CN113607521A - Preparation method of medicine/drug positive hair for scientific research - Google Patents
Preparation method of medicine/drug positive hair for scientific research Download PDFInfo
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- 210000004209 hair Anatomy 0.000 title claims abstract description 146
- 238000011160 research Methods 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- 231100000640 hair analysis Toxicity 0.000 claims abstract description 153
- 238000000034 method Methods 0.000 claims abstract description 23
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 6
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- 230000003647 oxidation Effects 0.000 claims description 5
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- 235000018417 cysteine Nutrition 0.000 claims description 4
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- 239000000523 sample Substances 0.000 description 18
- 230000000052 comparative effect Effects 0.000 description 14
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- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 12
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- 238000001514 detection method Methods 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 8
- 229960003299 ketamine Drugs 0.000 description 8
- 238000003908 quality control method Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 229960005181 morphine Drugs 0.000 description 6
- 239000004081 narcotic agent Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000012488 sample solution Substances 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000003648 hair appearance Effects 0.000 description 4
- 229940124811 psychiatric drug Drugs 0.000 description 4
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- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 3
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- 239000003053 toxin Substances 0.000 description 3
- 231100000765 toxin Toxicity 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000003638 chemical reducing agent Substances 0.000 description 2
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- 231100000614 poison Toxicity 0.000 description 2
- 239000002574 poison Substances 0.000 description 2
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- 239000012224 working solution Substances 0.000 description 2
- 206010013654 Drug abuse Diseases 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 1
- WJAJPNHVVFWKKL-UHFFFAOYSA-N Methoxamine Chemical compound COC1=CC=C(OC)C(C(O)C(C)N)=C1 WJAJPNHVVFWKKL-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N2001/2893—Preparing calibration standards
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the technical field of forensic science, drugs and drug analysis, in particular to a preparation method of drug/drug positive hair for scientific research, which carries out a controllable reduction-molecular fixation-oxidation closed chemical treatment path on a hair sample structure, closely combines drug/drug molecules with hair protein to obtain high-fidelity positive hair, has uniform distribution of the drug/drug molecules in the positive hair, can be washed by organic solvents, surfactants and other substances, has stable properties in the processes of storage and use, has high application value in the fields of judicial identification, laboratory capability authentication, scientific research and development and the like, and lays a foundation for the research and development of hair standard reference substances.
Description
Technical Field
The invention relates to the technical field of forensic drug analysis, in particular to a preparation method of drugs/drugs positive hair for scientific research.
Background
In judicial practice, analysis of the composition of organic toxicants, particularly drugs of abuse and metabolites, in hair is important and even the only scientific evidence for determining whether a subject is taking drugs and for inferring the history of abuse. The tenth regulation of drug related personnel hair sample detection regulation (official drug ban (2018) 938) (hereinafter, regulation) issued by the ministry of public security also indicates that the hair sample detection result within 3cm of the hair root end is positive, which indicates that the tested person takes drugs within 6 months before the day of hair sample extraction.
In the hair analysis, the positive judgment criterion is that the actual detection content value of the test substance is positive above a threshold value specified in "norm". Therefore, the quantitative analysis result of the hair plays a role in determining whether the suspect has the history of drug abuse, and certain requirements are made on the accuracy of the hair analysis result. As is well known, the accuracy and reliability of the analysis results depend on quality control and correction methods, and the determination of a suitable substrate quality control is of great importance. The existing preparation process of the hair quality control sample is to simply soak in a standard substance, and the soaked positive sample is subjected to concentration detection to compound to obtain positive hair with a certain concentration, but the solid property of the hair and the structural property of the hair are that the simply soaked solute is difficult to enter the hair, so that the sample is easily influenced by the surrounding environment in the storage and use processes, so that the sample is desorbed and deteriorated, the compound sample also has the problems in the aspect of mixing uniformity, and the actual concentration taken in the use process is inaccurate, so that the final detection result is influenced.
