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A method for simultaneously measuring the contents of nicotine, 3-(pyrrolidin-2-yl)pyridine, pyridylpyrrolidone and testosterone in serum

A pyridylpyrrolidone and nicotine technology is applied in the detection field to achieve the effects of high sensitivity, good repeatability and wide application range

Active Publication Date: 2020-08-25
WENZHOU VOCATIONAL COLLEGE OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In order to study the relationship between smoking behavior and testosterone secretion, it is necessary to simultaneously detect nicotine, 3-(pyrrolidin-2-yl)pyridine, pyridine pyrrolone and testosterone content. In the prior art, there is no simultaneous determination of serum nicotine in the world. , pyridinepyrrolone, 3-(pyrrolidin-2-yl)pyridine, testosterone content liquid phase tandem mass spectrometry standard

Method used

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  • A method for simultaneously measuring the contents of nicotine, 3-(pyrrolidin-2-yl)pyridine, pyridylpyrrolidone and testosterone in serum
  • A method for simultaneously measuring the contents of nicotine, 3-(pyrrolidin-2-yl)pyridine, pyridylpyrrolidone and testosterone in serum
  • A method for simultaneously measuring the contents of nicotine, 3-(pyrrolidin-2-yl)pyridine, pyridylpyrrolidone and testosterone in serum

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Experimental program
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Effect test

Embodiment 1

[0048]In the embodiment of the present invention, an ultra-high performance liquid phase tandem mass spectrometry method for determining the content of nicotine, 3-(pyrrolidin-2-yl)pyridine, pyridine pyrrolone and testosterone in serum, the steps are as follows:

[0049] 1.1 Preparations:

[0050] 4.1.1 Take 250uL of serum and treat it according to the pretreatment method, and increase the amount of pre-cooled acetonitrile according to the corresponding proportion to obtain more than 1.0mL of the test solution.

[0051] 4.1.2 Use a balance of 1 / 10,000 or 1 / 100,000 to weigh 1.0mL of the liquid to be tested by the difference method, thereby obtaining the density of the liquid to be tested.

[0052] 4.1.3 Repeat at least six times to obtain the average density of the liquid to be tested under the current experimental conditions. If the deviation is large, the stability of the experimental process should be considered.

[0053] 4.1.4 Calculate the approximate volume of the liquid...

Embodiment 2

[0081] Embodiment 2: Method accuracy and repeatability verification - add recovery experiment

[0082] Specific steps:

[0083] 1. Take 8 samples of 100uL serum from the same source, 2 of which are blank samples, and the other 6 samples are added with a certain amount of detection target.

[0084] 2. Add 200 μL of pre-cooled acetonitrile, vortex for 1 min, sonicate for 5 min, and then centrifuge at 10,000 rpm-5°C for 10 min.

[0085] 3. Pipette all the supernatant, add 200 μL of pre-cooled acetonitrile, L, vortex for 1 min, and then centrifuge at 10000 rpm-5 °C for 10 min.

[0086] 4. Pipette all the supernatant, pass through a 0.22 μm filter membrane, and wait for detection by liquid phase tandem mass spectrometry. Take one of the blank samples to be tested and add a certain concentration of the detection target to prepare a matrix standard solution.

[0087] 5. Liquid phase tandem mass spectrometry detection and calculation.

[0088] Using this method for ultra-high perf...

Embodiment 3

[0094] Example 3: Linear range and sensitivity verification (limit of quantification) of the method.

[0095] 1. Take 2 parts of 10.0mL after mixing and centrifuging serum from different sources, add 20mL of pre-cooled acetonitrile respectively, vortex for 1min, sonicate for 5min and then centrifuge at 10000rpm-5℃ for 10min.

[0096] 2. Pipette all the supernatants respectively, add 20 mL of pre-cooled acetonitrile, L, vortex for 1 min, and then centrifuge at 10,000 rpm-5°C for 10 min.

[0097] 3. Pipette all the supernatants separately, mix the two parts and pass through a 0.22 μm filter membrane to make a matrix blank solution.

[0098] 4. Use the matrix blank solution and target standard solution to prepare concentrations of 0.0001ng / mL, 0.0002ng / mL, 0.0005ng / mL, 0.001ng / mL, 0.002ng / mL, 0.005ng / mL, 0.01ng / mL, 0.02ng / mL, 0.05ng / mL, 0.1ng / mL, 0.2ng / mL, 0.5ng / mL, 1.0ng / mL, 2.0ng / mL, 5.0ng / mL, 10ng / mL, 20ng / mL, 50ng / mL mL, 100ng / mL matrix standard solution.

[0099] 5. Liqui...

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Abstract

The invention discloses a method for simultaneously measuring the content of nicotine, 3-(pyrrolidine-2-yl) pyridine, (-)-cotinine and testosterone in serum. The method has the advantages of high simplicity, great rapidity, high detection sensitivity, strong pertinency and wide application range. It is verified that the method has a relatively great recycling property, good linear relation and relatively low quantitation limit for the measurement of the four target objects, can simultaneously detect a plurality of kinds of target objects at one time, and is rapid and efficient. The pretreatment method, liquid phase method and some liquid-mass methods of the method of the invention are also applicable to the analysis of other object objects similar to the above four components in serum, andtherefore, the method of the invention has high applicability.

Description

technical field [0001] The invention belongs to the technical field of detection, and specifically refers to a method for simultaneously measuring the contents of nicotine, 3-(pyrrolidin-2-yl)pyridine, pyridylpyrrolidone and testosterone based on ultra-high performance liquid phase tandem mass spectrometry. Background technique [0002] Nicotine, commonly known as nicotine, is an important component of tobacco, which can cause addiction or dependence. Repeated use of nicotine also increases heart rate and blood pressure and reduces appetite. It is also the primary factor in tobacco product addiction. Nicotine can be metabolized into various substances in the human body, among which 3-(pyrrolidin-2-yl)pyridine and pyridine pyrrolidone are one of its main metabolites. The concentration level of nicotine and its metabolites in the blood is closely related to the exposure level of the human body in smoke, so its concentration is used to evaluate smoking behavior and smoke exposu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 张维一郑明华梁秀美徐静徐仁爱张井谢拾冰张辉林定鹏何如意高晴盈陈静静
Owner WENZHOU VOCATIONAL COLLEGE OF SCI & TECH
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