CN111982866B - Method for detecting ketamine by colorimetric fluorescence quenching dual-mode - Google Patents
Method for detecting ketamine by colorimetric fluorescence quenching dual-mode Download PDFInfo
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- CN111982866B CN111982866B CN202010503591.XA CN202010503591A CN111982866B CN 111982866 B CN111982866 B CN 111982866B CN 202010503591 A CN202010503591 A CN 202010503591A CN 111982866 B CN111982866 B CN 111982866B
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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"
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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
Abstract
The invention provides a colorimetric fluorescence quenching dual-mode ketamine detection method, which relates to a detection kit consisting of a first reagent bottle, a second reagent bottle, filter paper, a medicine spoon and an ultraviolet lamp, and is characterized in that based on a quantum dot energy matching transfer principle, a chemical colorimetric method and a fluorescence quenching method caused by quantum dot ligand energy matching transfer are adopted, a solution prepared by strong base and a quantum dot solution subjected to cobalt tetrasulfate ligand replacement are sequentially and directly dripped onto an extracted suspicious sample placed on the filter paper, if the suspicious sample contains ketamine, a suspicious object on the filter paper reacts with a reagent to generate lavender purple precipitate, and 365nm ultraviolet light is used for irradiating the precipitate to find that the fluorescence at the precipitate is weakened, so that the existence of ketamine poison is determined. The method can be used for rapid qualitative detection during field virus detection of customs, customs barriers, dance halls and the like, has the characteristics of simple and rapid operation, strong specificity, sensitive reaction, convenient use and low cost, and provides an effective technical means for field identification of ketamine by a drug prohibition department of a public security system.
Description
Technical Field
The invention relates to a colorimetric fluorescence quenching dual-mode detection method for ketamine, in particular to a rapid detection method for ketamine in on-site drugs.
Background
Since the 21 st century, the use of drugs has been on the rise. Ketamine as a third generation novel drug, namely a novel psychoactive substance, is originally a dissociation anesthetic with a systemic rapid effect. It has been used as anesthetic for human and animal, and has short effective time and stable drug effect. Also, because of its relatively good safety, ketamine is rapidly becoming the first anesthetic of choice for injured american soldiers during war-beyond. The intake of ketamine results in a decline in memory and learning, with symptoms of flotation, hallucinations, spatiotemporal distortion, and addiction. Ketamine is abused due to its anaesthetic and hallucinogenic properties, especially in bars, clubs, concerts and party parties. Therefore, in this case, we need to do a quick and accurate field test to obtain evidence. In addition, because of the difficulty in its synthesis, a major reason for its ubiquity in the market is the neglect of transport of drug stocks. It has powder, liquid and solid dosage forms, which have led drug vendors to often mix them with similar items (sugar and flour, etc.) to avoid the search of checkpoints and frontier police. Ketamine is increasingly gaining attention as an additive ingredient in ecstasy pill tablets. Other controlled substances that are occasionally encountered in combination with ketamine in the ecstasy class of tablets include (but are not limited to): amphetamine, methamphetamine, Methylenedioxymethylenepropylamine (MDA), methylenedioxymethamphetamine (MDMA), and phencyclidine (PCP). This poses a significant challenge to the sample preparation process of the detection method and the anti-interference capability of the detection method.
Currently, the Analytical techniques for ketamine detection are mainly mass spectrometry (Talanta 2015,143,50-55), chromatography (Talanta 2010,82(3), 969-. Such as ELISA, fluorescence, GC-MS, etc., are used to detect metabolic components in human blood, urine, hair, etc. The methods have the defects of long detection time, troublesome sample preparation, difficult field visual detection and the like. Raman spectroscopy and terahertz spectroscopy require comparison of standard sample spectra. Therefore, the mixed medicine sample is difficult to identify correctly, and the anti-interference capability is poor. Most importantly, these methods require a rigorous laboratory environment, expensive large equipment and trained professionals to operate, and are limited by the use of ketamine protocols and drug-vending processes. On-site detection of ketamine is difficult to achieve.
