Kit for detecting psychotropic drugs in hair and application thereof

Abstract

The invention provides a kit for detecting psychotropic drugs in hair and application of the kit, the kit comprises a hair cleaning agent and a hair digesting agent, the hair cleaning agent comprises Tween-20 and TXR-1, and the hair digesting agent comprises dithiothreitol, tris (2-carboxyethyl) phosphine hydrochloride and CHAPS. When the kit provided by the invention is used for pre-treating a hair sample, pollutants attached to the hair sample can be effectively removed, the operation steps are simplified, the accuracy and the stability of a detection result are improved, and a sample extracting solution can still be accurately detected after being stored for a certain time. Moreover, the mental drugs in the sample extracting solution are detected in combination with quantum dot immunofluorescence chromatography, various novel mental drugs can be fully extracted and accurately detected, the sensitivity and specificity of the detection method are greatly improved, and on-site rapid screening is facilitated.

Description

Technical Field

[0001] The present invention belongs to the field of identification technology, and particularly relates to a kit for detecting psychotropic drugs in hair and its use. Background Art

[0002] The abuse and taking of psychotropic drugs seriously endanger social security and human life and health. After a human body takes such drugs, the drugs and their metabolites mainly remain in blood, urine, hair, sweat, feces, nails, etc. Detecting the presence of psychotropic drugs in the test sample can reflect whether the taker has the behavior of taking psychotropic drugs. At present, the test samples for drug abuse are mainly urine, saliva, blood, and hair. The psychotropic drugs remaining on the hair sample can exist for at least more than 3 months, which can reflect whether the psychotropic drug taker has taken psychotropic drugs in the past 3 months or even half a year. Moreover, the hair sample is easy to collect and preserve, and is a commonly used sample for current detection.

[0003] Hair is rich in keratin, and the ingested drugs are embedded in the keratin. However, since hair is in contact with the external environment, it is easily contaminated by the external environment. In addition, the tested person often perm and dye their hair, and the hair dye will also affect the test results. The existing detection technologies are prone to false negatives or false positives. Patent document CN112525641B discloses a method for preparing a test solution of psychotropic drugs in hair, including multiple steps such as hair washing, softening, and lysis. The formulations of the softening solution and the lysis solution involve various substances, and the lysis process requires grinding to assist lysis, with complex steps; at the same time, the detected drugs are traditional psychotropic drugs, and the detection effect for new psychotropic drugs is unknown. Patent document CN108844922B discloses a rapid detection method for psychotropic drugs in hair, which uses an enzymatic method to extract psychotropic drugs in hair, and then combines a colloidal gold method and SPR to detect the target substance. The enzymatic method has limited digestion effect on hair, resulting in low release efficiency of the psychotropic drug substances to be detected in hair.

[0004] Therefore, there is an urgent need to develop a simpler, faster, and more effective reagent and method for extracting and detecting psychotropic drugs in hair, especially for the detection of some new psychotropic drugs, which can be more accurate and sensitive, while ensuring the stability and repeatability of the detection and extending the storage time of the test samples. Summary of the Invention

[0005] Aiming at the problems existing in the prior art, the present invention provides a kit for detecting psychotropic drugs in hair and its use, improves the cleaning and digestion reagents for hair, fully extracts psychotropic drugs in hair in two steps, and then combines immunochromatography to detect the sample to be tested. The reagents provided by the present invention have the same accuracy and stability in the detection of traditional and new psychotropic drugs, effectively eliminate the interference of various factors such as the oil, hair dye, and external pollution of the hair itself, simplify the operation steps, and are conducive to actual operation and application.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] On the one hand, the present invention provides a kit for extracting psychotropic drugs in hair, the kit includes a cleaning agent and a digestion agent, the cleaning agent includes Tween-20 and TXR-1, and the digestion agent includes dithiothreitol (DTT), tris(2-carboxyethyl)phosphine hydrochloride (TCEP) and CHAPS.

[0008] Currently, various detection methods for psychotropic drugs in hair generally involve multiple reagents, steps and treatments. On the one hand, the operation is cumbersome and complex. On the other hand, the processing of hair samples and the accuracy and sensitivity of the extraction and detection of new psychotropic drugs in hair are limited, and the false positive and false negative rates are high. Therefore, for the extraction and detection of new psychotropic drugs in hair, the present invention makes a creative improvement to the hair cleaning and digestion reagents, uses the composition of Tween-20 and TXR-1 to prepare a hair cleaning agent, effectively removes pollutants such as oil and hair dye on the hair surface, and can, to a certain extent, destroy the keratin structure of the hair, promoting the digestion and extraction of the subsequent digestion agent. Using the composition of DTT, TCEP and CHAPS to prepare a hair digestion agent can effectively improve the accuracy of the detection result, reduce the false negative rate, improve the sensitivity and stability of the detection method, and can accurately detect hair samples containing stubborn pollutants such as hair dye and bleach.

[0009] As a mild surfactant, Tween-20 mainly removes hydrophilic pollutants (such as sweat, saliva, environmental dust) and does not damage the hair keratin structure; while TXR-1, as a new generation of sustainable detergent, has excellent biocompatibility, can effectively remove the oil, sebaceous gland secretions and bound pollutants in the hair, and can also penetrate the hair medulla and release the embedded fat-soluble psychotropic drugs, such as synthetic cannabinoids and fentanyl analogs. When the two are combined to clean the hair sample, on the one hand, Tween-20 is used to remove hydrophilic pollutants, and on the other hand, TXR-1 is used to dissolve deep lipids, balancing the cleaning power and compatibility, which is suitable for the detection of most psychotropic drugs.

[0010] In some embodiments, the present invention verifies the necessity of preparing a hair cleaning agent by combining Tween-20 and TXR-1. The results show that when Tween-20 or TXR-1 is used in combination with other surfactants such as SDS or CHAPS, the false negative rate of the detection result is high, the method sensitivity is low, the stability is poor, and the cleaning and detection sensitivity for hair dyeing samples containing toluene-2,5-diamine and resorcinol are poor. The reason may be that only the combined cleaning agent of Tween-20 and TXR-1 can effectively remove the pollutants attached to the hair samples, reduce the interference of other factors, significantly promote the subsequent digestion and extraction efficiency, and improve the detection accuracy.

