Method for detecting concentration of psychotropic drugs by liquid-phase mass spectrum solid-phase extraction method

Abstract

The invention belongs to the technical field of drug concentration detection, particularly relates to a method for detecting concentration of psychiatric drugs by a liquid-phase mass spectrum solid-phase extraction method, and provides the following scheme aiming at the problems of matrix interference, low extraction efficiency and narrow application range in the existing psychiatric drug concentration detection process: S1, selecting reagents and materials; an eluent is prepared; s2, collecting a biological sample, and pretreating the biological sample to obtain a clarified sample extracting solution; s3, adding the treated sample extracting solution into a solid-phase extraction column for adsorption; s4, monitoring the adsorption process in real time; according to the method disclosed by the invention, the problems of matrix interference, low extraction efficiency and narrow application range are effectively avoided through a solid-phase extraction technology and a liquid-phase mass spectrum combined method, and an efficient, accurate and reliable analysis means is provided for clinical monitoring and drug research and development of mental drugs.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of drug concentration detection, and particularly relates to a method for detecting concentration of psychotropic drugs by liquid chromatography-mass spectrometry and solid phase extraction. BACKGROUND

[0002] Psychotropic drugs are widely used in clinical applications, and the concentration monitoring of the psychotropic drugs in the human body is of great significance for the evaluation of treatment effect and the guarantee of drug safety. The existing methods for detecting the concentration of psychotropic drugs have certain limitations. For example, although the traditional high performance liquid chromatography can detect some psychotropic drugs, it is easy to be interfered by matrix effect when dealing with complex biological samples, resulting in the decrease of detection sensitivity and accuracy. Some methods based on immune analysis have poor specificity, which may cause cross-reaction and affect the reliability of the detection results. The liquid chromatography-mass spectrometry technology combines the separation ability of liquid chromatography and the high sensitivity detection advantage of mass spectrometry, and becomes a powerful means for drug concentration detection. However, in the actual application, there are still many problems in the sample pretreatment process. For example, the commonly used liquid-liquid extraction method is complicated to operate, large amount of organic solvent is used, and emulsification phenomenon is easy to occur, which affects the subsequent detection efficiency and accuracy. Although the solid phase extraction as a sample pretreatment technology has certain advantages, the existing solid phase extraction method still needs to be improved in the extraction efficiency of psychotropic drugs, and the applicability is not wide enough for different structures and properties of psychotropic drugs, which is difficult to meet the demand of detecting the concentration of multiple psychotropic drugs at the same time.

[0003] The patent document with publication number CN112730686A discloses a method for detecting concentration of psychotropic drugs by liquid chromatography-mass spectrometry and solid phase extraction, which comprises the following steps: A, pretreating the sample to be detected; the pretreatment is specifically performed by the method of solid phase extraction; B, preparing standard samples of different concentrations of psychotropic drugs, then performing liquid chromatography-mass spectrometry detection, and drawing a standard curve; C, performing liquid chromatography-mass spectrometry detection on the sample to be detected obtained after step A, and obtaining the concentration of the psychotropic drugs. The liquid chromatography-mass spectrometry method used in the present application detects the concentration of drugs by the different mass ratios of various drugs, and has high specificity and sensitivity.

[0004] However, the above-mentioned patent document has the problems of matrix interference, low extraction efficiency and narrow application range in the process of detecting the concentration of psychotropic drugs. Therefore, the present application proposes a method for detecting concentration of psychotropic drugs by liquid chromatography-mass spectrometry and solid phase extraction to solve the above-mentioned problems. SUMMARY

[0005] The present application aims to solve the problems of matrix interference, low extraction efficiency and narrow application range in the process of detecting the concentration of psychotropic drugs in the prior art, and proposes a method for detecting concentration of psychotropic drugs by liquid chromatography-mass spectrometry and solid phase extraction.

