CN108037117B - Method and kit for rapidly detecting industrial hydrogen peroxide residue in food - Google Patents
Method and kit for rapidly detecting industrial hydrogen peroxide residue in food Download PDFInfo
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Abstract
The invention relates to a method for rapidly detecting industrial hydrogen peroxide residue in food, which is characterized by comprising the following steps of: preparation of a sample solution: soaking a food sample in a dichloromethane solution, shaking and taking a supernatant to obtain the food; preparation of a standard solution: dissolving appropriate amount of 2-ethyl anthraquinone standard substance in dichloromethane to obtain the product; dropwise adding concentrated sulfuric acid into the sample solution or the standard solution, and developing color; sequentially dropwise adding a sodium hydroxide solution and a phenolphthalein test solution, and developing; dropwise adding a sodium hydroxide solution into the sample solution or the standard solution, and standing after slight shaking; dripping 5% mixed solution of malonic ester and dichlorohexane, heating, and developing. The used extracting solution and reaction test solution are common chemical reagents, and the method is very safe, simple and convenient to operate, short in experimental process time and low in analysis cost.
Description
Technical Field
The invention relates to the field of food detection, in particular to a method and a kit for rapidly detecting industrial hydrogen peroxide residue in food.
Background
The food-grade hydrogen peroxide has the characteristics of broad spectrum, high efficiency, no toxicity and no residue, and can play the roles of disinfection, sterilization, bleaching and the like when being used as a production processing aid in the production process of foods, fruits, vegetables and beverages. In foreign countries, food grade hydrogen peroxide is also widely used as a food processing aid in the pore-making agent of cheese, the bleaching of meat products, tea leaves and nuts, the modification of proteins such as starch and milk, and the like. In addition, the food grade hydrogen peroxide is more used for disinfection and sterilization of food. However, if industrial hydrogen peroxide is used as a food processing aid and is used for whitening food such as chicken feet, pork tripe and the like, the industrial hydrogen peroxide can be left in the food, and heavy metals and impurities in the food are harmful to human bodies.
Whether the anthraquinone impurities are contained or not is one of the main differences between the industrial hydrogen peroxide and the food-grade hydrogen peroxide, so that the method for detecting the residues of the anthraquinone compounds in the food can be used for representing whether the food is treated by the industrial hydrogen peroxide or not. At present, the commonly used method for detecting anthraquinone compounds is an HPLC method, which comprises the steps of adding water and hydrochloric acid for hydrolysis, refluxing in boiling water, cooling and filtering. Extracting the filtrate with diethyl ether, volatilizing at room temperature, dissolving the residue with methanol, filtering, and detecting the content of anthraquinone components by liquid chromatography; or extracting food with methanol, hydrolyzing with acid, oxidizing bound anthraquinone to free anthraquinone, oxidizing reduced anthraquinone such as anthralin, anthrone, and dianthrone to oxidized anthraquinone, extracting with diethyl ether, and measuring with magnesium acetate methanol solution. The two common detection methods have high operation difficulty and long consumption time, and are not beneficial to on-site rapid detection and analysis during food quality detection.
Disclosure of Invention
In view of the above problems, an object of the present invention is to provide a method for rapidly detecting industrial hydrogen peroxide residue in food.
The specific technical scheme is as follows:
a method for rapidly detecting industrial hydrogen peroxide residue in food comprises the following steps:
preparation of a sample solution: soaking food sample with dichloromethane, shaking, and collecting supernatant;
preparation of a standard solution: dissolving appropriate amount of 2-ethyl anthraquinone standard substance in dichloromethane to obtain the product;
dropwise adding concentrated sulfuric acid into one part of the sample solution or the standard solution, and developing color; sequentially dropwise adding a sodium hydroxide solution and a phenolphthalein test solution, and developing;
dropwise adding a sodium hydroxide solution into the other sample solution or standard solution, slightly shaking and standing; dropwise adding a 5% malonate dichloromethane solution, heating and developing.
In some embodiments, the concentrated sulfuric acid is 98% concentrated sulfuric acid, the concentration of the sodium hydroxide solution is 10mol/L, and the concentration of the phenolphthalein test solution is 5 g/L.
