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Method for simultaneously detecting 16 drugs and metabolites thereof in blood by UPLC-MS/MS (ultra performance liquid chromatography-mass spectrometry/mass spectrometry) method

A metabolite and drug technology, applied in the field of drug detection, can solve the problems of occupying multiple instruments, increasing the detection time, and increasing the amount of blood collection, so as to reduce the use of internal standards, reduce the detection time, and ensure the detection accuracy.

Active Publication Date: 2022-07-22
BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the other hand, when multiple methods are required for detection, pretreatment and standard curve preparation also need to double the time, and it will take up multiple instruments and require a larger space
At the same time, for some patients, different types of drugs are taken at the same time. If different types of blood drug concentrations need to be detected, multiple methods are required for measurement, which will increase the detection time and require more blood collection.

Method used

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  • Method for simultaneously detecting 16 drugs and metabolites thereof in blood by UPLC-MS/MS (ultra performance liquid chromatography-mass spectrometry/mass spectrometry) method
  • Method for simultaneously detecting 16 drugs and metabolites thereof in blood by UPLC-MS/MS (ultra performance liquid chromatography-mass spectrometry/mass spectrometry) method
  • Method for simultaneously detecting 16 drugs and metabolites thereof in blood by UPLC-MS/MS (ultra performance liquid chromatography-mass spectrometry/mass spectrometry) method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] This embodiment provides a method for preparing a standard solution, including preparing a standard stock solution, preparing a standard working solution, preparing an internal standard stock solution, preparing an internal standard working solution, and preparing a standard solution. The specific steps are as follows:

[0054] (1) Prepare standard working solution

[0055] A. The standard stock solutions corresponding to the 16 drugs and their metabolites are diluted with diluent to obtain the standard product intermediate solution. The concentrations of the standard product stock solution and standard product intermediate solution are shown in Table 4;

[0056] Table 4

[0057] substance name Standard stock solution concentration (μg / mL) Standard intermediate concentration (μg / mL) methotrexate 1943 1080 Rosuvastatin 2274 41 Risperidone 3719 54 9-Hydroxyrisperidone 1800 54 Olanzapine 4811 65 perphenazine 1479 7 ...

Embodiment 2

[0072] This embodiment provides a method for sample detection using the instrument AB SCIEX; the model specification is: AB SCIEX Jasper HPLC MSTRIPLE QUAD 4500MD;

[0073] The mobile phases of the analytical chromatographic column used were: phase A: water (0.1% formic acid + 5 mmol / L ammonium formate), phase B: methanol (0.1% formic acid + 5 mmol / L ammonium formate), and the analytical column was washed with gradient The removal mode, the chromatographic condition parameters are shown in Table 7:

[0074] Table 7

[0075]

[0076] For mass spectrometry conditions, the parameters are shown in Table 8-9:

[0077] Table 8

[0078] parameter set value parameter set value CUR 20L / min TEM 550℃ CAD 9 L / min GS1 50L / min IS 5000 V GS2 80L / min

[0079] Table 9

[0080]

[0081]

[0082] The retention time (peak time) of each target in the chromatogram obtained in Example 2 is shown in Table 10:

[0083] Table 10

[0084] ...

Embodiment 3

[0087] The present embodiment provides a method for preparing a sample to be tested from serum blood collection tubes, EDTA blood collection tubes, and lithium heparin blood collection tubes, as follows:

[0088] (1) Take at least 2 mL of blood from the serum blood collection tube, EDTA blood collection tube or lithium heparin blood collection tube to be tested, centrifuge at a centrifugal speed of 3500 r / min for 10 min, and take the supernatant to obtain serum or plasma, which is placed in the Store in a freezer at -20°C until use before analysis;

[0089] (2) Pipette 10 μL of the internal standard working solution into a 1.5 mL centrifuge tube, then add 100 μL of serum or plasma, add 400 μL of methanol, vortex and mix at 2500 r / min for 5 min, 14000 Centrifuge at high speed at r / min for 10 min, transfer 50 µL of the supernatant to a 1.5 mL plastic centrifuge tube, add 100 µL of ultrapure water, vortex at 2500 r / min for 1 min, and take 150 µL of the supernatant to obtain the s...

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Abstract

The invention relates to a method for simultaneously detecting 16 drugs and metabolites thereof in blood through a UPLC-MS / MS method. The sixteen medicines and metabolites thereof comprise methotrexate, olanzapine, risperidone, 9-hydroxyrisperidone, amisulpride, ziprasidone, perphenazine, gliridazine, sertindole, chlorprothixol, rosuvastatin, theophylline, iscitalopram or citalopram, maprotiline, bupropion, hydroxybupropion and azithromycin. The method comprises the following steps: determining a standard curve, processing a blood sample to be detected, and detecting the sample to be detected by using a UPLC-MS / MS method. The detection method provided by the invention can be used for rapidly, accurately and simultaneously detecting the contents of 16 medicines and metabolites thereof in blood. A protein precipitation method is adopted for sample pretreatment, the used blood amount is small, and the treatment method is simple and easy to implement. The detection method is wide in linear range, short in analysis time, capable of analyzing 10 groups of samples within one hour, high in sensitivity and good in reproducibility.

Description

technical field [0001] The invention relates to the technical field of drug detection, in particular to a method for simultaneously detecting 16 drugs and their metabolites in blood by a UPLC-MS / MS method. Background technique [0002] Therapeutic drug monitoring (TDM) is a pharmacy clinical discipline that studies the mechanisms, techniques, methods and clinical standards of individualized drug treatment, and transforms the research results into clinical treatment to maximize rational drug use. Under the guidance of the principle of pharmacokinetics (PK), the concentration of drugs and their metabolites in the body fluids of patients after treatment is determined quantitatively by modern analytical techniques, and the relationship between them and the curative effect is analyzed, and individual drug regimens are adjusted so as to improve drug Efficacy, reduce toxic and side effects, and maximize drug treatment cost savings through rational drug use. In the TDM detection in...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72G01N30/06
CPCG01N30/02G01N30/72G01N30/06G01N2030/027
Inventor 任晓航贾永娟刘春冉赵金宝倪君君
Owner BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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