CN115728413A - Quantitative Analysis Method of Sertraline Plasma Concentration - Google Patents

Abstract

The invention discloses a quantitative analysis method for sertraline blood drug concentration, which adopts a two-dimensional liquid chromatography system and comprises the following steps: Step 1, preparation of a solution, the solution includes a test solution prepared from the blood plasma of a patient and sertraline standard solution; step 2, setting chromatographic parameters, the one-dimensional column is a solid phase extraction column, the one-dimensional eluent includes methanol and water, and the one-dimensional elution method is gradient elution; the two-dimensional column is a chromatographic column, The two-dimensional eluent includes acetonitrile and potassium dihydrogen phosphate solution, and the two-dimensional elution method is isocratic elution; Step 3, implement detection, inject standard solutions of different concentrations into the two-dimensional liquid chromatography system to draw a standard curve and Obtain a regression equation, inject the test solution into the two-dimensional liquid chromatography system, and use the obtained regression equation to calculate the content of sertraline in the test solution. The invention has the advantages of simple operation, strong specificity, high sensitivity, wide linear range, high accuracy and the like.

Description

Quantitative Analysis Method of Sertraline Plasma Concentration

technical field

The invention relates to the technical field of medical blood testing, in particular to a quantitative analysis method for sertraline blood drug concentration.

Background technique

Depression is a common affective disorder mental illness, characterized by high incidence, high recurrence rate and high burden rate. At present, sertraline is a relatively commonly used drug for the treatment of depression in the medical field. However, due to the large individual differences in the pharmacokinetics of sertraline in the human body, the difference between the drug concentration and efficacy of sertraline in the patient's body is relatively large. The action relationship is of great significance for guiding the clinical rational use of drugs.

At present, the methods for detecting blood drug concentration of sertraline are mainly high-performance liquid chromatography (HPLC) and immunoassay (IM). These methods have good specificity, and are convenient and efficient. The method is relatively general, and although it can effectively detect various antidepressants, it still has the following defects, for example: the detection sensitivity of sertraline is poor and cannot meet the clinical requirements; meanwhile, the current sertraline is available in The operation of the concentration detection is very complicated, and there are many interfering substances, so the detection of the blood drug concentration cannot be carried out very well.

Contents of the invention

Based on the above defects in the prior art, the present application proposes a highly targeted quantitative analysis method for sertraline blood concentration, which can quickly, accurately and efficiently detect the sertraline content in human blood samples.

The technical scheme adopted in the invention is: a quantitative analysis method for sertraline blood drug concentration, using a two-dimensional liquid chromatography system to detect sertraline blood drug concentration, and the detection method includes the following steps: step 1, preparation of the solution, Described solution comprises the need testing solution that is made by the blood plasma of the patient who is undergoing sertraline treatment and the standard solution that contains sertraline; The eluent includes methanol and water, and the one-dimensional elution method is gradient elution; the two-dimensional column is a column filled with octadecylsilane bonded silica gel, and the two-dimensional eluent includes acetonitrile and phosphoric acid at a concentration of 0.01M Potassium dihydrogen solution, the two-dimensional elution mode is isocratic elution; Step 3, implement detection, first inject the described standard solution of different concentrations into the two-dimensional liquid chromatography system to draw a standard curve and obtain a regression equation, and then The test solution is injected into the two-dimensional liquid chromatograph system, and a typical spectrogram is recorded, and the obtained regression equation is used to calculate the content of sertraline in the test solution.

