CN112147122A - Primary screening method for medicinal material quality - Google Patents
Primary screening method for medicinal material quality Download PDFInfo
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- CN112147122A CN112147122A CN202011203956.3A CN202011203956A CN112147122A CN 112147122 A CN112147122 A CN 112147122A CN 202011203956 A CN202011203956 A CN 202011203956A CN 112147122 A CN112147122 A CN 112147122A
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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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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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"
- G01N2021/6432—Quenching
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Abstract
The invention discloses a primary screening method for medicinal material quality, which comprises the following steps: firstly, preparing carbon quantum dots; secondly, establishing a quantitative equation of the concentration of a reference substance and the fluorescence quenching condition of the carbon quantum dots by adopting the reference substance of the index component M of the medicinal material to be detected; quenching the fluorescence of the carbon quantum dots by using the medicinal material extracting solution, and calculating to obtain a total phenol value in the medicinal material according to the fluorescence quenching condition; and step four, comparing the calculated total phenol value of the medicinal material with the standard of the index component, and judging that the quality of the medicinal material is unqualified if the total phenol value of the medicinal material does not reach the lower limit of the standard. The method takes the carbon quantum dots as a detection reagent, does not need to use a toxic solvent during detection, has simple solution preparation, short detection time, lower detection equipment cost and obviously better stability after long-term storage. Can be used for rapidly judging the quality of medicinal materials with phenol components as index components.
Description
Technical Field
The invention belongs to the technical field of medicine preparation, and particularly relates to a primary screening method for medicinal material quality.
Background
The quality of most Chinese patent medicines is mainly influenced by the quality of Chinese medicinal materials, so the quality control of the Chinese medicinal materials is very important. The factors influencing the quality of the traditional Chinese medicinal materials are many, and the traditional Chinese medicinal materials also comprise various conditions in a plurality of links such as planting, harvesting, processing, storing and transporting besides the production area and the foundation. Before use, the traditional Chinese medicine is generally required to be subjected to relevant inspection to judge whether the quality is qualified. In Chinese pharmacopoeia, liquid chromatography is often used to quantitatively determine the content of index components in addition to the judgment of the quality of medicinal materials by means of identification, inspection and the like. In most cases, the index component is an effective component, the content of the component in the medicinal material is not lower than the lower limit, otherwise, the quality of the medicinal material is considered unqualified. Common index components include flavone, organic acid, saponin, alkaloid, coumarin, lignin, etc. Many of the flavone, coumarin, organic acid, lignin and quinone components belong to phenolic compounds.
The index components are generally quantified by liquid chromatography, and the method has the advantages of accurate and reliable results, but has a plurality of defects. First, liquid chromatography is not only costly in equipment, but also has high requirements on the site where the chromatograph is placed and the analysts, and overall operating costs are high. Secondly, the components of part of the traditional Chinese medicine system are complex, and the sample analysis time is long, so that the analysis and detection efficiency is not high. Third, liquid chromatography uses toxic solvents such as methanol and acetonitrile in large amounts. Because of the problems of chromatographic analysis, it is still necessary to develop a method for discriminating the quality of medicinal materials with green, rapid and low cost.
The carbon quantum dots belong to zero-dimensional nano materials, the size of the carbon quantum dots is less than 10nm, the carbon quantum dots often have photoluminescence capability, and the fluorescence emission behavior of the carbon quantum dots can be influenced by various properties such as excitation wavelength, ion concentration, pH value, solution temperature and the like. The carbon quantum dots have the advantages of simple synthesis method, low cost, low toxicity, good long-term storage stability and wide synthetic raw material sources, and are widely concerned by academia and industry in the last decade. By utilizing the characteristic that the fluorescence of the carbon quantum dots can be rapidly quenched by specific chemical components, researchers have realized the detection of heavy metals and pesticide residues, and a new idea is provided for developing a low-cost rapid traditional Chinese medicine quality detection method.
Disclosure of Invention
The invention aims to provide a primary screening method for medicinal material quality, which takes a carbon quantum dot as a reagent, utilizes the quenching effect of an index component M on fluorescence of the carbon quantum dot to detect and obtain the total phenol content in the medicinal material, and can preliminarily judge whether the medicinal material quality is unqualified by comparing whether the total phenol content is lower than a standard lower limit.
The method comprises the following specific steps:
step one, preparing carbon quantum dots: and dissolving citric acid and ethylenediamine in water or a mixed solvent of water and tetraethyleneglycol to prepare a solution, and heating the solution to prepare the carbon quantum dots.