In some embodiments, the above-mentioned rapid detection method includes the following steps:
preparation of a sample solution: soaking a food sample in a dichloromethane solution, shaking and taking a supernatant to obtain the food;
preparation of a standard solution: dissolving appropriate amount of 2-ethyl anthraquinone standard substance in dichloromethane to obtain the product;
dropwise adding concentrated sulfuric acid into the sample solution or the standard solution, developing color, and measuring absorbance at the wavelength of 530 nm; sequentially dropwise adding a sodium hydroxide solution and a phenolphthalein test solution, and developing;
dropwise adding a sodium hydroxide solution into the sample solution or the standard solution, and standing after slight shaking; dripping 5% dichlorohexane solution of malonate, heating, and developing.
In some of these embodiments, the above-described standard solution preparation comprises: dissolving 1mg of 2-ethyl anthraquinone standard substance in 10ml of dichloromethane, and performing gradient dilution to obtain the product.
In some of these embodiments, the shaking is at room temperature for 10-20 s.
In some of these embodiments, the heating is: heating at 60-80 deg.C for 5-15 min.
The invention also aims to provide a rapid detection kit for industrial hydrogen peroxide residue in food. The specific technical scheme is as follows:
a kit for rapidly detecting industrial hydrogen peroxide residue in food comprises the dichloromethane solution, concentrated sulfuric acid, sodium hydroxide solution, phenolphthalein test solution and 5% malonate dichlorohexane solution, wherein the dichloromethane is sealed with water during storage.
In some embodiments, the rapid detection kit further comprises a 2-ethyl anthraquinone standard.
The rapid detection method and the kit for industrial hydrogen peroxide residue in food have the following beneficial effects:
the rapid detection method for the residual anthraquinone compounds in the food uses dichlorohexane to rapidly extract an object to be detected, the anthraquinone compounds have weak alkalinity due to the existence of oxygen ions, can be dissolved in concentrated sulfuric acid and salified, and is accompanied with color change, and the reaction is simple; and in addition, in combination with the neutralization reaction of sodium hydroxide and concentrated sulfuric acid, the excessive alkali is indicated by the color development of the phenolphthalein test solution, and the sample solution is further verified to contain substances which react with the concentrated sulfuric acid. Meanwhile, a new sample solution is taken, and the differential experiment of anthraquinone, benzoquinone and naphthoquinone is used to eliminate the interference of other quinone substances in the rapid detection of benzoquinone. The reaction test solution used by the rapid detection method is easy to obtain, the experimental operation is simple, convenient and rapid, the result can be judged through the color reaction, no complex instrument and equipment are needed, the cost is low, and the method is favorable for on-site analysis. And the persuasion is strong through two-step mutual verification.
Furthermore, the method adopts a method for measuring absorbance and a standard curve, utilizes the color reaction of anthraquinone and concentrated sulfuric acid to carry out preliminary quantitative test on the residue of the industrial hydrogen peroxide anthraquinones in the food, and is simple and easy to implement.
The kit for rapidly detecting the industrial hydrogen peroxide residue in the food integrates the reagents required for rapidly detecting the industrial hydrogen peroxide in the food, so that the detection is more convenient.
Drawings
FIG. 1 is a standard curve obtained by measurement of a 2-ethylanthraquinone standard in example 2;
FIG. 2 is a plot of the 2-ethylanthraquinone standard from example 3.
Detailed Description
The present invention provides a method and a kit for rapidly detecting hydrogen peroxide residue in food, which will be further described with reference to the following examples.
Example 1
The present embodiment is a rapid test kit for rapidly detecting industrial hydrogen peroxide residue in a sea cucumber sample by using the method of the present invention, and the rapid test kit comprises the following reagents:
reagent A: subpackaging small bottles, adding 6ml of dichloromethane into each small bottle, adding 1ml of distilled water along the wall, sealing, and performing one-time quick test by using a small bottle of reagent;
and (3) reagent B: 98% concentrated sulfuric acid (18.4 mol/L);
and (3) reagent C: 10mol/L sodium hydroxide solution;
and (3) reagent D: 5g/L phenolphthalein test solution;
and (3) reagent E: 5% malonic ester in dichloromethane (v/v).
The detection steps are as follows:
(1) sea cucumber samples: adding 1ml of the leacheate of the sea cucumber sample into a centrifugal tube;
(2) adding 1 bottle of reagent A into the centrifuge tube, covering the centrifuge tube with a cover, screwing down, fully shaking or manually shaking vigorously for 10-20s, and measuring. 2 parts of the solution to be tested should be prepared.