Further, the gradient elution program of the one-dimensional elution is: 0min to 7.0min, the flow rate of the one-dimensional eluent is 0.2mL/min, the volume ratio of methanol: water is 1:9; 7.0min to 7.5min , the flow rate of the one-dimensional eluent increased to 1.5mL/min, and the volume ratio of methanol:water became 8:2; from 7.5min to 10.0min, the flow rate of the one-dimensional eluent was maintained at 1.5mL/min, methanol:water The volume ratio of the eluent was maintained at 8:2; from 10.0min to 10.2min, the flow rate of the one-dimensional eluent was maintained at 1.5mL/min, and the volume ratio of methanol: water was changed to 1:9; from 10.2min to 11.0min, the flow rate of the one-dimensional eluent The flow rate of dehydration is maintained at 1.5mL/min, and the volume ratio of methanol:water is maintained at 1:9; from 11.0min to 11.5min, the flow rate of one-dimensional eluent is reduced to 0.2mL/min, and the volume ratio of methanol:water is maintained At 1:9; from 11.5 min to 15 min, the flow rate of the one-dimensional eluent was maintained at 0.2 mL/min, and the volume ratio of methanol: water was maintained at 1:9.

Further, the gradient elution program of the one-dimensional elution also includes: from 0 min to 7.0 min, the column temperature of the one-dimensional column is maintained at 20±0.5°C; from 7.0 min to 7.5 min, the column temperature of the one-dimensional column rises to 25±0.5°C; from 7.5min to 10.0min, the column temperature of the one-dimensional column was maintained at 25±0.5°C; from 10.0min to 10.2min, the column temperature of the one-dimensional column continued to rise to 30±0.5°C; from 10.2min to 11.0 min, the column temperature of the one-dimensional column is maintained at 30±0.5°C; from 11.0min to 11.5min, the column temperature of the one-dimensional column is maintained at 30±0.5°C; from 11.5min to 15min, the column temperature of the one-dimensional column is maintained at 30°C ±0.5°C.

Further, the isocratic elution program of the two-dimensional elution is: 0min-15min, the flow rate of the two-dimensional eluent is 1.0mL/min, and the volume ratio of acetonitrile: potassium dihydrogen phosphate is 2:3.

Further, during one-dimensional elution, inject water into the one-dimensional eluent for dilution, and the water injection dilution procedure is: 0min-3min, the flow rate of water is 1.2mL/min; 3min-3.1min, the flow rate of water drops to 0.1 mL/min; 3.1min-14.5min, the flow rate of water is maintained at 0.1mL/min; 14.5min-14.8min, the flow rate of water rises to 1.2mL/min; 14.8min-15min, the flow rate of water is maintained at 1.2mL/min .

Preferably, the temperature of the one-dimensional column and the two-dimensional column is 35°C.

Further, the one-dimensional column is an MFPh-1S-5 fixed extraction column with a specification of 4.0mmL.D.×20mm; the two-dimensional column is an AcclaimTM120C18 column with a specification of 250×4.6mm and 5 μm.

Further, the injection volume of the two-dimensional liquid chromatography system is 200 μL.

Further, the potassium dihydrogen phosphate solution contains 5% triethylamine, and phosphoric acid is used to adjust the pH value of the potassium dihydrogen phosphate solution to 3.5.

Further, the two-dimensional liquid chromatography system includes an ultraviolet detector, and the wavelength of the ultraviolet detector is 210nm.

Compared with the prior art, the quantitative analysis method of sertraline blood drug concentration proposed by the present invention has the advantages of simple operation, strong specificity, high sensitivity, wide linear range, high accuracy and high reproducibility. The detachment method can realize the enrichment and quantitative analysis of sertraline in a very short time, which improves the efficiency of the hospital in monitoring the plasma concentration of sertraline in patients, reduces the time for clinical diagnosis, and greatly saves time and cost. Achieve precision medicine.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the accompanying drawings that need to be used in the descriptions of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only of the present invention. For some embodiments, those of ordinary skill in the art can also obtain other drawings based on these drawings without paying creative efforts.

Fig. 1 is the standard curve figure of standard solution in the present invention;

Fig. 2 is a typical spectrogram of standard solution in the present invention;

Fig. 3 is a typical spectrogram of blank plasma solution in the present invention;

Fig. 4 is a typical spectrogram of system suitability solution in the present invention;

Fig. 5 is typical spectrogram of standard curve in the present invention;

Fig. 6 is the typical spectrogram of need testing solution repeatability experiment among the present invention.

