A hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers

Abstract

The present invention relates to a hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers. The steps include: cleaning the sample hair with an organic solvent, drying it at room temperature, transferring it to an internal standard working solution, successively grinding, centrifuging, and microfiltering, and then injecting it into an LC-MS / MS system. Among them, the chromatographic conditions include: the chromatographic column is an Allure PFPP column; mobile phase A is an ammonium acetate buffer solution containing formic acid and acetonitrile; mobile phase B is acetonitrile; the elution method is isocratic constant flow elution; the column temperature is room temperature. The mass spectrometry conditions include: the mass spectrometer is an electrospray ionization mass spectrometer; the detection mode is the positive ion mode; the monitoring mode is the multiple reaction monitoring mode; the ion spray voltage is 5000V to 6000V; the ion source temperature is 400°C to 600°C; the curtain gas pressure is 30psi to 50psi, the nebulizer gas pressure is 40psi to 60psi, and the auxiliary gas pressure is 40psi to 60psi. The present invention has a low detection limit, high resolution, high sensitivity, and high automation degree, and can be used as a reference evidence in court.

Description

Technical Field

[0001] The present invention relates to the field of monitoring and analysis of drug abuse, and particularly to a hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers. Background Art

[0002] Hallucinogens are a class of psychoactive substances that can cause hallucinations and change a person's consciousness. Although most hallucinogens do not cause death due to direct overdose, fatal accidents may occur during ingestion due to panic, mental confusion, and strange behaviors. The use of hallucinogens shows an increasing trend, and the identification, crackdown, and prevention of hallucinogens are important requirements for current drug abuse prevention and control.

[0003] Hair analysis is a powerful means for monitoring drug abuse, with unique advantages such as easy collection, easy preservation, non-invasive sampling, and a long detection window, and can record the drug-taking history over a long period. The growth rate of hair is generally 1 cm / month, and segmental hair analysis can estimate the drug intake time of an individual. However, due to the low oral dose of hallucinogens and their content in hair generally being below the picogram level, there is an urgent need for a hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers. Summary of the Invention

[0004] The object of the present invention is to provide, in view of the deficiencies in the prior art, a hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] The present invention provides a hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers. The hallucinogens include: lysergic acid diethylamide (LSD), 1-propionyl-lysergic acid diethylamide (1P-LSD), mescaline, 2,5-dimethoxy-4-bromoamphetamine (2-CB), 2-(8-bromo-2,3,6,7-tetrahydrobenzofuro[2,3-f][1]furan-4-yl)-ethylamine (2C-B-fly), 2-fluorodeschloroketamine (2-FDCK), methoxetamine (MXE), 2-phenyl-2-(methylamino)cyclohexanone (Deschloroketamine, DCK), and methylenedioxypyrovalerone (MDPV); the hallucinogen biomarkers include: lysergic acid diethylamide isomer, isolysergic acid diethylamide (iso-LSD), lysergic acid diethylamide metabolite, 2-oxo-3-hydroxy-lysergic acid diethylamide (O-H-LSD), and 2-fluorodeschloroketamine metabolite, 2-(2-fluorophenyl)-2-aminocyclohexanone (nor-2-FDCK);

[0007] The steps of the hair analysis method include: cleaning the sample hair with an organic solvent, drying it at room temperature, transferring it to the internal standard working solution, grinding, centrifuging and microfiltrating in sequence, and then injecting it into the LC-MS / MS system; wherein,

[0008] The internal standard working solution is a methanol solution of lysergic acid diethylamide-d3 (LSD-d3);

[0009] The chromatographic conditions include: the chromatographic column is an Allure PFPP column; mobile phase A is an ammonium acetate buffer solution containing formic acid and acetonitrile; mobile phase B is acetonitrile; the elution mode is isocratic constant flow elution; the column temperature is room temperature;

[0010] The mass spectrometry conditions include: the mass spectrometer is an electrospray ionization mass spectrometer; the detection mode is the positive ion mode; the monitoring mode is the multiple reaction monitoring mode; the ion spray voltage is 5000V - 6000V; the ion source temperature is 400°C - 600°C; the curtain gas pressure is 30psi - 50psi, the nebulizer gas pressure is 40psi - 60psi, and the auxiliary gas pressure is 40psi - 60psi.

[0011] Preferably, the organic solvent is acetone.

[0012] Preferably, the conditions for the cleaning include: the number of cleaning times is 2 - 4 times.