Secondly, establishing a quantitative equation of the concentration of a reference substance and the fluorescence quenching condition of the carbon quantum dots by adopting the reference substance of the index component M of the medicinal material to be detected, wherein the index component M is a phenolic compound; the quantitative equation is Y ═ a + bX, wherein Y is the ratio of fluorescence intensity before and after quenching, and X is the reference substance concentration of the index component M; a and b are parameters obtained by fitting;
quenching the fluorescence of the carbon quantum dots by using the medicinal material extracting solution, and calculating to obtain a total phenol content value in the medicinal material converted into an index component M according to the fluorescence quenching condition;
thirdly, quenching the fluorescence of the carbon quantum dots by using the medicinal material extracting solution to obtain a fluorescence intensity ratio, substituting the fluorescence intensity ratio into a quantitative equation to calculate the total phenol content value in the medicinal material,
the total phenol content value in the medicinal materials is converted into the total phenol content value of the index component M;
the fluorescence intensity ratio was calculated as shown in equation 1:
wherein A is0Fluorescence intensity of control solution as index component M, A1Fluorescence intensity of a mixed solution obtained after a control solution as an index component M was brought into contact with a carbon quantum dot solution, A2The fluorescence intensity of the mixture of the carbon quantum dot solution and the solvent used in the reference solution;
and step four, comparing the calculated total phenol value of the medicinal material with the index component standard, and judging that the quality of the medicinal material is unqualified if the total phenol value of the medicinal material does not reach the lower limit of the standard.
The invention also aims to apply the method to the quality judgment of the traditional Chinese medicinal materials taking the phenolic components as index components. The method does not need to use toxic solvent during detection, and the detection time is short.
The reagent for detecting the total phenol in the method is a carbon quantum dot, and has the advantages of strong fluorescence signal, low preparation cost, good storage stability, no toxicity and quenching by phenol components. The contact moment of the carbon quantum dots and the phenolic components in the solution can be quenched, and the time required for detection is short. When the degree of quenching of fluorescence by phenol components is detected, large-scale equipment such as an enzyme labeling instrument and the like can be used, and methods such as color extraction by a smart phone and the like can be used, and the equipment cost of the two methods is obviously lower than that of a liquid chromatograph. If the color is taken by the smart phone, the field detection can be realized, and the limitation that an experiment field with higher environmental control requirement is needed is broken through. The carbon quantum dots can be dissolved in water to be quenched, and toxic solvents such as methanol or acetonitrile are avoided. Compared with a conventional forskol reagent for measuring the total phenol content, the method takes the carbon quantum dots as the detection reagent, so that the solution preparation is simple, the cost is lower, and the stability of long-term storage is obviously better.
Drawings
FIG. 1 is a schematic flow chart of detecting the quality of medicinal materials by using carbon quantum dots as detection materials.
Fig. 2 is a photograph of carbon quantum dots under sunlight and ultraviolet light (365 nm).
Fig. 3 is a transmission electron microscope image of a carbon quantum dot.
Fig. 4 is a fluorescence spectrum of the carbon quantum dots.
Detailed Description
The invention is further explained by the accompanying drawings and examples.
Example 1
Referring to fig. 1, citric acid and ethylenediamine are respectively dissolved in a mixed solvent of water and tetraethylene glycol to prepare two solutions, and the two solutions are mixed and heated to prepare the carbon quantum dots. The properties of the carbon quantum dots were characterized as shown in fig. 2-4.
Honeysuckle is used as a medicinal material to be detected, and chlorogenic acid is used as an index component. Contacting chlorogenic acid with different concentrations with a carbon quantum dot, and detecting and determining the fluorescence quenching condition by adopting a method of combining a paper-based chip and a smart phone to obtain a quantitative equation of Y-0.917 + 3.62X. Taking honeysuckle of a batch to be detected, extracting with water to obtain an extracting solution, and diluting the extracting solution to quench the fluorescence of the carbon quantum dots. Substituting the result of quenching the fluorescence of the carbon quantum dots in the extracting solution into a quantitative equation, converting to obtain the total phenol concentration of 1.60mg/mL in the honeysuckle extracting solution by taking chlorogenic acid as equivalent, and converting to the total phenol content of 3.2% in the honeysuckle medicinal material. The chlorogenic acid content in the honeysuckle is not lower than 1.5% specified in the 2020 edition of Chinese pharmacopoeia. Because the obtained total phenol content value is more than 1.5%, the quality of the honeysuckle medicinal material of the batch cannot be considered unqualified.
Example 2
Referring to fig. 1, citric acid and ethylenediamine are respectively dissolved in a mixed solvent of water and tetraethylene glycol to prepare two solutions, and the two solutions are mixed and heated to prepare the carbon quantum dots. The red sage root is used as the medicinal material to be detected, and the salvianolic acid B is used as the index component. And (3) contacting salvianolic acid B with different concentrations with a carbon quantum dot, and detecting and determining the fluorescence quenching condition by adopting a method of combining a paper-based chip and a smart phone to obtain a quantitative equation of Y-1.23 + 2.86X. Extracting salvia miltiorrhiza to be tested with water to obtain an extracting solution, and diluting the extracting solution to quench the fluorescence of the carbon quantum dots. Substituting the result of quenching the fluorescence of the carbon quantum dots in the extracting solution into a quantitative equation, converting to obtain the total phenol concentration of 0.95mg/mL taking the salvianolic acid B as the equivalent in the salvia miltiorrhiza extracting solution, and converting to the total phenol content value of 1.9% in the salvia miltiorrhiza medicinal material. The content of salvianolic acid B in the salvia miltiorrhiza is not less than 3.0 percent specified in the China pharmacopoeia 2020 edition. And the obtained total phenol content value is lower than 3.0%, so that the quality of the salvia miltiorrhiza medicinal material in the batch is not qualified.