Colorimetric reactions have the competitive advantage of being cost-effective and readily detectable without the need for complex and expensive analytical instruments (Nature 2000,406(6797), 710-. Moreover, the colorimetric reaction has the advantages of interference resistance, strong specificity, strong visual identification capability and the like. However, due to strong chemical interactions between the active center of the colorimetric probe molecule and the substance to be measured, such as electron transfer and ion exchange, the structure of the chromophore molecule linked to the active center changes, and accumulation of the chromophore molecule is observed by the human eye or a microscope, and thus the color change of the colorimetric reaction depends on the concentration of the chromophore molecule. This means that colorimetric reactions have the disadvantage of low sensitivity. Therefore, in order to improve the sensitivity of the colorimetric reaction, the idea of colorimetric fluorescence quenching dual-mode detection is adopted, color change signals before and after the reaction are increased, and energy level matching change is generated between the quantum dots and the ligand, so that quantum dot fluorescence quenching is caused by energy transfer, the change of fluorescence intensity before and after the reaction is increased, the characteristic of low sensitivity of the colorimetric reaction is compensated, and the reliability and the sensitivity of ketamine detection are greatly improved by the double-signal response method.
Disclosure of Invention
The invention aims to provide a colorimetric fluorescence quenching dual-mode ketamine drug detection method, which is based on the quantum dot energy matching transfer principle and relates to a detection kit consisting of a first reagent bottle, a second reagent bottle, filter paper, a medicine spoon and an ultraviolet lamp.
The invention relates to a method for detecting ketamine by a colorimetric fluorescence quenching dual-mode, which relates to a detection kit comprising a first reagent bottle, a second reagent bottle, filter paper, a medicine spoon and an ultraviolet lamp, wherein the colorimetric method and the fluorescence quenching method are combined to obtain double signal change detection ketamine, and the specific operation is carried out according to the following steps:
preparing an alkalized solution:
a. weighing 1.2-4.8g of NaOH or KOH at room temperature, dissolving in 100ml of water, and sealing in a first reagent bottle (1);
preparation of cobalt tetrathiocyanate ligand:
b. weighing 0.145-0.58g CoCl at room temperature2Dissolving in 50-200ml water at 50 deg.CHeating for 15min to obtain solution A;
c. taking 0.152-0.608g NH4Dissolving SCN solid in 50-200ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 1-4h to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. b, melting 0.1-1g of CdSe green-light quantum dots into 50-200ml of water solution, adding 0.05-0.2g of cobalt ammonium thiocyanate obtained in the step d, stirring for 5min, purifying, melting into 50-200ml of water to obtain a detection reagent, and sealing into a second reagent bottle (2);
and (3) detection:
f. using a medicine spoon (4) to extract a sample from the suspicious powdery object, placing the sample on a filter paper (3), dropwise adding a first reagent bottle (1) onto the filter paper containing the sample, dropwise adding a reagent onto the filter paper containing the sample by using a second reagent bottle (2), after 3 seconds, forming lavender purple precipitate by ketamine in the suspicious powdery object and quantum dots with cobalt tetrathiocyanate, irradiating the precipitate by using a 365nm ultraviolet lamp (5) to find that green fluorescence at the precipitate is weakened, and determining the existence of the ketamine.
The invention relates to a detection method for detecting ketamine through a colorimetric fluorescence quenching dual-mode, which comprises the steps of extracting a sample from a drug suspicious source by using a medicine spoon (4), placing the sample on filter paper (3), dropwise adding a detection reagent onto the filter paper (3) containing the sample by using a first reagent bottle (1), enabling the ketamine to be alkalized, dropwise adding the detection reagent onto the filter paper (3) containing the sample by using a second reagent bottle (2), enabling the alkalized ketamine to perform a coordination reaction with a cobalt tetrasulfate ligand on CdSe subjected to ligand exchange to generate lavender purple precipitate, irradiating the lavender purple precipitate by using 365nm ultraviolet light (5), enabling the fluorescence intensity of green fluorescence quantum dots to be weakened compared with a surrounding unreacted area at the purple precipitate, and determining the existence of the ketamine through the lavender purple precipitate and a fluorescence quenching phenomenon.
The invention relates to a detection method for detecting ketamine by a colorimetric fluorescence quenching dual-mode, wherein a detection object is ketamine mixed in a suspicious substance, and two reagents are respectively an alkaline solution prepared at room temperature and a quantum dot solution subjected to ligand exchange; a sample is extracted from a suspicious substance on site and placed on a filter paper, and reagents are sequentially dripped on the filter paper, so that if ketamine exists in the substance, a special color development reaction and fluorescence quenching phenomena can occur with a detection reagent. Namely, the alkalized ketamine can react with CdSe with a cobalt tetrathiocyanate ligand to generate a purple precipitate substance, and fluorescence quenching of quantum dots is caused, so that the existence of the ketamine is determined.
