CN111122528B - SiO 2 Sol-gel encapsulated CsPbBr 3 Method for detecting cholesterol by quantum dots - Google Patents
SiO 2 Sol-gel encapsulated CsPbBr 3 Method for detecting cholesterol by quantum dots Download PDFInfo
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Abstract
The invention discloses a SiO 2 Sol-gel encapsulated CsPbBr 3 A method for detecting cholesterol by using quantum dots. The method comprises the steps of preparing the MTMOS encapsulated CsPbBr with the core-shell structure by adopting a thermal injection method 3 QDs as fluorescent signal unit to prepare MIPs/CsPbBr molecular imprinting fluorescent sensor 3 @SiO 2 QDs are used. The sensor takes TMOS as a cross-linking agent and APTESs as functional monomers, reacts with a template molecule CHO under the action of Van der Waals force, obtains a linear relation between fluorescence quenching intensity and cholesterol concentration through elution and recognition processes, measures a fluorescence peak value of human serum to be detected, and can calculate the content of cholesterol in the human serum by substituting the fluorescence peak value into the linear relation. The detection method has the advantages of high sensitivity, accurate precision, low cost, high efficiency, rapidness and the like, and the detection limit is as low as 2.05 multiplied by 10 ‑12 mol/L, has important application value in the detection of the content of human serum cholesterol.
Description
Technical Field
The invention belongs to the technical field of molecular imprinting fluorescent sensors, and relates to SiO 2 Sol-gel encapsulated CsPbBr 3 A method for detecting cholesterol by using quantum dots.
Background
Excess Cholesterol (CHO) in human serum can form plaques in vascular arteries to prevent blood circulation and cause cardiovascular disease. Therefore, the determination of the cholesterol content in serum is an important reference index in clinical diagnosis, and has extremely important effects on preventing cardiovascular diseases, stroke, peripheral arterial diseases, diabetes and hypertension.
At present, common methods for detecting the content of cholesterol in human serum comprise a colorimetric method, a gas-liquid chromatography-mass spectrometry method, a temperature measuring method, a molecular luminescence method, an electrochemical method and the like, but the detection methods generally have the problems of complex operation process, poor specificity, high cost and the like to a certain extent. Compared with the detection method, the detection method based on fluorescence has the advantages of simple operation, low cost, high sensitivity and strong specificity. The highly fluorescent probe plays a very important role in the Construction of high-efficiency fluorescence sensors, and different monometallic nanoparticles, bimetallic nanoparticles and Quantum Dots (QDs), such as carbon dot-hemoglobin complex, multi-walled carbon nanotube-gold nanoparticle composite material, b-cyclodextrin functionalized carbon dots, etc., are used as probes for detecting CHO by fluorescence (Li, Y.; Cai, J.; Liu, F.; et al, conjugation of a turn off-on fluoro luminescence nano sensor for fluorescence on fluorescence sensitivity gene transfer and emission, Talan 2019,201, 82-89). However, these probes often require immobilization using a large amount of cholesterol oxidase to achieve highly sensitive detection of cholesterol. However, the use of enzymes has the disadvantages of high cost, enzyme denaturation, low storage temperature and the like. Furthermore, most reported fluorometric detection of CHO generally requires a very complex derivatization process (Batcz, O.; Tomoskozi, S., Determination of total Cholesterol content in food by flow information analysis with immobilized Cholesterol oxidase enzyme reaction, NaHrun 2002,46(1), 46-50; Kalaiyaras, G., Joseph, J., Cholester derived carbon fiber dots as fluorescent detection for the specific detection of hemoglobin in dispersed human samples Mater Sci Eng C Mater Biol 2019,94, 580).
Disclosure of Invention
The invention aims to provide SiO 2 Sol-gel encapsulated CsPbBr 3 A method for detecting cholesterol by using quantum dots. The method is simple and rapid, has high sensitivity and strong specificity, and can rapidly and sensitively detect the cholesterol content in human serum by the molecularly imprinted fluorescent sensor.