Detailed ways

In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

The invention provides a quantitative analysis method for sertraline blood drug concentration. In the detection method, a two-dimensional liquid chromatography system is used to detect the blood drug concentration of sertraline, and the blood drug concentration of sertraline is quantitatively analyzed , the specific detection steps include:

Step 1. Prepare the solution:

1. Prepare sertraline standard solution

Transfer all the 10mg sertraline standard substance to a 100ml volumetric flask, dilute with methanol, and finally set the volume to 100ml to obtain a sertraline standard substance mother solution with a concentration of 0.1mg/ml. Sertraline standard solutions with different concentrations were obtained by diluting the mother solution of the standard product according to the requirement.

2. Prepare blank plasma solution

Take 2 mL of normal human blank plasma, add protein precipitant to dilute to 10 mL, mix and sonicate for 5 min, transfer the above solution to an EP tube, then place it in a centrifuge and centrifuge at 10,000 r/min for 10 min, and take the supernatant , the obtained supernatant was filtered through a nylon filter membrane (0.22 μm), and the filtered filtrate was used as a blank plasma solution.

3. Preparation of system suitability solution

Dilute to obtain a sertraline standard solution with a concentration of 2ug/ml, accurately take 40ul of a sertraline standard solution with a concentration of 2ug/ml, then add 2ml of plasma, mix well, dilute to 10ml with acetonitrile, mix and sonicate for 5min ; The sample in the 10ml volumetric flask is transferred to the EP tube of 10ml, then placed in a centrifuge with 10000r/min and centrifuged for 10min, the supernatant is taken, and the obtained supernatant is filtered through a nylon filter membrane (0.22 μm) , and the filtrate obtained by filtration can be used as the system suitability solution.

4. Prepare the test solution

Take 2mL to 10mL of plasma from a patient undergoing sertraline treatment into a volumetric flask, add a protein precipitant to mix, dilute to volume, mix and sonicate for 5 minutes, transfer the above solution to an EP tube, and spin in a centrifuge at 10000r /min centrifugal 10min, get the supernatant and filter it through a nylon filter membrane (0.22 μm), and the filtrate obtained by filtering is the test solution.

Step 2. Set chromatographic parameters

Set the parameters according to the characteristics of sertraline. Setting the chromatographic parameters includes setting the test conditions, including: setting the parameters of the one-dimensional column chromatography conditions on the two-dimensional liquid chromatography system, the parameters of the two-dimensional column chromatography conditions and the large-volume process. parameters of sample conditions and parameters of valve switching conditions.

The two-dimensional liquid chromatograph system used in this embodiment is the ThermoUltimate3000SD system, and the system includes a dual-gradient standard pump DGP-3600SD with an online degassing unit, an auxiliary pump, an autosampler (WPS-3000SL), and a The column thermostat (TCC-3000RS) and detector (model DAD-3000) of the six-way valve, the chromatographic software adopts Chromeleon ChromatographyDataSystem7.2; The position of the intermediate chromatographic column in the flow channel realizes the storage function of the intermediate chromatographic column. The one-dimensional eluent is driven by the one-dimensional pump to pass through the one-dimensional column to realize the enrichment and concentration of the analyte. The intercepted part after separation is temporarily stored by The two-dimensional pump is transported to the two-dimensional column. The two-dimensional column separates the analyte and detects it through the detector. The detector displays the corresponding typical spectrum on the computer through signal transmission and conversion, and the analyte is qualitative and quantitative.

Further, the filling material of the one-dimensional column in this application requires good hydrophilicity, because blood plasma contains blood cells and macromolecular proteins, which requires chromatographic purification and separation with molecular exclusion and chromatographic distribution. Modes exist at the same time, combined with the interaction between sertraline and filler materials, in this embodiment, the one-dimensional column is preferably a MFPh-1S-5 fixed extraction column, and its specific specification is 4.0mmL.D.×20mm.