Disclosure of Invention
Aiming at the technical problems of uneven distribution of drugs/drugs, easy desorption and deterioration and difficult storage of the drugs/drugs positive hair for scientific research and examination in the prior art, the invention provides the preparation method of the drugs/drugs positive hair for scientific research.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
the embodiment of the invention provides a preparation method of medicine/drug positive hair for scientific research, which specifically comprises the following steps:
s1: cleaning and drying a blank hair sample in deionized water, putting the blank hair sample into a disulfide bond reduction solution for reaction until the elongation at break of the blank hair sample is more than or equal to 20%, taking the blank hair sample out, cleaning the blank hair sample with deionized water, and drying the blank hair sample to obtain a pretreated hair sample;
s2: soaking the pretreated hair sample in a solution of a target immersed in a medicine/drug, and drying to obtain a soaked hair sample;
s3: and putting the soaked hair sample into a disulfide bond oxidation solution for reaction until the fracture elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample with deionized water, and drying the hair sample to obtain the medicine/drug positive hair for scientific research.
Compared with the prior art, the preparation method of the drug/drug positive hair for scientific research disclosed by the application firstly opens the disulfide bonds in the hair protein through the disulfide bond reducing agent, so that the structure of the hair becomes loose, and the entry and combination of target drug/drug molecules in the subsequent soaking process are facilitated; then, the target is soaked in the solution of the medicine/drug, so that the medicine/drug molecules are uniformly distributed in the hair protein and combined with the hair protein, and the uniformity of the medicine/drug components in the obtained positive hair is ensured; after soaking, the disulfide bonds in the hair protein are closed again through the disulfide bond oxidizing agent, so that the hair is restored to a compact state, and the target drug/drug molecules are tightly wrapped in the hair protein and are more tightly combined with the hair protein. Through inspection, the positive hair prepared by the preparation method of the medicine/drug positive hair for scientific research disclosed by the application is stable in property, can be washed by an organic solvent and a surfactant, and has a longer shelf life. The preparation method has important significance for developing standard reference substances for hair.
Preferably, the disulfide bond reducing solution is an aqueous or organic solution of at least one of thiol, dithiothreitol, cysteine, thioglycolic acid, tris (2-carboxyethyl) phosphine, dimercaptoethanol and sodium sulfite at a concentration of 0.1 wt% to 1 wt%.
The optimal disulfide bond reduction solution has moderate reduction capability, can selectively destroy chemical bonds of hair protein molecules, protects other chemical bonds among the hair proteins while destroying the disulfide bonds among the hair proteins, avoids the situation that the hair proteins are thoroughly disintegrated and cannot be restored, does not generate side reactions with drugs and medicines immersed in a target, and protects the stability of the drugs/medicines immersed in the target.
Preferably, the target immersion drug/drug solution in S2 is an aqueous solution or an organic solution of one drug/drug in the national food and drug administration, the ministry of public security, the appendix of the national institutes of health "notice on the catalogues of published narcotics and psychiatric drug (food and drug administration (2013) 230) and the catalogues of non-pharmaceutical narcotics and psychiatric drug administration supplementary catalogues published by the above-mentioned departments over the years, and the immersion time of the pretreated hair sample in the target immersion drug/drug solution is between 6h and 12 h.
The preparation method of the medicine/drug positive hair for scientific research has good applicability, and can ensure the rate of medicine/drug molecules entering hair protein and ensure a more excellent fixing effect by combining the optimal concentration and soaking time of the target medicine/drug solution. The preferred target immersion concentration and drug/drug solution concentration, immersion time relationship is: when the target immersion concentration of the drugs/drugs in the positive hair is 0.2ng/mg, 1-3ppm of the target is selected to be immersed in the drug/drug solution for 6-8 h; when the target immersion concentration of the traditional Chinese medicine/drug for the positive hair is 0.4ng/mg, 3-6ppm of target immersion medicine/drug solution is selected for immersion for 8-10 h; when the target immersion concentration of the drug/drug in the positive hair is 0.6ng/mg, the drug/drug solution is immersed in the target immersion concentration of 6-10ppm for 10-12 h.
Preferably, the disulfide bond oxidation solution in S3 is an aqueous solution of one of hydrogen peroxide, peracetic acid or sodium percarbonate, and the concentration is 1 wt% -5 wt%.
Preferably, the drying temperature of the hair in S1 is 10-40 ℃, and the drying temperature of the hair in S2 and S3 is 40-70 ℃.
The drying temperature of 10-40 ℃ in S1 can ensure the basic structure and properties of hair protein and simultaneously dry the hair, and the drying temperature of 40-70 ℃ in S2 and S3 can ensure that the hair protein does not deteriorate and the hair is dried under the condition that medicines/drugs are not attached.
