Method for separating and assaying organic acid and fatty acid substances in saliva

A technology for organic acids and fatty acids, applied in the field of analytical chemistry, can solve problems such as no reports, and achieve the effects of simple pretreatment, low detection limit, and high methyl esterification efficiency.

Active Publication Date: 2016-09-21
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 many reports on the determination of organic acids or fatty acids in different m

Method used

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  • Method for separating and assaying organic acid and fatty acid substances in saliva
  • Method for separating and assaying organic acid and fatty acid substances in saliva
  • Method for separating and assaying organic acid and fatty acid substances in saliva

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A method for separating and measuring organic acids and fatty acid substances in the saliva of tobacco smokers, comprising the following steps:

[0042]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.

[0043] Step (2), salting-out extraction: Pipette 2.0 mL of saliva sample into a centrifuge tube, add 2.0 mL of methanol, 1 g of anhydrous magnesium sulfate, 0.2 g of sodium chloride, 0.2 g of sodium citrate, 0.1 g of dihydrogen citrate Sodium, vortex at 500 rpm for 2.0 min, then centrifuge at 10000 rpm for 5 min, and take the extract layer;

[0044] Step (3), methyl esterification: Add sulfuric acid with 10% volume of the extract to the extract, move it into a microwave extraction tank and seal...

Embodiment 2

[0052] A method for separating and measuring organic acids and fatty acid substances in the saliva of liquor drinkers, comprising the following steps:

[0053] 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.

[0054] Step (2), salting-out extraction: pipette 8.0 mL of saliva sample into a centrifuge tube, add 8.0 mL of methanol, 5 g of anhydrous sodium sulfate, 0.8 g of sodium chloride, 10 μL of formic acid, and vortex at 15000 rpm for 10.0 min, then centrifuge at 10000 rpm for 5 min, and take the extract layer;

[0055] Step (3), methyl esterification: add boron trifluoride with 30% volume of the extract to the extract, and carry out methyl esterification reaction at 75°C in a water bath for 5 mi...

Embodiment 3

[0062] A method for separating and measuring organic acids and fatty acid substances in saliva, comprising the following steps:

[0063] Step (1), sample collection: put 0.2 g of absorbent cotton in the mouth for 0.2 min, 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, after high-speed centrifugation, collect the supernatant to obtain a saliva sample; the centrifugation speed of saliva should not be lower than 10,000 rpm, and the centrifugation time should not be less than 10 min;

[0064] Step (2), salting-out extraction: pipette 0.5 mL of saliva sample into a centrifuge tube, add methanol, sodium chloride, extraction salt and pH regulator, vortex until well mixed, and then centrifuge, take the extraction solution containing methanol liquid layer to obtain the first extract; wherein, the vortex oscillation speed is 100 ...

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Abstract

The invention relates to a method for separating and assaying organic acid and fatty acid substances in saliva, belongs to the technical field of analytical chemistry and particularly relates to the method dedicated to separating and assaying the organic acid and fatty acid substances in the saliva. The method comprises the steps of collecting a saliva sample by adopting absorbent cotton, then, putting the absorbent cotton into a needle cylinder of a syringe, carrying out pushing squeezing by using a push rod of the syringe so as to obtain the saliva sample, carrying out high-speed centrifugation, then, extracting supernatant, then, carrying out salting-out extraction, a methyl esterification reaction and solvent extraction separation sequentially, then, concentrating an extract until the extract is approximately dry, then, carrying out redissolving, carrying out filtrating by a microporous filtrating membrane, assaying the organic acid and fatty acid substances in the saliva by adopting a gas chromatography-mass spectrometry method by using filtrate, making a standard curve, and carrying out quantification by an external standard method. The method provided by the invention can be effectively used for assaying the organic acid and fatty acid substances in the saliva, is feasible, accurate and reliable, is low in detection limit and good in accuracy and has popularization and 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 organic acids and fatty acids in saliva. Background technique [0002] Organic acids and fatty acids in food are the main sources of food flavor and taste, and their content and mutual ratio have a direct impact on color, aroma, taste, and even quality and style. The research on organic acids and fatty acids in food usually directly determines the content of these components in food. However, in the process of food and diet, the ingredients selectively dissolved and absorbed by oral saliva are the substances that stimulate the human body to produce sensory sensations, and determine the style and flavor of food. Research on the absorption and dissolution of organic acids and fatty acids in the saliva of eaters can more directly and objectively obtain the chemical composition data that determine the flavor of food, whi...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88
Inventor 司晓喜张凤梅朱瑞芝刘志华杨光宇苏美玲何沛刘春波申钦鹏王昆淼苏钟璧
Owner CHINA TOBACCO YUNNAN IND
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