Further, in view of the compound structure of sertraline, the surface of the filler material matrix of the two-dimensional column is required to have good hydrophobicity, the two-dimensional column requires excellent double-end capping technology, and the residual silanol on the surface of the silica gel does not exceed 1%. According to the thermodynamic and kinetic characteristics of sertraline in the two-dimensional column, sertraline has a better resolution. In this embodiment, the preferred packing agent for the two-dimensional column is octadecylsilane-bonded silica gel. The chromatographic column, the specific two-dimensional column is ChromCoreTM120C18 chromatographic column, the specification is 250×4.6mm, 5μm, this chromatographic column has excellent stability and durability, and has a long service life.

The working principle of the above-mentioned two-dimensional elution is: the large-volume sample first flows through the one-dimensional column to enrich at the top of the column, and then passes through the one-dimensional eluent to elute the macromolecular protein components and other impurities in the plasma sample. The slit valve flows into a 2D column for separation and quantification.

Moreover, since the sample solvent contains a high proportion of organic solvents, this application also uses a large-volume pump to dilute with water to ensure that the measured substance is enriched on the one-dimensional column and does not flow out, and an auxiliary pump is designed to concentrate on the one-dimensional column. Purified water is injected during the purification to fully retain the components to be measured in the one-dimensional column, which can avoid the phenomenon of broadening and tailing of the chromatographic peak shape during large-volume injection, and effectively improve the problem of peak asymmetry.

Further, the one-dimensional eluent used in the one-dimensional elution in this application includes two mobile phases, which are mobile phase A methanol and mobile phase B water, and the one-dimensional elution is performed in a gradient elution manner. For forest extraction, the specific procedure of the gradient elution is: 0min to 7.0min, the flow rate of one-dimensional eluent is 0.2mL/min, the volume ratio of methanol: water is 1:9; 7.0min to 7.5min, one-dimensional The flow rate of the eluent is increased to 1.5mL/min, and the volume ratio of methanol:water becomes 8:2; from 7.5min to 10.0min, the flow rate of the one-dimensional eluent is maintained at 1.5mL/min, and the volume ratio of methanol:water Maintained at 8:2; from 10.0min to 10.2min, the flow rate of the one-dimensional eluent was maintained at 1.5mL/min, and the volume ratio of methanol: water became 1:9; from 10.2min to 11.0min, the flow rate of the one-dimensional eluent The flow rate was maintained at 1.5mL/min, and the volume ratio of methanol:water was maintained at 1:9; from 11.0min to 11.5min, the flow rate of the one-dimensional eluent decreased to 0.2mL/min, and the volume ratio of methanol:water was maintained at 1: 9; From 11.5 min to 15 min, the flow rate of the one-dimensional eluent was maintained at 0.2 mL/min, and the volume ratio of methanol: water was maintained at 1:9. By eluting sertraline in the above way, sertraline can be extracted and enriched in a very short time, which can greatly shorten the analysis cycle of sertraline, and the effect of extraction and purification is good, which can effectively improve sertraline Lin's peak shape increases the sensitivity of detection.