[0011] Further, the concentration of Tween-20 is 0.005% - 0.01% (v / v), and the concentration of TXR-1 is 0.01% - 0.05% (v / v).

[0012] Further, the concentration of dithiothreitol is 0.015 g / 100 mL - 2.0 g / 100 mL, the concentration of tris(2-carboxyethyl)phosphine hydrochloride is 0.025 g / 100 mL - 3.0 g / 100 mL, and the concentration of CHAPS is 0.03 g / 100 mL - 3.0 g / 100 mL.

[0013] During the process of digesting hair samples, dithiothreitol (DTT) and tris(2-carboxyethyl)phosphine hydrochloride (TCEP) act as reducing agents to break the disulfide bonds in hair keratin, open the tight structure of hair fibers, and release the embedded drug molecules. As an amphoteric ion surfactant, CHAPS acts simultaneously with its hydrophilic end (sulfonic acid group) and hydrophobic end (cholic acid ring) to dissolve the hair lipid layer and membrane proteins, destroy the hydrophobic barrier on the hair surface, and enhance the penetration of the reducing agents (DTT, TCEP) into the internal structure. In some embodiments, the present invention verifies the necessity of combining DTT, TCEP, and CHAPS to digest hair samples. The results show that compared with using a single reducing agent or CHAPS, the combination of DTT and CHAPS or TCEP and CHAPS can increase the release amount of psychotropic drugs in hair samples, but the effect is limited compared with the combination of the three. The combination of two reducing agents, DTT and TCEP, and CHAPS can synergistically increase the drug release amount, effectively remove interfering substances, improve the detection accuracy and stability, and have the same detection sensitivity for hair samples using toluene-2,5-diamine and resorcinol hair dyes.

[0014] On the other hand, the present invention provides a kit for detecting psychotropic drugs in hair, the kit comprising a cleaning agent, a digestive agent and a test strip, the cleaning agent comprising Tween-20 and TXR-1, and the digestive agent comprising dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride and CHAPS.

[0015] Further, the concentration of the Tween-20 is 0.005% - 0.01% (v / v), and the concentration of the TXR-1 is 0.01% - 0.05% (v / v).

[0016] Further, the concentration of the dithiothreitol is 0.015 g / 100 mL to 2.0 g / 100 mL, the concentration of the tris(2-carboxyethyl)phosphine hydrochloride is 0.025 g / 100 mL to 3.0 g / 100 mL, and the concentration of the CHAPS is 0.03 g / 100 mL to 3.0 g / 100 mL.

[0017] On yet another aspect, the present invention provides a method for extracting psychotropic drugs in hair, using the kit as described in any one of the above to extract psychotropic drugs in hair.

[0018] Further, the method comprises the following steps:

[0019] (1) Hair cleaning: After cutting the hair into pieces, use the cleaning agent to clean it;

[0020] (2) Hair digestion and extraction: Use the digestive agent to digest and extract the cleaned hair for 5 - 15 min to obtain a sample extract.

[0021] Using the hair cleaning agent and digestive reagent pretreatment method provided by the present invention, there is no need for physical means such as heating and ultrasonic treatment, which greatly simplifies the operation steps, and effectively removes pollutants such as grease and hair dye in different hair samples, promotes the release of psychotropic drugs, and improves the accuracy of the detection result and the stability of the method.

[0022] On yet another aspect, a method for determining psychotropic drugs in hair comprises the following steps:

[0023] (1) Extract psychotropic drugs in hair using the kit as described above;

[0024] (2) Use immunochromatography for detection.

[0025] In some embodiments, in step (2), the immunochromatography method specifically uses a quantum dot immunofluorescence chromatography method based on the competition principle. The quantum dot immunofluorescence chromatography method is a rapid detection method that combines the high fluorescence characteristics of quantum dots (QDs) with immunochromatography technology. Quantum dots are used to label antibodies or antigens, and through chromatography, a test line (T line) and a quality control line (C line) are formed on the test strip, and the target psychotropic drug molecules are quantitatively or qualitatively analyzed through fluorescence signals. Compared with ordinary fluorescent dyes, quantum dots have a wide excitation spectrum range, a relatively narrow and symmetrically distributed emission spectrum range. At the same time, quantum dots have high fluorescence intensity, good stability, and better biocompatibility.

[0026] In some embodiments, the principle of the immunochromatography method based on the competition method is to label a certain amount of target drug with quantum dots and fix it on the conjugate pad. The T line and the C line are respectively sprayed with an antibody (primary antibody) that specifically binds to the target drug and an "anti-antibody" against species-specific IgG: for a negative sample, the quantum dot-labeled antigen flows laterally with the sample, binds to the primary antibody at the T line, and binds to the secondary antibody at the C line, resulting in fluorescence signals on both the T line and the C line; for a positive sample, the unlabeled target antigen in the sample and the quantum dot-labeled target antigen on the conjugate pad compete to bind to the primary antibody at the T line. The more target antigen in the sample, the less quantum dots bind to the T line, and the weaker the fluorescence signal. Excess quantum dots bind to the secondary antibody at the C line, so only the C line shows a fluorescence value. The test samples are respectively detected and analyzed according to the above detection principle.

[0027] Further, the hair samples include the hair, fine hair, armpit hair, or pubic hair of a person who has taken psychotropic drugs.

[0028] The research of the present invention proves that using the hair cleaning reagent and hair digestion reagent provided by the present invention to process various samples with keratinized structures such as human hair, fine hair, armpit hair, or pubic hair can achieve equally excellent extraction effects; at the same time, for hair samples containing stubborn interfering substances such as bleach and hair dye, the cleaning and digestion reagents provided by the present invention can also effectively remove the interfering substances and achieve excellent extraction and detection effects.

[0029] Further, the psychotropic drugs include any one or more of fentanyl analogs, synthetic cannabinoids, synthetic cathinones, etomidate, and GHB.

[0030] In some embodiments, the research results of the present invention prove that the kit and the extraction and detection methods provided by the present invention have significantly excellent technical effects for the extraction and detection of new psychoactive substances in hair. The new psychoactive substances include, but are not limited to, fentanyl analogs, synthetic cannabinoids, synthetic cathinones, etomidate, and GHB, improving the accuracy and stability of the detection of psychoactive substances, and improving the detection efficiency in practical applications, and being applicable to rapid screening sites such as traffic police drug detection and hospital diagnosis.