The application also provides the medicine/drug positive hair for scientific research prepared by the preparation method of the medicine/drug positive hair for scientific research.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Compared with biological test materials such as blood, urine, bile and the like, drugs/medicines are metabolized in hair slowly and exist for a long time, so that hair analysis has unique advantages in forensic science toxicant analysis, and the analysis of the components of drugs/drugs and metabolites thereof abused in hair is one of important scientific evidences for judging whether a person is drug addict or not. In hair analysis, it is very important to prepare high-quality positive hair quality control products in order to eliminate bias and enhance the accuracy of analysis results. The existing preparation process of the positive hair is to simply soak in a medicine/drug sample solution, the soaked positive sample is compounded with a blank sample after accurate concentration detection to obtain a positive hair quality control product with standard concentration, but due to the solid property of the hair and the protein structure of the hair, the medicine in the standard sample solution is difficult to enter the hair through simple soaking, but is adhered to the surface of the hair through physical adsorption, so that the medicine is easy to desorb and deteriorate in the storage and use processes, and the uniformity of the sample concentration is difficult to ensure because the quality control product is compounded, so that adverse factors influence the quality of the hair quality control product, and the stability of a detection result is also damaged to a certain extent.
In order to enable the medicine/drug in the medicine/drug positive hair to be distributed more uniformly and combined with the hair more tightly, improve the stability of the positive hair in the storage and use processes, prolong the quality guarantee period of the positive hair and provide technical support for the development of hair standard reference substances, the application provides a preparation method of the medicine/drug positive hair for scientific research: the method specifically comprises the following steps:
s1: washing a blank hair sample in deionized water, drying, putting the blank hair sample into a disulfide bond reduction solution for reaction, taking out the blank hair sample until the elongation at break of the blank hair sample is more than or equal to 20%, washing the blank hair sample with the deionized water, and drying to obtain a pretreated hair sample;
s2: soaking the pretreated hair sample in a solution of a target immersed in a medicine/drug, and drying to obtain a soaked hair sample;
s3: and putting the soaked hair sample into a disulfide bond oxidation solution for reaction until the fracture elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample with deionized water, and drying the hair sample to obtain the medicine/drug positive hair for scientific research.
According to the preparation method of the drug/drug positive hair for scientific research, firstly, the disulfide bonds in the hair protein are opened through the disulfide bond reducing agent, so that the structure of the hair is loosened, and the entry and combination of target drug/drug molecules in the subsequent soaking process are facilitated; then, the target is soaked in the solution of the medicine/drug, so that the medicine/drug molecules are uniformly distributed in the hair protein and combined with the hair protein, and the uniformity of the medicine/drug components in the obtained positive hair is ensured; after soaking, the disulfide bonds in the hair protein are closed again through the disulfide bond oxidizing agent, the hair is restored to a compact state, target medicine/drug molecules are tightly wrapped in the hair protein and are combined with the hair protein more tightly, and through inspection, the positive hair prepared by the preparation method of the medicine/drug positive hair for scientific research disclosed by the application is stable in property, can be washed by organic solvents, surfactants and other substances, is longer in quality guarantee period, and has important significance for development of hair standard reference substances.
In the reduction process of the disulfide bond, the reduction capability of the reduction solution is moderate, and the reduction solution needs to be selective when chemical bonds in the hair protein are destroyed, namely the disulfide bond is destroyed as much as possible without damaging other chemical bonds, the basic structure of the hair protein is protected from being destroyed, the hair protein does not react with a target immersion drug/drug to avoid the hair protein from being degraded, so that the aqueous solution or the organic solution of one or more of mercaptan, dithiothreitol, cysteine, thioglycolic acid, tris (2-carboxyethyl) phosphine, dimercaptoethanol and sodium sulfite is selected, and the concentration is kept between 0.1 and 1 percent by weight.
The preparation method of the positive hair of the medicines/drugs for scientific research provided by the application has good applicability, and is suitable for the aqueous solution or inorganic solution of all the medicines/drugs in the appendix of the State food and drug administration, the Ministry of public Security, the Notification of catalogues of narcotics and psychiatric drugs of the national institutes of health (food and drug administration, No. 230) and the supplement catalogues of non-medicinal narcotics and psychiatric drug control varieties, which are published by the departments all the year round.
The immersion rate and the bonding degree of the drugs/drugs are key factors for determining the bonding of the target drugs/drugs and the hair proteins, and the inventors have long studied that the concentration of the target immersion drug/drug solution in S2 is 1ppm-10ppm, and the immersion time of the pretreated hair sample in the target immersion drug/drug solution is between 6h and 12h, so that the effect is the best.