Furthermore, in a specific embodiment, when carrying out one-dimensional gradient elution, the present application further cooperates with temperature changes to improve the extraction and separation effect of sertraline. In this embodiment, a temperature gradient change and The combination of gradient elution procedures is used to optimize the extraction and separation efficiency of sertraline, so that the detection efficiency and test results of sertraline are optimal. The specific combination of temperature gradient and gradient elution procedures is as follows: From 0min to 7.0min, the flow rate of the one-dimensional eluent is 0.2mL/min, and the volume ratio of methanol: water is 1:9. At this time, the column temperature of the one-dimensional column is maintained at 20±0.5°C; from 7.0min to 7.5min, The flow rate of the one-dimensional eluent increases to 1.5mL/min, the volume ratio of methanol: water becomes 8:2, and the column temperature of the one-dimensional column rises to 25±0.5°C; 7.5min to 10.0min, one-dimensional elution The flow rate of the liquid was maintained at 1.5mL/min, the volume ratio of methanol: water was maintained at 8:2, and the column temperature of the one-dimensional column was maintained at 25±0.5°C; from 10.0min to 10.2min, the flow rate of the one-dimensional eluent was maintained at At 1.5mL/min, the volume ratio of methanol:water becomes 1:9, and the column temperature of the one-dimensional column continues to rise to 30±0.5°C; from 10.2min to 11.0min, the flow rate of the one-dimensional eluent is maintained at 1.5mL/min, the volume ratio of methanol: water is maintained at 1:9, and the column temperature of the one-dimensional column is maintained at 30±0.5°C; from 11.0min to 11.5min, the flow rate of the one-dimensional eluent is reduced to 0.2mL/min, The volume ratio of methanol:water is maintained at 1:9, and the column temperature of the one-dimensional column is maintained at 30±0.5°C; from 11.5min to 15min, the flow rate of the one-dimensional eluent is maintained at 0.2mL/min, methanol:water The volume ratio of the column was maintained at 1:9, and the column temperature of the one-dimensional column continued to be maintained at 30±0.5°C. In order to avoid the problem of band broadening due to the temperature difference between the one-dimensional mobile phase and the one-dimensional column, it is also necessary to preheat the one-dimensional mobile phase in advance in this embodiment, so that the temperature of the one-dimensional mobile phase can be compared with that of the one-dimensional mobile phase. The temperature of the one-dimensional column is similar, so that the problem of peak broadening is well avoided. The gradient elution program cooperates with the gradually rising temperature gradient of the one-dimensional column, which shortens the analysis time of sertraline exponentially, and at the same time maintains a very good The analysis effect has greatly improved the detection efficiency of serum drug concentration of sertraline.

In another embodiment, the temperature of the one-dimensional column and the two-dimensional column in this application can also be set at normal temperature of 35° C., and the column efficiency can be better guaranteed at this temperature.

Further, during one-dimensional elution, inject water into the one-dimensional eluent for dilution, and the water injection dilution procedure is: 0min-3min, the flow rate of water is 1.2mL/min; 3min-3.1min, the flow rate of water drops to 0.1 mL/min; 3.1min-14.5min, the flow rate of water is maintained at 0.1mL/min; 14.5min-14.8min, the flow rate of water rises to 1.2mL/min; 14.8min-15min, the flow rate of water is maintained at 1.2mL/min . In this way, it can be well ensured that the analyte is enriched on the one-dimensional column and does not flow out, so that the analyte in the one-dimensional column can be fully retained.

Further, the two-dimensional eluent used in the two-dimensional elution in the present application includes two-dimensional mobile phase A acetonitrile and two-dimensional mobile phase B potassium dihydrogen phosphate solution, wherein the two-dimensional mobile phase B potassium dihydrogen phosphate The concentration of the solution is 0.01M, and the two-dimensional elution adopts isocratic elution to separate sertraline. The isocratic elution program of the two-dimensional elution is: 0min-15min, and the flow rate of the two-dimensional eluent is The volume ratio of acetonitrile: potassium dihydrogen phosphate solution is 1.0mL/min, the volume ratio of acetonitrile: potassium dihydrogen phosphate solution is 2:3, the setting of the mobile phase ratio and flow rate in the above-mentioned two-dimensional elution can realize the rapid separation of sertraline, and improve the sertraline detection efficiency.

Preferably, in order to ensure the separation effect of sertraline, the potassium dihydrogen phosphate solution in the present application also contains 5% triethylamine, and phosphoric acid is used to adjust the pH value of the potassium dihydrogen phosphate solution to 3.5. The measured sertraline is less affected by impurities, and the peak shape of sertraline is good, which can realize the accurate determination of sertraline.