[0006] The method for detecting the concentration of psychotropic drugs by liquid chromatography-mass spectrometry solid phase extraction provided by the application adopts the following technical scheme:

[0007] The method for detecting the concentration of psychotropic drugs by liquid chromatography-mass spectrometry solid phase extraction comprises the following steps:

[0008] S1: selecting reagents and materials, and configuring an eluent;

[0009] S2: collecting a biological sample and pretreating the biological sample to obtain a clear sample extract;

[0010] S3: adding the pretreated sample extract to a solid phase extraction column for adsorption;

[0011] S4: monitoring the adsorption process in real time;

[0012] S5: eluting the solid phase extraction column after adsorption and collecting the eluent;

[0013] S6: analyzing and monitoring the collected eluent and evaluating the effect;

[0014] S7: injecting the concentrated sample solution into a liquid chromatography-mass spectrometry instrument for detection;

[0015] S8: calculating the accurate concentration of psychotropic drugs in the biological sample and performing data processing and analysis.

[0016] Further, in S1, a filler with high purity and a specific particle size (such as 3-10 μm) is selected, and a mixed mode functional group (such as an ion exchange group and a reversed phase chromatography group) is chemically bonded to the surface of the filler, for example, the filler surface contains carboxylic acid groups (for ion exchange) and C18 hydrophobic chains (for reversed phase adsorption), which can capture the ionized and hydrophobic parts of psychotropic drugs under specific pH conditions, an internal standard (such as a deuterated psychotropic drug) similar in structure to the target drug but different in mass spectrometry characteristic ion is selected, and a 100 μg / mL stock solution is prepared, and the storage temperature is-20℃.

[0017] Further, in S1, a special eluent is configured, which contains the following components: methanol or acetonitrile: 70-90% v / v, used to destroy the hydrophobic interaction between the drug and the filler; ammonium formate: 10 mM, used to adjust the ionic strength; formic acid: 0.1%, used to maintain pH 3.0-4.0 and ensure partial ionization of the drug; surfactant: 0.05%, used to assist in desorption of strongly adsorbed drugs, the eluent is filtered and sterilized (0.22 μm filter membrane) and ultrasonically degassed.

[0018] Further, in S2, the biological sample is collected by a collection mechanism, placed in a centrifuge tube containing an anticoagulant (such as heparin sodium), and stored at a temperature of 4°C to prevent degradation. To 1 mL of the biological sample, 3 mL of acetonitrile (containing 0.1% formic acid) is added, and the mixture is mixed by a vortex oscillator. The supernatant is collected, the vortex oscillator is set to 10,000 rpm, and the centrifugation time is 10 min. The supernatant is filtered through a 0.45 μm filter to obtain a clear sample extract, which is placed in an ice bath for standby. Phosphate buffer (pH 7.0-8.0) is added to the sample extract to adjust the pH of the mixture to 6.5-7.5, ensuring that the drug is in an ionized state suitable for adsorption.

[0019] Further, in S3, 5 mL of methanol and 5 mL of equilibration liquid (such as 0.1% formic acid aqueous solution) are sequentially passed through the extraction column to activate the filler. The pretreated sample extract is slowly added to the solid-phase extraction column at a flow rate of 1-2 mL / min to ensure that the drug molecules are in full contact with the filler for adsorption. The adsorption temperature is 4°C-25°C to avoid high temperature-induced drug degradation or non-specific adsorption.

[0020] Further, in S4, a UV detector is connected to the outlet end of the solid-phase extraction column, and the wavelength is 210-280 nm to monitor the absorbance change of the effluent. When the absorbance drops to the baseline level (indicating that no drug is flowing out), the adsorption process is complete.

[0021] Further, in S5, the column is quickly rinsed with 0.5 mL of pretreatment liquid (such as 5% methanol aqueous solution) to remove weakly adsorbed impurities. 1.5 mL of prepared eluent is slowly injected at a flow rate of 0.5-1 mL / min. In the initial stage, a low flow rate is maintained to fully desorb the drug. According to the UV signal of the effluent, the temperature (25°C-40°C) and flow rate of the eluent are dynamically adjusted to avoid drug peak broadening. Subsection collection (each 0.5 mL as a subsection) is adopted, and the eluent is collected in a labeled centrifuge tube.