The technical scheme for realizing the purpose of the invention is as follows:
SiO 2 sol-gel encapsulated CsPbBr 3 The method for detecting cholesterol by using the quantum dots comprises the following steps:
step 2, according to the molar ratio of 1: 5-1: 6 of cholesterol to 3-aminopropyl triethoxysilane (APTEs), adding cholesterol, octadecene, 3-aminopropyl triethoxysilane (APTEs) and CsPbBr 3 @SiO 2 After QDs are stirred and mixed uniformly, Tetramethoxysilane (TMOS) is added according to the molar ratio of 1: 9-1: 10 between Tetramethoxysilane (TMOS) and 3-Aminopropyltriethoxysilane (APTEs), and the mixture is stirred and reacted in a closed manner at room temperature to obtain MIPs/CsPbBr 3 @SiO 2 QDs composites;
Preferably, in the step 1, the volume ratio of oleic acid to octadecene is 1: 12-1: 13; the molar ratio of the lead bromide to the cesium carbonate is 1: 5-1: 6; the volume ratio of octylamine to oleic acid to methyltrimethoxysilane is 1: 1:1 to 1:2: 2.
Preferably, in the step 1, the reaction time is 2-3 h at 150-160 ℃.
Preferably, in the step 1, the stirring reaction time is 30-40 min.
Preferably, in the step 2, the stirring and mixing time is 30-40 min.
Preferably, in the step 2, the reaction time is 12-14 h under closed stirring at room temperature.
Compared with the prior art, the invention has the following advantages:
(1) the invention uses CsPbBr 3 QDs encapsulation in SiO 2 In the matrix, a typical core-shell structure is formed, and the formed protective layer is largeGreatly improves CsPbBr 3 Stability of QDs and as imprinting matrices for CsPbBr 3 The good selectivity of QDs, the good selectivity to the detection of cholesterol, and the more accurate detection result;
(2) the fluorescent probe CsPbBr of the invention 3 @SiO 2 The synthesis process of QDs is simple, a complex derivation process is not needed, cholesterol oxidase is not needed for immobilization, and the enzyme has the defects of changeability, high cost, low storage temperature and the like in use, so that the fluorescent probe is selected to ensure that the detection process is simpler and more convenient, and the detection cost is greatly reduced;
(3) the introduction of MIPs which are polymers with higher molecular weight greatly improves CsPbBr 3 @SiO 2 Stability of QDs, CsPbBr 3 @SiO 2 The QDs as the fluorescent probe can detect the biomolecules, thereby improving the dual guarantee;
(4) the detection method has the advantages of high sensitivity, accurate precision, low cost, high efficiency, rapidness and the like, and the detection limit is as low as 2.05 multiplied by 10 -12 mol/L, has important application value in the detection of the content of the human serum cholesterol.
Drawings
FIG. 1(A) is a graph showing the effect of the amount of TMOS added as a crosslinking agent on the fluorescence peak of the sensor in example 3, FIG. 1(B) is a graph showing the effect of the molar ratio of CHO to APTES in example 4 on the fluorescence peak of the sensor, FIG. 1(C) is a graph showing the effect of the elution time on the fluorescence peak of the sensor in example 5, and FIG. 1(D) is a graph showing the effect of the discrimination time on the fluorescence peak of the sensor in example 6.
FIG. 2 shows CsPbBr in example 1 3 QDs (A) and CsPbBr 3 @SiO 2 QDs (B) Transmission Electron microscopy images.
FIG. 3A is a graph showing the fluorescence peaks of cholesterol at different concentrations in example 1, and B is a graph showing the linear relationship between the cholesterol concentration and the fluorescence quenching intensity.
Detailed Description
The present invention will be described in more detail with reference to the following examples and the accompanying drawings.
