CN118090690A - Detection method of amylose in grains - Google Patents
Detection method of amylose in grains Download PDFInfo
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- CN118090690A CN118090690A CN202410289884.0A CN202410289884A CN118090690A CN 118090690 A CN118090690 A CN 118090690A CN 202410289884 A CN202410289884 A CN 202410289884A CN 118090690 A CN118090690 A CN 118090690A
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- 229920000856 Amylose Polymers 0.000 title claims abstract description 56
- 238000001514 detection method Methods 0.000 title abstract description 12
- 239000000243 solution Substances 0.000 claims abstract description 36
- 238000000034 method Methods 0.000 claims abstract description 17
- 239000007864 aqueous solution Substances 0.000 claims abstract description 12
- 229910052761 rare earth metal Inorganic materials 0.000 claims abstract description 12
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 70
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 claims description 48
- 239000002244 precipitate Substances 0.000 claims description 44
- 238000010438 heat treatment Methods 0.000 claims description 33
- 239000012043 crude product Substances 0.000 claims description 31
- 239000011259 mixed solution Substances 0.000 claims description 26
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 22
- 238000001816 cooling Methods 0.000 claims description 22
- 229920002472 Starch Polymers 0.000 claims description 13
- KTUQUZJOVNIKNZ-UHFFFAOYSA-N butan-1-ol;hydrate Chemical compound O.CCCCO KTUQUZJOVNIKNZ-UHFFFAOYSA-N 0.000 claims description 13
- 235000019698 starch Nutrition 0.000 claims description 13
- 239000008107 starch Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 11
- 238000003756 stirring Methods 0.000 claims description 11
- 238000005406 washing Methods 0.000 claims description 11
- 229910016644 EuCl3 Inorganic materials 0.000 claims description 4
- NNMXSTWQJRPBJZ-UHFFFAOYSA-K europium(iii) chloride Chemical compound Cl[Eu](Cl)Cl NNMXSTWQJRPBJZ-UHFFFAOYSA-K 0.000 claims description 4
- 230000005284 excitation Effects 0.000 claims description 4
- SNMVRZFUUCLYTO-UHFFFAOYSA-N n-propyl chloride Chemical compound CCCCl SNMVRZFUUCLYTO-UHFFFAOYSA-N 0.000 claims description 3
- 229910004664 Cerium(III) chloride Inorganic materials 0.000 claims description 2
- 229910017544 NdCl3 Inorganic materials 0.000 claims description 2
- 229910019328 PrCl3 Inorganic materials 0.000 claims description 2
- VYLVYHXQOHJDJL-UHFFFAOYSA-K cerium trichloride Chemical compound Cl[Ce](Cl)Cl VYLVYHXQOHJDJL-UHFFFAOYSA-K 0.000 claims description 2
- ATINCSYRHURBSP-UHFFFAOYSA-K neodymium(iii) chloride Chemical compound Cl[Nd](Cl)Cl ATINCSYRHURBSP-UHFFFAOYSA-K 0.000 claims description 2
- LHBNLZDGIPPZLL-UHFFFAOYSA-K praseodymium(iii) chloride Chemical compound Cl[Pr](Cl)Cl LHBNLZDGIPPZLL-UHFFFAOYSA-K 0.000 claims description 2
- 239000000047 product Substances 0.000 claims description 2
- 235000013305 food Nutrition 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 150000002910 rare earth metals Chemical class 0.000 abstract 1
- 235000013339 cereals Nutrition 0.000 description 7
- 241000209094 Oryza Species 0.000 description 4
- 235000007164 Oryza sativa Nutrition 0.000 description 4
- 235000009566 rice Nutrition 0.000 description 4
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 229920000945 Amylopectin Polymers 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention belongs to the field of food detection, and particularly relates to a detection method of amylose in grains; according to the invention, the amylose in the grains is extracted by using the rare earth metal aqueous solution as the extracting solution and detected by using the fluorescence spectrophotometer, and compared with the method, the method has a good detection effect.
Description
Technical Field
The invention belongs to the field of food detection, and particularly relates to a detection method of amylose in grains.
Background
Starch is a mixture of different types of starch, one being amylose and the other being amylopectin.
Wherein, the amylose is the main nutrition component of crops such as wheat, corn, rice and the like, and has great influence on the quality and the taste of food. Rice with a high amylose content absorbs more moisture during cooking, resulting in a fluffy and dark rice. The amylose content of wheat directly affects the softness, viscosity, smoothness of the final flour. While corn starch is one of the main sources of amylose.
Since amylose has a great influence on the quality of foods, the method has important theoretical value and practical significance for detecting the content of the amylose.
Disclosure of Invention
The invention aims to provide a method for detecting amylose in grains.
The invention is realized by the following technical scheme:
a method for detecting amylose in grains, comprising the steps of:
Extraction of crude product: crushing a sample to be detected, adding the crushed sample into a rare earth element aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the crude amylose product, wherein; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain amylose;
and detecting the content of the amylose by using a fluorescence spectrophotometer.
Further, the volume ratio of butanol to isoamyl alcohol in the butanol and isoamyl alcohol mixed solution is 1:3.
Further, the volume ratio of the n-butanol to the water in the n-butanol water mixed solution is 1:9.