[0013] Preferably, after drying at room temperature, the sample hair is cut into segments of 2mm - 3mm, and then 20mg - 30mg of the accurately weighed sample hair is transferred to 300μL - 500μL of the internal standard working solution.

[0014] Preferably, the mass concentration of lysergic acid diethylamide-d3 in the internal standard working solution is 20ng / mL - 30ng / mL.

[0015] Preferably, the conditions for the grinding include: the grinding temperature is -40°C - -30°C; the grinding speed is 10m / s - 20m / s; the running time is 30s - 50s, the residence time is 10s - 30s, and the number of cycles is 15 times.

[0016] Preferably, the conditions for the centrifuging include: the centrifuging speed is 11000×g - 13000×g; the centrifuging time is 2min - 4min.

[0017] Preferably, the conditions for the microfiltrating include: the pore size of the filter membrane is 0.2μm - 0.3μm.

[0018] Preferably, the chromatographic conditions further include: the injection volume is 4μL - 6μL; the temperature of the autosampler is 3°C - 5°C.

[0019] Preferably, the chromatographic conditions further include: the volume fraction of formic acid in the mobile phase A is 0.1% - 0.2%, the volume fraction of acetonitrile is 4% - 6%, and the molar concentration of ammonium acetate is 10 mmol / L - 30 mmol / L; the volume ratio of the mobile phase A to the mobile phase B in the eluent is 30:70; the flow rate is 0.2 mL / min - 0.3 mL / min; the elution time is 10 min - 20 min.

[0020] The present invention adopts the above technical solutions, and compared with the prior art, has the following technical effects:

[0021] The hair analysis method of the present invention constructs an LC-MS / MS method for quantitatively analyzing 1P-LSD and 2C-B-fly in hair, and constructs an LC-MS / MS method for simultaneously quantitatively analyzing various hallucinogens and hallucinogen biomarkers in hair. The pretreatment method is simple and rapid, with a low detection limit, high resolution, high sensitivity, and high automation degree. It can systematically and standardizedly monitor the prevalence of hallucinogen abuse, and at the same time, the analysis results can be used as reference evidence in court. Description of the Drawings

[0022] Figure 1 It is a chromatogram when the mass fraction of LSD in hair is LOQ;

[0023] Figure 2 It is a chromatogram when the mass fraction of Iso-LSD in hair is LOQ;

[0024] Figure 3 It is a chromatogram when the mass fraction of O-H-LSD in hair is LOQ;

[0025] Figure 4 It is a chromatogram when the mass fraction of 1P-LSD in hair is LOQ;

[0026] Figure 5 It is a chromatogram when the mass fraction of 2-CB in hair is LOQ;

[0027] Figure 6 It is a chromatogram when the mass fraction of 2C-B-fly in hair is LOQ;

[0028] Figure 7 It is a chromatogram when the mass fraction of MDPV in hair is LOQ;

[0029] Figure 8 It is a chromatogram when the mass fraction of MXE in hair is LOQ;

[0030] Figure 9 It is a chromatogram when the mass fraction of DCK in hair is LOQ;

[0031] Figure 10 It is the chromatogram when the mass fraction of 2-FDCK in hair is at the LOQ;

[0032] Figure 11 It is the chromatogram when the mass fraction of Nor-2-FDCK in hair is at the LOQ;

[0033] Figure 12 It is the chromatogram when the mass fraction of Mescaline in hair is at the LOQ;

[0034] Figure 13 It is the chromatogram of blank hair. Detailed implementation manners

[0035] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0036] It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other.

[0037] Next, the present invention will be further described in conjunction with the accompanying drawings and specific embodiments, but it is not a limitation of the present invention.

[0038] Reagent conditions

[0039] In the present invention, lysergic acid diethylamide, iso-lysergic acid diethylamide, 2-oxo-3-hydroxy-lysergic acid diethylamide, mescaline, 2,5-dimethoxy-4-bromophenethylamine, 2-(8-bromo-2,3,6,7-tetrahydrobenzofuro[2,3-f][1]furan-4-yl)-ethylamine, fluanisone, 2-(2-fluorophenyl)-2-aminocyclohexanone, methoxetamine, 2-phenyl-2-methylaminocyclohexanone, methylenedioxypyrovalerone and lysergic acid diethylamide-d3 standard products are purchased from Cerilliant Corporation in the United States;