(3) Detecting the first part of liquid to be detected:
adding 3 drops of reagent B into a centrifugal tube of the liquid to be detected, and observing the phenomenon after shaking slightly;
8 drops of reagent C and 4 drops of reagent D were added dropwise to the above reagents, and the reaction was observed after shaking gently.
(4) And (3) detecting the second to-be-detected liquid:
adding 4 drops of reagent C into a centrifugal tube of the liquid to be detected, slightly shaking and standing for 1 minute;
4 drops of reagent E were added dropwise to the above reagent, and after heating at 60 ℃ for 5 minutes, the solution phenomenon was observed.
The qualitative determination of the results is shown in table 1:
TABLE 1
Since the reagent A contains dichlorohexane and water, the layering phenomenon occurs, and the color reaction in the table is analyzed as follows:
the first part of the solution to be detected, namely the step (3): the upper layer and the lower layer of the qualified sample are initially colorless after concentrated sulfuric acid is dripped, the liquid surface of the upper layer is initially red after sodium hydroxide and phenolphthalein are dripped, and the upper layer and the lower layer are colorless after being lightly shaken and uniformly mixed; and after the concentrated sulfuric acid is dripped into the unqualified sample, the upper layer is colorless, the lower layer is initially yellow or yellowish brown, after the sodium hydroxide and the phenolphthalein are dripped, the liquid level of the upper layer is initially red, the red color does not disappear or only becomes a little after the mixture is shaken and uniformly mixed, and the solution of the lower layer is changed from yellow or yellowish brown to colorless. The darker the red color, the higher the content of the target component in the solution.
The second part of the solution to be detected, namely the step (4): the upper and lower layers of the qualified sample are initially colorless, and do not change after sodium hydroxide is added dropwise, and the solution is colorless after 5% of malonic ester solution is added dropwise and heated. Unqualified samples are colorless at first, sodium hydroxide is dripped without change, 5% malonic ester dichloromethane solution is dripped, and the solution is blue-green or blue-purple after heating.
The qualitative detection of the above samples gave a color reaction with the following results:
and (3) dropwise adding concentrated sulfuric acid into the first to-be-detected liquid, then enabling the upper layer to be colorless, enabling the lower layer to be yellow or yellowish brown, dropwise adding sodium hydroxide and phenolphthalein, enabling the liquid surface of the upper layer to be red, shaking gently, uniformly mixing, and enabling the red not to disappear, wherein the solution of the lower layer is changed from yellow or yellowish brown to colorless.
The second test solution was initially colorless, sodium hydroxide was added dropwise unchanged, a dichloromethane solution of 5% malonic ester was added dropwise, and after heating, the solution … … was blue-green.
And (3) taking the 2-ethyl anthraquinone standard solution with the concentration of 10ppm, and detecting according to the steps, wherein the color reaction in the result is consistent with that of the sample solution.
In addition, the sample solution was confirmed to contain 5mg/kg of 2-ethylanthraquinone by the method of GB/T23672-.
Example 2
In this embodiment, the method of the present invention is used to rapidly detect the industrial hydrogen peroxide residue in the sea cucumber sample, and the following reagents are used:
reagent A: dichloromethane; sealing with water liquid during storage;
and (3) reagent B: 98% concentrated sulfuric acid (18.4 mol/L);
and (3) reagent C: 10mol/L sodium hydroxide solution;
and (3) reagent D: 5g/L phenolphthalein test solution;
and (3) reagent E: 5% malonic ester in dichloromethane (v/v).
The detection steps are as follows:
(1) preparation of a sample solution: taking 5g of sea cucumber sample, shearing, soaking in 20ml of dichloromethane, shaking vigorously at room temperature for 20s, standing, and taking supernatant to obtain the sea cucumber extract;
(2) preparation of a standard solution: taking a proper amount of 2-ethyl anthraquinone standard substance, dissolving with dichloromethane, and performing gradient dilution to obtain 0.625, 3.125, 12.5, 35.25, 45, 62.5ppm 2-ethyl anthraquinone standard substance solution;
(3) taking 7ml of the sample solution and the standard solution with each concentration, dropwise adding 0.92ml of 98% concentrated sulfuric acid, sufficiently shaking, observing, and measuring absorbance at 530nm wavelength, namely OD 530; and (3) sequentially dropwise adding 1ml of sodium hydroxide solution and 4 drops of phenolphthalein test solution, fully shaking up, and observing.
