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Enrichment detection method and application of five antitumor drugs in a blood sample

A technology of anti-tumor drugs and detection methods, which is applied in the application field of therapeutic drug concentration monitoring, can solve the problems of inability to achieve accurate measurement of samples, inconsistent detection methods and standards, long detection time of liquid quality analysis, etc., to avoid matrix effect , Good impurity removal effect, good sensitivity effect

Active Publication Date: 2021-03-26
大连润生康泰医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] So far, most of these methods are aimed at the development of detection methods for one structural type of antineoplastic drugs, or for taxanes, or for TKIs, or for methotrexate. A method that can simultaneously detect methotrexate, The detection methods of the three structural types of taxanes and TKIs; at the same time, the detection methods and standards of these methods are not uniform. This increases the workload and increases the waste of experimental resources, and it is impossible to achieve accurate determination of high-throughput samples.

Method used

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  • Enrichment detection method and application of five antitumor drugs in a blood sample
  • Enrichment detection method and application of five antitumor drugs in a blood sample
  • Enrichment detection method and application of five antitumor drugs in a blood sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Establishment and verification of the enrichment detection method for five antitumor drugs in plasma

[0041] 1. Materials and reagents

[0042] Methotrexate (Solarbio, purity>99%), paclitaxel and docetaxel (purchased from China Institute for the Control of Pharmaceutical and Biological Products), imatinib (MACKUN, purity>99%), gefitinib (J&K, purity 99%), D8-imatinib (cmass, purity 99%), D3-methotrexate (cmass, purity 99.4%); formic acid (HPLC grade, purity ≥ 96%, sigma), ethanol, KOH, methanol (chromatographically pure, purity 99.9%, Merck), water (self-made ultrapure water).

[0043] 2. Instruments and equipment

[0044] AB SCIEX 4500MD high performance liquid chromatography-tandem mass spectrometer, the detector is a triple quadrupole tandem mass spectrometer; one ten-thousandth electronic analytical balance; MiliQ pure water instrument; solid phase extraction 96-well plate device; vortex mixer.

[0045] 3. Preparation of standard curve plasma samples ...

Embodiment 2

[0085] Example 2: Detection of the concentration of methotrexate in plasma samples of clinical patients

[0086] 1. Materials and reagents: the same as in Example 1.

[0087] 2. Apparatus and equipment: the same as in Example 1.

[0088] 3. Operation steps for establishing standard curve

[0089] According to the operation of item 3 and item 4 in Example 1, after processing the standard curve plasma sample, according to the conditions of item 5 in Example 1, carry out HPLC-MS / MS analysis, according to the peak area As of each drug and the internal standard The ratio Y (Y=As / Ai) of the peak area Ai carries out weighted linear regression to the drug concentration X, and fits the standard curve, and the weight is 1 / X 2 (X is the concentration value), the standard curve represented by Y=slope*X+intercept.

[0090] 4. Detection of methotrexate concentration in plasma samples

[0091] Take 100 μL of plasma sample in a 1.5 mL tube, add 10 μL of 2% KOH solution (prepared in 95% et...

Embodiment 3

[0095] Example 3: Investigation on the extraction recovery of five drugs with different solid-phase extraction fillers

[0096] Take 5 blank plasma samples from different sources, prepare medium-concentration plasma QC samples according to item 3 in Example 1, and use 4 kinds of SPE chromatographic cartridges with solid phase extraction fillers C18, SAX, C18 SAX, and Phenyl, all with a specification of 100 mg / 1mL for enrichment, according to the method of extraction recovery in item 6 "methodological evaluation" in Example 1, and measure the extraction recovery.

[0097] Table 8 Investigation of the extraction recovery (REC) of five drugs with different solid-phase extraction packing materials

[0098]

[0099]

[0100] It can be seen from Table 7 that the extraction of C18 and Phenyl to methotrexate is low and unstable, and the extraction effect on paclitaxel, docetaxel, imatinib and gefitinib is better; SAX has a better effect on the extraction of paclitaxel, Docetax...

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Abstract

The invention discloses an enrichment detection method and application of five antitumor drugs in a blood sample. The method comprises the following steps: adjusting the pH value of the blood sample by using an alkaline solution; enriching a target object by adopting a mixed mode filler with C8 (C18) and anion exchange capacity, then using liquid chromatography-tandem mass spectrometry for analysis, and establishing a standard curve by an internal standard method to calculate the concentration of the drugs in the blood sample, wherein the concentrations of anti-tumor drugs related to three structural types can be simultaneously determined. The method has the advantages of high flux, quickness, high extraction recovery rate and no matrix effect, meets the quantitative requirements of various mass spectrometric devices, and can be applied to blood concentration monitoring of the five drugs clinically.

Description

technical field [0001] The invention belongs to the technical field of inspection and analysis, and in particular relates to an enrichment detection method for five antitumor drugs including methotrexate, paclitaxel, docetaxel, imatinib and gefitinib, and the method is used in the monitoring of therapeutic drug concentration. in the application. [0002] technical background [0003] Antineoplastic drug therapy is one of the commonly used treatments for cancer. Most antineoplastic drugs have low therapeutic index, many adverse reactions, and large differences in pharmacokinetics among individuals. At the same time, combined use of antineoplastic drugs is common, and there may be drug-drug interactions that affect drug absorption and metabolism. [0004] Therapeutic drug monitoring (TDM) is guided by the principle of pharmacokinetics, analyzes and measures the concentration of drugs in the blood, and individualizes clinical dosage regimens to improve efficacy and avoid or red...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/08G01N30/30G01N30/32G01N30/34G01N30/72
CPCG01N30/02G01N30/08G01N30/30G01N30/32G01N30/34G01N30/72G01N2030/324
Inventor 曹云峰刘明莉王璐张继民王爽曹冉
Owner 大连润生康泰医学检验实验室有限公司
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