[0022] Further, in S6, the drug concentration and purity of each subsection of the eluent are rapidly analyzed by liquid chromatography-mass spectrometry, the pH value (should be 3.0-4.0) and conductivity (<10 mS / cm) of the eluent are measured, the elution efficiency is evaluated, the drug recovery rate (target value >85%) and purity (impurity content <5%) are calculated, and if the recovery rate is low, the eluent composition or flow rate is checked; if the purity is insufficient, the pretreatment step is optimized.

[0023] Further, in S7, the eluent is concentrated to 100 mu L at 40 DEG C with a nitrogen blowing instrument to avoid high temperature damage to the drug, and the liquid chromatography conditions are as follows: a chromatographic column is a C18 column (150 mm*2.1 mm, 3 mu m); a mobile phase is that A phase is 0.1% formic acid aqueous solution, and B phase is acetonitrile; gradient elution is that 0-2 min (20% B), 2-8 min (20%-80% B), a flow rate is 0.3 mL / min; mass spectrometry conditions are that an electrospray ion source (ESI+ mode) is used, a spraying voltage is 3.5 kV; a quantitative ion is selected [M+H]+ ion (such as m / z 300) of a drug molecule; and an internal standard ion is selected characteristic ion (such as m / z 305) of an internal standard molecule.

[0024] Further, in S8, according to the mass spectrometry signal ratio (such as peak area ratio) of the drug to be detected and the internal standard substance, a standard curve equation y=kx+b, R 2 >0.99 is used to calculate the drug concentration in the sample, original data are integrated, corrected and statistically analyzed by using Waters QuanLynx, a detection limit (LOD, signal-to-noise ratio 3:1) and a quantitative limit (LOQ, signal-to-noise ratio 10:1) are evaluated, and it is ensured that the sensitivity of the method meets the clinical requirements.

[0025] In summary, the present application includes at least one of the following beneficial technical effects:

[0026] 1. The present application can effectively improve the extraction efficiency and purity of psychotropic drugs by optimizing the chemical composition and structure of the solid phase extraction column filler and carefully preparing the special eluent, the component design of the eluent can fully destroy the interaction between the drug and the filler, ensure the efficient elution of the drug molecule, and improve the sensitivity and accuracy of the detection;

[0027] 2. The present application can effectively cope with the polarity and binding characteristics of different drugs by adjusting the pH value, ionic strength and organic solvent ratio of the eluent, and realize the simultaneous detection of multiple psychotropic drugs;

[0028] 3. The present application reduces the complexity and error of manual operation by automatically monitoring the adsorption process and dynamically adjusting the elution conditions, the segmented collection and real-time monitoring design of the eluent not only improves the recovery rate and purity of the drug, but also facilitates rapid evaluation and optimization of the elution effect, in addition, the nitrogen blowing concentration step and the automatic analysis function of the liquid chromatography-mass spectrometry instrument further improve the efficiency of the entire detection process, making it more suitable for high-throughput clinical sample analysis.

[0029] The present application effectively avoids the problems of matrix interference, low extraction efficiency and narrow application range by using solid phase extraction technology and liquid chromatography-mass spectrometry method, and provides an efficient, accurate and reliable analysis means for clinical monitoring and drug research and development of psychotropic drugs. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 A flow chart of a method for detecting the concentration of psychotropic drugs by liquid chromatography-mass spectrometry solid phase extraction according to the present application. DETAILED DESCRIPTION

[0031] The technical solutions in the embodiments of the present application will be clearly and completely described with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments.

