CN112345658A - Quality control product for analyzing organic toxicants in human hair and preparation method thereof - Google Patents
Quality control product for analyzing organic toxicants in human hair and preparation method thereof Download PDFInfo
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- CN112345658A CN112345658A CN202010790606.5A CN202010790606A CN112345658A CN 112345658 A CN112345658 A CN 112345658A CN 202010790606 A CN202010790606 A CN 202010790606A CN 112345658 A CN112345658 A CN 112345658A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses an analysis and quality control product for organic toxicants in human hair and a preparation method thereof. According to the quality control product for analyzing the organic toxicants in the human hair, the raw materials are selected from fresh and real human hair which is collected in a controlled manner and subjected to strict quality control, and after the raw materials are added externally or mixed internally, the quality control product containing the trace organic toxicants is formed through quality control evaluation, so that the accurate assignment, high traceability and low uncertainty of the quality control product can be effectively guaranteed, and the regulation requirements of comparison among laboratories are met; solves the technical and legal problems that a plurality of inspection data lack comparability and traceability due to the lack of corresponding matrix quality control products. The invention is applied to the practice of forensic poison analysis, so that each laboratory has human hair quality control products for organic poison analysis with unified traceability, unified quantity value and unified specification, thereby the analysis results can obtain the measurement traceability, the reliability of the results can be improved by orders of magnitude, and the fairness and the authority of the inspection results can be ensured.
Description
Technical Field
The invention relates to the field of court science, in particular to an analysis quality control product for organic toxicants in human hair and a preparation method thereof.
Background
Quality control products are always valued by countries and industries in the world due to their measurement traceability, reliability and comparability. Among them, chemical quality control materials are widely used in laboratory approval, capability verification, and laboratory quality control activities. The analysis and detection material in the court science field has the characteristics of complex matrix, multiple interference factors and the like, so the requirement on the matrix quality control product is particularly strong. At present, corresponding matrix-containing quality control products are lacked in the field of forensic science internationally, and especially, the matrix quality control products using solid matrixes such as hair are lacked, so that a lot of test data lack comparability and traceability, the reliability of analysis results meets serious challenges, the identification conclusions often contradict each other, the repeated identification phenomenon is common, and the proving efficacy of the identification conclusions is seriously threatened.
The matrix quality control product can avoid the difference of matrix effect on the analysis of the substance components in the analysis test; for the analysis of organic toxicants in hair, quality control of the substrate is of outmost importance. In judicial practice, analysis of organic toxicants, especially drug components, in hair is important or even the only scientific evidence for determining whether a party is taking poison. In the "hair sample detection standard for drug-related personnel" (official drug ban [ 2018 ] 938), the ministry of public security has strict regulations on the detection result limit (namely threshold): the threshold value of the detection content of O6-monoacetylmorphine, morphine, methamphetamine, amphetamine, 3, 4-Methylenedioxyamphetamine (MDA), 3, 4-methylenedioxymethamphetamine, ketamine, norketamine and methcathinone in the hair sample is 0.2 ng/mg; the threshold value of the detection content of cocaine is 0.5 ng/mg; the threshold value of the content of the benzoyl ikangning and the tetrahydrocannabinol is 0.05 ng/mg. And (4) determining that the detection result is positive if the actual detection content value is above the threshold value. However, in the current domestic judicial investigation laboratory, due to the lack of related quality control products, the threshold value is difficult to control in the analysis process, so that the detection results in different laboratories are possibly inconsistent.
Disclosure of Invention
The invention aims to solve the problems of the prior art and provides an analysis and quality control product for organic toxicants in human hair and a preparation method thereof.