Furthermore, the inventor finds that the immersion effect of the drugs/drugs is not greatly related to the types of the drugs/drugs, but has a great relationship with the concentration of the target drugs/drugs and the time in the research process, so the inventor researches the target immersion concentration of the drugs/drugs in the positive hair, the optimal concentration and the optimal immersion time of the target drug/drug solution, and the specific result is that when the target immersion concentration is 0.2ng/mg, 1-3ppm of sample solution is selected to be immersed for 6-8 hours; when the target immersion concentration is 0.4ng/mg, 3-6ppm of sample solution is selected for immersion for 8-10 h; when the target immersion concentration is 0.6ng/mg, 6-10ppm of sample solution is selected and soaked for 10-12 h.
The whole preparation process should keep the structure and properties of hair protein from being damaged excessively, so the drying temperature in S1 is selected to be 10-40 ℃, the hair samples in S2 and S3 are combined with drugs/narcotics, the properties are stable, and the drying temperature in S2 and S3 is selected to be 40-70 ℃.
The present invention will be further described below by way of examples, in which the hair samples of the same 30-year-old healthy male are selected to demonstrate the effect of the method for preparing drug/drug positive hair for scientific research.
Example 1
The embodiment provides a preparation method of ice toxin positive hair for scientific research, which specifically comprises the following steps:
s1: washing a blank hair sample in deionized water, drying the blank hair sample in a constant temperature environment at 30 ℃, putting the blank hair sample into an ethanol solution of tris (2-carboxyethyl) phosphine with the concentration of 0.5 wt% for reaction, taking out 5 samples every 1 hour in the reaction process for tensile test until the average breaking elongation of the blank hair sample is more than or equal to 20%, taking out the samples, washing the samples with deionized water, and drying the samples in a constant temperature environment at 60 ℃ to obtain a pretreated hair sample;
s2: taking 400mg of the pretreated hair sample, soaking the hair sample in 250ml of 1ppm glacial acetic acid aqueous solution for 7.5h, taking out the hair sample, and drying the hair sample in a constant temperature environment at 50 ℃ to obtain a soaked hair sample;
s3: and putting the soaked hair sample into a 3 wt% hydrogen peroxide solution for reaction, taking out 3 samples every 5min in the reaction process for tensile test until the average breaking elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample by deionized water, and drying the hair sample in a constant temperature environment at 50 ℃ to obtain the ice toxicity positive hair for scientific research, wherein the ice toxicity content in the positive hair is 0.215ng/mg through inspection.
Example 2
The embodiment provides a preparation method of ice toxin positive hair for scientific research, which specifically comprises the following steps:
s1: cleaning a blank hair sample in deionized water, drying the blank hair sample in a constant temperature environment at 25 ℃, putting the blank hair sample into an ethanol solution of dimercaptoethanol and sodium sulfite with the concentration of 0.5 wt% to react, taking out 5 samples every 1 hour in the reaction process to perform a tensile test until the average breaking elongation of the blank hair sample is more than or equal to 20%, taking out the samples, cleaning the samples with deionized water, and drying the samples in a constant temperature environment at 60 ℃ to obtain a pretreated hair sample;
s2: taking 400mg of the pretreated hair sample, soaking the hair sample in 250ml of an glacial acetic acid aqueous solution with the concentration of 3ppm for 9.5h, taking out the hair sample, and drying the hair sample in a constant temperature environment at 50 ℃ to obtain a soaked hair sample;
s3: and putting the soaked hair sample into a 3 wt% peracetic acid aqueous solution for reaction, taking 3 samples out every 5min in the reaction process for tensile test until the average breaking elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample by deionized water, and drying the hair sample in a constant temperature environment at 50 ℃ to obtain the ice toxicity yang hair for scientific research, wherein the ice toxicity content in the positive hair is 0.413ng/mg through inspection.