Example 1
(1) Preparation of CsPbBr 3 @SiO 2 QDs：
12mL of octadecene, 1mL of oleic acid, and 0.32g of cesium carbonate were mixed, and N was introduced 2 And (4) deoxidizing, and reacting for 2h at 150 ℃ to synthesize the cesium oleate precursor. Then 0.068g of lead bromide, 5mL of octadecene, 500. mu.L of octylamine, 500. mu.L of oleic acid and 500. mu.L of methyltrimethoxysilane (MTMOS) in N 2 Heating to 170 ℃ under protection, rapidly adding 700 mu L of cesium oleate solution (preheating to 100 ℃), reacting for 5 seconds, immediately performing ice bath, exposing the reaction solution to air, and stirring for 30min to generate CsPbBr 3 @SiO 2 QDs are used. FIG. 2 shows CsPbBr 3 QDs (A) and CsPbBr 3 @SiO 2 Transmission electron microscopy of QDs (B). As can be seen from FIG. 2(A), CsPbBr 3 The QDs particles have smooth and flat surfaces, uniform particle size distribution, substantially uniform size, average diameter of about 7.9nm, and CsPbBr 3 QDs have good dispersibility and dispersability. As can be seen in FIG. 2(B), CsPbBr 3 @SiO 2 The surface of QDs is coated with a silica gel layer, which shows that CsPbBr is generated after the silicon substrate MTMOS is hydrolyzed 3 QDs are successfully encapsulated and embedded in larger particles, CsPbBr 3 The QDs and the silica gel layer form a typical core-shell structure, so that the fluorescent quantum dots (CsPbBr) can be effectively protected 3 QDs)。
(2) Preparation of MIPs/CsPbBr 3 @SiO 2 QDs composites:
0.389g cholesterol, 10mL octadecene, 0.90mL 3-Aminopropyltriethoxysilane (APTES), and 100. mu.L CsPbBr 3 @SiO 2 Stirring QDs for 30min, mixing, adding 100 μ L Tetramethoxysilane (TMOS), stirring under sealed condition at room temperature for 12 hr to obtain MIPs/CsPbBr 3 @SiO 2 QDs composites.
(3) MIPs/CsPbBr 3 @SiO 2 And adding the QDs composite material into a mixed solution of ethyl acetate and n-hexane in a volume ratio of 1:3 for ultrasonic elution, wherein the elution time is 15 min.
(4) Eluting MIPs/CsPbBr 3 @SiO 2 The QDs composite material is dispersed in ethyl acetate solution and then respectively identified with cholesterol solution with different concentrationsThe solution concentrations were 1.0X 10 respectively -11 ，5.0×10 -11 ， 1.0×10 -10 ，5.0×10 -10 ，1.0×10 -9 ，5.0×10 -9 ，1.0×10 -8 And 5.0X 10 -8 And (3) mol/L, incubating for 30min, respectively measuring eight groups of fluorescence peak values F by using a fluorescence spectrometer, and drawing a linear relation graph of fluorescence quenching intensity and cholesterol concentration. FIG. 3A shows the fluorescence peak of cholesterol at different concentrations, and B shows the linear relationship between the concentration of cholesterol and the fluorescence quenching intensity. As can be seen from FIG. 3(B), the linear relationship between the cholesterol concentration and the fluorescence quenching intensity is (F) 0 -F)/F＝ 2.6549+0.2352lg C(mol/L)(R 2 ＝0.9937)。
Example 2
Detection of cholesterol content in human serum:
diluting human serum by a certain multiple, and then eluting the MIPs/CsPbBr 3 @SiO 2 The QDs composite material was dispersed in ethyl acetate solution and mixed with diluted human serum for 30min to determine the fluorescence peak F'. Substituting the F' into a linear relation between the concentration of the cholesterol and the fluorescence quenching intensity to calculate the corresponding concentration, and multiplying the corresponding concentration by the dilution multiple of the human serum to obtain the content of the cholesterol in the human serum, wherein the obtained result is about 3.38mmol/L and is basically consistent with the known concentration of the human serum sample of 3.45 mmol/L. The result shows that the MIPs/CsPbBr of the invention 3 @SiO 2 The QDs fluorescence sensor has good application effect on the detection of the cholesterol content in the human serum, is simple to operate, high in sensitivity, low in cost, efficient and rapid, and has important application value in the detection process of the cholesterol concentration in the human serum.
