Liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs

Abstract

The invention discloses a liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs, relates to the field of analysis and detection, and solves the problem that a traditional mass spectrometry analysis method can only detect a small number of drugs and needs multiple times of analysis. The method solves the problems that in the prior art, the detection response of a useful ultraviolet detector for psychotropic drugs is weak, pretreatment with a gas chromatography-mass spectrometry analysis method is complex, and a volatile toxic solvent is used, and comprises the following steps: S1, preparing a blank sample: mixing methanol and water; s2, preparation of a test solution: adding the blank sample in the step S1 into a to-be-detected sample, centrifuging, taking supernate, and filtering to obtain the to-be-detected test solution. 55 kinds of positive and negative ions can be scanned at the same time, whether the sample contains 55 kinds of target drugs or not can be judged after one-time detection is carried out for five minutes, and the accuracy is high. The method can more efficiently and quickly separate and detect psychotropic drugs in blood, and has the advantages of simplicity, high specificity and sensitivity.

Description

Technical Field

[0001] The present invention relates to the field of analytical detection, and particularly relates to a liquid chromatography-mass spectrometry combined detection method for simultaneously detecting multiple drugs. Background Art

[0002] Traditional mass spectrometry analysis methods can usually only detect a few drugs and require multiple analyses. At the same time, psychotropic drugs have weak detection responses when detected by ultraviolet detectors, and the pretreatment of gas chromatography-mass spectrometry combined analysis methods is complex and volatile toxic solvents are used. For this reason, we have proposed a liquid chromatography-mass spectrometry combined detection method for simultaneously detecting multiple drugs. Summary of the Invention

[0003] In order to solve the above deficiencies in the prior art, the purpose of the present invention is to provide a liquid chromatography-mass spectrometry combined detection method for simultaneously detecting multiple drugs, which has the advantages of simplicity, strong specificity, high sensitivity, and high accuracy.

[0004] The technical solution adopted by the present invention to solve its technical problems is as follows: A liquid chromatography-mass spectrometry combined detection method for simultaneously detecting multiple drugs, comprising the following steps:

[0005] S1: Prepare a blank sample: Mix methanol and water.

[0006] S2: Prepare a test sample solution: Add the blank sample in S1 to the sample to be detected, centrifuge and take the supernatant for filtration, and then obtain the test sample solution to be detected.

[0007] S3: Prepare a sample solution: Take the test sample solution in S2, add a blank sample for dilution and quantification to make a sample solution.

[0008] S4: Liquid chromatography-mass spectrometry combined detection: The chromatographic column is a C18 column with a particle size of 1.7 μm and a size of 2.1*50 mm, using 0.1% formic acid aqueous solution as mobile phase A and methanol as mobile phase B, and performing gradient elution detection on the sample solution in S3.

[0009] In at least some embodiments, the volume ratio of methanol to water in step S1 is set to 1:9.

[0010] In at least some embodiments, the concentration of the test sample solution in S2 is 0.01 μg / ml - 10 μg / ml.

[0011] In at least some embodiments, positive and negative ions are simultaneously scanned in S4.

[0012] In at least some embodiments, the flow rate in S4 is 0.4 - 0.6 ml per minute.

[0013] In at least some embodiments, the column temperature of the chromatographic column in S4 is set to 30 - 50 °C.

[0014] In at least some embodiments, the sample injection volume in S4 is 1 μl.

[0015] The present invention can simultaneously scan a total of 55 positive and negative ions. It only takes five minutes for a single detection to determine whether 55 target drugs are contained in a sample, and it can more efficiently and quickly separate and detect psychotropic drugs in blood. The present invention has the advantages of simplicity, strong specificity, high sensitivity, and high accuracy. Detailed implementation manners

[0016] The following further elaborates on the present application in conjunction with embodiments. It can be understood that the specific embodiments described herein are only used to explain the related invention and not to limit the invention.

[0017] It should be noted that, without conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The following will describe the present application in detail with reference to the embodiments.

[0018] To make the objectives, technical solutions, and advantages of the embodiments of the present disclosure clearer, the technical solutions of the embodiments of the present disclosure will be clearly and completely described below. Obviously, the described embodiments are some but not all of the embodiments of the present disclosure. All other embodiments obtained by those of ordinary skill in the art based on the described embodiments of the present disclosure without creative efforts fall within the scope of protection of the present disclosure.

[0019] Unless otherwise defined, the technical terms or scientific terms used herein shall have the ordinary meanings understood by those of ordinary skill in the art to which the present disclosure pertains. The terms "first", "second", and similar terms used in the specification and claims of the present patent application of the present disclosure do not indicate any order, quantity, or importance, but are only used to distinguish different components. The terms "including" or "comprising" and similar terms mean that the elements or items appearing before "including" or "comprising" cover the elements or items listed after "including" or "comprising" and their equivalents, and do not exclude other elements or items. The terms "connected" or "coupled" and similar terms are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. The terms "upper", "lower", "left", "right", etc. are only used to represent relative positional relationships, and when the absolute position of the object being described changes, the relative positional relationships may also change accordingly.