[0031] In another aspect, the present invention provides the use of a composition for preparing a reagent for extracting psychoactive substances from hair, and the composition includes dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride, and CHAPS.

[0032] In another aspect, the present invention provides the use of a composition for preparing a reagent for improving the detection sensitivity and stability of psychoactive substances in hair, and the composition includes dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride, and CHAPS.

[0033] In another aspect, the present invention provides the use of a composition for preparing a reagent for improving the detection accuracy of psychoactive substances in dyed hair, and the composition includes dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride, and CHAPS.

[0034] In some embodiments, the present invention proves through experiments that treating hair samples dyed with toluene-2,5-diamine and resorcinol with a digestion reagent prepared from dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride, and CHAPS can significantly improve the accuracy and sensitivity of the detection method.

[0035] The present invention has the following beneficial effects:

[0036] 1. Optimize the formula of the hair cleaning reagent, and use the combination of Tween-20 and TXR-1 to clean the hair samples, effectively removing interfering substances such as oil and hair dye in the hair, and improving the accuracy of the detection results.

[0037] 2. Use a composition of dithiothreitol (DTT), tris(2-carboxyethyl)phosphine hydrochloride (TCEP), and CHAPS as the hair digestion reagent. Through the synergistic effect among the three substances, the hair cutin structure and protein are effectively destroyed, and the psychoactive substance molecules in the hair samples are fully released. Without physical means such as heating and ultrasonic treatment, the operation steps are simplified, and the extraction efficiency and the accuracy of the detection results are improved.

[0038] 3. The hair cleaning agent and digestive provided by the present invention are combined with quantum dot immunofluorescence chromatography to detect psychotropic drugs in the sample extract. Utilizing the advantages of high fluorescence intensity and good stability of quantum dots, the detection efficiency is further improved, which is applicable to the rapid screening sites of traffic police drug detection and emergency diagnosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 It is a schematic structural diagram of a test strip, where serial number 1 represents the sample pad, serial number 2 represents the conjugate pad, serial number 3 represents the test pad, serial number 4 represents the absorbent pad, serial number 5 represents the support bottom plate, serial number 31 represents the test line (T line), serial number 32 represents the control line (C line), and the black dots represent the labeling material.

[0040] Figure 2 It is a schematic diagram of the 4PL curve of fentanyl.

[0041] Figure 3 It is a schematic diagram of the 4PL curve of 5F-ADB. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0042] To make the above objects, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below with reference to the embodiments. The illustrative embodiments and descriptions thereof of the present invention are only used to explain the present invention and are not intended to limit the present invention.

[0043] Unless otherwise specified, the raw materials used in the embodiments are all commercially purchased.

[0044] Embodiment 1: Kit and Method for Determining Psychotropic Drugs in Hair Provided by the Present Invention

[0045] The kit provided in this embodiment includes a cleaning agent, a digestive, and a test strip.

[0046] I. Reagent Preparation

[0047] (1) Preparation of the cleaning agent

[0048] Take about 80 mL of ultrapure water into a beaker, slowly add 50 μL of TXR-1 (purchased from Croda Healthcare) while stirring; add 10 μL of Tween-20 (purchased from Sigma), and continue stirring until completely transparent; make up the ultrapure water to 100 mL, and after mixing, measure the pH to be 7.0.

[0049] (2) Preparation of the digestive

[0050] Weigh 0.03 - 3.0 g of CHAPS (1.0 g is preferred in this example, purchased from Sangon Biotech Co., Ltd.) and add it to 80 mL of 50 mM Tris-HCl buffer (pH 7.0), stir until completely transparent; weigh 0.025 g - 3.0 g of TCEP-HCl (2.5 g is preferred in this example, purchased from Sangon Biotech Co., Ltd.) and add it to the above solution, stir to dissolve; weigh 0.015 - 2.0 g of DTT (2.0 g is preferred in this example, purchased from Sigma) and add it to the solution, stir to dissolve in the dark; supplement the buffer to 100 mL, mix well to obtain a hair digestant with a DTT concentration of 2.0 g / 100 mL, a TCEP concentration of 2.5 g / 100 mL, and a CHAPS concentration of 1.0 g / 100 mL. Filter and sterilize with a 0.22 μm filter membrane, dispense into light-proof EP tubes, and store at -20°C.

[0051] (3)Preparation of test strip

[0052] As Figure 1 shown, the test strip 10 includes a sample pad 1, a conjugate pad 2, a detection pad 3, and an absorbent pad 4. The sample pad 1, conjugate pad 2, detection pad 3, and absorbent pad 4 are sequentially overlapped and adhered to the support bottom plate 5.

[0053] Preparation method of the absorbent pad 4: Place the filter paper in an oven at 50°C and dry it for later use.

[0054] Preparation method of the conjugate pad 2: For the conjugate pad for detecting fentanyl analogs, dilute the quantum dot-labeled fentanyl antigen analog (Fentanyl-BSA-QDs) with a coating diluent and spray it on glass fiber, and dry it overnight in a drying oven at 37°C; for the conjugate pad for detecting synthetic cannabinoids (5F-ADB), dilute the quantum dot-labeled 5F-ADB antigen analog (5F-ADB-OVA-QDs) with a coating diluent and spray it on glass fiber, and dry it overnight in a drying oven at 37°C.

[0055] The detection pad 3 is provided with a test line and a control line. For detecting fentanyl analogs, the test line 31 (T line) is coated with anti-fentanyl monoclonal antibody (Anti-Fentanyl mAb), and the control line 32 (C line) is coated with goat anti-mouse IgG antibody; for detecting synthetic cannabinoids, the test line 31 (T line) is coated with anti-5F-ADB monoclonal antibody (Anti-5F-ADB mAb). The control line 32 (C line) is coated with goat anti-rabbit IgG antibody. The material of the detection pad is selected from nitrocellulose membrane.

[0056] During detection, the sample treated with the hair cleaning solution and the hair digestion solution is dropped onto the sample pad of the test strip. The target antigen fentanyl or 5F-ADB in the sample and the quantum dot-labeled fentanyl antigen analog (Fentanyl-BSA-QDs) or 5F-ADB antigen analog (5F-ADB-OVA-QDs) flow along the nitrocellulose membrane (detection pad) towards the absorbent pad under the action of chromatography, and bind to the antibody that specifically binds to the target antigen at the position of the test line to form a "target antigen-target antigen-specific antibody" complex; when passing through the quality control line (C line), the target antigen binds to the coated IgG antibody to form the final quality control reaction complex. The fluorescence signal is read by a portable fluorescence reader (Suzhou Hemai Precision Instrument Co., Ltd., model FIC-H2), and the result is analyzed and displayed.