Example 3
This example was substantially the same as example 1 except that the amounts of the crosslinking agents TMOS added in step (2) were adjusted to 60. mu.L, 80. mu.L, 100. mu.L, 120. mu.L and 140. mu.L, respectively, and the effect of the amount of TMOS added on the CHO measurement was examined. FIG. 1(A) is a graph showing the effect of the amount of TMOS added as a crosslinking agent on the fluorescence peak of the sensor. In the molecular imprinting polymerization, the amount of the cross-linking agent has an important influence on the selectivity and binding ability of MIPs. Its main role is to immobilize the functional monomers around the template molecule, thus forming a highly crosslinked rigid polymer even after template removal. As is clear from FIG. 1(A), the crosslinking effect is best when the amount of TMOS added is 100. mu.L. Too high a concentration of the cross-linking agent may result in excessive cross-linking and agglomeration of the polymer, while too low a concentration may result in insufficient cross-linking and insufficient formation of imprinted sites, which may affect the experimental results.
Example 4
This example is essentially the same as example 1, except that the molar ratios of CHO to APTES in step (2) were adjusted to 1:3, 1:4, 1:5, 1:6 and 1:7, respectively, and the effect of the molar ratios of CHO to APTES on the blotting effect was investigated. FIG. 1(B) is a graph showing the effect of the molar ratio of CHO to APTES on the fluorescence peak of the sensor. As shown in FIG. 1(B), as the amount of APTES increases, the peak value of the fluorescence after identification gradually increases and reaches a maximum at a ratio of 1:5, and then starts to decrease. This indicates that the template to monomer molar ratio is 1:3 or 1:4, the combinatorial ability is reduced due to fewer recognition sites for the target analyte. And when the molar ratio of the template to the monomer is more than 1:5, the MIP/CsPbBr is added 3 @SiO 2 The QDs molecularly imprinted membrane is too thick, so that the elution and re-adsorption functions of the recognition sites are hindered, and the fluorescence peak value is reduced. Therefore, the optimal molar ratio of CHO to APTEs is 1: 5.
Example 5
This example is substantially the same as example 1, except that the elution times in step (3) were adjusted to 5min, 10min, 15min, 20min and 25min, respectively, and the influence of the elution times on the detection effect of the sensor was investigated. FIG. 1(C) is a graph showing the effect of elution time on the fluorescence peak of the sensor. As shown in FIG. 1(C), when MIPs/CsPbBr 3 @SiO 2 The fluorescence peak value of the QDs composite material reaches the maximum value after being eluted for 15min and then gradually becomes stable, which indicates that the template is basically and completely eluted. Therefore the optimal elution time for MIPs should be 15 min.
Example 6
The present embodiment is substantially the same as embodiment 1, except that the identification time in step (4) is respectively adjusted to 10min, 20min, 30min, 40min and 50min, and the influence of the identification time on the identification effect of the sensor is studied. FIG. 1(D) is a graph showing the effect of the identification time on the fluorescence peak of the sensor. As shown in fig. 1(D), the fluorescence peak increased dramatically within the first 30min, indicating that most of the binding cavity was occupied by the template molecule CHO. After 30min of identification, the fluorescence peak value is slightly reduced and even tends to be stable, which indicates that the identification process reaches a saturation state. Therefore, 30min was selected as the optimal recognition time.