The one-dimensional eluent in this application flows through the one-dimensional column with a temperature range of 30°C-40°C at a flow rate of 0.2mL/min-1.5mL/min. With a flow rate of 1.5mL/min and a one-dimensional column temperature within 35°C-45°C, sertraline in the purified sample can be quickly enriched. Preferably, the temperature of the one-dimensional column in this embodiment is 40°C.

The gradient elution program of the above one-dimensional eluent is: 0min-7min, the flow rate is 0.2mL/min, the volume ratio of one-dimensional mobile phase A methanol: one-dimensional mobile phase B water is 1:9; 7min-7.5min, The flow rate was increased to 1.5mL/min, the volume ratio of one-dimensional mobile phase A methanol: one-dimensional mobile phase B water was changed to 8:2; 7.5min-11.0min, the flow rate was maintained at 1.5mL/min, one-dimensional mobile phase A The volume ratio of methanol: one-dimensional mobile phase B water is maintained at 8:2; 11.0min-11.2min, the flow rate is maintained at 1.5mL/min, and the volume ratio of one-dimensional mobile phase A methanol: one-dimensional mobile phase B water is changed to 1 : 9; 11.2min-12.0min, the flow rate is maintained at 1.5mL/min, the volume ratio of one-dimensional mobile phase A methanol: one-dimensional mobile phase B water is maintained at 1:9; 12.0min-12.5min, the flow rate drops to 0.2mL /min, the volume ratio of one-dimensional mobile phase A methanol: one-dimensional mobile phase B water is changed to 1:9; 12.5min-15min, the flow rate is maintained at 0.2mL/min, one-dimensional mobile phase A methanol: one-dimensional mobile phase B The volume ratio of water was maintained at 1:9.

Preferably, the injection volume of the two-dimensional liquid chromatography system in this application is 200 μL.

Preferably, the two-dimensional liquid chromatography system in the present application includes an ultraviolet detector, and the wavelength of the ultraviolet detector is 210nm.

Step 3. Implement detection

After setting the system parameters, the detection work can be carried out. In this step, the need testing solution and the standard solution of different volumes need to be injected into the two-dimensional liquid chromatograph system respectively, and a typical spectrum is recorded; Draw a standard curve in the system and obtain a regression equation, then inject the test solution into the two-dimensional liquid chromatograph system, and record the typical spectrum, and use the obtained regression equation to calculate the sertraline in the test solution. content.

The specific operation method in this embodiment is: according to the mother solution of the sertraline standard substance, standard solutions of different concentrations are injected five times, and the mass concentrations of the five injections are 5 ng/mL, 10 ng/mL, and 15 ng/mL in turn. , 20ng/mL and 30ng/mL, record the typical spectrum for 15min, and draw a standard curve with the mass concentration (abscissa) and chromatographic peak area (ordinate) of sertraline to obtain the relevant linear equation, which is used to calculate the The content of sertraline in the test solution. The standard curve diagram drawn by it is shown in Figure 1, wherein the equation of the standard curve is Y=0.0079X+0.0067, the linear correlation coefficient R=0.9975, and the corresponding chromatographic peak area can be calculated by bringing the test solution into the standard curve , through the formula: concentration of sertraline in the blood sample = peak area of sertraline in the test product * concentration of sertraline standard * dilution factor / peak area of sertraline standard, the measured plasma concentration can be obtained The concentration of sertraline in the samples.

Step 4. Methodological verification

1. Verification method specificity

In order to test the blank plasma solution, standard solution, and system suitability solution in advance, compare the measured results with the requirements in the method description. If the measured results meet the requirements, the subsequent test product test Analysis can then be performed. As shown in Figure 2 to 4, concentration is the typical spectrogram of sertraline standard solution of 20ng/mL in this application, the typical spectrogram of blank plasma solution, the typical spectrogram of system suitability solution; Analysis typical spectrogram shows that the blank plasma solution There is no sertraline interference peak in the typical spectrum, the resolution of the impurity peak closest to sertraline in the typical spectrum of the system suitability solution is greater than 1.5, and the retention time of sertraline in the typical spectrum of the two standard solutions is It is consistent, and it is verified that the two-dimensional liquid chromatography system and parameter settings have strong specificity and good applicability, and subsequent detection and analysis of the test product can be carried out.