[0032] Embodiment one

[0033] Reference Figure 1 A method for detecting the concentration of psychotropic drugs by liquid chromatography-mass spectrometry solid phase extraction, comprising the following steps:

[0034] S1: Select a filler with high purity and a specific particle size (such as 3-10 μm), and the surface of the filler is chemically bonded with mixed mode functional groups (such as ion exchange groups and reverse phase chromatography groups). For example, the surface of the filler contains carboxylic acid groups (for ion exchange) and C18 hydrophobic chains (for reverse phase adsorption), which can simultaneously capture the ionized part and the hydrophobic part of psychotropic drugs under specific pH conditions. Select an internal standard (such as a deuterated psychotropic drug) similar in structure to the target drug but different in mass spectrometry characteristic ion, prepare a 100 μg / mL stock solution, store at -20℃, and prepare a special eluent containing the following components: methanol or acetonitrile: 70-90% v / v, used to destroy the hydrophobic interaction between the drug and the filler; ammonium formate: 10 mM, used to adjust the ionic strength; formic acid: 0.1%, used to maintain pH 3.0-4.0 and ensure the ionization of the drug; surfactant: 0.05%, used to assist in desorption of strongly adsorbed drugs. Filter and sterilize the eluent (0.22 μm filter) and ultrasonic degassing;

[0035] S2: Collect biological samples through a collection mechanism, place them in a centrifuge tube containing an anticoagulant (such as heparin sodium), store them at a temperature of 4℃ to avoid degradation, add 3 mL of acetonitrile (containing 0.1% formic acid) to 1 mL of the biological sample, mix well with a vortex oscillator, collect the supernatant, the vortex oscillator speed is 10,000 rpm, the centrifugation time is 10 min, pass the supernatant through a 0.45 μm filter to obtain a clear sample extract, place it in an ice bath for standby, add a phosphate buffer (pH 7.0-8.0) to the sample extract, adjust the pH of the mixture to 6.5-7.5 to ensure that the ionization state of the drug is suitable for adsorption;

[0036] S3: Activate the packing of the column with 5 mL of methanol and 5 mL of equilibration buffer (e.g. 0.1% formic acid in water) at a flow rate of 1-2 mL / min. Slowly inject the pretreated sample extract into the solid phase extraction column, ensuring that the drug molecules are in full contact with the packing for adsorption. The adsorption temperature should be between 4°C and 25°C to avoid drug degradation or non-specific adsorption caused by high temperatures.

[0037] S4: Connect a UV detector to the outlet end of the solid phase extraction column, with a wavelength of 210-280 nm. Monitor the absorbance changes of the effluent. When the absorbance drops to the baseline level (indicating that no more drug is flowing out), the adsorption process is complete.

[0038] S5: Rinse the column with 0.5 mL of pretreatment solution (e.g. 5% methanol in water) to remove weakly adsorbed impurities. Slowly inject 1.5 mL of prepared elution solution at a flow rate of 0.5-1 mL / min. Maintain a low flow rate in the initial stage to fully desorb the drug. Dynamically adjust the temperature (25°C-40°C) and flow rate of the elution solution based on the UV signal of the effluent to avoid drug peak broadening. Use segmented collection (0.5 mL per segment) and collect the elution solution in labeled centrifuge tubes.

[0039] S6: Rapidly analyze each segment of the elution solution for drug concentration and purity using liquid chromatography-mass spectrometry. Measure the pH value (should be 3.0-4.0) and conductivity (<10 mS / cm) of the elution solution. Evaluate the elution efficiency, calculate the drug recovery (target value >85%) and purity (impurity content <5%), and optimize the pretreatment steps if the recovery is low or the purity is insufficient.

[0040] S7: Concentrate the elution solution to 100 μL at 40°C using a nitrogen blow instrument to avoid high-temperature damage to the drug. Liquid chromatography conditions: column: C18 column (150 mm x 2.1 mm, 3 μm); mobile phase: A phase: 0.1% formic acid in water, B phase: acetonitrile; gradient elution: 0-2 min (20% B), 2-8 min (20%-80% B), flow rate 0.3 mL / min; mass spectrometry conditions: electrospray ion source (ESI+ mode), spray voltage 3.5 kV; quantitative ion: select the [M+H]+ ion of the drug molecule (e.g. m / z 300); internal standard ion: select the characteristic ion of the internal standard molecule (e.g. m / z 305);

[0041] S8: Calculate the drug concentration in the sample based on the mass spectrometry signal ratio (e.g. peak area ratio) of the drug to be tested and the internal standard substance, combined with the standard curve equation y=kx+b, R 2 >0.99. Use Waters QuanLynx to integrate, correct, and statistically analyze the raw data to evaluate the detection limit (LOD, signal-to-noise ratio 3:1) and the quantification limit (LOQ, signal-to-noise ratio 10:1), ensuring that the method sensitivity meets clinical requirements.