Claims (7)
1.SiO 2 Sol-gel encapsulated CsPbBr 3 The method for detecting cholesterol by using the quantum dots is characterized by comprising the following steps of:
step 1, octadecene, oleic acid and cesium carbonate are placed in N 2 Under protection, reacting at 150-160 ℃ to synthesize a cesium oleate precursor, and then reacting lead bromide, octylamine, oleic acid and methyltrimethoxysilane in N 2 Heating to 170-180 ℃ under protection, quickly injecting a cesium oleate solution, forming quantum dots in ice bath, finally exposing in air, stirring and reacting to generate CsPbBr 3 @SiO 2 The QDs composite material is characterized in that the volume ratio of oleic acid to octadecene is 1: 12-1: 13;
step 2, according to the molar ratio of 1: 5-1: 6 of cholesterol to 3-aminopropyltriethoxysilane, adding cholesterol, octadecene, 3-aminopropyltriethoxysilane and CsPbBr 3 @SiO 2 After QDs are stirred and mixed uniformly, tetramethoxysilane is added according to the molar ratio of 1: 9-1: 10 of tetramethoxysilane to 3-aminopropyltriethoxysilane, and the mixture is stirred and reacted in a closed manner at room temperature to obtain MIPs/CsPbBr 3 @SiO 2 QDs composites;
step 3, MIPs/CsPbBr is added 3 @SiO 2 Adding the QDs composite material into a mixed solution of ethyl acetate and normal hexane in a volume ratio of 1:3 for ultrasonic elution, wherein the elution time is 15-25 min, and then carrying out MIPs/CsPbBr after elution 3 @SiO 2 And dispersing the QDs composite material in an ethyl acetate solution, adding human serum to be detected, identifying for 30-50 min, measuring the fluorescence quenching intensity, and calculating to obtain the cholesterol concentration in the human serum to be detected according to the linear relationship between the fluorescence quenching intensity and the cholesterol concentration.
2. The method according to claim 1, wherein in step 1, the molar ratio of the lead bromide to the cesium carbonate is 1:5 to 1: 6.
3. The method as claimed in claim 1, wherein in step 1, the volume ratio of octylamine, oleic acid and methyltrimethoxysilane is 1: 1:1 to 1:2: 2.
4. The method of claim 1, wherein in step 1, the reaction time is 2-3 hours at 150-160 ℃.
5. The method according to claim 1, wherein in the step 1, the stirring reaction time is 30-40 min.
6. The method according to claim 1, wherein in the step 2, the stirring and mixing time is 30-40 min.
7. The method according to claim 1, wherein in the step 2, the reaction time is 12-14 h under closed stirring at room temperature.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107698719A (en) * | 2017-10-26 | 2018-02-16 | 江南大学 | Method for preparing water-dispersible quantum dot labeled molecularly imprinted nanoparticles based on macromolecular self-assembly |
| CN108801990A (en) * | 2018-04-19 | 2018-11-13 | 华南师范大学 | One kind being based on CsPbBr3The detection method of the flolimat of perovskite quantum dot-molecular engram fluorescent optical sensor |
| CN109520979A (en) * | 2018-11-02 | 2019-03-26 | 闽南师范大学 | A kind of detection method of cholesterol in serum |
| CN109897633A (en) * | 2019-03-20 | 2019-06-18 | 南宁师范大学 | The method of the preparation method and cholesterol detection of cobalt doped magnetism carbon quantum dot |
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| US10859560B2 (en) * | 2015-10-25 | 2020-12-08 | Adriel Sumathipala | Biosensors for detecting cholesterol and OxLDL in blood sample |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107698719A (en) * | 2017-10-26 | 2018-02-16 | 江南大学 | Method for preparing water-dispersible quantum dot labeled molecularly imprinted nanoparticles based on macromolecular self-assembly |
| CN108801990A (en) * | 2018-04-19 | 2018-11-13 | 华南师范大学 | One kind being based on CsPbBr3The detection method of the flolimat of perovskite quantum dot-molecular engram fluorescent optical sensor |
| CN109520979A (en) * | 2018-11-02 | 2019-03-26 | 闽南师范大学 | A kind of detection method of cholesterol in serum |
| CN109897633A (en) * | 2019-03-20 | 2019-06-18 | 南宁师范大学 | The method of the preparation method and cholesterol detection of cobalt doped magnetism carbon quantum dot |
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