Example 3
The embodiment provides a preparation method of ketamine positive hair for scientific research, which specifically comprises the following steps:
s1: cleaning blank hair samples in deionized water, drying in a constant temperature environment at 20 ℃, putting into a dithiothreitol aqueous solution with the concentration of 0.5 wt% for reaction, taking out 5 samples every 1h in the reaction process for tensile test until the average breaking elongation of the blank hair samples is more than or equal to 20%, taking out the samples, cleaning with deionized water, and drying in a constant temperature environment at 40 ℃ to obtain pretreated hair samples;
s2: taking 400mg of the pretreated hair sample, soaking the hair sample in 250ml of ketamine aqueous solution with the concentration of 6ppm for 8.5h, taking out the hair sample, and drying the hair sample in a constant-temperature environment at 40 ℃ to obtain a soaked hair sample;
s3: and putting the soaked hair sample into 2 wt% of peroxyacetic acid solution for reaction, taking 3 samples out every 5min in the reaction process for tensile test until the average breaking elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample by deionized water, and drying the hair sample in a constant temperature environment at 50 ℃ to obtain the ice toxicity yang hair for scientific research, wherein the content of ketamine in the positive hair is 0.433ng/mg through inspection.
Example 4
The embodiment provides a preparation method of ketamine positive hair for scientific research, which specifically comprises the following steps:
s1: cleaning a blank hair sample in deionized water, drying the blank hair sample in a constant temperature environment of 40 ℃, putting cysteine with the concentration of 1 wt% and a dimercaptoethanol aqueous solution for reaction, taking out 5 samples every 1 hour in the reaction process, carrying out a tensile test until the average breaking elongation of the blank hair sample is more than or equal to 20%, taking out the samples, cleaning the samples with deionized water, and drying the samples in a constant temperature environment of 30 ℃ to obtain a pretreated hair sample;
s2: taking 400mg of the pretreated hair sample, soaking the hair sample in 250ml of ketamine aqueous solution with the concentration of 10ppm for 10.5h, taking out the hair sample, and drying the hair sample in a constant-temperature environment at 50 ℃ to obtain a soaked hair sample;
s3: putting the soaked hair sample into 4 wt% of sodium percarbonate solution for reaction, taking 3 samples out every 5min in the reaction process for tensile test until the average breaking elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample by deionized water, and drying the hair sample in a constant temperature environment at 60 ℃ to obtain the ice toxicity yang hair for scientific research, wherein the content of ketamine in the positive hair is 0.608ng/mg through inspection.
Example 5
The embodiment provides a preparation method of morphine-positive hair for scientific research, which specifically comprises the following steps:
s1: cleaning a blank hair sample in deionized water, drying the blank hair sample in a constant temperature environment at 35 ℃, putting the blank hair sample into a dithiothreitol aqueous solution with the concentration of 0.5 wt% for reaction, taking out 5 samples every 1 hour in the reaction process for tensile test until the average breaking elongation of the blank hair sample is more than or equal to 20%, taking out the blank hair sample, cleaning the blank hair sample with deionized water, and drying the blank hair sample in a constant temperature environment at 40 ℃ to obtain a pretreated hair sample;
s2: taking 400mg of the pretreated hair sample, soaking the hair sample in 250ml of morphine aqueous solution with the concentration of 2ppm for 7 hours, taking out the hair sample, and drying the hair sample in a constant temperature environment at 50 ℃ to obtain a soaked hair sample;
s3: and putting the soaked hair sample into a 3 wt% hydrogen peroxide solution for reaction, taking out 3 samples every 5min in the reaction process for tensile test until the average breaking elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample by deionized water, and drying the hair sample in a constant temperature environment at 60 ℃ to obtain the ice toxicity positive hair for scientific research, wherein the content of morphine in the positive hair is 0.218ng/mg through inspection.
Example 6
The embodiment provides a preparation method of morphine-positive hair for scientific research, which specifically comprises the following steps:
s1: cleaning a blank hair sample in deionized water, drying the blank hair sample in a constant temperature environment of 30 ℃, putting the blank hair sample into dithiothreitol aqueous solution with the concentration of 1 wt% for reaction, taking out 5 samples every 1 hour in the reaction process for tensile test until the average breaking elongation of the blank hair sample is more than or equal to 20%, taking out the blank hair sample, cleaning the blank hair sample with deionized water, and drying the blank hair sample in the constant temperature environment of 30 ℃ to obtain a pretreated hair sample;
s2: taking 400mg of the pretreated hair sample, soaking the hair sample in 250ml of morphine aqueous solution with the concentration of 10ppm for 11h, taking out the hair sample, and drying the hair sample in a constant temperature environment at 50 ℃ to obtain a soaked hair sample;
s3: and putting the soaked hair sample into 5 wt% of hydrogen peroxide solution for reaction, taking out 3 samples every 5min in the reaction process for tensile test until the average breaking elongation of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample by deionized water, and drying the hair sample in a constant temperature environment at 60 ℃ to obtain the ice toxicity positive hair for scientific research, wherein the content of morphine in the positive hair is 0.625ng/mg through inspection.