[0040] In the present invention, 1-propionyl-lysergic acid diethylamide standard product is purchased from MedChemExpress;

[0041] In the present invention, acetone (analytical pure, 99.5%) is purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd. in China;

[0042] In the present invention, methanol (HPLC) and acetonitrile (HPLC) are purchased from Sigma-Aldrich in the United States;

[0043] In the present invention, ammonium acetate (HPLC, ≥98.0%) and formic acid (HPLC, ≥98.0%) were purchased from Anpel Laboratory Technologies (Shanghai) Inc., China;

[0044] In the present invention, ultrapure water was prepared by the AFS-10 water purification system of Millipore Corporation, USA.

[0045] Instrument conditions

[0046] In the present invention, the AFS-10 ultrapure water preparation system was purchased from Millipore Corporation, USA;

[0047] In the present invention, the BSA124S electronic balance was purchased from Sartorius Scientific Instruments (Beijing) Co., Ltd.;

[0048] In the present invention, the JXFSTPRP-CLN cryogenic grinder was purchased from Shanghai Jingxin Industrial Development Co., Ltd.;

[0049] In the present invention, the MiniSpin high-speed centrifuge was purchased from Eppendorf AG, Germany;

[0050] In the present invention, the polytetrafluoroethylene filter membrane (0.22 μm) was purchased from Sinopharm Chemical Reagent Co., Ltd.;

[0051] In the present invention, the LC-20AD LC high-performance liquid chromatograph was purchased from Shimadzu Corporation, Japan;

[0052] In the present invention, the AB Sciex API 4000 triple quadrupole linear ion trap mass spectrometer was purchased from Applied Biosystems, USA.

[0053] Example

[0054] This example provides a hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers, and the hallucinogens and hallucinogen biomarkers include: lysergic acid diethylamide, iso-lysergic acid diethylamide, 2-oxo-3-hydroxy-lysergic acid diethylamide, 1-propionyl-lysergic acid diethylamide, mescaline, 2,5-dimethoxy-4-bromophenethylamine, 2-(8-bromo-2,3,6,7-tetrahydrobenzofuro[2,3-f][1]furan-4-yl)-ethylamine, fluanisone, 2-(2-fluorophenyl)-2-aminocyclohexanone, methoxymethamphetamine, 2-phenyl-2-methylaminocyclohexanone, and methylenedioxypyrovalerone;

[0055] The steps of the hair analysis method include: placing the sample hair in a stoppered test tube, washing it 3 times with acetone, drying it at room temperature, cutting the sample hair into segments of 2 mm to 3 mm, and then transferring 25 mg of the accurately weighed sample hair to 400 μL of the internal standard working solution. After grinding, centrifugation, and microfiltration in sequence, it is injected into the LC-MS / MS system; among them,

[0056] The internal standard working solution is a methanol solution of lysergic acid diethylamide-d3, and the mass concentration of lysergic acid diethylamide-d3 is 25 ng / mL;

[0057] The conditions for grinding include: the grinding temperature is -39°C; the grinding speed is 18 m / s; the running time is 40 s, the residence time is 20 s, and the number of cycles is 15 times;

[0058] The conditions for centrifugation include: the centrifugation speed is 12000×g; the centrifugation time is 3 min;

[0059] The conditions for microfiltration include: the pore size of the filter membrane is 0.22 μm;

[0060] The chromatographic conditions include: the chromatographic column is an Allure PFPP column (100 mm×2.1 mm, 5 μm), preceded by an Agilent ZORBAX Extend-C18 narrow-bore guard column (12.5 mm×2.1 mm, 5 μm); data analysis is performed using Analyst 1.5.2 software and a MultiQuant 3.0.2 workstation; mobile phase A is an ammonium acetate buffer solution containing formic acid and acetonitrile, the volume fraction of formic acid is 0.1%, the volume fraction of acetonitrile is 5%, and the molar concentration of ammonium acetate is 20 mmol / L; mobile phase B is acetonitrile; the elution method is isocratic constant flow elution, and the volume ratio of mobile phase A to mobile phase B in the eluent is 30:70, the flow rate is 0.25 mL / min, the elution time is 12 min; the column temperature is room temperature; the injection volume is 5 μL; the temperature of the autosampler is 4°C;