As shown in Figures 1 and 5, it is a standard curve figure in the embodiment of the present invention, which is obtained by using a standard curve of different concentrations. After measuring, this method has a good linear relationship when the concentration of Traserline is 20ng/mL-120ng/mL, and can Meet the needs of clinical testing.

2. Verification of the detection precision of the test solution

In order to prove the precision of the method in this example, a repetitive experiment was carried out, that is, five parts of the system suitability solution of the same volume were taken for testing, and five injections of the system suitability solution were injected. The typical spectrum obtained is shown in Figure 6 , the test results are shown in Table 1

Table 1. Repeatability experiment test results table

Retention time/min Peak area/mAU*min first stitch 12.420 0.2334 second stitch 12.420 0.2505 third stitch 12.430 0.2459 fourth stitch 12.440 0.2378 fifth stitch 12.433 0.2314 RSD/% 0.07 3.40

According to the above table, the result is that the relative standard deviation of the sertraline content of the obtained 5 parts of test solution is 0.07%, and the number of impurities in the typical spectrograms is basically the same.

3. Verify the accuracy of the detection method

For recovery experiments, configure system suitability solutions with low, medium and high concentrations, such as sertraline system suitability solutions with three concentrations of 8ng/mL, 15ng/mL and 30ng/mL, each concentration system suitability solution Take five needles for sample detection, take 200uL for each needle, and get the spectrum through detection, obtain the peak area according to the spectrum, and calculate the actual concentration value in the system suitability solution according to the calculation formula, and calculate the recovery rate; the concentration is 8ng /mL system suitability solution data for illustration, the specific data of the system suitability solution with a concentration of 5ng/mL is shown in Table 2:

Table 2, recovery rate experimental result table

According to the above table, the average recovery rate of the system suitability solution with a concentration of 8 ng/mL in this example is 99.5%, and the system suitability solutions with a concentration of 15 ng/mL and 30 ng/mL are respectively detected by the above method and the average recovery rate is calculated. Recovery rate (since the detection methods are all the same, so omitted, here you can directly refer to the detection of 8ng/mL), the average recovery rate range of this application is 95%-108% after testing, so the detection method of this application conforms to the bioanalysis methodology Verification required.

In summary, the method proposed by the present invention has extremely strong specificity, is very suitable for the detection of blood drug concentration of sertraline, and the method has high sensitivity, high accuracy and high precision, and the detection method is very reliable , and the linear range is wide, which can meet the clinical monitoring of sertraline blood concentration, and the detection time does not exceed 15 minutes, which can greatly improve the detection efficiency of sertraline blood concentration.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention within.

Claims (10)

1. A quantitative analysis method for blood concentration of sertraline is characterized in that a two-dimensional liquid chromatograph system is adopted for detecting the blood concentration of sertraline, and the detection method comprises the following steps:

preparing a solution comprising a test sample solution prepared from plasma of a patient undergoing sertraline treatment and a standard solution containing sertraline;

setting chromatographic parameters, wherein the one-dimensional column is a solid phase extraction column, the one-dimensional eluent comprises methanol and water, and the one-dimensional elution mode is gradient elution; the two-dimensional column is a chromatographic column with a filler of octadecylsilane chemically bonded silica, the two-dimensional eluent comprises acetonitrile and a potassium dihydrogen phosphate solution with the concentration of 0.01M, and the two-dimensional elution mode is isocratic elution;

and step three, performing detection, namely injecting the standard solutions with different concentrations into a two-dimensional liquid chromatograph system to draw a standard curve and obtain a regression equation, injecting the test solution into the two-dimensional liquid chromatograph system, recording a typical spectrogram, and calculating the content of sertraline in the test solution by using the obtained regression equation.