The colorimetric method is used for generating lavender purple precipitate by reacting a cobalt tetrathiocyanate ligand with alkalized ketamine, the fluorescence quenching method is used for generating lavender purple precipitate by reacting a quantum dot bridging cobalt tetrathiocyanate ligand with ketamine, energy is transferred to the cobalt tetrathiocyanate ligand after the reaction of the cobalt tetrathiocyanate ligand with ketamine so that the fluorescence of a system is quenched, a mechanism diagram is shown in figure 1, and the sensitivity and the reliability of ketamine detection can be improved by double signal response of the colorimetric and fluorescence quenching methods.
Drawings
FIG. 1 is a diagram of the mechanism for detecting ketamine according to the present invention;
FIG. 2 is a schematic view of the kit of the present invention, wherein 1 is a first reagent bottle, 2 is a second reagent bottle, 3 is filter paper, 4 is a spoon, and 5 is an ultraviolet lamp.
Detailed Description
The invention will now be further illustrated by means of specific examples, without the invention being restricted to these examples
Example 1
Preparing an alkalized solution:
a. 2.4g of NaOH is weighed by a tray balance at room temperature, dissolved in 100ml of water and sealed in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.29g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 100ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.304g NH4Dissolving SCN solid in 100ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 2 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. taking 0.5g of CdSe green quantum dots, melting into 100ml of water solution, adding 0.1g of cobalt ammonium thiocyanate obtained in the step d, stirring for 5min, purifying, melting into 100ml of water, obtaining a detection reagent, and sealing into a second reagent bottle 2;
and (3) detection:
f. using a medicine spoon 4 to extract a sample from a suspicious powdery object obtained from a singing and dancing hall, placing the sample on a filter paper 3, dropwise adding a first reagent bottle 1 on the filter paper containing the sample, dropwise adding a second reagent bottle 2 on the filter paper containing the sample, alkalifying ketamine in the suspicious powdery object after 3 seconds, generating lavender purple precipitate with quantum dots with cobalt tetrathiocyanate, irradiating the precipitate with a 365nm ultraviolet lamp 5 to find that green fluorescence at the precipitate is weakened, and determining the existence of the ketamine.
Example 2
Preparing an alkalized solution:
a. weighing 4.8g of NaOH by using a tray balance at room temperature, dissolving in 200ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.58g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 200ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.608g NH4Dissolving SCN solid in 200ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 4 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. b, melting 1.0g of CdSe green-light quantum dots into 200ml of water solution, adding 0.2g of cobalt ammonium thiocyanate obtained in the step d, stirring for 5min, purifying, melting into 200ml of water to obtain a detection reagent, and sealing into a second reagent bottle 2;
and (3) detection:
f. using a medicine spoon 4 to extract a sample from the suspicious powdery object obtained from the checkpoint, placing the sample on a filter paper 3, dropwise adding the sample on the filter paper containing the sample by using a first reagent bottle 1, dropwise adding a reagent on the filter paper containing the sample by using a second reagent bottle 2, alkalifying the ketamine in the suspicious powdery object, generating lavender purple precipitate with quantum dots with cobalt tetrathiocyanate after 3 seconds, irradiating the precipitate by using a 365nm ultraviolet lamp 5 to find that the green fluorescence at the precipitate is weakened, and determining the existence of the ketamine.
Example 3
Preparing an alkalized solution:
a. weighing 1.2g of NaOH by using a tray balance at room temperature, dissolving in 50ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.145g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 50ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.152g NH4Dissolving SCN solid in 50ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 1h to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. b, melting 0.25g of CdSe green light quantum dots into 50ml of water solution, adding 0.05g of cobalt ammonium thiocyanate obtained in the step d, stirring for 5min, purifying, melting into 50ml of water to obtain a detection reagent, and sealing the detection reagent into a second reagent bottle 2;
and (3) detection:
f. using a medicine spoon 4 to extract a sample from a suspicious powdery object obtained from a virus-removal site, placing the sample on a filter paper 3, dropwise adding a first reagent bottle 1 onto the filter paper containing the sample, dropwise adding a reagent onto the filter paper containing the sample by using a second reagent bottle 2, alkalifying ketamine in the suspicious powdery object, then generating lavender purple precipitate with quantum dots with cobalt tetrathiocyanate, irradiating the precipitate by using a 365nm ultraviolet lamp 5 to find that green fluorescence at the precipitate is weakened, and determining the existence of the ketamine.