[0061] The mass spectrometry conditions include: the mass spectrometer is an electrospray ionization mass spectrometer (Applied Biosystems / MDS SCIEX, Toronto, Canada); the detection mode is the positive ion mode; the monitoring mode is the multiple reaction monitoring (MRM) mode; data collection and analysis are performed using Analyst 1.5.2 software and a MultiQuant 3.0.2 workstation; the ion spray voltage is 5500 V; the ion source temperature is 500°C; the curtain gas pressure is 40 psi, the nebulizing gas (GS1) pressure is 50 psi, and the auxiliary gas (GS2) pressure is 50 psi;

[0062] After injection, precursor ions, fragment ions, declustering potential (DP), and collision energy (CE) were directly screened to obtain the maximum ion intensity, and the collision dissociation energy was kept stable.

[0063] As a preferred embodiment, the preparation steps of the internal standard working solution include: taking an appropriate amount of the internal standard stock solution, diluting it with methanol to a mass concentration of 25 ng / mL of lysergic acid diethylamide-d3, and then obtaining the internal standard working solution, which needs to be stored in a refrigerator at -20°C after sealing for standby.

[0064] As a preferred embodiment, the preparation steps of the mobile phase A include: weighing 3.08 g of ammonium acetate, placing it in a 2000 mL volumetric flask, dissolving it with ultrapure water, adding 2 mL of formic acid solution with a volume fraction of 98%, and 100 mL of acetonitrile, and diluting it to 2000 mL with ultrapure water to obtain the mobile phase A.

[0065] Verification example

[0066] In this validation example, by directly injecting each target analyte into the mass spectrometer, the fragment ions, declustering voltage, and collision energy of the target compound and the internal standard compound were established in turn (see Table 1). To obtain the maximum response value of the fragment ions, each compound was calculated through two fragment ions;

[0067] Table 1

[0068]

[0069]

[0070] * represents the quantitative ion pair

[0071] This validation example carried out methodological validation according to the guidelines of the Society of Hair Testing (SoHT) and international guidelines. The evaluation parameters mainly included: selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, matrix effect, and extraction recovery rate, etc.

[0072] ① Selectivity

[0073] Blank hair samples from 8 healthy volunteers were collected for selectivity detection to ensure that there was no interference from other substances within the peak emergence time of the target component;

[0074] Such as Figures 1-13As shown, by comparing the chromatograms of blank hair and the hair with the mass fraction of each target substance at LOQ, it was confirmed that the endogenous substances had no interference on the target substances and the internal standard, and the peak emergence times of each target substance were between 2.96 min and 10.65 min.

[0075] ② LOD and LOQ

[0076] Blank hair was taken and mixed reference substance solution was added to prepare hair samples containing all target substances with different mass fractions; the mass fraction when the signal-to-noise ratio S / N≥3 was taken as the detection limit, and the mass fraction when S / N≥10 was taken as the quantification limit, and the quantification limit was used as the minimum content of the linear range. The precision and accuracy experiments of the hair samples at LOQ were investigated.

[0077] Hair samples with different added contents were investigated separately. The detection limit was selected as the signal-to-noise ratio S / N≥3, and the quantification limit was S / N≥10. The accuracy and precision of the hair samples at LOQ were both within ±20%, as shown in Table 2 and Table 3; the results showed that the detection limit was 0.2 pg / mg to 5 pg / mg, and the quantification limit was 0.5 pg / mg to 10 pg / mg.

[0078] Table 2

[0079]

[0080] Table 3

[0081]

[0082]

[0083] ③ Linearity

[0084] Blank hair was taken and an appropriate amount of mixed reference substance solution was added to obtain hair added samples with different mass fractions; the sample pretreatment and injection analysis were carried out according to the examples; with the mass fraction of the target substance in the hair as the abscissa and the peak area ratio of the target substance to the internal standard as the ordinate, the weighted (W = 1 / x) least squares method was used for regression calculation to obtain the linear equation and calculate the correlation coefficient (R2); the lowest content was LOQ (S / N≥10), and it was ensured that the content of the actual sample fell within the linear range.

[0085] By examining hair samples with mass fractions of 0.5 pg / mg, 1 pg / mg, 2 pg / mg, 5 pg / mg, 10 pg / mg, 20 pg / mg, 50 pg / mg, 100 pg / mg, 200 pg / mg, 500 pg / mg, and 1000 pg / mg, the peak area ratios and contents of the target compounds and internal standards were regressed by weighted (1 / x) least squares method to obtain linear data and correlation coefficient R2. The results showed that each compound in the hair samples had good linearity within the corresponding content range, and R2 was greater than 0.99. The linear data results are shown in Table 2.

