CN115144515A - Indigo standard solution and preparation method and detection method thereof - Google Patents
Indigo standard solution and preparation method and detection method thereof Download PDFInfo
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- CN115144515A CN115144515A CN202210928935.0A CN202210928935A CN115144515A CN 115144515 A CN115144515 A CN 115144515A CN 202210928935 A CN202210928935 A CN 202210928935A CN 115144515 A CN115144515 A CN 115144515A
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Abstract
The invention discloses an indigo standard solution, a preparation method and a detection method thereof, and particularly relates to the field of chemical detection. The preparation method comprises the steps of dissolving an indigo standard substance in water to obtain an indigo water solution; adding a sulfuric acid solution to the indigo aqueous solution; carrying out ultrasonic treatment to prepare a uniform large-concentration indigo blue sulfuric acid aqueous solution; diluting the large-concentration indigo blue sulfuric acid aqueous solution into the required indigo blue standard solution by using a sulfuric acid solution. According to the invention, the detection environment and the storage environment are researched by adding the sulfuric acid solution, so that compared with the national standard method GB 28317-2012 which uses a spectrophotometer for detection, the complexity of preparing the standard solution is reduced; the invention adopts the high performance liquid chromatography for detection, and the detection environment is widened compared with the detection environment; the test condition is easy to master and operate. The indigo solution standard substance has the characteristics of good stability, no need of refrigeration, convenient transportation and use, and can be stored at normal temperature, and the stabilization period is as long as 2 years.
Description
Technical Field
The invention relates to the field of chemical detection, and particularly relates to an indigo standard solution and a preparation method and application thereof.
Background
Indigo pigments are one of the oldest pigments known to man and are widely used in the food, pharmaceutical and printing industries. It can be used for coloring food, medicine and daily cosmetics. Edible indigo is an edible synthetic pigment. Food colors are a class of additives used in food coloring, including synthetic and natural colors, totaling over 60.
The indigo (CAS: 860-22-0) is unstable in property, and is not light-resistant and heat-resistant, the reason that the yield of the indigo in the experiment is unstable is mainly that the indigo standard product is degraded or the indigo in the sample is degraded in the pretreatment process, in order to avoid the loss of the indigo, a solid indigo standard product is needed in each experiment, the indigo standard product is prepared at present, the standard products used in the experiment comprise a machine-mounted product and a labeling product, the indigo standard product and the labeling product are needed to be prepared at the same time and stored under the same conditions, and the indigo is protected from light in the whole experiment process.
However, the standard substance of the indigo solution on the market at present has variable values in storage period and poor stability, and particularly, the standard substance of the indigo solution has to be prepared at present.
With the rapid development of food industry detection, the standard substance of the indigo solution is in short supply. Therefore, it is urgent to develop and prepare a standard substance that can be stabilized for one year or more.
The existing detection method mostly adopts a titanium trichloride titration method, the method needs more manpower consumption, and the requirement on the titration proficiency of operators is higher; the spectrophotometric colorimetric method has high requirement on environment, harsh test conditions and relatively complex standard solution preparation, and needs multiple measurements to calculate the average value.
Disclosure of Invention
Therefore, the invention provides an indigo standard solution, and a preparation method and application thereof, so as to solve the problems that the existing indigo standard solution is unstable and needs to be prepared at present.
The invention finds that the stability of the indigo solution can be improved by adding a sulfuric acid solution to the indigo solution. According to the invention, the research on the detection environment and the storage environment is carried out by adding the sulfuric acid solution, the stability of the indigo standard solution is prolonged, and the indigo detection can be carried out by using a high performance liquid chromatography. Researches show that the stability of the standard solution can be improved by adding the sulfuric acid solution, and compared with a national standard method GB 28317-2012, the method adopts a spectrophotometer for detection, thereby not only reducing the complexity of preparing the standard solution, but also widening the detection environment relative to the standard solution; and the test condition is easy to master and operate.
In order to achieve the above purpose, the invention provides the following technical scheme:
according to a first aspect of the present invention, there is provided a method for preparing an indigo standard solution, comprising: dissolving an indigo standard substance in water to obtain an indigo aqueous solution; adding a sulfuric acid solution to the indigo aqueous solution; carrying out ultrasonic treatment to prepare a uniform large-concentration indigo blue sulfuric acid aqueous solution; diluting the large-concentration indigo sulfuric acid aqueous solution into the required indigo standard solution by using a sulfuric acid solution.
Further, the purity of the indigo standard substance is more than or equal to 98.5%.
Further, the concentration of the sulfuric acid solution is 1.0% -2.0%.
Further, the concentration of the large-concentration indigo blue aqueous solution is 1000 mug/mL.
Furthermore, the concentration of the indigo standard solution is 5-50 mu g/mL.
According to a second aspect of the present invention there is provided an indigo standard solution prepared as described above.
According to a third aspect of the present invention, there is provided the method for detecting indigo blue standard solution as described above, wherein the detection is performed by a method combining liquid chromatography and ultraviolet detection.
