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Method for separating and determining pyrazine substances and pyridine substances in saliva

A technology of pyrazines and pyridines, which is applied in the field of analytical chemistry, can solve the problems of unreported determination of pyrazines and pyridines, and achieve the effects of simple operation, low detection limit and feasible method

Active Publication Date: 2016-09-07
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there have been a large number of reports on the determination of pyrazines and pyridines in samples such as flavors and fragrances, food, liquor, and smoke. However, the determination of pyrazines and pyridines in saliva samples has not been reported yet.

Method used

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  • Method for separating and determining pyrazine substances and pyridine substances in saliva
  • Method for separating and determining pyrazine substances and pyridine substances in saliva
  • Method for separating and determining pyrazine substances and pyridine substances in saliva

Examples

Experimental program
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Embodiment 1

[0034] A method for separating and measuring pyrazines and pyridines in the saliva of tobacco smokers, comprising the following steps:

[0035] Step (1), sample collection: put 0.2 g of absorbent cotton in the mouth of tobacco smokers, leave it for 0.3 min, take it out, put it in the syringe barrel, push and squeeze it with the syringe push rod, and collect the extruded liquid in a centrifuge tube, Saliva samples were obtained by centrifugation at 10000 rpm for 10 min.

[0036] Step (2), extraction: Pipette 2.0 mL of saliva sample into a centrifuge tube, add internal standard pyridine-D5 until the internal standard concentration is 2.0 mg / L; pass through a silica gel solid-phase extraction column with a filler of 300 mg, first add 6 mL of dichloromethane, 6 mL of methanol and 10 mL of deionized water were pre-washed sequentially; the sample was transferred to the pre-washed solid phase extraction column, and the flow rate was controlled at 0.5 drops / s, and 6 mL of dichlorometh...

Embodiment 2

[0045] A method for separating and measuring pyrazines and pyridines in the saliva of liquor drinkers, comprising the following steps:

[0046] Step (1), sample collection: 1.5 g of absorbent cotton was placed in the mouth of tobacco smokers for 4.0 min, then taken out, placed in the syringe barrel, pushed and squeezed with the syringe push rod, the extruded liquid was collected in a centrifuge tube, Saliva samples were obtained by centrifugation at 15000 rpm for 12 min.

[0047] Step (2), extraction: Pipette 10.0 mL of saliva sample into a centrifuge tube, add internal standard pyridine-D5 until the internal standard concentration is 1.0 mg / L; pass through a 600 mg HLB solid-phase extraction column, first add 10 mL of dichloromethane, 10 mL of methanol and 12 mL of deionized water were pre-washed sequentially; the sample was transferred to the pre-washed solid phase extraction column, and the flow rate was controlled at 0.25 drops / s, and 10 mL of dichloromethane was added for...

Embodiment 3

[0055] A method for separating and measuring pyrazines and pyridines in saliva, comprising the following steps:

[0056] Step (1), sample collection: put 0.2 g of absorbent cotton in the mouth for 0.2 minutes, then take out the absorbent cotton that has absorbed saliva, put it in the syringe barrel, push and squeeze it with the syringe push rod, and collect the extruded liquid in a centrifuge tube In the process, the supernatant is collected after high-speed centrifugation to obtain a saliva sample; the centrifugation speed is not lower than 10,000 rpm, and the centrifugation time is not less than 10 min;

[0057] Step (2), extraction: pipette 0.5 mL of saliva sample, add internal standard pyridine-D5 to the saliva until the internal standard concentration is 1 mg / L, and then transfer to a pre-washed solid-phase extraction column with 200 mg filler , control the flow rate not greater than 0.5 drops / s, the pyrazines and pyridines are adsorbed by the solid phase extraction colum...

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Abstract

The invention relates to a method for separating and determining pyrazine substances and pyridine substances in saliva, and belongs to the technical field of analytical chemistry. A saliva sample is collected through absorbent cotton, is centrifuged and undergoes solid phase extraction, and the pyrazine substances and pyridine substances in saliva are determined through gas chromatography-mass spectrometry. The method comprises the following steps: 1, collecting the saliva sample by using absorbent cotton, placing the absorbent cotton in a syringe needle cylinder after collection ends, pushing a syringe push rod to carry out extrusion in order to obtain a saliva sample, carrying out high speed centrifuge, and taking the obtained supernatant; 2, adding an internal standard substance to the saliva sample, carrying out solid extraction column enrichment, and eluting the obtained material with a solvent; and 3, concentrating the obtained eluate, re-dissolving the obtained concentrate, allowing the re-dissolved concentrate to go through a millipore filter membrane, determining the filtered solution through gas chromatography-mass spectrometry, making a standard curve, and carrying out internal standard method quantification. The method can be effectively used to determine pyrazine substances and pyridine substances in saliva, and has the advantages of feasibility, low detection limit, good precision, high accuracy and reliability, and promotion application values.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a method for separating and measuring pyrazines and pyridines in saliva. Background technique [0002] Pyrazines and pyridines are important aroma components in food, tobacco, etc. During the eating process, the aroma components in the food are selectively dissolved and absorbed by the oral saliva, and those absorbed components are the substances that stimulate the human body to produce sensory sensations, which determine the style and flavor of the food. At present, the evaluation of food aroma is usually evaluated by the perception of mouth, nose, tongue, throat, etc., but it is difficult to rule out human subjectivity and the resulting uncertainties. By studying and measuring the flavor components absorbed and dissolved in saliva, the data of chemical components that determine the flavor of food can be directly and objectively obtained. The analysis ...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/025
Inventor 司晓喜刘志华朱瑞芝张凤梅石倩倩刘春波申钦鹏王昆淼何沛苏钟璧
Owner CHINA TOBACCO YUNNAN IND
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