Example 3
Referring to fig. 1, citric acid and ethylenediamine are dissolved in water, and the carbon quantum dots are prepared after microwave heating. Folium mori is used as a medicinal material to be detected, and rutin is used as an index component. And (3) contacting rutin solutions with different concentrations with carbon quantum dots on a 96-well plate, and detecting by using an enzyme-labeling instrument to determine the fluorescence quenching condition to obtain a quantitative equation of Y ═ 1.15+ 2.97X. Extracting the mulberry leaves of the batch to be detected with water, and diluting the obtained extracting solution to quench the fluorescence of the carbon quantum dots. Substituting the result of quenching the fluorescence of the carbon quantum dots in the extracting solution into a quantitative equation, converting to obtain the total phenol concentration of 0.17mg/mL taking rutin as equivalent in the mulberry leaf extracting solution, and converting to the total phenol value of 0.68% in the mulberry leaf medicinal material. The rutin content in mulberry leaves is not less than 0.1 percent specified in the China pharmacopoeia 2020 edition. The obtained total phenol content value is more than 0.1%, so the quality of the mulberry leaf medicinal material in the batch cannot be considered unqualified.
Claims (6)
1. A primary screening method for medicinal material quality is characterized by comprising the following steps: firstly, preparing carbon quantum dots; secondly, establishing a quantitative equation of the concentration of a reference substance and the fluorescence quenching condition of the carbon quantum dots by adopting the reference substance of the index component M of the medicinal material to be detected; quenching the fluorescence of the carbon quantum dots by using the medicinal material extracting solution to obtain a fluorescence intensity ratio, and substituting the fluorescence intensity ratio into a quantitative equation to calculate a total phenol content value in the medicinal material; and step four, comparing the calculated total phenol value of the medicinal material with the standard of the index component M, and judging that the quality of the medicinal material is unqualified if the total phenol value of the medicinal material does not reach the lower limit of the standard.
2. The primary screening method for medicinal material quality according to claim 1, wherein the raw materials for preparing the carbon quantum dots in the first step comprise citric acid and ethylenediamine, and specifically, the citric acid and the ethylenediamine are dissolved in water or a mixed solvent of water and tetraethylene glycol to prepare a solution, and the solution is heated to prepare the carbon quantum dots.
3. The method for primary screening of the quality of medicinal materials according to claim 1, wherein in the second step, the index component M is phenolic compounds, and the quantitative equation is a linear equation in the form of Y ═ a + bX. Wherein, Y is the fluorescence intensity ratio before and after quenching, and X is the reference substance concentration of the index component M; and a and b are parameters obtained by fitting.
4. The primary screening method for the quality of medicinal materials according to claim 3, wherein the step of calculating the fluorescence intensity ratio is shown in formula 1:
wherein A is0The fluorescence intensity of a control solution as an index component M; a. the1The fluorescence intensity of the mixed solution obtained after the reference substance solution which is the index component M contacts with the carbon quantum dot solution; a. the2The fluorescence intensity of the mixture of the carbon quantum dot solution and the solvent used in the control solution.
5. The method for preliminary screening of medicinal material quality according to claim 1, wherein the total phenol value in the medicinal material of the third step is a total phenol value converted to the index component M.
6. The preliminary screening method according to claim 1 is applied to quality discrimination of a Chinese medicinal material using a phenol component as an index component.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115368891A (en) * | 2022-08-05 | 2022-11-22 | 扬州大学 | Fluorescent carbon quantum dot and application thereof in rapid detection of chlorogenic acid |
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Non-Patent Citations (6)
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| （加）塞萨尔丁•阿拉萨尔瓦等著，尚德军等主译: "《干果的植物化学成分及其保健作用》", 28 February 2018 * |
| LIZHEN LIUET.AL: "Sensitive determination of kaempferol using carbon dots as a fluorescence probe", 《TALANTA》 * |
| 姚荣林等: "《中药鉴定技术 第3版》", 31 January 2017, 中国医药科技出版社 * |
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| 李海红等: "碳量子点荧光猝灭法检测药物中的异鼠李素", 《分析测试学报》 * |
| 解军波等: "《中药材及饮片鉴定知识与方法》", 31 October 2015, 中国科学技术出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115368891A (en) * | 2022-08-05 | 2022-11-22 | 扬州大学 | Fluorescent carbon quantum dot and application thereof in rapid detection of chlorogenic acid |
| CN115368891B (en) * | 2022-08-05 | 2023-10-24 | 扬州大学 | Fluorescent carbon quantum dot and application thereof in rapid chlorogenic acid detection |
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Application publication date: 20201229 |