Example 4
Preparing an alkalized solution:
a. 2.4g of KOH is weighed by a tray balance at room temperature, dissolved in 100ml of water and sealed in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.29g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 100ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.304g NH4Dissolving SCN solid in 100ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 2 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. taking 0.5g of CdSe green quantum dots, melting into 100ml of water solution, adding 0.1g of cobalt ammonium thiocyanate obtained in the step d, stirring for 5min, purifying, melting into 100ml of water, obtaining a detection reagent, and sealing into a second reagent bottle 2;
and (3) detection:
f. using a medicine spoon 4 to extract a sample from a suspicious powdery object obtained from customs, placing the sample on a filter paper 3, dropwise adding a first reagent bottle 1 onto the filter paper containing the sample, dropwise adding a reagent onto the filter paper containing the sample by using a second reagent bottle 2, alkalifying ketamine in the suspicious powdery object, then generating lavender purple precipitate with quantum dots with cobalt tetrathiocyanate, irradiating the precipitate by using a 365nm ultraviolet lamp 5 to find that green fluorescence at the precipitate is weakened, and determining the existence of the ketamine.
Example 5
Preparing an alkalized solution:
a. weighing 4.8g of KOH by using a tray balance at room temperature, dissolving in 200ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.58g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 200ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.608g NH4Dissolving SCN solid in 200ml water, heating at 85 deg.C for 20min to obtainSolution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 4 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. taking 0.1g of CdSe green light quantum dots, melting into 200ml of water solution, adding 0.2g of ammonium cobalt thiocyanate, stirring for 5min to ensure that sufficient ligand exchange occurs between the ligand and the CdSe green light quantum dots, then purifying the quantum dots, melting into 200ml of water solution to obtain a detection reagent, and sealing into a second reagent bottle 2;
and (3) detection:
f. using a medicine spoon 4 to extract a sample from a suspicious powdery object obtained from a checkpoint, placing the sample on a filter paper 3, dropwise adding the sample on the filter paper containing the sample by using a first reagent bottle 1, dropwise adding a reagent on the filter paper containing the sample by using a second reagent bottle 2, alkalifying the ketamine in the suspicious powdery object, generating a lavender purple precipitate with quantum dots with cobalt tetrathiocyanate after 3 seconds, and irradiating the precipitate by using a 365nm ultraviolet lamp 5 to find that the green fluorescence at the precipitate is weakened, so that the existence of the ketamine can be determined.
Example 6
Preparing an alkalized solution:
a. weighing 1.2g of KOH by using a tray balance at room temperature, dissolving in 50ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.145g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 50ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.152g NH4Dissolving SCN solid in 50ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 1h to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. taking 0.25g of CdSe green quantum dots, melting into 50ml of aqueous solution, adding 0.05g of ammonium cobalt thiocyanate, stirring for 5min to ensure that sufficient ligand exchange occurs between the ligand and the CdSe green quantum dots, then purifying the quantum dots, melting into 50ml of aqueous solution to obtain a detection reagent, and sealing into a second reagent bottle 2;
f. a medicine spoon 4 is used for extracting a sample from a suspicious powdery object obtained from a virus-removal site, the sample is placed on a filter paper 3, a first reagent bottle 1 is used for dropwise adding the sample on the filter paper containing the sample, a second reagent bottle 2 is used for dropwise adding a reagent on the filter paper containing the sample, ketamine in the suspicious powdery object is alkalized and then generates lavender purple precipitate together with quantum dots with cobalt tetrathiocyanate after 3 seconds, the precipitate is irradiated by a 365nm ultraviolet lamp 5, green fluorescence at the precipitate is found to be weakened, and the existence of ketamine can be determined.
Example 7
Preparing an alkalized solution:
a. 2.4g of NaOH is weighed by a tray balance at room temperature, dissolved in 100ml of water and sealed in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.29g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 100ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.304g NH4Dissolving SCN solid in 100ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 2 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
e. ligand exchange: taking 0.5g of CdSe green quantum dots, blending into 100ml of water solution, adding 0.1g of cobalt ammonium thiocyanate, stirring for 5min to ensure that sufficient ligand exchange occurs between the ligand and the CdSe quantum dots, then purifying the quantum dots, blending into 100ml of water solution to obtain a detection reagent, and sealing into a second reagent bottle 2;
f. the method comprises the steps of extracting a sample from a suspicious powdery object obtained from a singing and dancing hall by using a medicine spoon 4, placing the sample on a filter paper 3, dropwise adding a first reagent bottle 1 onto the filter paper containing the sample, dropwise adding a second reagent bottle 2 onto the filter paper containing the sample, wherein after 3 seconds, no purple precipitate is generated, and when the suspicious object is irradiated by a 365nm ultraviolet lamp 5, green fluorescence at the position is not weakened, so that the sample does not contain ketamine.