[0086] ④ Precision and accuracy

[0087] The precision and accuracy of four contents of LOQ, low, medium, and high were investigated. An appropriate amount of mixed reference solution was added to blank hair. Six samples were taken for each content and continuously determined for 4 days. The samples were pretreated and injected for analysis according to the examples to calculate the within-day precision and accuracy. The accuracy was expressed by bias. The content was calculated linearly, and the percentage of the difference between the value obtained by linear calculation and the added value and the added value was compared. The precision was expressed by relative standard deviation (RSD). It was continuously determined for 4 days and simultaneously determined with the standard curve on the same day. The content of the quality control sample was calculated using the standard curve of the same day to calculate the between-day precision. The precision investigated the changes in repeated experiments on the same day and different days.

[0088] The precision and accuracy of four contents of LOQ, low, medium, and high were investigated. Six samples were taken for each content and continuously determined for four days. The results showed that the within-day precision of each compound was 0.6% - 12.0%, and the between-day precision was 2.7% - 12.0%. The ranges of within-day and between-day accuracy were -10.7% - 12.5% and -4.3% - 7.3% respectively. The results showed that the present invention had good accuracy and precision for the target compounds. The precision and accuracy results are shown in Table 3.

[0089] ⑤ Matrix effect and extraction recovery

[0090] According to the method proposed by Matuszewski et al., the samples were divided into three groups to calculate the extraction recovery and matrix effect. Two contents were used, and eight blank hair samples from different sources were used for each content, and three groups of experiments were carried out respectively. Group I: A certain mass concentration of mixed reference solution was added to eight hair samples from different sources before extraction. Group II: The corresponding mass concentration of mixed reference solution was added to eight hair samples from different sources after extraction. Group III: A mixed reference solution with the corresponding mass concentration was prepared. The samples were injected for analysis according to the examples, and the peak area (A) was recorded. Extraction recovery = AⅠ / AⅡ, Matrix effect = AⅡ / AⅢ.

[0091] The results showed that the extraction recovery rates of the target analytes ranged from 69.3% to 112.2%, and the matrix effects ranged from 33.7% to 157.4%. Among them, methoxymethamphetamine, 2-phenyl-2-(methylamino)cyclohexanone, fluanisone, and 2-(2-fluorophenyl)-2-aminocyclohexanone had obvious ion suppression effects, and the relative standard deviation RSD of hair from different sources was above 20%. Therefore, the influence of ion suppression on other key validation parameters (such as LOD and LOQ) should also be investigated. According to the international standard guidelines, 9 hair matrices from different sources were used to evaluate the LOD and LOQ of these 4 analytes, and the analysis was carried out continuously for 3 days. The results showed that the hair matrices from different sources had no influence on the LOD and LOQ of these 4 analytes. The matrix effect and extraction recovery rate results are shown in Table 4.

[0092] Table 4

[0093]

[0094]

[0095] Application example

[0096] The examples were applied to the hair analysis of 18 LSD users and 2 2-FDCK users.

[0097] When the hair length was greater than 3 cm, segmental hair analysis was performed on 0 cm - 3 cm (S1), 3 cm - 6 cm (S2), and 6 cm - 9 cm (S3); otherwise, the full length was taken for analysis; each hair segment was analyzed 2 times. The results showed that LSD was detected in the hair of all 18 suspected LSD users. The mass fractions of LSD in the hair were <LOQ - 4.0 pg / mg (n = 18, median 1.5 pg / mg), <LOQ - 1.8 pg / mg (n = 8), and <LOQ - 0.6 pg / mg (n = 4) in the 0 cm - 3 cm, 3 cm - 6 cm, and 6 cm - 9 cm segments, respectively. The mass fraction of iso-LSD detected in the S1 segment was <LOQ - 1.4 pg / mg (n = 4), and in the S2 segment was <LOQ (n = 1); DCK, 2-FDCK, and nor-2-FDCK were simultaneously detected in the hair of 2 2-FDCK users.