In order to achieve the purpose, the invention adopts the technical scheme that:
the invention provides a preparation method of an organic poison analysis quality control product in human hair, which comprises the following steps:
step one, human hair preparation
Taking each volunteer individual as a human hair source, respectively carrying out LC-MS/MS analysis on the human hair provided by the volunteer individual, and respectively carrying out quantitative analysis according to the industry standard SF/T0063-; the human hair source with negative analysis result is mixed and cut into 5-8cm sections for standby;
step two, preparing a soaking solution
Accurately weighing 5-40mg of each reference substance of the target organic poison, sequentially adding 250-300mL of deionized water, 250-300mL of dimethyl sulfoxide reagent and 425-1000 mu L of concentrated hydrochloric acid, and uniformly mixing for later use;
step three, sample preparation
Adding 30-40g of negative human hair in the first step into the soaking solution prepared in the second step, uniformly stirring, sealing and storing by using a sealing film, and fully soaking for at least two weeks at the temperature of 1-7 ℃ and under the humidity of less than 80%; taking out the hair soaked sample after soaking, washing the hair soaked sample with methanol, drying the hair soaked sample in the air, cutting the hair soaked sample into pieces of 0.2-0.5mm to obtain a hair soaked sample, and performing quantitative analysis according to an industry standard SF/T0063-;
setting the content of the target organic poison in the batch of products as a planned fixed value; adding different positive source hair samples into the hair soaking samples to obtain mixed samples, carrying out quantitative analysis on the mixed samples according to the industry standard SF/T0063-2020, and obtaining the organic poison analysis quality control product in human hair after packaging and detection are qualified if the content of the target organic poison in the mixed samples is measured to be equal to a set value.
Further, the set-up value is from 0.01 to 4.0 ng/mg.
Further, LC-MS/MS analysis of human hair is specifically: according to technical specification SF/Z JD0107025-2018, washing human hair to be detected by sequentially oscillating with water and acetone, cutting into 1-1.5mm sections after drying, crushing in a cold grinding instrument, weighing 20mg of hair powder, adding 1.0mL of 1ng/mL internal standard methoxamine working solution, carrying out ultrasonic treatment in an ice bath for 30min, centrifuging, removing supernatant, drying by blowing under 60 ℃ water bath air flow, redissolving the residue with 100uL of methanol, and carrying out sample injection analysis by LC-MS/MS.
Further, the second step is specifically as follows: accurately weighing 5-40mg of each reference substance of the target organic poison, adding 250mL of deionized water, stirring and dissolving uniformly, placing in an ice bath environment, sequentially adding 250mL of dimethyl sulfoxide reagent and 425 mu L of concentrated hydrochloric acid, and mixing uniformly for later use.
Further preferably, the concentration of the concentrated hydrochloric acid is 36%.
Further, the target organic poison is one or more of the appendix of the State food and drug administration, the Ministry of public Security, Notification of catalogs on publishing narcotic drugs and psychiatric drugs by the State Ministry of health and Ministry of health (food and drug administration and chemical administration (2013) 230) and the supplement catalogs on non-medicinal narcotic drugs and psychiatric drug control varieties published by the departments all the year round.
Further, the package was nitrogen blanketed and controlled to ambient humidity < 30%.
The second aspect of the invention is to provide a quality control product for analyzing organic toxicants in human hair prepared by the preparation method.
By adopting the technical scheme, compared with the prior art, the invention has the following technical advantages:
according to the quality control product for analyzing the organic poison in the human hair, the raw material is selected from fresh and real human hair which is collected in a controlled manner and subjected to strict quality control, and the trace organic poison which is evaluated by the quality control is added, so that the high traceability and low uncertainty of the quality control product can be effectively guaranteed, and the regulation requirement of comparison between laboratories is realized; solves the technical and legal problems that a plurality of inspection data lack comparability and traceability due to the lack of corresponding matrix quality control products.
The invention fills the blank of human hair quality control products in China court science and related fields; by applying the method to forensic toxicant analysis practice, each laboratory has human hair quality control products for organic toxicant analysis with unified traceability, unified quantity value and unified specification, so that the analysis results can obtain measurement traceability, the reliability of the results can be improved by a magnitude level, and the fairness and the authority of the inspection results can be ensured. Meanwhile, the technology progress and the specification of the judicial identification industry can be promoted, the identification quality and the proof effectiveness of the identification conclusion can be guaranteed, and the justice of the judicial can be maintained.