(4) Taking another 7ml of sample solution and standard solution with various concentrations, dripping 1ml of sodium hydroxide solution into the sample solution by using a rubber head dropper, shaking slightly, and standing for 1 min; 4 drops of 5% malonic ester dichlorohexane solution are added dropwise, heated at 60 ℃ for 5min and the color is observed.
(5) And drawing a standard curve, and calculating the anthraquinone content in the sample.
The qualitative determination of the results is shown in table 1, the quantitative determination is shown in table 2, and the standard curve is drawn according to the standard solution as shown in fig. 1:
TABLE 2
In the detection process of the embodiment, the observation result of the sample solution in the step (3) is colorless after concentrated sulfuric acid is dripped, the liquid level is red after phenolphthalein is dripped, the red color disappears after the phenolphthalein is uniformly shaken, the solution becomes colorless, and the solution becomes blue-green after the heating in the step (4).
In this example, the absorbance of the standard solution was measured between 0.209 and 2.158 according to the concentration, and the standard curve was plotted as y 10.611x2+7.0878 x-1.9325. When the absorbance is more than 0.2, the error is small, so the detection limit of the method can reach 0.625 ppm.
In this example, the OD530 value of the sample solution was measured to be 0.028, and the absorbance thereof was much lower than 0.2, which is not in the range measurable by the standard curve, so that the sample contained no 2-ethylanthraquinone exceeding the detection limit, but contained benzoquinone or naphthoquinone.
The sample solution is confirmed to contain no 2-ethyl anthraquinone by using a detection method of GB/T23672-.
Example 3
In this example, the method of the present invention is used to rapidly detect the industrial hydrogen peroxide residue in the chicken feet with pickled peppers, and the reagents used in the method are the same as those in example 1.
The method comprises the following steps:
(1) preparation of a sample solution: taking 5g of chicken feet with pickled peppers, cutting into pieces, immersing the chicken feet with pickled peppers in 50ml of dichloromethane solution, shaking vigorously at room temperature for 20s, filtering, and taking supernatant fluid to obtain the chicken feet with pickled peppers;
(2) preparation of a standard solution: taking a proper amount of 2-ethyl anthraquinone standard substance, dissolving with dichloromethane, and performing gradient dilution to obtain 0.625, 3.125, 12.5, 35.25, 45, 62.5ppm 2-ethyl anthraquinone standard substance solution;
(3) taking 5ml of the sample solution and the standard solution with each concentration, dropwise adding 0.46ml of 98% concentrated sulfuric acid into the sample solution, sufficiently shaking, observing, and measuring absorbance at 530nm, namely OD 530; dripping 0.5ml sodium hydroxide solution and 3 drops phenolphthalein test solution in turn, shaking up fully, observing;
(4) taking another 5ml of sample solution and standard solution with various concentrations, dripping 0.5ml of sodium hydroxide solution into the sample solution by using a rubber head dropper, shaking slightly, and standing for 1 min; 6 drops of 5% malonic ester dichlorohexane mixed solution are added dropwise, heated at 60 ℃ for 5min and observed.
(5) And drawing a standard curve, and calculating the anthraquinone content in the sample.
The qualitative determination of the results is shown in table 1, the quantitative determination is shown in table 3, and the standard curve is drawn according to the standard solution as shown in fig. 2:
TABLE 3
In the detection process of the embodiment, the observation result of the sample solution in the step (3) is colorless after concentrated sulfuric acid is dropwise added, the liquid level is red after phenolphthalein is dropwise added, the red color disappears after the phenolphthalein is uniformly shaken, the solution is colorless after the heating in the step (4), and the interference of benzoquinone and naphthoquinone substances is eliminated.
In this example, the absorbance of the standard solution was measured in the range of 0.21 to 2.157 depending on the concentration, and the standard curve was plotted as y-10.652 x2+6.9885 x-1.8858. When the absorbance is more than 0.2, the error is small, so the detection limit of the method can reach 0.625 ppm.
In this example, the OD530 value of the sample solution was measured to be 1.128, and the concentration of the anthraquinone compound (in terms of 2-ethylanthraquinone) in the sample solution was calculated to be 19.55ppm by substituting the above standard curve formula, so that the anthraquinone (in terms of 2-ethylanthraquinone) content in the chicken feet with pickled peppers could be calculated.
