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Method for measuring phenylephrine concentration by lc-ms/ms and sample pretreatment method

A technology for phenylephrine and samples, which is applied in the field of pretreatment of biological sample analysis, can solve problems such as inability to accurately determine the concentration of phenylephrine, incomplete separation of target compounds and impurities, and high quantitative limits of detection methods, achieving fast and effective The effect of detecting, removing the interference of plasma matrix, and reducing the detection limit

Active Publication Date: 2021-07-30
CHINA RESOURCES SANJIU MEDICAL & PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] For this reason, the technical problem to be solved by the present invention is that the pretreatment method of the sample adopted by the concentration detection method in the prior art does not completely separate the target compound and impurities, which leads to high quantification limit and low sensitivity of the detection method, which cannot be accurate. Determination of the concentration of low-level phenylephrine in plasma samples, thereby providing a pretreatment method for samples of phenylephrine in plasma, especially suitable for the method of LC-MS / MS determination of phenylephrine in plasma

Method used

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  • Method for measuring phenylephrine concentration by lc-ms/ms and sample pretreatment method
  • Method for measuring phenylephrine concentration by lc-ms/ms and sample pretreatment method
  • Method for measuring phenylephrine concentration by lc-ms/ms and sample pretreatment method

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Embodiment 1

[0067] Collection of plasma samples to be tested: 12 subjects were fasted overnight for at least 10 hours the day before the administration, and at 0 min, the subjects took renin Namin tablets (batch number: 20170401, each containing hydrochloric acid to Oxynephrine 5mg), each subject was given one tablet, and the subject’s forearm vein was embedded with an indwelling needle. 1 mL of blood was collected at different time points at 40, 60, 90, 120, 240, 360, and 480 min, and placed in a 2 Put in vacuum blood collection tubes with EDTA anticoagulant, mix well, let stand, centrifuge, take 200 μL of the upper layer of plasma, cover the tube tightly, and store in an ultra-low temperature freezer at -60°C to -90°C.

[0068] The method for measuring the concentration of phenylephrine in plasma by LC-MS / MS described in this embodiment specifically includes the following steps:

[0069] (a) Sample pretreatment

[0070] (1) Preparation of standard curve samples: Accurately weigh pheny...

Embodiment 2

[0085] Collection of plasma samples to be tested: 12 subjects were fasted overnight for at least 10 hours the day before the administration, and at 0 min, the subjects took renin Namin tablets (batch number: 20170301, each containing hydrochloric acid to Oxynephrine 10mg), each subject took one tablet, and the subjects were respectively before administration (0min) and after administration 10, 15, 20, 25, 30, 40, 60, 90, 120, 240, 1 mL of blood was collected at different time points at 360 and 480 min, and placed in a 2 Put in vacuum blood collection tubes with EDTA anticoagulant, mix well, let stand, centrifuge, take 200 μL of the upper layer of plasma, cover the tube tightly, and store in an ultra-low temperature freezer at -60°C to -90°C.

[0086] The method for measuring the concentration of phenylephrine in plasma by LC-MS / MS described in this embodiment specifically includes the following steps:

[0087] (a) Sample pretreatment

[0088] (1) Preparation of standard curv...

Embodiment 3

[0101] Collection of plasma samples to be tested: 12 subjects were fasted overnight for at least 10 hours the day before the administration, and at 0 min, the subjects took renin Namin tablets (batch number: 20170402, each containing hydrochloric acid to Oxynephrine 20mg), each subject took one tablet, and the subjects were respectively before administration (0min) and after administration 10, 15, 20, 25, 30, 40, 60, 90, 120, 240, 1 mL of blood was collected at different time points at 360 and 480 min, and placed in a 2 Put in vacuum blood collection tubes with EDTA anticoagulant, mix well, let stand, centrifuge, take 200 μL of the upper layer of plasma, cover the tube tightly, and store in an ultra-low temperature freezer at -60°C to -90°C.

[0102] The method for measuring the concentration of phenylephrine in plasma by LC-MS / MS described in this embodiment specifically includes the following steps:

[0103] (a) Sample pretreatment

[0104] (1) Preparation of standard curv...

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Abstract

The present invention relates to the technical field of medical testing, in particular to a method for measuring the concentration of phenylephrine by LC-MS / MS, which includes one extraction: adding an internal standard working solution and an extracting solution to the sample, mixing them uniformly and then centrifuging and stratifying , collect the supernatant to obtain the first supernatant; derivation reaction: after the first supernatant is concentrated and dried, add NaHCO 3 The buffer salt is dissolved, and then the acetonitrile solution of dansyl chloride is added to carry out the derivatization reaction; secondary extraction: adding the extract solution to the reaction solution after the derivatization reaction for secondary extraction, centrifuging to obtain the second supernatant, and the second supernatant After the liquid is concentrated and dried, it is redissolved to obtain a sample solution; the sample pretreatment method of the present invention can maximize the extraction rate of the low-content phenylephrine by the pretreatment method, effectively remove the interference of the plasma matrix, and improve the specificity. The minimum detection limit is reduced to 0.02ng / ml, and the sensitivity is increased to 0.1nM, so as to adapt to the detection requirements of low-concentration phenylephrine in drug clinical research.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a method for measuring the concentration of phenylephrine by LC-MS / MS and an applicable pretreatment method for analyzing biological samples. Background technique [0002] Phenylephrine is a catecholamine selective α1 receptor agonist, mainly used to prevent and treat hypotension caused by spinal anesthesia, general anesthesia, application of chlorpromazine, etc. It is also used for supraventricular tachycardia and mydriasis Check, phenylephrine is an over-the-counter drug that can be substituted for pseudoephedrine. [0003] Currently, the phenylephrine-like nasal decongestants used clinically usually need to be administered every four hours to relieve nasal congestion due to the short plasma half-life of the active ingredient phenylephrine in the body. Therefore, in order to delay the release time, improve the therapeutic effect and reduce the side effect...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 陈光磊李黎王勇马鹏岗杨彦佳
Owner CHINA RESOURCES SANJIU MEDICAL & PHARMA
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