[0057] II. Hair pretreatment

[0058] (1) Hair cleaning

[0059] Take about 5 mg of hair samples. After cutting the hair to 0.5 - 1 cm with medical scissors, wash the hair alternately with the hair cleaning agent and pure water 3 - 5 times, and vortex for 3 - 5 min each time.

[0060] (2) Hair digestion

[0061] The hair after washing in step (1) is added with the hair digestive provided by the present invention and soaked for digestion for 5 - 10 min. The ratio of hair to digestive is 1 mg:100 μL. Vortex 3 times during digestion, and the hair test solution is obtained after digestion.

[0062] III. Determination of psychotropic drugs in hair

[0063] (1) Standard curve establishment

[0064] Take blank samples (hair of personnel without a history of taking psychotropic drugs), and obtain sample extracts through the cleaning agent and digestive provided in this example. Different concentrations of psychotropic drugs fentanyl and synthetic cannabinoid (5F-ADB) are added to the extracts to prepare 8 kinds of concentration standards S1 - S8, as shown in Table 1:

[0065] Table 1 Standard concentrations of fentanyl and 5F-ADB

[0066]

[0067] The test strips prepared in this embodiment were used to measure the fentanyl and 5F-ADB standard samples at 8 different concentrations in Table 1 through quantum dot competitive immunochromatographic assay. Each concentration was tested 3 times, and the mean value of the fluorescence signal was calculated respectively. Taking the dilution concentration as the independent variable and the mean value of the fluorescence signal as the dependent variable, and calibrating the background matrix with blank samples, a fitted four-parameter logistic regression (4PL) curve was obtained. The 4PL curve of fentanyl is: , such as Figure 2 ; The 4PL curve of 5F-ADB is: , such as Figure 3 shown.

[0068] (2) Accuracy verification

[0069] Refer to the industry standard method to prepare 36 positive hair samples of fentanyl with a concentration of 2 ng / mL and 40 positive hair samples of synthetic cannabinoids with a concentration of 1 ng / mL respectively, and collect 50 mg of hair from 50 people without a history of taking psychotropic drugs (negative samples), and take 5 mg as samples respectively. The specific steps for preparing positive hair samples are as follows:

[0070] S1: Wash the blank hair samples in deionized water, dry them in a constant temperature environment at 25°C, put them into an ethanol solution of 0.5 wt% dithiothreitol and sodium sulfite for reaction. During the reaction, take out 5 samples every 1 h for tensile testing until the average breaking elongation of the blank hair samples ≥ 20% and then take them out, wash them with deionized water and dry them in a constant temperature environment at 60°C to obtain pretreated hair samples;

[0071] S2: Take 400 mg of the pretreated hair samples, soak them in 250 ml of a fentanyl aqueous solution with a concentration of 10 ppm for 10 h and then take them out, dry them in a constant temperature environment at 50°C to obtain soaked hair samples;

[0072] S3: Put the soaked hair samples into a 3 wt% peracetic acid aqueous solution for reaction. During the reaction, take out 3 samples every 5 min for tensile testing until the average breaking elongation of the hair samples ≤ 5% and then take them out, wash them with deionized water and dry them in a constant temperature environment at 50°C to obtain a fentanyl-positive hair. After testing, the content of fentanyl in this positive hair is 2 ng / mL.

[0073] The preparation steps of the positive hair samples of synthetic cannabinoids with a concentration of 1 ng / mL are basically the same. Use the kit and test strips provided in this embodiment for sample pretreatment and quantum dot competitive immunochromatographic assay, observe whether the quality control line (C line) and the test line (T line) are clear and whether there is tailing, and count the test results according to the display of the C line and T line, as shown in Table 2.

[0074] Table 2 Hair test results

[0075]

[0076] The test results show that among the 36 fentanyl-positive hair samples, only 1 sample had a clear test line, and the positive detection rate was 97.22%; for the 40 synthetic cannabinoid-positive hair samples, the test lines all disappeared, and synthetic cannabinoids were detected in all samples, with a positive detection rate of 100%; no psychotropic drugs were detected in the 50 negative hair samples. It can be seen from the test results that the method for detecting psychotropic drugs in hair provided by the present invention can efficiently and accurately detect new psychotropic drugs such as fentanyl and 5F-ADB. Among them, the positive detection rate of 5F-ADB can be as high as 100%, with higher accuracy, which is conducive to the detection of psychotropic drugs and the application in rapid screening on-site.

[0077] (3)Sensitivity verification

[0078] Take two batches I and II of the kits for detecting fentanyl analogs and synthetic cannabinoids, with 10 copies in each batch, and operate according to the steps of this example. Repeat the detection of the standards S3 of fentanyl and 5F-ADB respectively, and test each sample 3 times. Substitute the fluorescence signal values (T / C values) into the above standard curves to obtain the concentrations, and the results are shown in Table 3.

[0079] Table 3 Sensitivity detection of the kit provided by the present invention

[0080]

[0081] As shown in the results of Table 3, when using the kits of different batches of the present invention to detect the same fentanyl standard, the detected values are basically consistent with its detection limit (1 ng / ml). When detecting the 5F-ADB standard, the detected values can reach its detection limit (0.5 ng / ml), indicating that the detection kit provided by the present invention has high sensitivity, and the effect is better when used for the detection of 5F-ADB.

[0082] (4)Consistency verification

[0083] In order to further verify the high sensitivity and high accuracy of the kit and method provided by the present invention, in this example, fentanyl-positive hair samples with a concentration of 2 ng / ml and synthetic cannabinoid-positive hair samples with a concentration of 1 ng / ml were further prepared. The method was the same as the preparation steps in (2) of this example. Then, the kit and detection method provided by this example were respectively used for comparative detection with the liquid chromatography-tandem mass spectrometry method (LC-MS / MS) in the forensic technical specification "Liquid Chromatography-Tandem Mass Spectrometry Method for 15 Drugs and Metabolites in Hair". The results are shown in Table 4.

