to adsorb impurities of the dark products in the fermented alcoholic product in the sample pretreatment.
4. The method for detecting aspartame in wine according to claim 1, characterized in that: in the step 1), the precipitant is potassium ferrocyanide and zinc acetate, and the fermented wine product with high protein content in the pretreatment of the sample adopts potassium ferrocyanide and zinc acetate to precipitate protein to obtain supernatant.
5. The method for detecting aspartame in wine according to claim 1, characterized in that: and when the liquor product is subjected to water bath ultrasound, the water bath ultrasound time is 8-12 min at the water bath temperature of 55-65 ℃.
6. The method for detecting aspartame in wine according to claim 1, characterized in that: in the step 1), accurately weighing 4g to 6g of sample in a beaker, carrying out ultrasonic treatment in a water bath at 55 ℃ to 65 ℃ for 8min to 12min for no alcohol smell, transferring the sample liquid to a 22mL to 28mL volumetric flask, continuously adding 4mL to 6mL of distilled water into the beaker, carrying out ultrasonic treatment for 4min to 6min, transferring the liquid to the same volumetric flask, repeating the operation, and carrying out volume fixing with water to a scale mark.
7. The method for detecting aspartame in wine according to claim 1, characterized in that: in the step 1), accurately weighing 4g to 6g of sample in a beaker, carrying out ultrasonic treatment in a water bath at 55 ℃ to 65 ℃ for 8min to 12min without alcohol smell, adding 8mL to 12mL of 50% methanol water and 0.8g to 1.2g of graphitized carbon, shaking up, continuing to carry out ultrasonic treatment for 8min to 12min, transferring the sample solution to a 22mL to 28mL volumetric flask, continuing to add 4mL to 6mL of 50% methanol water in the beaker, carrying out ultrasonic treatment for 4min to 6min, transferring the liquid to the same volumetric flask, repeating the above operations, fixing the volume to a scale line by using water, carrying out vortex mixing, pouring into a centrifuge tube, and centrifuging at 3000r/min for 2min to 4 min.
8. The method for detecting aspartame in wine according to claim 1, characterized in that: in the step 1), accurately weighing 4-6 g of sample of the fermented wine product with high protein content, putting the sample into a beaker, carrying out water bath ultrasonic treatment at 55-65 ℃ for 8-12 min without alcohol smell, transferring the sample liquid into a 50mL colorimetric tube, continuously adding 4-6 mL of distilled water into the beaker, carrying out ultrasonic treatment for 4-6 min, transferring the liquid into the same volumetric flask, repeating the operation, adding a potassium ferrocyanide solution and a zinc acetate solution into the colorimetric tube, then carrying out constant volume treatment on the colorimetric tube to a scale line by using water, carrying out vortex mixing, pouring into a centrifuge tube, and centrifuging at 3000r/min for 2-4 min.
9. The method for detecting aspartame in wine according to claim 8, characterized in that: when the protein of the fermented wine product with higher protein content is removed, 0.8-1.2 mL of potassium ferrocyanide solution is added, 0.8-1.2 mL of zinc acetate solution is added, vortex is carried out, and the supernatant is obtained by centrifugation after constant volume.
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| CN202210402658.XA CN114778723A (en) | 2022-04-18 | 2022-04-18 | Method for detecting aspartame in wine |
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