2. The method for quantitative analysis of blood concentration of sertraline as claimed in claim 1, wherein said one-dimensional elution is performed by a gradient elution procedure comprising: 0min to 7.0min, the flow rate of the one-dimensional eluent is 0.2mL/min, methanol: the volume ratio of water is 1; 7.0min to 7.5min, the flow rate of the one-dimensional eluent is increased to 1.5mL/min, and the ratio of methanol: the volume ratio of water was changed to 8; 7.5min to 10.0min, the flow rate of the one-dimensional eluent is maintained at 1.5mL/min, methanol: the volume ratio of water is maintained at 8; 10.0min to 10.2min, the flow rate of the one-dimensional eluent was maintained at 1.5mL/min, methanol: the volume ratio of water becomes 1; 10.2min to 11.0min, the flow rate of the one-dimensional eluent was maintained at 1.5mL/min, methanol: the volume ratio of water is maintained between 1 and 9; from 11.0min to 11.5min, the flow rate of the one-dimensional eluent was reduced to 0.2mL/min, methanol: the volume ratio of water is maintained at 1; 11.5min to 15min, the flow rate of the one-dimensional eluent was maintained at 0.2mL/min, methanol: the volume ratio of water is maintained at 1.

3. The method of claim 2, wherein the one-dimensional elution gradient further comprises: 0min to 7.0min, wherein the column temperature of the one-dimensional column is maintained at 20 +/-0.5 ℃; the temperature of the one-dimensional column rises to 25 +/-0.5 ℃ from 7.0min to 7.5 min; 7.5min to 10.0min, and maintaining the column temperature of the one-dimensional column at 25 +/-0.5 ℃; the temperature of the one-dimensional column is continuously increased to 30 plus or minus 0.5 ℃ from 10.0min to 10.2 min; the column temperature of the one-dimensional column is maintained at 30 plus or minus 0.5 ℃ for 10.2min to 11.0 min; keeping the column temperature of the one-dimensional column at 30 +/-0.5 ℃ for 11.0min to 11.5 min; the column temperature of the one-dimensional column is continuously maintained at 30 +/-0.5 ℃ from 11.5min to 15 min.

4. The method for quantitative analysis of blood concentration of sertraline according to claim 1, wherein said isocratic elution procedure of two-dimensional elution is: 0min-15min, the flow rate of the two-dimensional eluent is 1.0mL/min, and the ratio of acetonitrile: the volume ratio of the potassium dihydrogen phosphate is 2.

5. The method for quantitative analysis of blood concentration of sertraline according to claim 1, wherein during one-dimensional elution, water is injected into the one-dimensional eluent for dilution, and the water injection and dilution procedures are as follows: 0min-3min, the flow rate of water is 1.2mL/min;3min-3.1min, the flow rate of the water is reduced to 0.1mL/min;3.1min-14.5min, and the flow rate of water is kept at 0.1mL/min; the flow rate of water is increased to 1.2mL/min after 14.5min to 14.8 min;14.8min-15min, and the flow rate of water is kept at 1.2mL/min.

6. The method of claim 1 wherein the temperature of the one-dimensional column and the two-dimensional column is 35 ℃.

7. A method for quantitative analysis of blood concentration of sertraline according to claim 1, wherein said one-dimensional column is MF Ph-1S-5 fixed extraction column, specification is 4.0mm l.d. x 20mm; the two-dimensional column is an Acclaim 120C18 chromatographic column with the specification of 250 multiplied by 4.6mm and 5 mu m.

8. The method of claim 1, wherein the sample injection volume of the two-dimensional liquid chromatograph system is 200 μ L.

9. A method for quantitative analysis of blood concentration of sertraline according to claim 1, wherein said potassium dihydrogen phosphate solution contains 5% triethylamine, and pH of the potassium dihydrogen phosphate solution is adjusted to 3.5 with phosphoric acid.

10. The method of claim 1 wherein the two-dimensional liquid chromatograph system comprises an ultraviolet detector having a wavelength of 210nm.

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