Detailed Description
The invention provides a preparation method of an analysis and quality control product for organic toxicants in human hair, which comprises the following steps:
step one, human hair preparation
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/Z JD0107025-2018, sequentially oscillating and washing the human hair to be detected with water and acetone, cutting the human hair into 1-1.5mm sections after drying in the air, crushing in a freeze grinding instrument, weighing 20mg of hair powder, adding 1.0mL of 1ng/mL internal standard methoxamine standard working solution, performing ice bath ultrasound for 30min, centrifuging, removing supernatant, drying under 60 ℃ water bath air flow, re-dissolving the residue with 100uL of methanol, and performing LC-MS/MS sampling analysis); analyzing the human hair source with a positive analysis result, respectively carrying out quantitative analysis by referring to the industry standard SF/T0063-2020 to obtain the content value of the target organic poison, respectively recording, and respectively cutting to about 0.2-0.5mm to obtain a positive hair source sample for later use; the human hair source with negative analysis result is mixed and cut into 5-8cm sections for standby;
step two, preparing a soaking solution
Accurately weighing 5-40mg of each reference substance of the target organic poison, sequentially adding 250-300mL of deionized water, 250-300mL of dimethyl sulfoxide reagent and 425-1000 mu L of concentrated hydrochloric acid, and uniformly mixing for later use;
step three, sample preparation
Adding 30-40g of negative human hair in the first step into the soaking solution prepared in the second step, uniformly stirring, sealing and storing by using a sealing film, and fully soaking for at least two weeks at the temperature of 1-7 ℃ and under the humidity of less than 80%; taking out after soaking, washing with methanol, drying in the air, cutting into pieces of 0.2-0.5mm to obtain a hair soaking sample, and performing quantitative analysis according to industry standard SF/T0063-;
setting the content of the target organic poison in the batch of products as a planned value; adding different positive hair samples into the hair soaking sample to obtain a mixed sample, carrying out quantitative analysis on the mixed sample according to the industry standard SF/T0063-2020, and obtaining an organic poison analysis quality control product in human hair after packaging and qualified detection if the content of the target organic poison in the mixed sample is detected to be equal to a set value.
In another preferred embodiment, the set value is from 0.05 to 0.8 ng/mg.
In another preferred example, the second step is specifically: accurately weighing 5-40mg of each reference substance of the target organic poison, adding 250mL of deionized water, stirring and dissolving uniformly, placing in an ice bath environment, sequentially adding 250mL of dimethyl sulfoxide reagent and 425 mu L of concentrated hydrochloric acid, and mixing uniformly for later use; the concentration of concentrated hydrochloric acid was 36%.
In another preferred example, the target organic poison is one or more of the postscript of the national food and drug administration, the ministry of public security, the national institutes of health, the "Notification about the List of publishing narcotics and psychiatric products" (Institution of food and drug administration [ 2013 ] 230) and the "supplementary List of non-medicinal narcotics and psychiatric products regulated varieties" published by the above departments over the years.
In another preferred embodiment, the package is filled with nitrogen for protection, and the ambient humidity is controlled to be less than 30%.
The invention also provides a freeze-dried quality control product for analyzing organic toxicants in human blood, which is prepared by the preparation method.
The present invention will be described in detail and specifically with reference to the following examples to facilitate better understanding of the present invention, but the following examples do not limit the scope of the present invention.
(1) Environmental and appliance requirements:
the environmental temperature is 5-18 ℃, and the relative humidity is less than 80%;
the environment is required to be clean, dustless, smokeless, non-corrosive gas, non-volatile organic solvent and non-volatile substances which may interfere with the preparation;
the operation room is preferably provided with an outward unidirectional air flow;
the countertop and the appliances should be cleaned and sterilized prior to preparation.