Comparative example 1
The comparative example provides ice poison positive hair for scientific research, which is prepared by adopting a preparation method of an organic poison analysis quality control product in human hair provided by the application with the application number of 202010790606.5, and comprises the following specific steps:
step 1: preparation of human hair:
taking each volunteer individual as a human hair source, respectively performing LC-MS/MS analysis on human hair provided by the volunteer individual (referring to technical specification SF/ZJD0107025-2018, washing the human hair to be detected by sequentially oscillating water and acetone, cutting the hair into 1-1.5mm sections after drying, crushing the hair sections in a freeze grinder, weighing 20mg of hair powder, adding 10mL of 1ng/mL internal metametamine standard working solution, centrifuging the hair for 30min by ice bath ultrasound, removing supernatant, drying the supernatant under 60 ℃ water bath air flow, redissolving the residue by 100uL of methanol, and analyzing the sample injection by LC-MS/MS); analyzing the human hair source with positive analysis result, then respectively carrying out quantitative analysis according to industry standard SF/T0063-2020 to obtain the content value of the target organic toxicant, respectively recording, and respectively cutting to about 0.2-0.5mm to obtain a positive hair source sample for later use; and (4) mixing human hair sources with negative analysis results, and shearing into 5-8cm sections for later use.
Step 2: preparing a soaking solution:
taking one beaker with the capacity of 1L, adding 15mg of an ice toxin reference substance and 40mg of a ketamine reference substance, adding 250mL of deionized water, stirring and dissolving uniformly, placing in an ice bath environment, and sequentially adding 250mL of dimethyl sulfoxide reagent and 425uL of concentrated hydrochloric acid to obtain a soaking solution.
And step 3: sample preparation:
adding 30g of negative human hair into the soaking solution, stirring uniformly, sealing by using a sealing film, and fully soaking for two weeks at the temperature of 1-7 ℃ and under the humidity of less than 80%; taking out after soaking, washing with methanol, air drying, cutting into pieces of 0.2-0.5mm to obtain hair soaking sample according to industry standard SF/T0063-2020. And respectively carrying out quantitative analysis to measure the content of the ice toxicity reference substance and the ketamine reference substance in the hair soaking sample.
Setting the preset value to be 0.4ng/mg, adding different positive hair samples into the hair soaking sample to obtain a mixed sample, and carrying out quantitative analysis on the mixed sample according to an industry standard SF/T0063-2020 to finally obtain the positive hair with the ice toxicity content of 0.403 ng/mg.
Comparative example 2
This comparative example provides a method for producing drug/drug positive hair for scientific research, which replaces the ethanol solution of tris (2-carboxyethyl) phosphine with a mass concentration of 0.5% used in S1 with an ethanol solution of tris (2-carboxyethyl) phosphine with a mass concentration of 2% on the basis of example 2, and the rest of the process is the same as in example 2.
Through the test, the structure of the hair protein is seriously damaged in S1, and finally, the positive hair meeting the requirement cannot be obtained.
Comparative example 3
The comparative example provides a preparation method of medicine/drug positive hair for scientific research, the preparation method changes the concentration of the glacial toxic aqueous solution used in S2 to 10ppm on the basis of the example 2, the soaking time is 4 hours, other process steps are consistent with the example 2, and the content of the glacial toxic in the positive hair prepared by the comparative example is 0.418ng/mg through inspection.
Comparative example 4
The comparative example provides a preparation method of medicine/drug positive hair for scientific research, the preparation method changes the concentration of the glacial toxic aqueous solution used in S2 to 0.5ppm on the basis of the example 2, the soaking time is changed to 24 hours, the other process steps are consistent with the example 2, and the content of the glacial toxic in the positive hair prepared by the comparative example is 0.406ng/mg through inspection.