Furthermore, the chromatographic column adopted by the detection method is C18, and the size of the chromatographic column is 250mm multiplied by 4.6mm multiplied by 0.5 mu m;
and/or the mobile phase is an ammonium acetate solution: methanol = 60;
and/or, detecting wavelength: 610nm-630nm;
and/or, flow rate: 1.0ml/min;
and/or, sample size: 10 μ L.
Further, the concentration of the ammonium acetate solution is 0.01mol/L-0.05mol/L.
The invention has the following advantages:
the raw materials of the invention adopt the purity standard substance with the purity of 98.5-100%, and the sulfuric acid aqueous solution is added, thus greatly improving the stability of the product.
The indigo solution standard substance has the characteristics of good stability, no need of refrigeration, and convenient transportation and detection.
The indigo solution standard substance has good uniformity, has little change of magnitude when standing for a long time, can be stored at normal temperature, and has a stabilization period of 2 years.
The detection method of the indigo solution utilizes the high performance liquid chromatography for detection, and compared with the national standard method GB 28317-2012, the detection method of the indigo solution utilizes a spectrophotometer for detection, thereby not only reducing the complexity of preparing the standard solution, but also widening the detection environment relative to the detection environment; and the test condition is easy to master and operate.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary and that other implementation drawings may be derived from the provided drawings by those of ordinary skill in the art without inventive effort.
The structures, the proportions, the sizes, and the like shown in the specification are only used for matching with the contents disclosed in the specification, so that those skilled in the art can understand and read the present invention, and do not limit the conditions for implementing the present invention, so that the present invention has no technical essence, and any modifications of the structures, changes of the proportion relation, or adjustments of the sizes, should still fall within the scope of the technical contents disclosed in the present invention without affecting the efficacy and the achievable purpose of the present invention.
FIG. 1 is an HPLC chromatogram of an indigo standard solution of example 2 provided by the present invention;
FIG. 2 is an HPLC chromatogram of an indigo standard solution of comparative example 1 provided by the present invention;
FIG. 3 is a stability test chart of indigo standard solutions of example 2, example 3, example 4 and comparative example 1 provided by the present invention;
in the figure: a-example 2; b-example 3; c-example 4; d-comparative example 1.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. 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.
Example 1
This example provides a method for preparing an indigo standard solution:
step one, putting 0.5g of indigo into a 500mL volumetric flask, adding 400mL of water for dissolving to prepare an indigo water solution;
wherein the purity of the indigo is 98.5-100%; can be obtained directly through purchase or can be prepared by reaction.
Adding a sulfuric acid solution with the concentration of 1.0-2.0% into the indigo blue aqueous solution, fixing the volume to 500mL, dissolving and performing ultrasonic treatment to prepare a uniform solution with the volume of 1000 mu g/mL;
step three, dilution: diluting the prepared standard solution with the value of 1000 mug/mL by using a sulfuric acid aqueous solution according to a proportion to obtain an indigo standard solution with the nominal value of 5-50 mug/mL.
Example 2
This example provides a method for preparing a 10 μ g/mL indigo standard solution:
step one, putting 0.50403g of indigo into a 500mL volumetric flask, adding 400mL of water for dissolving to prepare an indigo water solution;
wherein the purity of the indigo is 99.2%; can be obtained directly through purchase or can be prepared by reaction.
Adding a 1.4% sulfuric acid solution into the indigo blue aqueous solution, diluting to a constant volume of 500mL, and dissolving and ultrasonically treating to prepare a uniform solution of 1000 mug/mL;
step three, dilution: the prepared standard solution with the value of 1000 mug/mL is diluted by 1.4 percent of sulfuric acid aqueous solution according to a proportion to obtain an indigo standard solution with the nominal value of 10 mug/mL.
Example 3
This example provides a method for preparing a 20 μ g/mL indigo standard solution:
step one, placing 0.50403g of indigo into a 500mL volumetric flask, adding 400mL of water for dissolving to prepare an indigo water solution;
wherein the purity of the indigo is 99.2%; can be obtained directly through purchase or can be prepared by reaction.
Adding a 1.4% sulfuric acid solution into the indigo blue aqueous solution, diluting to a constant volume of 500mL, and dissolving and ultrasonically treating to prepare a uniform solution of 1000 mug/mL;
step three, dilution: the prepared standard solution with the value of 1000 mug/mL is diluted by 1.4 percent of sulfuric acid aqueous solution according to a proportion to obtain the indigo standard solution with the nominal value of 20 mug/mL.
Example 4
This example provides a method for preparing 40 μ g/mL indigo standard solution:
step one, placing 0.50403g of indigo into a 500mL volumetric flask, adding 400mL of water to dissolve the indigo into the volumetric flask to prepare an indigo aqueous solution;
wherein the purity of the indigo is 99.2%; can be obtained directly through purchase or can be prepared by reaction.
Step two, adding a 1.4% sulfuric acid solution into the indigo blue aqueous solution, fixing the volume to 500mL, dissolving and ultrasonically treating to prepare a uniform solution of 1000 mu g/mL;
step three, dilution: the prepared standard solution with the value of 1000 mug/mL is diluted by 1.4 percent of sulfuric acid aqueous solution according to a proportion to obtain an indigo standard solution with the nominal value of 40 mug/mL.