Example 8
Preparing an alkalized solution:
a. weighing 4.8g of NaOH by using a tray balance at room temperature, dissolving in 200ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.58g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 200ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.608g NH4Dissolving SCN solid in 200ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 4 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
e. ligand exchange: 1g of CdSe green quantum dots are taken to be blended into 200ml of water solution, then 0.2g of ammonium cobalt tetrathiocyanate is added, stirring is carried out for 5min, so that sufficient ligand exchange occurs between the ligand and the CdSe quantum dots, then the quantum dots are purified and blended into 200ml of water solution, and a detection reagent is obtained and sealed into a second reagent bottle 2;
f. the method comprises the steps of extracting a sample from a suspicious powder object obtained by checkpoint by using a medicine spoon 4, placing the sample on a filter paper 3, dropwise adding a first reagent bottle 1 onto the filter paper containing the sample, dropwise adding a second reagent bottle 2 onto the filter paper containing the sample, wherein no purple precipitate is generated after 3 seconds, and when the suspicious object is irradiated by a 365nm ultraviolet lamp 5, green fluorescence at the suspicious object is not weakened, so that the sample does not contain ketamine.
Example 9
Preparing an alkalized solution:
a. weighing 1.2g of NaOH by using a tray balance at room temperature, dissolving in 50ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.145g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 50ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.152g NH4Dissolving SCN solid in 50ml water, heating at 85 deg.C for 20min to obtainObtaining a solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 1h to obtain blue-green cobalt ammonium tetrasulfate crystals;
e. ligand exchange: taking 0.25g of CdSe green quantum dots, melting into 50ml of aqueous solution, adding 0.05g of ammonium cobalt thiocyanate, stirring for 5min to ensure that sufficient ligand exchange occurs between the ligand and the CdSe green quantum dots, then purifying the quantum dots, melting into 50ml of aqueous solution to obtain a detection reagent, and sealing into a second reagent bottle 2;
f. a medicine spoon 4 is used for extracting a sample from a suspicious powdery object obtained in a virus-sweeping field, the sample is placed on a filter paper 3, a first reagent bottle 1 is used for dropwise adding the sample on the filter paper containing the sample, a second reagent bottle 2 is used for dropwise adding a reagent on the filter paper containing the sample, after 3 seconds, no purple precipitate is generated, and when the suspicious object is irradiated by a 365nm ultraviolet lamp 5, green fluorescence at the position of the suspicious object is not weakened, and the sample does not contain ketamine.
Example 10
Preparing an alkalized solution:
a. 2.4g of KOH is weighed by a tray balance at room temperature, dissolved in 100ml of water and sealed in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.29g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 100ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.304g NH4Dissolving SCN solid in 100ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 2 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. taking 0.5g of CdSe green light quantum dots, melting into 100ml of water solution, adding 0.1g of ammonium cobalt thiocyanate, stirring for 5min to ensure that sufficient ligand exchange occurs between the ligand and the CdSe green light quantum dots, then purifying the quantum dots, melting into 100ml of water solution to obtain a detection reagent, and sealing into a second reagent bottle 2;
f. a medicine spoon 4 is used for extracting a sample from a suspicious powdery object obtained from customs, the sample is placed on a filter paper 3, a first reagent bottle 1 is used for dropwise adding the sample on the filter paper containing the sample, a second reagent bottle 2 is used for dropwise adding a reagent on the filter paper containing the sample, after 3 seconds, no purple precipitate is generated, and when the suspicious object is irradiated by a 365nm ultraviolet lamp 5, green fluorescence at the position is not weakened, and the sample does not contain ketamine.
