Method for detecting content of urea in brewed wine by using high performance liquid chromatography

A technology of high performance liquid chromatography and urea content, which is applied in the field of detection of urea content in fermented wine, can solve the problems of instability, high level requirements, increased colorimetric measurement error, etc., and achieves high recovery rate and simple processing method. Effect

Inactive Publication Date: 2014-04-30
CENT TESTING INT GRP CO LTD
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Problems solved by technology

The sensitivity of the chemical analysis method is relatively low, and there are amino acids and ammonium ions similar in structure and properties to urea in rice wine, which have a great influence on the determination results
Diacetyl-oxime spectrophotometry uses urea in the sample and the color developing reagent to develop color in a boiling water bath for 10-15 minutes and then colorimetrically determines, but the error of colorimetric determination increases with the increase of urea content
It is also reported in China that the urea content in yellow rice wine is determined by high-performance liquid chromatography-ultraviolet detector (HPLC-UV) using the characteristic of urea's ultraviolet absorption at λ=203nm. Susceptible to disturbances causing baseline instability

Method used

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  • Method for detecting content of urea in brewed wine by using high performance liquid chromatography
  • Method for detecting content of urea in brewed wine by using high performance liquid chromatography

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Experimental program
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Effect test

Embodiment 1

[0028] Accurately draw 2.0mL of the sample, put it in a 10mL volumetric flask, and dilute to the mark with absolute ethanol. Take 2 mL of the diluted sample solution and place it in a test tube with a stopper, add 0.5 mL of hydrochloric acid solution (concentration: 1.5 mol / L) and 2 mL of 9-hydroxyzanthene solution (concentration: 0.03 mol / L), mix well, and place it at room temperature in the dark The derivatization was carried out under the condition of 30min. Do this three times, combine the derivatized solution into a 100mL round bottom flask, concentrate the solution to about 5mL by a rotary evaporator to form a concentrated solution, transfer the concentrated solution to a 10mL volumetric flask, dilute to the mark with ethanol, pass 0.45 um organic filter membrane to form a pre-treated solution to be tested for liquid chromatography testing. Five parallel samples were made for the sample. According to the derivation method, the fermented wine sample was derivatized by ad...

Embodiment 2

[0032] Derivative time is set as 50min, other conditions are the same as embodiment 1, the relative standard deviation of gained parallel sample<8%, the relative standard deviation of repeatability test<5%, the standard addition recovery rate of sample is between 85%-110%. between.

Embodiment 3

[0034] Column temperature is set as 40 ℃, and other conditions are the same as embodiment 1, and the result peak time slightly changes, and other is constant, the relative standard deviation of gained parallel sample<8%, the relative standard deviation<5% of repeatability test, The spiked recoveries of the samples ranged from 85% to 110%.

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Abstract

The invention relates to a method for detecting the content of urea in brewed wine by using high performance liquid chromatography. The method comprises the following steps: weighing an appropriate sample, selecting a proper solvent, and performing pre-treatment on the sample by using precolumn derivatization, rotary evaporation concentration and the like; and detecting urea derivatives by using a high performance liquid chromatograph connected with a fluorescence detector, wherein the organic flow phase and the inorganic flow phase are in variable ratio, the flow speed is set as 1mL / min, the sampling amount is set as 10 microliters, the chromatographic column is selected as the C18 chromatographic column, the column temperature is 40-50 DEG C, and the detection wavelength of the fluorescence detector as follows: the lambda ex is 208nm, and the lambda em is 308nm. The detection method provided by the invention has the characteristics of being simple, fast and high in recycling rate.

Description

technical field [0001] The invention relates to a method for detecting urea content in fermented wine, in particular to a method for detecting and detecting urea content in fermented wine by using high-performance liquid chromatography technology connected with a fluorescence detector. Background technique [0002] In fermented beverages, ethyl carbamate (IARC group2A, referred to as EC) is a potentially genotoxic and carcinogenic risk species, which is produced by arginine through yeast intracellular arginase- Produced by breakdown of the urease (AU) pathway. Internationally, the Food and Agriculture Organization of the United Nations (FAO) has included it as a key monitoring species in 2002. Many countries have formulated limits for ethyl carbamate in alcoholic beverages out of consideration for the health of their consumers and the interests of the industry. This undoubtedly increases the possibility of trade friction. At the same time, among the fermented alcoholic bev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 董宁彭琳婧戴煦
Owner CENT TESTING INT GRP CO LTD
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