Further, the rare earth element in the aqueous rare earth element solution is selected from one of PrCl 3、CeCl3、PrCl3、NdCl3、EuCl3.
Further, the rare earth element in the aqueous rare earth element solution is selected from EuCl3.
Further, the concentration of the EuCl 3 solution is 0.5M-1.0M.
Further, the concentration of the EuCl 3 solution was 0.5M.
Further, the volume of the EuCl 3 solution is 70-150 mL.
Further, the volume of EuCl 3 solution was 100mL.
Further, when the fluorescence spectrophotometer is used for detecting the content of the amylose, the maximum excitation wavelength and the emission wavelength are scanned and determined within the wavelength range of 300-500 nm.
Compared with the prior art, the invention has the following advantages and beneficial effects: see examples.
Detailed Description
The present invention will be described in further detail with reference to the following examples, for the purpose of making the objects, technical solutions and advantages of the present invention more apparent, and the description thereof is merely illustrative of the present invention and not intended to be limiting.
Example 1
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 100mL of 0.5M CeCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 2
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 100mL of 0.5M PrCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 3
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 100mL of 0.5M NdCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 4
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 100mL of 0.5M EuCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 5
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 100mL of 0.5M LaCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 6
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 70mL of 0.5M EuCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 7
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 80mL of 0.5M EuCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 8
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 120mL of 0.5M EuCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Example 9
Extraction of crude product: taking 10g of a sample to be detected, adding the crushed sample into 150mL of 0.5M EuCl 3 aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol, wherein the volume ratio of butanol to isoamyl alcohol is 1:3; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the amylose crude product, wherein the volume ratio of n-butanol to water is 1:9; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; then immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain the amylose.
Test example 1
The method in examples 1-9 is used for detecting amylose in the same rice, the detection method is to use water as a solvent, detect the content of the amylose by adopting a fluorescence spectrophotometer, determine the maximum excitation wavelength and the emission wavelength within the range of 300-500 nm, and the concentration range of a standard curve is 0.1 mug/mL-1.0 mug/mL. The detection value of NY/T55-1987 was also used as a standard. Each measurement was performed twice and the average was taken.
Table 1 performance test
As can be seen from the data in Table 1, the extracts of the different rare earth elements all have a certain effect, but the best effect in example 4 is closest to the result measured by national standard NY/T55-1987.
Test example 2
The method of example 4 was used for detecting the content of different grains, the detection method was to detect the content of amylose using a fluorescence spectrophotometer with water as a solvent, and the maximum excitation wavelength and emission wavelength were determined in the range of 300 to 500nm, with the standard curve concentration ranging from 0.1 μg/mL to 1.0 μg/mL. The detection value of NY/T55-1987 was also used as a standard. Each measurement was performed twice and the average was taken.
Table 2 amylose content determination in different cereals.
As can be seen from the data in table 2, the detection method of the present application can be used for the determination of different types of grains.
The foregoing is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art, who is within the scope of the present invention, should make equivalent substitutions or modifications according to the technical scheme of the present invention and the inventive concept thereof, and should be covered by the scope of the present invention.
Claims (10)
1. A method for detecting amylose in grains, comprising the steps of:
Extraction of crude product: crushing a sample to be detected, adding the crushed sample into a rare earth element aqueous solution, heating the solution to 60 ℃, and keeping the temperature at 60 ℃ for 90min; then adding 100mL of mixed solution of n-butanol and isoamyl alcohol; then heating to 100 ℃ and stirring for 10min; cooling to 4 ℃ and maintaining at 4 ℃ for 24 hours; finally, centrifuging for 20min at room temperature to obtain an amylose crude product;
Purifying: adding 100mL of n-butanol water mixed solution into the crude amylose product, wherein; heating the solution to 100 ℃ until starch is completely dissolved, cooling to 25 ℃, placing in an environment of 4 ℃ for 24 hours, centrifuging to obtain a precipitate, and repeatedly treating the precipitate for 3-4 times; immersing the precipitate in absolute ethyl alcohol for 24 hours, washing the precipitate with absolute ethyl alcohol, and then drying in vacuum to obtain amylose;
and detecting the content of the amylose by using a fluorescence spectrophotometer.
2. The method according to claim 1, wherein the volume ratio of butanol to isoamyl alcohol in the butanol and isoamyl alcohol mixed solution is 1:3.
3. The method according to claim 1, wherein the volume ratio of n-butanol to water in the n-butanol-water mixed solution is 1:9.
4. The method of claim 1, wherein the aqueous rare earth element solution contains a rare earth element selected from the group consisting of PrCl 3、CeCl3、PrCl3、NdCl3、EuCl3.
5. The method of claim 4, wherein the aqueous rare earth element solution contains a rare earth element selected from the group consisting of EuCl3.
6. The method of claim 5, wherein the concentration of the EuCl 3 solution is 0.5M to 1.0M.
7. The method of claim 6, wherein the concentration of EuCl 3 solution is 0.5M.
8. The method of claim 7, wherein the volume of EuCl 3 solution is 70-150 mL.
9. The method of claim 8, wherein the volume of the EuCl 3 solution is 100mL.
10. The method according to claim 1, wherein the maximum excitation wavelength and emission wavelength are determined by scanning in a wavelength range of 300 to 500nm when detecting the amylose content using a fluorescence spectrophotometer.
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