Example 11
Preparing an alkalized solution:
a. weighing 4.8g of KOH by using a tray balance at room temperature, dissolving in 200ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.58g CoCl was weighed out at room temperature using an analytical balance2Dissolving in 200ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.608g NH4Dissolving SCN solid in 200ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 4 hours to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. taking 0.1g of CdSe green light quantum dots, melting into 200ml of water solution, adding 0.2g of ammonium cobalt thiocyanate, stirring for 5min to ensure that sufficient ligand exchange occurs between the ligand and the CdSe green light quantum dots, then purifying the quantum dots, melting into 200ml of water solution to obtain a detection reagent, and sealing into a second reagent bottle 2;
f. the method comprises the steps of extracting a sample from a suspicious powder object obtained by checkpoint by using a medicine spoon 4, placing the sample on a filter paper 3, dropwise adding a first reagent bottle 1 onto the filter paper containing the sample, dropwise adding a second reagent bottle 2 onto the filter paper containing the sample, wherein no purple precipitate is generated after 3 seconds, and when the suspicious object is irradiated by a 365nm ultraviolet lamp 5, green fluorescence at the suspicious object is not weakened, so that the sample does not contain ketamine.
Example 12
Preparing an alkalized solution:
a. weighing 1.2g of KOH by using a tray balance at room temperature, dissolving in 50ml of water, and sealing in a first reagent bottle 1;
preparation of cobalt tetrathiocyanate ligand:
b. preparation of cobalt tetrathiocyanate ligands: 0.145g CoCl was weighed out at room temperature with an analytical balance2Dissolving in 50ml water, heating at 50 deg.C for 15min to obtain solution A;
c. take 0.152g NH4Dissolving SCN solid in 50ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 1h to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. taking 0.25g of CdSe green quantum dots, melting into 50ml of aqueous solution, adding 0.05g of ammonium cobalt thiocyanate, stirring for 5min to ensure that sufficient ligand exchange occurs between the ligand and the CdSe green quantum dots, then purifying the quantum dots, melting into 50ml of aqueous solution to obtain a detection reagent, and sealing into a second reagent bottle 2;
f. a medicine spoon 4 is used for extracting a sample from a suspicious powdery object obtained in a virus-sweeping field, the sample is placed on a filter paper 3, a first reagent bottle 1 is used for dropwise adding the sample on the filter paper containing the sample, a second reagent bottle 2 is used for dropwise adding a reagent on the filter paper containing the sample, after 3 seconds, no purple precipitate is generated, and when the suspicious object is irradiated by a 365nm ultraviolet lamp 5, green fluorescence at the position of the suspicious object is not weakened, and the sample does not contain ketamine.
Claims (1)
1. A method for detecting ketamine in a colorimetric fluorescence quenching dual-mode is characterized in that the method relates to a kit for detection, the kit comprises a first reagent bottle, a second reagent bottle, filter paper, a medicine spoon and an ultraviolet lamp, the colorimetric method and the fluorescence quenching method are combined to obtain the ketamine for detecting double signal changes, and the specific operation is carried out according to the following steps:
preparing an alkalized solution:
a. weighing 1.2-4.8g of NaOH or KOH at room temperature, dissolving in 100ml of water, and sealing in a first reagent bottle (1);
preparation of cobalt tetrathiocyanate ligand:
b. weighing 0.145-0.58g CoCl at room temperature2Dissolving in 50-200ml water, heating at 50 deg.CHeating for 15min to obtain solution A;
c. taking 0.152-0.608g NH4Dissolving SCN solid in 50-200ml water, heating at 85 deg.C for 20min to obtain solution B;
d. mixing the solution A obtained in the step B with the solution B obtained in the step c, and heating for 1-4h to obtain blue-green cobalt ammonium tetrasulfate crystals;
ligand exchange:
e. b, melting 0.1-1g of CdSe green-light quantum dots into 50-200ml of water solution, adding 0.05-0.2g of cobalt ammonium thiocyanate obtained in the step d, stirring for 5min, purifying, melting into 50-200ml of water to obtain a detection reagent, and sealing into a second reagent bottle (2);
and (3) detection:
f. using a medicine spoon (4) to extract a sample from the suspicious powdery object, placing the sample on a filter paper (3), dropwise adding a first reagent bottle (1) onto the filter paper containing the sample, dropwise adding a reagent onto the filter paper containing the sample by using a second reagent bottle (2), after 3 seconds, forming lavender purple precipitate by ketamine in the suspicious powdery object and quantum dots with cobalt tetrathiocyanate, irradiating the precipitate by using a 365nm ultraviolet lamp (5) to find that green fluorescence at the precipitate is weakened, and determining the existence of the ketamine.
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