[0042] Embodiment Two

[0043] With reference to Figure 1 A method for detecting the concentration of psychoactive drugs by liquid chromatography-mass spectrometry solid phase extraction, comprising the following steps:

[0044] S1: selecting reagents and materials, and configuring eluent;

[0045] S2: collecting biological samples and pretreating the biological samples to obtain clear sample extract;

[0046] S3: adding the pretreated sample extract to a solid phase extraction column for adsorption;

[0047] S4: real-time monitoring of the adsorption process;

[0048] S5: eluting the adsorbed solid phase extraction column and collecting the eluent, using a gradient elution program to gradually increase the proportion of organic solvents in the eluent or change the pH value of the eluent to achieve simultaneous elution of multiple psychoactive drugs in complex samples, and through gradient elution, different drug components with different elution behaviors can be effectively separated and collected, thereby improving the universality and application range of the method;

[0049] S6: analyzing and monitoring the collected eluent, and evaluating the effect;

[0050] S7: injecting the concentrated sample solution into a liquid chromatography-mass spectrometry instrument for detection;

[0051] S8: calculating the accurate concentration of psychoactive drugs in the biological sample, and performing data processing and analysis.

[0052] Embodiment Three

[0053] With reference to Figure 1 A method for detecting the concentration of psychoactive drugs by liquid chromatography-mass spectrometry solid phase extraction, comprising the following steps:

[0054] S1: selecting reagents and materials, and configuring eluent;

[0055] S2: collecting biological samples and pretreating the biological samples to obtain clear sample extract;

[0056] S3: adding the pretreated sample extract to a solid phase extraction column for adsorption;

[0057] S4: real-time monitoring of the adsorption process;

[0058] S5: Elute the solid phase extraction column after adsorption, collect the eluent, and add a specific ion strength regulator (such as a sodium salt or an ammonium salt) to the eluent to affect the electrostatic interaction between the drug and the solid phase extraction column by adjusting the ion strength of the eluent, thereby optimizing the elution effect, especially for psychoactive drugs with ionized functional groups;

[0059] S6: Analyze and monitor the collected eluent, and evaluate the effect;

[0060] S7: Inject the concentrated sample solution into a liquid chromatography-mass spectrometry instrument for detection;

[0061] S8: Calculate the accurate concentration of psychoactive drugs in the biological sample, and perform data processing and analysis.

[0062] Example Four

[0063] Reference Figure 1 A method for detecting the concentration of psychoactive drugs by liquid chromatography-mass spectrometry solid phase extraction, comprising the following steps:

[0064] S1: Select reagents and materials, and prepare an eluent;

[0065] S2: Collect biological samples and pretreat the biological samples to obtain a clear sample extract;

[0066] S3: Add the pretreated sample extract to a solid phase extraction column for adsorption;

[0067] S4: Monitor the adsorption process in real time;

[0068] S5: Elute the solid phase extraction column after adsorption, collect the eluent, and adjust the temperature of the eluent using a temperature control device, and elute at a higher temperature (such as 40°C to 60°C) to improve the elution efficiency, especially for psychoactive drugs that are tightly bound to the filler at room temperature. At the same time, the effect of temperature change on drug stability needs to be verified, and a suitable elution temperature range needs to be selected;

[0069] S6: Analyze and monitor the collected eluent, and evaluate the effect;

[0070] S7: Inject the concentrated sample solution into a liquid chromatography-mass spectrometry instrument for detection;

[0071] S8: Calculate the accurate concentration of psychoactive drugs in the biological sample, and perform data processing and analysis.