[0084] Table 4 Comparison between the kit provided in this example and the mass spectrometry detection results

[0085]

[0086] As can be seen from the results in Table 4, for the hair samples of two types of people taking psychotropic drugs, the detection values of the kit provided by the present invention are basically consistent with the detection results of the gold standard LC-MS / MS, indicating that the kit and detection method provided by the present invention can achieve the same sensitivity and accuracy as LC-MS / MS.

[0087] (5)Verification of repeatability and stability

[0088] In practical applications, reagents or methods for extracting and detecting hair samples are often required to have good repeatability and long-term stability to ensure that hair samples containing psychotropic drugs can still be accurately retested after being stored for a certain period of time. Therefore, the present invention further verifies the stability of the detection method.

[0089] One fentanyl-positive hair sample with a concentration of 2 ng / ml and one synthetic cannabinoid-positive hair sample with a concentration of 1.5 ng / ml prepared according to the preparation steps in (2) of this example were each repeatedly detected 20 times using the kit and determination method provided by the present invention. The fluorescence signal values were recorded each time using a portable fluorescence reader, and the corresponding psychotropic drug concentrations were calculated according to the standard curve. The average value M and standard deviation SD of the 20 measurement results were calculated, and the coefficient of variation CV was calculated. The test results are shown in Table 5.

[0090] Table 5 Results of the inter-batch difference test

[0091]

[0092] As can be seen from Table 5, when using the kit and method provided by the present invention to process and detect hair samples containing different new psychotropic drugs, the coefficients of variation are all within 5%, indicating good stability.

[0093] When the kit prepared by the present invention was stored for 1, 3, 6, 12, and 24 months respectively, the above-mentioned fentanyl-positive hair sample with a concentration of 2 ng / ml and synthetic cannabinoid-positive hair sample with a concentration of 1.5 ng / ml were measured using it, and the results are shown in Table 6.

[0094] Table 6 Stability test results

[0095]

[0096] As shown in the results of Table 6, after the kit provided by the present invention was stored for a certain period of time, the detection values of the same hair sample did not decrease significantly, indicating that the detection kit and method provided by the present invention have good long-term stability.

[0097] (6)Specificity verification

[0098] To further verify that the kit provided by the present invention can be used for specifically detecting hair samples containing fentanyl analogs and synthetic cannabinoids, referring to the preparation steps of the positive hair samples in (2) of Part Three of this Example, positive hair samples of fentanyl, synthetic cannabinoids, morphine, ketamine, cocaine, methamphetamine, methcathinone, pseudoephedrine, diazepam, phenobarbital, methadone, tramadol, ranitidine, gatifloxacin, and procaine with a concentration of 1 ng / mL were respectively prepared, and the above positive hair samples were respectively processed by the kit provided by this Example to obtain sample extracts. The above sample extracts were all detected by test strips containing antibodies against fentanyl and 5F-ADB, and the results were compared with the detection results of the LC-MS / MS method in the judicial expertise technical specification "Liquid Chromatography-Tandem Mass Spectrometry Test Method for 15 Drugs and Their Metabolites in Hair", and the results are shown in Table 7.

[0099] Table 7 Specificity verification results

[0100]

[0101] As shown in Table 7, the kit provided by the present invention can be used for specifically detecting two types of psychotropic drugs, fentanyl analogs and synthetic cannabinoids, and morphine, ketamine, cocaine, methamphetamine, methcathinone, pseudoephedrine, diazepam, phenobarbital, methadone, tramadol, ranitidine, gatifloxacin, and procaine do not cross-react with this product.

[0102] Example 2: Screening of hair cleaning agents

[0103] To verify the necessity of using a composition of Tween-20 and TXR-1 as a cleaning agent to improve the accuracy, sensitivity, and stability of the detection of psychotropic drugs in hair, the present invention also evaluated the differences in the effects of several commonly used reagents for hair cleaning.

[0104] I. Accuracy verification

[0105] First, 40 positive hair samples of synthetic cannabinoids with a concentration of 1.5 ng / mL were respectively prepared by referring to the preparation steps of the positive hair samples in (2) of Part Three of Example 1. After that, 40 positive hairs were respectively cleaned with 5 hair cleaning reagents shown in Table 8, and then the same hair digestion reagent and immunochromatography method as in Example 1 were used for drug extraction and detection, and the results are shown in Table 9.

[0106] Table 8 Formulations of 5 hair cleaning agents

[0107]

[0108] Table 9 Influence of Cleaners with Different Components on the Accuracy of Detection Results

[0109]

[0110] As shown in the results of Table 9, after cleaning the same sample of hair with cleaners of different components and then using the same digestion, extraction and detection methods, there are still significant differences in the accuracy of the detection results. Among them, the combined formula of 0.1% Tween-20 and 0.5% TXR-1 can accurately detect all positive samples, and the positive detection rate is 100%; after cleaning with Tween-20 or TXR-1 alone, the false negative rate of the detection results is high, and when using the combination of Tween-20 and CHAPS for cleaning or the combination of SDS and TXR-1, it will also cause the inability to detect all positive samples and negative results will appear. Similarly, after using the above three cleaning treatments on the hair of fentanyl analogue users and then performing digestion and detection, the results are the same as above.

[0111] II. Sensitivity Verification

[0112] Similarly, referring to the preparation steps of the positive hair samples in item (2) of part three in Example 1, one fentanyl positive hair sample with a concentration of 2 ng / mL and one synthetic cannabinoid positive sample with a concentration of 1.5 ng / mL were prepared, and were respectively cleaned with the cleaners shown in Table 8. The remaining reagents and steps were the same as those of the kit and detection method provided in Example 1. At the same time, LC-MS / MS of the Judicial Expertise Technical Specification "Liquid Chromatography-Tandem Mass Spectrometry Test Method for 15 Drugs and Metabolites in Hair" was used to extract and detect the same samples, and the results are shown in Table 10.