(2) Preparation of raw materials human hair and donor requirements:
human hair should be taken from healthy volunteers with informed consent or from drug-addicted (drug-use) populations and kept at ambient temperature 4 ℃ and ambient humidity < 65% before use in the preparation.
The provided hair should be used for hairdressing operation without hair waving and dyeing in the first half year.
Example 1
The embodiment provides a preparation method of an organic poison analysis quality control product in human hair, which comprises the following steps:
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/Z JD0107025-2018, sequentially oscillating and washing the human hair to be detected with water and acetone, cutting the human hair into 1-1.5mm sections after drying in the air, crushing in a freeze grinding instrument, weighing 20mg of hair powder, adding 1.0mL of 1ng/mL internal standard methoxamine standard working solution, performing ice bath ultrasound for 30min, centrifuging, removing supernatant, drying under 60 ℃ water bath air flow, re-dissolving the residue with 100uL of methanol, and performing LC-MS/MS sampling analysis); analyzing the human hair source with a positive analysis result, respectively carrying out quantitative analysis by referring to the industry standard SF/T0063-2020 to obtain the content value of the target organic poison, 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.
Preparing a soaking solution:
group A: adding 12mg morphine reference substance and 30mg monoacetylmorphine reference substance into one beaker with the volume of 1L, adding 250mL deionized water, stirring and dissolving uniformly, placing in an ice bath environment, and sequentially adding 250mL dimethyl sulfoxide reagent and 425 μ L concentrated hydrochloric acid.
Group B: taking one beaker with the capacity of 1L, adding 15mg of methamphetamine reference substance and 40mg of 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 425 μ L of concentrated hydrochloric acid.
Group C: taking one beaker with the capacity of 1L, adding 10mg of cocaine reference substance and 5mg of benzoyl ekanin 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 425 μ L of concentrated hydrochloric acid.
Respectively adding 30g of negative human hair into the soaking solutions of the group A, the group B and the group C, stirring uniformly, sealing with a sealing film, and fully soaking for two weeks at the temperature of 1-7 ℃ and the humidity of less than 80%; taking out after soaking, washing with methanol, drying in the air, cutting into pieces of 0.2-0.5mm to obtain a group A hair soaking sample, a group B hair soaking sample and a group C hair soaking sample, and respectively carrying out quantitative analysis according to industry standard SF/T0063-.
Setting a predetermined value A to 0.4 ng/mg, a predetermined value B to 0.2 ng/mg, and a predetermined value C to 0.05 ng/mg;
adding different positive source hair samples into the group A hair soaking samples to obtain mixed samples, carrying out quantitative analysis on the mixed samples according to an industry standard SF/T0063-; if the content of morphine and the content of monoacetylmorphine in the mixed sample are both 0.2 ng/mg, obtaining a product batch B and subpackaging; and if the content of the morphine and the monoacetylmorphine in the mixed sample is measured to be 0.05 ng/mg, obtaining a product batch C and subpackaging. And (4) obtaining an organic poison analysis quality control product in human hair after the batch of products are detected to be qualified.
Adding different positive source hair samples into the group B hair soaking samples to obtain mixed samples, carrying out quantitative analysis on the mixed samples according to an industry standard SF/T0063-2020, and obtaining and subpackaging a product batch A if the contents of methamphetamine and ketamine in the mixed samples are both 0.4 ng/mg; if the contents of methamphetamine and ketamine in the mixed sample are both 0.2 ng/mg, obtaining a product batch B and subpackaging; and if the contents of the methamphetamine and the ketamine in the mixed sample are both 0.05 ng/mg, obtaining a product batch C and subpackaging. And (4) obtaining an organic poison analysis quality control product in human hair after the batch of products are detected to be qualified.
