Method for quantitative detection of gluconic acids and glucono lactone

A quantitative detection, glucaric acid technology, applied in the field of separation and detection, can solve the problems of complicated methods, low detection efficiency, and no detection, and achieve the effect of simple method and good stability

Active Publication Date: 2016-04-20
JIANGNAN UNIV
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  • Application Information

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Problems solved by technology

The detection methods mainly include high-performance liquid chromatography and gas chromatography. Among them, gas chromatography is relatively complicated for the pre-treatment process of samples, and the detection efficiency is low; high-performance liquid chromatography mainly uses C18 column and SB-Aq chromatographic column. Easy to handle, high sensitivity, good detection effect
However, there is no report on the method of simultaneously detecting and separating gluconic acid, glucuronic acid, glucaric acid and glucaric acid-1,4-lactone at home and abroad. Therefore, it is necessary to explore a new, simple and efficient separation method to separate The effective separation and determination of these four glucose series acids and lactones can effectively solve the existing problems of gluconic acid separation, undetectability, complicated methods, and inability to quantify, and promote the development of kombucha fermented beverage industry

Method used

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  • Method for quantitative detection of gluconic acids and glucono lactone
  • Method for quantitative detection of gluconic acids and glucono lactone
  • Method for quantitative detection of gluconic acids and glucono lactone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Joint detection of glucose series acids and lactones in mixed samples by SB-Aq chromatographic column and amino chromatographic column

[0023] This is achieved by:

[0024] (1) Use mobile phase 0.025mol / LKH respectively 2 PO 4 (pH2.5) solution to prepare standard samples of gluconic acid, glucuronic acid, calcium gluconate and glucaric acid-1,4-lactone at 5 mg / mL each, and after filtering through a 0.22 μm microporous filter head, 4 standard samples Take 500 μL of each product and mix, that is, the mixed standard product with a concentration of 1.25 mg / mL, and perform high performance liquid chromatography determination respectively. Using AgilentSB-Aq chromatographic column (4.6mm×250mm, 5μm), chromatographic conditions: mobile phase is 0.025mol / LKH 2 PO 4 (pH2.5), flow rate 0.5mL / min, column temperature 30°C, injection volume 10μL, UV detection wavelength 210nm. Under this detection condition, the retention times of gluconic acid, glucaric acid, and g...

Embodiment 2

[0034] Determination of glucaric acid-1,4-lactone content in black tea fermented liquid by SB-Aq chromatographic column.

[0035] (1) Bacterial solution sample pretreatment: Take 500 μL of bacterial solution, place it in a 3K ultrafiltration tube for centrifugal ultrafiltration to remove bacteria, protein and other macromolecular substances, collect the filtrate and place it in a 2 mL centrifuge tube.

[0036] (2) Adopt AgilentSB-Aq chromatographic column (4.6mm×250mm, 5μm), chromatographic conditions: mobile phase is 0.025mol / LKH 2 PO 4 (pH2.5), flow rate 0.5mL / min, column temperature 30°C, injection volume 10μL, UV detection wavelength 210nm. The treated bacterial liquid is loaded into the sample to obtain the SB-Aq chromatogram of the black tea fermented liquid ( image 3 ), retention time 6.867min is glucaric acid-1,4-lactone, and its peak area is 3234720, according to the regression equation that glucaric acid-1,4-lactone records on the SB-Aq chromatographic column calc...

Embodiment 3

[0038] Amino column / SB-Aq column combined separation and determination of gluconic acid, glucuronic acid, glucaric acid and glucaric acid-1,4-lactone in sweet potato fermentation broth.

[0039] (1) Bacterial solution sample pretreatment: Take 500 μL of bacterial solution, place it in a 3K ultrafiltration tube for centrifugal ultrafiltration to remove bacteria, protein and other macromolecular substances, collect the filtrate and place it in a 2 mL centrifuge tube.

[0040] (2) Adopt AgilentSB-Aq chromatographic column, chromatographic conditions: mobile phase is 0.025mol / LKH 2 PO 4 (pH2.5), flow rate 0.5mL / min, column temperature 30°C, injection volume 10μL, UV detection wavelength 210nm. The processed bacterial liquid is loaded to obtain the SB-Aq chromatogram of the sweet potato fermentation liquid ( Figure 4 ), retention time 6.877min is glucaric acid-1,4-lactone, and its peak area is 863069, according to the regression equation that glucaric acid-1,4-lactone records on...

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Abstract

The invention discloses a method for quantitative detection of gluconic acids and glucono lactone, and belongs to the technical field of separation and detection. The method mainly adopts an SB-Aq chromatographic column and an amino chromatographic column in a combined manner to qualitatively and quantitatively detect the gluconic acids and the glucono lactone which comprise gluconic acid, glucuronic acid, d-saccharic acid and d-saccharic acid 1,4-lactone. According to the method, firstly, the SB-Aq chromatographic column is adopted to perform direct quantitative analysis on the d-saccharic acid 1,4-lactone in a sample; secondly, the amino chromatographic column is adopted to perform direct quantitative analysis on the glucuronic acid and the d-saccharic acid in the sample; finally, the content of the gluconic acid in the sample is obtained through calculation. The method is simple and convenient in pretreatment, high in sensitivity and capable of determining multiple components, and has important significance of improvement of detection working efficiency, control of kombucha production quality and increase of the functional factor yield.

Description

technical field [0001] The invention relates to a method for quantitatively detecting glucose series acids and lactones, in particular to high performance liquid chromatography for the detection of gluconic acid, glucuronic acid, glucaric acid and glucaric acid-1,4-lactone ( The separation detection of DSL) belongs to the technical field of separation detection. Background technique [0002] During the fermentation process of kombucha, the glucose in the black tea culture solution is oxidized by acetic acid bacteria into metabolites such as gluconic acid and acetic acid. -1,4-lactone. [0003] Glucuronic acid is one of the most important detoxification substances in the human liver. It can be combined with exogenous toxins or endogenous toxic substances produced by human metabolism to become water-soluble glucuronide and excreted together, making it possible The toxins that cause various diseases in the body or have already occurred are removed in time, which has a good ef...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/46
CPCG01N30/02G01N30/46G01N2030/027
Inventor 王淼邓莉川吕梦娴
Owner JIANGNAN UNIV
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