[0113] Table 10 Hair Detection Results of Different Cleaners

[0114]

[0115] As shown in the results of Table 10, when using one component of Tween-20 or TXR-1 as the cleaner (cleaners ④ and ⑤), the drug concentration detected after digestion is significantly lower than the mass spectrometry detection result, and the concentration of psychotropic drugs detected from the hair samples cleaned with cleaner ② or ③ is also significantly lower than that of the LC-MS / MS method. Only the detection results of the hair samples treated with cleaner ① prepared with 0.01% Tween-20 and 0.05% TXR-1 by the kit and test strip can reach the detection value consistent with or even higher than that of the mass spectrometry. The reason may be that when a surfactant is used in combination with the surfactant substances in the other two cleaners, the effect is too mild to effectively destroy the protein and keratin structure in the hair or there are pollutant residues, thus interfering with the subsequent detection. Therefore, the detection results are significantly lower and the sensitivity decreases.

[0116] III. Stability Verification

[0117] Furthermore, the present invention evaluated the stability differences of the kits prepared with the 5 cleaning agents shown in Table 8. After the kits prepared with the 5 cleaning agents completed the collection, cleaning, and digestion of the synthetic cannabinoid-positive hair samples with a concentration of 1.5 ng / mL, the concentration of 5F-ADB in the sample extract was measured by the quantum dot competitive immunochromatographic assay using the test strip prepared in Example 1. Then, when the 5 kits were stored for 1, 3, 6, 12, and 24 months, the concentration of 5F-ADB in the same sample was detected again, and the measurement results are shown in Table 11.

[0118] Table 11 Comparison of the Stability of Cleaning Agents with Different Components

[0119]

[0120] It can be seen from the results in Table 11 that different cleaning agents have obvious effects on the extraction and detection of drugs in hair samples. By comparing the initial drug concentrations under the three cleaning conditions, it can be seen that the contents of 5F-ADB extracted and detected by cleaning agents ②, ④, and ⑤ are significantly lower than the other two. At the same time, after the 5 kits are stored for 1, 3, 6, 12, and 24 months respectively, when the same sample extract is detected again using the kit prepared with cleaning agent ①, its detection value does not change significantly, and 5F-ADB close to the initial content can still be detected. However, after the kits containing cleaning agents ② to ⑤ are stored for several months, the concentrations of substances all decrease significantly. This may be because the effective substances in the other several reagents cannot fully play their roles after being stored for a certain period of time, or the reagents are contaminated after being used for a period of time, thereby affecting the detection accuracy. When Tween-20 and TXR-1 are used as a composition, the above problems are avoided. At the same time, TXR-1 fully dissolves lipids and membrane proteins, and Tween-20 gently removes pollutants. The two work together to promote drug release and are not easily contaminated during use and storage, improving the long-term stability of the detection method.

[0121] IV. Detection of Dyed Hair Samples

[0122] In addition, considering the actual situation that some users of psychotropic drugs may dye their hair, and bleach, hair dyes, etc. pose great challenges to the full extraction of drugs in hair and have a significant impact on the test results, therefore, the present invention further compares the effect differences when 5 kinds of cleaning agents shown in Table 8 are used to treat the dyed hair samples and undyed hair samples. Fentanyl positive hair samples with a concentration of 2 ng / mL and synthetic cannabinoid positive samples with a concentration of 1.5 ng / mL prepared according to the preparation steps of the positive hair samples in (2) of Reference Example 1 are respectively dyed. The hair dye used is a hair dye containing substances such as toluene-2,5-diamine and resorcinol (purchased from Suzhou Shangmei International Cosmetics Co., Ltd., L'Oréal, special national cosmetics approval number 20231478) for hair dyeing and color development. The dyed samples are respectively subjected to the same pre-treatment method and immunochromatographic test as in Example 1, except that the test kits prepared with 5 kinds of cleaning agents shown in Table 8 are used respectively. The dyed hair samples are all tested and compared with the LC-MS / MS test results in the forensic technical specification "Liquid Chromatography-Tandem Mass Spectrometry Test Method for 15 Kinds of Drugs and Metabolites in Hair" at the same time, as shown in Table 12.

[0123] Table 12 Detection Sensitivity of Different Cleaning Agents for Dyed Hair Samples

[0124]

[0125] As shown in the results of Table 12, for the extraction and detection of two psychotropic drugs in hair dye samples, only the results of the kit containing cleaning agent ① can reach the same detection level as LC-MS / MS. The concentrations of fentanyl and 5F-ADB in the samples treated with the kits containing cleaning agents ②-⑤ are significantly lower than the LC-MS / MS detection values. After treating the hair samples of fentanyl analog users with the kits containing cleaning agents ④ and ⑤, the detected fentanyl content is below the detection limit, resulting in false negative results. This shows that for hair samples with the same use of hair dye, the effects of cleaning agents with different compositions are significantly different, and significantly affect the accuracy and sensitivity of the detection results. The reason may be that for stubborn pollutants such as hair dye cream attached to the hair, the cleaning effect of a single surfactant is very limited. The combination of Tween-20 and CHAPS in cleaning agent ② and the combination of SDS and TXR-1 in cleaning agent ③ also cannot completely remove the pollutants. The residue of hair dye affects the full release of psychotropic drug molecules and the sensitivity of subsequent immunoassay. Only the cleaning agent ① prepared with Tween-20 and TXR-1 can effectively wash off the hair dye on the hair and fully release the drugs in the hair samples, achieving the same detection accuracy and sensitivity as the LC-MS / MS method. The treatment and detection effects for hair samples containing 5F-ADB are better. Therefore, the combination of Tween-20 and TXR-1 is preferably used to prepare the hair cleaning agent.

[0126] Example 3: Screening of hair digestants

[0127] I. Verification of accuracy and sensitivity

[0128] In the detection method of psychotropic drugs in hair, in the pre-treatment step of hair, hair cleaning is for more thorough digestion and extraction in the follow-up, and the effect of the hair digestion reagent in the extraction process is more crucial. Therefore, the present invention further verifies the necessity of using a combination of dithiothreitol (DTT), tris(2-carboxyethyl)phosphine hydrochloride (TCEP) and CHAPS as the hair digestant.

[0129] Referring to the preparation steps of the positive hair samples in item (2) of Example 1, prepare a fentanyl positive hair sample with a concentration of 2 ng / mL and a synthetic cannabinoid positive sample with a concentration of 1.5 ng / mL. Use the same hair pre-treatment method as in Example 1 and the quantum dot competitive immunochromatographic assay to detect whether 40 synthetic cannabinoid positive hair samples contain 5F-ADB. The difference is that different hair digestants shown in Table 13 are used as the digestants, and then the detection results are statistically analyzed, as shown in Table 13.