Adding different positive source hair samples into the group C hair soaking samples to obtain mixed samples, carrying out quantitative analysis on the mixed samples according to an industry standard SF/T0063-; if the content of cocaine and benzoyl ekanin in the mixed sample is measured to be 0.2 ng/mg, obtaining a product batch B and subpackaging; and if the content of cocaine and benzoyl ikangning in the mixed sample is measured to be 0.05 ng/mg, obtaining a product batch C and subpackaging. And (4) obtaining an organic poison analysis quality control product in human hair after the batch of products are detected to be qualified.
In this embodiment, the package is filled with nitrogen for protection, the ambient humidity is controlled to be less than 30%, and each 200mg of the package is packaged into a 4mL plastic light-proof small tube and stored at room temperature; each group and each batch are respectively packaged into 350 tubes, and the numbers of the tubes are 1 to 350 in sequence.
In this embodiment, the inspection process after packaging includes visual inspection and quality evaluation.
And (3) appearance inspection: the packaged human hair matrix quality control product is subjected to appearance inspection by referring to recommended analysis guidelines of the international hair analysis association, with emphasis on defects in sealability, filling amount and dimensions.
Taking a total of 9 transparent small tubes reserved in three stages of before the subpackaging stage (the first 2% of the total subpackaging amount, in the embodiment, 350 bottles are subpackaged in total, namely the 7 th, 8 th and 9 th bottles in the subpackaging process), in (the 50 th% of the total subpackaging amount, in the embodiment, the 175 th, 176 th and 177 th bottles in the subpackaging process) and after (the last 98% of the total subpackaging amount, in the embodiment, the 343 th, 344 th and 345 th bottles in the subpackaging process), and opening bottles to check quality control products, wherein the finished products are uniform in size and constant in volume.
And (3) quality evaluation:
(1) homogeneity test
The uniformity test was conducted by taking 10% (e.g., less than 20 bottles, based on 20 bottles) of the total amount of the preparation at equal intervals.
Sampling the quality control product according to the specified amount of the analysis method. The quantification is carried out by reference to a related method, and the RSD value of the quantification result should not exceed 3%.
(2) Short term test
20% (e.g., less than 120 bottles, based on 120 bottles) of the total preparation was randomly taken for short-term testing.
The sample is divided into 3 parts, and stored in three conditions of-20 deg.C + -3 deg.C, humidity < 65%, temperature 4 deg.C + -3 deg.C, humidity < 80%, 20 deg.C + -3 deg.C, and humidity < 85%.
On days 0, 1, 5, 10, and 15 of the storage period; 1/5 of each condition preservation amount is taken out and quantified by referring to the correlation method, the RSD value of the quantitative result should not exceed 3%, and the change of the quantitative value compared with the last time should not exceed 2%.
(3) Long term test
20% of the total preparation (e.g., less than 192 vials, based on 192 vials) was randomly taken for long-term testing.
The sample is divided into 3 parts, and stored in three conditions of-20 deg.C + -3 deg.C, humidity < 65%, temperature 4 deg.C + -3 deg.C, humidity < 80%, 20 deg.C + -3 deg.C, and humidity < 85%.
