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HILIC analysis method for determining amino acids and Amadori compounds in tobacco

A chromatographic analysis and amino acid technology, applied in the field of chemical composition analysis in tobacco, can solve problems such as lack of a liquid chromatography-tandem mass spectrometry method, etc., and achieve the effects of high sensitivity, simple pretreatment process and high resolution

Inactive Publication Date: 2019-09-03
SHANGHAI TOBACCO GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above-mentioned shortcoming of the prior art, the object of the present invention is to provide a kind of HILIC chromatographic analysis method of amino acid and Amadori compound in measuring tobacco, be used to solve the deficiency in the prior art that can simultaneously measure 16 kinds of amino acids and 5 kinds of Amadori compounds Problems with the HILIC-based liquid chromatography-tandem mass spectrometry method

Method used

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  • HILIC analysis method for determining amino acids and Amadori compounds in tobacco
  • HILIC analysis method for determining amino acids and Amadori compounds in tobacco
  • HILIC analysis method for determining amino acids and Amadori compounds in tobacco

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] 1. Sample pretreatment

[0078]The tobacco sample was dried and pulverized, and passed through a 70-mesh sieve to obtain a powder sample. Weigh 0.2g of tobacco powder sample, accurate to 0.1mg, into a 50mL capped centrifuge tube, add 20mL of 0.1mol / mL hydrochloric acid aqueous solution and 20μL of internal standard solution, sonicate for 30min and centrifuge at 4000r / min for 5min, take the supernatant After passing through a 0.22 μm PTFE filter membrane, the obtained sample solution 1#. Wherein, the ratio of the mass g added to the tobacco sample to the volume mL added to the hydrochloric acid aqueous solution is 2:200. The internal standard solution is hydrochloric acid aqueous solution containing norvaline, which is to weigh 30mg norvaline, accurate to 0.1mg, in a 10mL volumetric flask, dilute with 0.1mol / mL hydrochloric acid aqueous solution to the mark. That is, the concentration of norvaline in the internal standard solution is 3 mg / mL, the concentration of aqueo...

Embodiment 2

[0099] According to the preparation steps of the mixed standard working solution in Example 1, a mixed standard working solution was established for 16 kinds of amino acids and 5 kinds of Amadori compounds, and the corresponding concentrations in the mixed standard working solution were 1 pmol / μL, 2.5 pmol / μL, 5 pmol / μL, 12.5pmol / μL, 50pmol / μL, 125pmol / μL, 250pmol / μL. The above series of mixed standard working solutions with different concentrations were analyzed by LC-MS / MS to obtain the linear relationship between the chromatographic peak area ratios of various amino acids and Amadori compounds / internal standards and the concentration ratios of corresponding amino acids and Amadori compounds / internal standards, The corresponding standard working curves were drawn, and the regression equations of the standard working curves of various amino acids and Amadori compounds were calculated, and the results are shown in Table 2. As can be seen from Table 2, the peak areas measured i...

Embodiment 3

[0103] According to the pretreatment and measurement steps in Example 1, select tobacco samples and carry out 5 parallel measurements within 1 day to investigate the intraday repeatability of the method. Tobacco samples were also selected and measured for three consecutive days, with 5 parallel measurements per day to investigate the day-to-day repeatability of the method. The specific results are shown in Table 3. It can be seen from Table 3 that the relative standard deviation (RSD) of the 16 kinds of amino acids and the 5 kinds of Amadori compounds are all below 10%, indicating that the repeatability of the method is good.

[0104] Table 3 Intra-day and inter-day repeatability of tobacco samples

[0105]

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Abstract

The invention provides an HILIC (Hydrophilic Interaction Liquid Chromatography) analysis method for determining amino acids and Amadori compounds in tobacco. The method comprises the steps that LC-MS / MS is adopted to detect a sample solution obtained after a tobacco sample is subject to extraction; the amino acids and the Amadori compounds are separated through an HILIC column; and qualification is performed according to retention time and the mass-to-charge ratio of parent ions to daughter ions of all the amino acids and the Amadori compounds, quantification is performed through a standard curve method, and the content of the amino acids and the content of the Amadori compounds in the sample solution are determined. Through the HILIC analysis method for determining the amino acids and theAmadori compounds in the tobacco, 16 amino acids and five Amadori compounds in the tobacco can be subject to qualitative and quantitative analysis simultaneously, and the method has the advantages that the pretreatment process is easy and convenient to implement, detection is fast, the separation degree is high, and sensitivity is high.

Description

technical field [0001] The invention belongs to the technical field of chemical component analysis in tobacco, and relates to a HILIC chromatographic analysis method for determining amino acids and Amadori compounds in tobacco, in particular to a HILIC chromatographic analysis method for 16 kinds of amino acids and 5 kinds of Amadori compounds in tobacco. Background technique [0002] Amino acid is an important chemical component in tobacco leaves, and it is also an important class of aroma-inducing substances in tobacco. An appropriate amount of amino acids can help improve the strength and fullness of the smoke, but too high a content will cause the smoke to be spicy and irritating. [0003] During tobacco conditioning and curing, a series of non-enzymatic browning reactions (Maillard reactions) may occur between amino acids and reducing sugars, which can have a significant impact on the quality and color of tobacco leaves. Amadori compounds (Amadori products: 1-amino-1-d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/045G01N2030/062
Inventor 王先颖戚大伟罗辰周妍费婷王嘉乐吴达
Owner SHANGHAI TOBACCO GRP CO LTD
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