[0130] Table 13 Influence of digestants with different components on the accuracy of detection results

[0131]

[0132] The results showed that after using a single reducing agent, DTT or TCEP, for digesting and extracting hair samples, the number of positive samples detected significantly decreased, with false negative results and an increased false negative rate. In the detection results of CHAPS (an amphoteric ion surfactant) alone, the positive detection rate was significantly reduced and the false negative rate was significantly increased. When using the combination of DTT and TCEP (digestant ④), or the combination of DTT and CHAPS (digestant ⑤), or the combination of TCEP and CHAPS (digestant ⑥), the positive detection rate of the samples could be improved, but there were still false negative results, indicating that the combination of two reducing agents or the combination of a reducing agent and a surfactant could improve the detection accuracy, but the effect was limited. By comparing the results of digestants ⑦ and ⑧, it was found that the composition of DTT, TCEP, and CHAPS could accurately detect the drugs in the hair of synthetic cannabinoid users, with a positive detection rate reaching 100%, while there were still false negative results in the detection results of the composition of SDS, TCEP, and CHAPS. Similarly, when using the 8 digestants described in Table 13 of the present invention to extract the hair samples of fentanyl analogue users, the accuracy trend of the results was the same as above. In summary, only by using the composition of DTT, TCEP, and CHAPS to prepare the digestant could the detection accuracy of psychotropic drugs in hair be effectively improved and the false negative rate be reduced.

[0133] Furthermore, for 1 case each of a fentanyl-positive hair sample with a concentration of 2 ng / mL and a synthetic cannabinoid-positive sample with a concentration of 1.5 ng / mL, the psychotropic drugs in the hair samples were extracted using the kits prepared with the 8 digestants shown in Table 13 respectively. The remaining reagents and steps were the same as those of the kit and detection method provided in Example 1. At the same time, the LC-MS / MS method of the forensic technical specification "Liquid Chromatography-Tandem Mass Spectrometry Test Method for 15 Drugs and Metabolites in Hair" was used to detect the same samples. The results are shown in Table 14.

[0134] Table 14 Detection Sensitivity of Different Digestants

[0135]

[0136] As shown in the results of Table 14, there are obvious differences in the effects of different digestion reagents. Compared with the LC-MS / MS detection results, the hair samples treated with the digestion agent ⑦ prepared with 2.0 g / mL DTT, 2.5 g / mL TCEP, and 1 g / mL CHAPS can achieve the same results as the LC-MS / MS detection values after being detected by the kit and the test strip. However, the detection values of a single reducing reagent or surfactant (digestion agents ① - ③) are significantly lower than the LC-MS / MS detection values. Among them, for the samples treated with the single CHAPS reagent, the detection results of fentanyl and 5F-ADB in the sample extraction solution are both false negatives. The combination of DTT and TCEP or the combination of DTT / TCEP and CHAPS (digestion agents ④ - ⑥) helps to improve the detection sensitivity and accuracy, but the effect is limited and still cannot reach the same detection sensitivity as LC-MS / MS. Similarly, the detection results of the digestion agent ⑧ composed of SDS, TCEP, and CHAPS are also not as good as those of digestion agent ⑦ and LC-MS / MS. The reason may be that CHAPS destroys the hydrophobic interaction and helps release the drug molecules embedded in the hair medulla. DTT and TCEP reduce the disulfide bonds (—S—S—) in keratin, open the tight structure of the hair fiber, and release the deeply bound psychotropic drugs. At the same time, DTT, TCEP, and CHAPS act synergistically on the hydrophilic and hydrophobic regions of the hair, effectively destroying the keratin structure of the hair and promoting drug release, while other digestion reagents cannot achieve the same effect. Among them, in the digestion and detection effects of digestion agent ⑦ on the hair samples of two types of psychotropic drug users, the detection sensitivity for 5F-ADB is higher. Therefore, only the digestion reagent composed of DTT, TCEP, and CHAPS can effectively improve the sensitivity and accuracy of the detection method.

[0137] II. Stability Verification and Detection of Dyed Hair Samples

[0138] Similarly, the present invention also investigated the long-term stability of the kits prepared with the 8 digestion agents in Table 13, as well as the differences in the effects of sample extraction and detection for dyed hair samples. Referring to the preparation steps of the positive hair samples in item (2) of Part III of Example 1, fentanyl positive hair samples with a concentration of 2 ng / mL and synthetic cannabinoid positive samples with a concentration of 1.5 ng / mL were prepared. The hair pretreatment and immunochromatography method were basically the same as those in Example 1, where the digestion reagents were respectively the 8 digestion agents in Table 13, and at the same time, the LC-MS / MS method in the forensic technical specification "Liquid Chromatography-Tandem Mass Spectrometry Test Method for 15 Drugs and Metabolites in Hair" was used for extraction and detection.

[0139] In the stability assessment experiment, after collecting, cleaning, and detecting the same synthetic cannabinoid-positive hair sample using kits prepared with 8 kinds of digestive agents, the 5F-ADB content in the same sample was measured again when the 8 kits were stored for 1, 3, 6, 12, and 24 months respectively. The results are shown in Table 15.

[0140] In the experiment for evaluating the extraction effect of hair dyeing samples, hair dyes containing substances such as toluene-2,5-diamine and resorcinol (purchased from L'Oréal (China) Co., Ltd., Suzhou Shangmei International Cosmetics Co., Ltd., special national cosmetics approval number: 20231478) were used to dye fentanyl-positive hair samples and synthetic cannabinoid-positive samples. 8 kinds of digestive agents were used to clean, digest, and detect the two positive samples respectively. The results are shown in Table 16.

[0141] Table 15 Effects of digestive agents with different components on sample stability

[0142]

[0143] As shown in the results of Table 15, only the kits prepared with the combined digestive agent of DTT, TCEP, and CHAPS (digestive agent ⑦) could accurately detect the content of psychotropic drugs in the samples after being stored for a certain period. When using a single component (digestive agents ① - ③) or a combination of two components (digestive agents ④ - ⑥), on the one hand, the initial content of 5F-ADB digested and extracted from the hair samples was low, and all the kits prepared with CHAPS reagent showed false negative results; on the other hand, after these 6 kits were stored for a period of time, the detectable 5F-ADB content decreased significantly. After being stored for 12 months, the detection results for the positive samples were all false negative; although using digestive agent ⑧ improved the drug extraction efficiency, the drug content that could be detected from the samples by the kit still decreased after being stored for a period of time. The reason may be that digestive agents ① - ⑥, ⑧ degraded or volatilized to a certain extent as the storage time prolonged, or were easily contaminated as the number of uses increased. However, the combined digestive agent of DTT, TCEP, and CHAPS effectively overcame the volatilization of the reagent itself and was not easily contaminated, ensuring its detection accuracy and stability within a certain time range. After re-detecting and verifying the hair samples of fentanyl analogue users treated with 8 kinds of digestive agents, the results were the same as above, indicating that the digestive agent prepared with the combination of DTT, TCEP, and CHAPS can effectively improve the stability of the detection method for new psychotropic drugs.