[0072] Example Five

[0073] Reference Figure 1A method for detecting the concentration of psychotropic drugs by liquid chromatography-mass spectrometry solid phase extraction, comprising the following steps:

[0074] S1: Select reagents and materials, and configure eluent;

[0075] S2: Collect biological samples and pretreat the biological samples to obtain clear sample extract;

[0076] S3: Add the pretreated sample extract to a solid phase extraction column for adsorption;

[0077] S4: Real-time monitoring of the adsorption process;

[0078] S5: Elute the solid phase extraction column after adsorption and collect the eluent, using a stepwise elution method, first using a low-strength eluent (such as a low-concentration organic solvent) for preliminary elution, collecting the first part of the eluent; then using a high-strength eluent (such as a high-concentration organic solvent or a solution containing a specific additive) for secondary elution, collecting the second part of the eluent. By adjusting the strength of the eluent, different polar or different binding strength of psychotropic drug components can be enriched;

[0079] S6: Analyze and monitor the collected eluent, and evaluate the effect;

[0080] S7: Inject the concentrated sample solution into a liquid chromatography-mass spectrometry instrument for detection;

[0081] S8: Calculate the accurate concentration of psychotropic drugs in the biological sample, and perform data processing and analysis.

[0082] Experimental example

[0083] I. Experimental purpose: To verify the performance advantages of the liquid chromatography-mass spectrometry solid phase extraction method (hereinafter referred to as "new method") compared to traditional detection methods in detecting the concentration of psychotropic drugs, including extraction efficiency, sensitivity, accuracy, anti-matrix interference ability and application range;

[0084] II. Experimental samples: Collect plasma samples containing risperidone, quetiapine and sertraline, set multiple concentration gradients (low, medium and high), and each concentration sample quantity is not less than 10, and parallel control;

[0085] III. Experimental method

[0086] According to the operation steps of the liquid chromatography-mass spectrometry solid phase extraction method described in the document, the following experimental groups and control groups are set:

[0087] (I) Experimental group

[0088] The experimental group uses the five example methods in the document for detection, as follows:

[0089] Example One: using mixed-mode packing and special eluent;

[0090] Example Two: using gradient elution procedure;

[0091] Example Three: adding ion strength adjuster in eluent;

[0092] Example Four: adjusting eluent temperature using temperature control device;

[0093] Example Five: using stepwise elution method;

[0094] (B) Control group

[0095] The control group corresponds to the five examples respectively, and the detection is carried out after deleting the innovation points of each example:

[0096] Control group one: deleting mixed-mode packing and special eluent, using traditional single-mode packing and ordinary eluent;

[0097] Control group two: deleting gradient elution procedure, using fixed elution conditions;

[0098] Control group three: deleting ion strength adjuster, using basic eluent;

[0099] Control group four: elution at room temperature, without adjusting eluent temperature;

[0100] Control group five: using single strength eluent, without stepwise elution;

[0101] Four, detection index

[0102] The detection index includes peak area, peak purity, recovery rate, matrix effect, detection limit and quantitative limit;

[0103] Five, experimental data

[0104]

[0105]

[0106] Six, data analysis

[0107] Through the above experimental data comparison table, the advantages of liquid chromatography-mass spectrometry solid phase extraction method compared with traditional high performance liquid chromatography in detecting the concentration of psychoactive drugs can be directly shown, such as shorter detection time, higher peak purity and recovery rate, smaller matrix effect, wider application range and lower quantitative limit, etc., so as to verify the effectiveness and superiority of the method.

[0108] The above merely describes preferred specific embodiments of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art, according to the technical solution and inventive concept of the present application, makes equivalent replacement or change within the technical range disclosed by the present application, which should be covered within the protection scope of the present application.

Claims

1. A method for detecting the concentration of a psychotropic drug by liquid phase mass spectrometry solid phase extraction, characterized by: The method comprises the following steps: S1: selecting reagents and materials, and preparing eluent; S2: collecting biological samples and pretreating the biological samples to obtain clear sample extract; S3: adding the pretreated sample extract to a solid-phase extraction column for adsorption; S4: monitoring the adsorption process in real time; S5: eluting the solid-phase extraction column after adsorption and collecting the eluent; S6: analyzing and monitoring the collected eluent and evaluating the effect; S7: injecting the concentrated sample solution into a liquid chromatography-mass spectrometry instrument for detection; S8: calculating the accurate concentration of the psychotropic drug in the biological sample and performing data processing and analysis.