On days 0, 15, 30, 60, 90, 120, 150, 180 of the shelf life; 1/8 of each condition preservation amount is taken out and quantified by referring to the correlation method, the RSD value of the quantitative result should not exceed 3%, and the change of the quantitative value compared with the last time should not exceed 2%.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (8)
1. A preparation method of a quality control product for analyzing organic toxicants in human hair is characterized by comprising the following steps:
step one, human hair preparation
Taking each volunteer individual as a human hair source, respectively carrying out LC-MS/MS analysis on the human hair provided by the volunteer individual, and respectively carrying out quantitative analysis according to the industry standard SF/T0063-; the human hair source with negative analysis result is mixed and cut into 5-8cm sections for standby;
step two, preparing a soaking solution
Accurately weighing 5-40mg of each reference substance of the target organic poison, sequentially adding 250-300mL of deionized water, 250-300mL of dimethyl sulfoxide reagent and 425-1000 mu L of concentrated hydrochloric acid, and uniformly mixing for later use;
step three, sample preparation
Adding 30-40g of negative human hair in the first step into the soaking solution prepared in the second step, uniformly stirring, sealing and storing by using a sealing film, and fully soaking for at least two weeks at the temperature of 1-7 ℃ and under the humidity of less than 80%; taking out after soaking, washing with methanol, drying in the air, cutting into pieces of 0.2-0.5mm to obtain a hair soaking sample, and performing quantitative analysis according to industry standard SF/T0063-2020 to obtain the content of the target organic toxicant in the hair soaking sample;
setting the content of the target organic poison in the batch of products as a planned fixed value; adding different positive source hair samples into the hair soaking samples to obtain mixed samples, carrying out quantitative analysis on the mixed samples according to the industry standard SF/T0063-2020, and obtaining the organic poison analysis quality control product in human hair after packaging and detection are qualified if the content of the target organic poison in the mixed samples is measured to be equal to a set value.
2. The method of claim 1, wherein the predetermined value is in the range of 0.01-4.0 ng/mg.
3. The method of claim 1, wherein the LC-MS/MS analysis of human hair is specifically: according to technical specification SF/Z JD0107025-2018, washing human hair to be detected by sequentially oscillating with water and acetone, cutting into 1-1.5mm sections after drying, crushing in a cold grinding instrument, weighing 20mg of hair powder, adding 1.0mL of 1ng/mL internal standard methoxamine working solution, carrying out ultrasonic treatment in an ice bath for 30min, centrifuging, removing supernatant, drying by blowing under 60 ℃ water bath air flow, redissolving the residue with 100uL of methanol, and carrying out sample injection analysis by LC-MS/MS.
4. The preparation method according to claim 1, wherein the second step is specifically: accurately weighing 5-40mg of each reference substance of the target organic poison, adding 250mL of deionized water, stirring and dissolving uniformly, placing in an ice bath environment, sequentially adding 250mL of dimethyl sulfoxide reagent and 425 mu L of concentrated hydrochloric acid, and mixing uniformly for later use.
5. The method according to claim 4, wherein the concentration of the concentrated hydrochloric acid is 36%.
6. The method according to claim 1, wherein the target organic toxicant is one or more of the national food and drug administration, the ministry of public Security, the appendix of Notification of catalogs of narcotics and psychiatric drugs by the national institutes of health (Institution for food and drug administration [ 2013 ] 230) and the supplementary catalogs of non-medicinal narcotics and psychiatric drug regulated varieties published by the departments over the years.
7. The method of claim 1, wherein the package is nitrogen blanketed and controlled to have an ambient humidity of < 30%.
8. A quality control product for analyzing organic toxicants in human hair prepared by the preparation method according to any one of claims 1 to 7.
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| CN113607521A (en) * | 2021-08-02 | 2021-11-05 | 中国刑事警察学院 | Preparation method of medicine/drug positive hair for scientific research |
| 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 |
| CN114705780A (en) * | 2022-04-07 | 2022-07-05 | 中国计量科学研究院 | Preparation method and application of fentanyl drug hair standard substance |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113311084A (en) * | 2021-05-24 | 2021-08-27 | 公安部物证鉴定中心 | Hair drug quality control product for criminal investigation field and rapid preparation method thereof |
| CN113607521A (en) * | 2021-08-02 | 2021-11-05 | 中国刑事警察学院 | Preparation method of medicine/drug positive hair for scientific research |
| CN113607521B (en) * | 2021-08-02 | 2024-02-09 | 中国刑事警察学院 | Preparation method of scientific research medicine/drug positive hair |
| 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 |
| CN114705780A (en) * | 2022-04-07 | 2022-07-05 | 中国计量科学研究院 | Preparation method and application of fentanyl drug hair standard substance |
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| CN112345658B (en) | 2022-06-24 |
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