[0098] In summary, the hair analysis method of the present invention constructs an LC-MS / MS method for quantitatively analyzing 1P-LSD and 2C-B-fly in hair, and constructs an LC-MS / MS method for simultaneously quantitatively analyzing multiple hallucinogens and hallucinogen biomarkers in hair. The pretreatment method is simple and rapid, with one-step extraction. The extraction solution is used to wet-mill the hair at low temperature to extract the target substances. It has a low detection limit, a low quantification limit, high resolution, high sensitivity, and high automation. It can systematically and standardizedly monitor the prevalence of hallucinogen abuse, and at the same time, the analysis results can be used as reference evidence in court.

[0099] The above are only the preferred embodiments of the present invention, and do not limit the implementation manners and protection scope of the present invention. For those skilled in the art, it should be realized that all the equivalent replacements and obvious changes made by using the description and illustrations of the present invention should be included in the protection scope of the present invention.

Claims

1. A hair analysis method for simultaneously detecting multiple hallucinogens and hallucinogen biomarkers, characterized in that, The hallucinogens and hallucinogen biomarkers include: lysergic acid diethylamide, iso-lysergic acid diethylamide, 2-oxo-3-hydroxy-lysergic acid diethylamide, 1-propionyl-lysergic acid diethylamide, mescaline, 2,5-dimethoxy-4-bromophenethylamine, 2-(8-bromo-2,3,6,7-tetrahydrobenzofuro[2,3-f][1]benzofuran-4-yl)-ethylamine, fluanisone, 2-(2-fluorophenyl)-2-aminocyclohexanone, methoxetamine, 2-phenyl-2-(methylamino)cyclohexanone, and methylenedioxypyrovalerone; The steps of the hair analysis method include: cleaning the sample hair with an organic solvent, drying it at room temperature, transferring it to the internal standard working solution, grinding, centrifuging, and microfiltrating it in sequence, and then injecting it into the LC-MS / MS system; where The internal standard working solution is a methanol solution of lysergic acid diethylamide-d3; The chromatographic conditions include: the chromatographic column is an Allure PFPP column; mobile phase A is an ammonium acetate buffer solution containing formic acid and acetonitrile; mobile phase B is acetonitrile; the elution method is isocratic constant flow elution; the column temperature is room temperature; In the mobile phase A, the volume fraction of formic acid is 0.1% - 0.2%, the volume fraction of acetonitrile is 4% - 6%, and the molar concentration of ammonium acetate is 10 mmol / L - 30 mmol / L; the volume ratio of the mobile phase A to the mobile phase B in the eluent is 30:70; the flow rate is 0.2 mL / min - 0.3 mL / min; the elution time is 10 min - 20 min; The mass spectrometry conditions include: the mass spectrometer is an electrospray ionization mass spectrometer; the detection mode is the positive ion mode; the monitoring mode is the multiple reaction monitoring mode; the ion spray voltage is 5000 V - 6000 V; the ion source temperature is 400 °C - 600 °C; the curtain gas pressure is 30 psi - 50 psi, the nebulizing gas pressure is 40 psi - 60 psi, and the auxiliary gas pressure is 40 psi - 60 psi.

2. The hair analysis method according to claim 1, wherein The organic solvent is acetone.

3. The hair analysis method according to claim 1, characterized in that, The conditions for the cleaning include: the number of cleaning times is 2 - 4 times.

4. The hair analysis method according to claim 1, wherein After drying at room temperature, cut the sample hair into segments of 2 mm - 3 mm, and then transfer 20 mg - 30 mg of the accurately weighed sample hair to 300 μL - 500 μL of the internal standard working solution.

5. The hair analysis method according to claim 4, characterized in that, The mass concentration of lysergic acid diethylamide-d3 in the internal standard working solution is 20 ng / mL - 30 ng / mL.

6. The hair analysis method according to claim 1, wherein The conditions for the grinding include: the grinding temperature is -40 °C - -30 °C; the grinding speed is 10 m / s - 20 m / s; the running time is 30 s - 50 s, the residence time is 10 s - 30 s, and the number of cycles is 15 times.

7. The hair analysis method according to claim 1, wherein The conditions for the centrifugation include: the centrifugation speed is 11000×g - 13000×g; the centrifugation time is 2 min - 4 min.

8. The hair analysis method according to claim 1, characterized in that, The conditions for the microfiltration include: the pore size of the filter membrane is 0.2 μm - 0.3 μm.

9. The hair analysis method according to claim 1, characterized in that, The chromatographic conditions also include: the injection volume is 4 μL - 6 μL; the temperature of the autosampler is 3 °C - 5 °C.

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