[0020] In the embodiments of the present disclosure, the term "proximal end" refers to the side close to the operator, the term "distal end" refers to the side far from the operator, and the term "plurality" refers to two or more.

[0021] The present invention discloses a liquid chromatography - mass spectrometry detection method for simultaneously detecting multiple drugs, comprising the following steps:

[0022] S1: Prepare a blank sample: Mix methanol and water;

[0023] S2: Prepare a test sample solution: Add the blank sample in S1 to the sample to be detected, centrifuge, and take the supernatant for filtration, then obtain the test sample solution to be detected;

[0024] S3: Prepare a sample solution: Take the test sample solution in S2, add the blank sample for dilution and quantification to make a sample solution;

[0025] S4: Liquid chromatography - mass spectrometry detection: Use a C18 reverse - phase chromatography column with a particle size of 1.7 um and a size of 2.1 * 50 mm. Use 0.1% formic acid aqueous solution as mobile phase A and methanol as mobile phase B, and then perform gradient elution detection on the sample solution in S3.

[0026] According to an embodiment of the present disclosure, in step S1, the volume ratio of methanol to water is set to 1:9.

[0027] According to an embodiment of the present disclosure, the concentration of the test sample solution in S2 is 0.01 ug / ml - 10 ug / ml.

[0028] According to an embodiment of the present disclosure, positive and negative ions are simultaneously scanned in S4.

[0029] According to an embodiment of the present disclosure, the flow rate in S4 is 0.4 - 0.6 ml per minute.

[0030] According to an embodiment of the present disclosure, the column temperature of the chromatography column in S4 is set to 30 - 50 °C.

[0031] According to an embodiment of the present disclosure, the injection volume in S4 is preferably 1 ul, which can effectively avoid oversaturation or insufficient response of some drugs.

[0032] Among them, the mass spectrometry conditions in step S4 are as follows:

[0033] Ionization mode: ESI+

[0034] Spray voltage: 3 kv

[0035] Desolvation gas: Nitrogen, 1000 L / Hr, 500 °C

[0036] Cone - hole gas: Nitrogen, 50 L / Hr

[0037] Source temperature: 150 °C

[0038] Acquisition mode: Multiple reaction monitoring (MRM)

[0039] Collision gas: argon, pressure 3.5x10-3 mBar

[0040] For example, according to the embodiments of the present disclosure, in step S4, a specific pair of precursor ions and product ions can be selected as quantitative ions, and at the same time, one or more specific pairs of precursor ions and product ions can be selected as qualitative ions. The mass spectrometry parameters of the target substance are shown in the following table (monitoring ion pairs, reference values of declustering potential (DP) and collision energy (CE) for each compound):

[0041]

[0042]

[0043]

[0044] In the present invention, all detected drugs have obvious peak shapes, and the mass spectrometry detection method of the present invention can effectively integrate the peak areas.

[0045] The above description is only the preferred embodiments of the present application and the description of the applied technical principles. Those skilled in the art should understand that the scope of the invention involved in the present application is not limited to the technical solutions formed by the specific combination of the above technical features, and should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the inventive concept. For example, the technical solutions formed by mutually replacing the above features with the (but not limited to) technical features with similar functions disclosed in the present application.

[0046] Except for the technical features described in the specification, the remaining technical features are well-known to those skilled in the art. To highlight the innovative features of the present invention, the remaining technical features are not described herein again.

Claims

1. A liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs, characterized in that, It includes the following steps: S1: Configure a blank sample: Mix methanol and water; S2: Prepare a test sample solution: Add the blank sample in S1 to the sample to be detected, centrifuge, take the supernatant for filtration, and then obtain the test sample solution to be detected; S3: Prepare a sample solution: Take the test sample solution to be detected in S2, dilute it with a blank sample, and quantitatively prepare a sample solution; S4: Liquid chromatography - mass spectrometry detection: Use a C18 column with a particle size of 1.7 μm and a size of 2.1 * 50 mm as the reversed - phase chromatography column. Use 0.1% formic acid aqueous solution as mobile phase A and methanol as mobile phase B. Inject the sample solution in S3, and then perform gradient elution detection.

2. The liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs according to claim 1, wherein In the step S1, the volume ratio of methanol to water is set to 1:

9.

3. A liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs according to claim 1, characterized in that, In S2, the concentration of the test sample solution is 0.01 μg / ml - 10 μg / ml.

4. The liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs according to claim 1, wherein In S4, positive and negative ions are scanned simultaneously.

5. The liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs according to claim 1, wherein In S4, the flow rate is 0.4 - 0.6 ml per minute.

6. The liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs according to claim 1, characterized in that, In S4, the column temperature of the chromatography column is set to 30 - 50 °C.

7. A liquid chromatography-mass spectrometry detection method for simultaneously detecting multiple drugs according to claim 1, characterized in that, In S4, the injection volume is 1 μl.

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