Example of detection
Taking the positive hair samples prepared in example 2, comparative example 1, comparative example 3 and comparative example 4, respectively marking as sample 1, sample 2, sample 3 and sample 4, and respectively detecting the stability of 4 hairs by adopting a hair sample treatment method in 'liquid chromatography-tandem mass spectrometry detection method of 15 drugs and metabolites thereof in SFZJD0107025-2018 hairs', wherein the specific scheme is as follows:
(1) washing a hair sample with water, washing with sodium dodecyl sulfate, washing with water, washing with acetone, drying in the air, cutting into small sections, and crushing in a freezing grinder to obtain hair powder;
(2) adding hair powder into internal standard methoxamine standard working solution, performing ultrasonic centrifugation in ice bath, removing supernatant, drying under 60 deg.C water bath air flow, re-dissolving residue with methanol, and analyzing concentration with instrument.
Each hair sample is provided with 5 groups of parallel experiments, except that the time for washing and oscillating the hair sample by sodium dodecyl sulfate and the time for washing and oscillating the hair sample by acetone in the step (1) are different, namely 2h +2h, 4h +4h, 6h +6h, 8h +8h and 10h +10h, other processes are the same, the content of the ice toxicity in the hair sample before and after comparison treatment is carried out by combining the analysis result of an instrument, and the result is shown in a table 1:
TABLE 1
As can be seen from the data in Table 1, the positive hair of the medicines/drugs for scientific research prepared by the method of the present invention is more closely combined with hair, can resist the action of organic solvents and surfactants, and has excellent stability in use compared with the positive hair provided in comparative examples 1, 3 and 4.
The above description is only exemplary of the present invention and should not be taken as limiting, and any modifications, equivalents, improvements, etc. that are made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (8)
1. A preparation method of medicine/drug positive hair for scientific research is characterized by comprising the following steps:
s1: washing a blank hair sample in deionized water, drying, putting the blank hair sample into a disulfide bond reduction solution for reaction, taking out the blank hair sample until the breaking elongation of the blank hair sample is more than or equal to 20%, washing the blank hair sample with the deionized water, and drying to obtain a pretreated hair sample;
s2: soaking the pretreated hair sample in a solution of a target immersed in a medicine/drug, and drying to obtain a soaked hair sample;
s3: and putting the soaked hair sample into a disulfide bond oxidation solution for reaction until the elongation at break of the hair sample is less than or equal to 5%, taking out the hair sample, washing the hair sample with deionized water, and drying to obtain the medicine/drug positive hair for scientific research.
2. The method of claim 1, wherein the disulfide bond reducing solution in S1 is an aqueous or organic solution of at least one of thiol, dithiothreitol, cysteine, thioglycolic acid, tris (2-carboxyethyl) phosphine, dimercaptoethanol, and sodium sulfite, and has a concentration of 0.1 wt% to 1 wt%.
3. The method of claim 1, wherein the solution of the target immersed in the drug/drug is an aqueous solution or an organic solution of the target immersed in the drug/drug at a concentration of 1ppm to 10ppm in S2.
4. The method of claim 3, wherein the pre-treated hair sample is immersed in the drug/drug solution at step S2 for a period of time ranging from 6 hours to 12 hours.
5. The method for preparing the drug/drug positive hair for scientific research according to claim 4, wherein when the target immersion concentration of the drug/drug in the positive hair is 0.2ng/mg, 1-3ppm of the target is immersed in the drug/drug solution for 6-8 hours; when the target immersion concentration of the drugs/drugs in the positive hair is 0.4ng/mg, 3-6ppm of the target is selected to be immersed in the drug/drug solution for 8-10 h; when the target immersion concentration of the drugs/drugs in the positive hair is 0.6ng/mg, the drug/drug solution is immersed in the target immersion concentration of 6-10ppm for 10-12 h.
6. The method for preparing medicine/drug positive hair for scientific research according to claim 1, wherein the disulfide bond oxidation solution in S3 is an aqueous solution of one of hydrogen peroxide, peracetic acid or sodium percarbonate with a concentration of 1 wt% to 5 wt%.
7. The method of claim 1, wherein the drying temperature of the hair in S1 is 10 ℃ to 40 ℃, and the drying temperature of the hair in S2 and S3 is 40 ℃ to 70 ℃.
8. A scientific drug/drug positive hair, which is prepared by the preparation method of the scientific drug/drug positive hair according to any one of claims 1 to 7.
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| CN114689757A (en) * | 2022-04-07 | 2022-07-01 | 中国计量科学研究院 | Preparation method and application of amphetamine-type drug hair standard substance |
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