The concentration of the 2-ethyl anthraquinone in the sample solution is 19.28ppm by using a detection method of GB/T23672-.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A method for rapidly detecting industrial hydrogen peroxide residue in food is characterized by comprising the following steps:
preparation of a sample solution: soaking food sample with dichloromethane, shaking, and collecting supernatant;
preparation of a standard solution: dissolving appropriate amount of 2-ethyl anthraquinone standard substance in dichloromethane to obtain the product;
dropwise adding concentrated sulfuric acid into one part of the sample solution and the standard solution, and developing color; sequentially dropwise adding a sodium hydroxide solution and a phenolphthalein test solution, and developing; the molar ratio of the sodium hydroxide to the concentrated sulfuric acid is 2: 1;
dropwise adding a sodium hydroxide solution into the other sample solution or standard solution, slightly shaking and standing; dripping 5% malonate dichloromethane solution or 5% malonate dichlorohexane solution, heating, and developing; the concentration of the sodium hydroxide solution is 10 mol/L.
2. The rapid detection method according to claim 1, wherein the concentrated sulfuric acid is 98% concentrated sulfuric acid, and the concentration of the phenolphthalein test solution is 5 g/L.
3. The rapid detection method according to claim 2, comprising the steps of:
preparation of a sample solution: soaking food sample with dichloromethane, shaking, and collecting supernatant;
preparation of a standard solution: dissolving appropriate amount of 2-ethyl anthraquinone standard substance in dichloromethane to obtain the product;
dropwise adding concentrated sulfuric acid into one part of the sample solution or the standard solution, developing color, and measuring absorbance at the wavelength of 530 nm; sequentially dropwise adding a sodium hydroxide solution and a phenolphthalein test solution, and developing;
dropwise adding a sodium hydroxide solution into the other sample solution or standard solution, slightly shaking and standing; dripping 5% malonic ester dichlorohexane solution or 5% malonic ester dichloromethane solution, heating, and developing.
4. The rapid detection method according to claim 3, wherein the standard solution is prepared by: dissolving 1mg of 2-ethyl anthraquinone standard substance in 10ml of dichloromethane, and performing gradient dilution to obtain the product.
5. The rapid detection method according to any one of claims 1 to 4, wherein the shaking is performed at room temperature for 10 to 20 seconds.
6. The rapid detection method according to any one of claims 1 to 4, wherein the heating is: heating at 60-80 deg.C for 5-15 min.
7. A kit for rapidly detecting industrial hydrogen peroxide residue in food is characterized by comprising dichloromethane, concentrated sulfuric acid, a sodium hydroxide solution, a phenolphthalein test solution, a 5% malonic ester dichlorohexane solution and a 5% malonic ester dichloromethane solution, wherein the dichloromethane is sealed with water during storage, and the concentration of the sodium hydroxide solution is 10 mol/L.
8. The rapid detection kit according to claim 7, further comprising a 2-ethylanthraquinone standard.
9. The rapid detection kit according to claim 7 or 8, wherein the concentrated sulfuric acid is 98% concentrated sulfuric acid.
10. The rapid detection kit according to claim 7 or 8, wherein the concentration of the phenolphthalein test solution is 5 g/L.
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| CN101451957A (en) * | 2008-12-17 | 2009-06-10 | 上海和黄药业有限公司 | Method for detecting anthraquinone content in medicaments |
| CN103760160A (en) * | 2014-01-25 | 2014-04-30 | 福州大学 | Colorimetric detection method of hydrogen peroxide |
| CN104280516A (en) * | 2014-09-20 | 2015-01-14 | 中山鼎晟生物科技有限公司 | Rapid detection kit and detection method for phenolphthalein in diet food |
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| CN101451957A (en) * | 2008-12-17 | 2009-06-10 | 上海和黄药业有限公司 | Method for detecting anthraquinone content in medicaments |
| CN103760160A (en) * | 2014-01-25 | 2014-04-30 | 福州大学 | Colorimetric detection method of hydrogen peroxide |
| CN104280516A (en) * | 2014-09-20 | 2015-01-14 | 中山鼎晟生物科技有限公司 | Rapid detection kit and detection method for phenolphthalein in diet food |
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