2. The method according to claim 1, wherein the method is characterized by: In S1, a filler with high purity and a specific particle size range is selected, the surface of which is chemically bonded with a mixed mode functional group, and an internal standard with a different mass spectrometry characteristic ion but a similar structure to the target drug is selected, a stock solution of 100 μg / mL is prepared, and the storage temperature is-20℃.

3. The method according to claim 2, wherein the method is characterized by: In S1, a special eluent is prepared, which contains the following components: methanol or acetonitrile: 70-90% v / v, which is used to destroy the hydrophobic interaction between the drug and the filler; ammonium formate: 10 mM, which is used to adjust the ionic strength; formic acid: 0.1%, which is used to maintain pH 3.0-4.0 and ensure partial ionization of the drug; surfactant: 0.05%, which is used to assist in desorption of strongly adsorbed drugs, the eluent is filtered and sterilized, and ultrasonic degassing is performed.

4. The method according to claim 3, wherein the method is characterized by: In S2, the biological sample is collected by a collection mechanism and placed in a centrifuge tube containing an anticoagulant, which is stored at a temperature of 4℃ to prevent degradation, 3 mL of acetonitrile is added to 1 mL of the biological sample, which is mixed by a vortex oscillator, the supernatant is collected, the vortex oscillator rotates at a speed of 10,000 rpm, and the centrifugation time is 10 min, the supernatant is filtered through a 0.45 μm filter to obtain a clear sample extract, which is placed in an ice bath for standby, and phosphate buffer is added to the sample extract to adjust the pH of the mixture to 6.5-7.5 to ensure that the ionization state of the drug is suitable for adsorption.

5. The method according to claim 4, wherein the method is characterized by: In S3, 5 mL of methanol and 5 mL of equilibrium liquid are sequentially passed through the extraction column to activate the filler, and the pretreated sample extract is slowly added to the solid-phase extraction column at a flow rate of 1-2 mL / min and an adsorption temperature of 4℃-25℃ to avoid high temperature-induced drug degradation or non-specific adsorption.

6. The method according to claim 5, wherein the method is characterized by: In S4, a UV detector is connected to the outlet end of the solid-phase extraction column, the wavelength is 210-280 nm, and the absorbance change of the effluent is monitored.

7. The method according to claim 6, wherein the method is characterized by: In S5, the column is quickly rinsed with 0.5 mL of pretreatment liquid to remove weakly adsorbed impurities, 1.5 mL of prepared eluent is slowly injected, the flow rate is 0.5-1 mL / min, the initial stage maintains a low flow rate to fully desorb the drug, the temperature and flow rate of the eluent are dynamically adjusted according to the UV signal of the effluent to avoid broadening of the drug peak, and the eluent is collected in a marked centrifuge tube by using a segmented collection method.

8. The method according to claim 7, wherein the method is characterized by: In S6, the drug concentration and purity of each eluent segment are rapidly analyzed by a liquid chromatography-mass spectrometry instrument, the pH value and conductivity of the eluent are measured, the elution efficiency is evaluated, and the drug recovery rate and purity are calculated.

9. The method according to claim 8, wherein the method is a liquid chromatography-mass spectrometry solid phase extraction method for detecting the concentration of psychotropic drugs. In S7, the eluate was concentrated to 100 μL at 40 °C using a nitrogen blow down apparatus to avoid drug destruction by high temperature.

10. The method according to claim 9, wherein the method is a liquid chromatography-mass spectrometry solid phase extraction method for detecting the concentration of psychotropic drugs. In the S8, according to the mass spectrum signal ratio of the drug to be tested and the internal standard substance, combined with the standard curve equation y=kx+b, R 2 >0.99, calculate the drug concentration in the sample, use Waters QuanLynx to integrate, correct and statistically analyze the original data, evaluate the detection limit and the quantification limit, and ensure that the method sensitivity meets the clinical needs.

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