Example 5
In this example, the indigo standard solution prepared in example 1 was detected by Agilent1200 hplc.
Detection conditions are as follows:
the chromatographic column is C18, 250mm multiplied by 4.6mm multiplied by 0.5 μm;
the mobile phase is ammonium acetate solution: methanol = 60;
detection wavelength: 610nm-630nm;
flow rate: 1.0ml/min;
sample introduction amount: 10 μ L.
The concentration of the ammonium acetate aqueous solution is 0.01mol/L-0.05mol/L.
The methanol was filtered through an organic phase filter and the aqueous ammonium acetate solution was filtered through an aqueous phase filter.
Example 6
This example uses a liquid chromatography-ultraviolet detector to detect the indigo standard solutions of example 2, example 3 and example 4.
Detection conditions are as follows:
and (3) chromatographic column: c18 250mm × 4.6mm × 0.5 μm;
mobile phase: mobile phase a (0.02 mol/L ammonium acetate solution): mobile phase B (methanol) = 80;
detection wavelength: 620nm;
flow rate: 1.0ml/min;
sample injection amount: 10uL.
Filtering methanol with organic phase filter membrane, filtering ammonium acetate water solution with water phase filter membrane
Comparative example 1
This comparative example was prepared in exactly the same manner as example 2, except that the sulfuric acid solution was changed to water.
Test example 1
The change of the indigo concentration in 24 months was examined in each of example 2, example 3, example 4 and comparative example 1 using the examination method and examination conditions of example 6, as shown in table 1.
TABLE 1
As can be seen from table 1, the indigo standard solution prepared by the method of the present invention has very good stability. The HPLC spectrum of example 2 is shown in fig. 1, and the HPLC spectrum of comparative example 1 is shown in fig. 2, from which it can be seen that the spectrum bandwidth range of the addition of sulfuric acid is free from interference.
As shown in a stability test chart of FIG. 3, as can be seen from the curves in FIG. 3 and the data in Table 1, the standard substance of the present invention has very good stability, and the magnitude value can be stabilized for 24 months under the condition of normal temperature storage; the standard substance without the addition of the sulfuric acid aqueous solution was stable for only 6 months, and the amount was in a downward trend thereafter.
Therefore, the indigo standard solution prepared by the method can be stored at normal temperature, and the quantity value is stable for 2 years.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (9)
1. A method for preparing an indigo standard solution, which is characterized by comprising the following steps:
dissolving an indigo standard substance in water to obtain an indigo aqueous solution;
adding a sulfuric acid solution to the indigo aqueous solution;
carrying out ultrasonic treatment to prepare a uniform large-concentration indigo blue sulfuric acid aqueous solution;
diluting the large-concentration indigo blue sulfuric acid aqueous solution into the required indigo blue standard solution by using a sulfuric acid solution.
2. The method for preparing the indigo standard solution according to claim 1, wherein the purity of the indigo standard substance is 98.5% or more.
3. The method for preparing the indigo standard solution according to claim 1, wherein the concentration of the sulfuric acid solution is 1.0% to 2.0%.
4. The method for preparing the indigo standard solution according to claim 1, wherein the concentration of the large-concentration indigo aqueous solution is 1000 μ g/mL.
5. The method for preparing the indigo standard solution according to claim 1, wherein the concentration of the indigo standard solution is 5 μ g/mL to 50 μ g/mL.
6. An indigo standard solution prepared by the method of any one of claims 1 to 5.
7. The detection method of the indigo standard solution is characterized in that a liquid chromatography-ultraviolet detection combined method is adopted for detection.
8. The method for detecting indigo standard solution according to claim 7, wherein the chromatographic column used in the method is C18, 250mm x 4.6mm x 0.5 μm;
and/or the mobile phase is an ammonium acetate solution: methanol = 60;
and/or, detecting wavelength: 610nm-630nm;
and/or, flow rate: 1.0ml/min;
and/or, sample size: 10 μ L.
9. The method for detecting an indigo standard solution according to claim 8, wherein a concentration of the ammonium acetate solution is 0.01mol/L to 0.05mol/L.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105319301A (en) * | 2015-12-05 | 2016-02-10 | 中华人民共和国赤峰出入境检验检疫局 | Method for determining non-pigmented organic matter in food additive indigo |
| CN112898800A (en) * | 2021-01-18 | 2021-06-04 | 山东云卿生物科技有限公司 | Method for extracting indigo |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105319301A (en) * | 2015-12-05 | 2016-02-10 | 中华人民共和国赤峰出入境检验检疫局 | Method for determining non-pigmented organic matter in food additive indigo |
| CN112898800A (en) * | 2021-01-18 | 2021-06-04 | 山东云卿生物科技有限公司 | Method for extracting indigo |
Non-Patent Citations (2)
| Title |
|---|
| 曾林泉: "纤维织物上靛蓝浓度的测定", 染料工业, vol. 39, no. 5, pages 46 * |
| 杨昌彪 等: "高效液相色谱检测冷饮中的靛蓝色素", 贵州科学, vol. 30, no. 6, pages 60 - 62 * |
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