[0144] Table 16 Detection sensitivity of different digestive agents for hair dyeing samples

[0145]

[0146] As can be seen from the test results in Table 16, there are obvious differences in the results of treating hair dye samples with different digestants. Among them, only the digestant prepared with DTT, TCEP and CHAPS can have excellent sensitivity for extracting and detecting two kinds of psychotropic drugs in hair samples, reaching the same detection accuracy as mass spectrometry detection, indicating that the composition of DTT, TCEP and CHAPS can effectively remove the pollutants attached to the hair and fully release the psychotropic drugs in the hair. The drug contents detected by digestants ① - ⑥ for hair dye samples are significantly lower than the mass spectrometry results, indicating that the use of a reducing agent or a surfactant or the combination of any two substances has poor effects and cannot eliminate the influence of residual pollutants such as bleaching agents and hair dyes, resulting in a decrease in the sensitivity of the detection method. And using digestant ⑧ also cannot solve the influence of hair dye residue, resulting in a decrease in the detected drug content. Digestant ⑦ is used to treat the hair dye samples of fentanyl analogue users and synthetic cannabinoid users, and the detected values are all close to the mass spectrometry detection values, and the detection sensitivity for 5F-ADB is higher. In summary, it is preferred to use the composition of DTT, TCEP and CHAPS to prepare a hair digestant, which helps to improve the extraction, detection accuracy and sensitivity of drugs in hair samples containing pollutants such as hair dyes.

[0147] At the beginning of the present invention, the washed hair samples were added to the digestant and placed in a water bath at 55°C - 65°C for heating digestion. However, it was found that for different positive hair samples of psychotropic drugs, the detection results could not always reach the same level as mass spectrometry detection and were significantly lower than the mass spectrometry results. Therefore, a method without water bath heating was further adopted, and it was found that the detection level was significantly improved. Therefore, in order to verify the necessity of not heating for digestion assistance during the pretreatment of samples by the kit provided by the present invention, the present invention further prepared 20 synthetic cannabinoid positive hair samples (concentration: 1.5 ng / mL) according to the positive hair sample preparation steps in item (2) of Reference Example 1, used the kit provided by Example 1 for pretreatment, and determined the content of 5F-ADB by immunochromatographic test with a test strip, with the difference that the digestion process adopted heating digestion and non-heating digestion, and the heating condition was a water bath at 55°C - 65°C. At the same time, it was compared with the detection results of the LC-MS / MS method in the forensic identification technical specification "Liquid Chromatography-Tandem Mass Spectrometry Test Method for 15 Kinds of Drugs and Metabolites in Hair". The results are shown in Table 17.

[0148] Table 17 5F-ADB content under different digestion conditions

[0149] According to the data in Table 17, it can be seen that for the same sample, the psychotropic drug detection values of the extract obtained by digesting hair with the kit provided by the present invention under the condition of not heating are almost the same as the mass spectrometry detection results. However, after digesting the hair sample under the heating condition, the drug content detected in the extract is significantly lower than the mass spectrometry detection value. This may be because the digestion process becomes too intense after heating, resulting in the volatilization loss of the drugs in the sample and the decrease in the detected content. Therefore, only by adopting the non-heating digestion method can the digestion reagent in the kit play the most suitable role and extract the maximum drug content, thereby improving the accuracy of the detection method.

[0150] Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Any person familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention should be defined by the claims.

Claims

1. A kit for extracting psychotropic drugs from hair, characterized in that: The kit comprises a cleaning agent and a digestion agent, wherein the cleaning agent comprises Tween-20 and TXR-1, and the digestion agent comprises dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride and CHAPS.

2. The kit according to claim 1, wherein The concentration of the Tween-20 is 0.005%-0.01% (v / v), and the concentration of the TXR-1 is 0.01%-0.05% (v / v).

3. The kit according to claim 1, wherein The concentration of the dithiothreitol is 0.015 g / 100 mL to 2.0 g / 100 mL, the concentration of the tris(2-carboxyethyl)phosphine hydrochloride is 0.025 g / 100 mL to 3.0 g / 100 mL, and the concentration of the CHAPS is 0.03 g / 100 mL to 3.0 g / 100 mL.

4. A kit for detecting psychotropic drugs in hair, characterized in that: The kit comprises a cleaning agent, a digestion agent and a test strip, wherein the cleaning agent comprises Tween-20 and TXR-1, and the digestion agent comprises dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride and CHAPS.

5. A method for extracting psychotropic drugs from hair, characterized in that: Psychotropic drugs in hair are extracted using the kit as described in any one of claims 1 to 3.

6. The method according to claim 5, characterized in that The steps include: (1) Hair cleaning: Use a cleaning agent to clean the hair after cutting it into pieces; (2) Hair digestion and extraction: The cleaned hair is digested and extracted using a digestive agent for 5-15 minutes to obtain a sample extract.

7. A method for determining psychotropic drugs in hair, characterized in that: The steps include: (1) Extracting psychotropic drugs from hair using the kit as described in any one of claims 1 to 3; (2) Detection using immunochromatography.

8. The method according to claim 7, characterized in that The hair sample may be hair, sweat hair, armpit hair or pubic hair of a person taking psychotropic drugs.

9. The method according to claim 7, characterized in that The psychotropic drugs include any one or more of fentanyl analogs, synthetic cannabinoids, synthetic cathinones, etomidate, and GHB.

10. Use of a composition for preparing a reagent for improving the accuracy of detecting psychotropic drugs in dyed hair, characterized in that: The composition includes dithiothreitol, tris(2-carboxyethyl)phosphine hydrochloride and CHAPS.

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