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Quantitative method of short-chain fatty acid

A technology for the quantitative detection of short-chain fatty acids, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problem of no detection method found, and achieve the effect of low detection cost, high precision and convenient detection

Inactive Publication Date: 2020-05-19
HKUST SHENZHEN RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, no similar detection method has been searched; especially the method for detecting short-chain fatty acids in feces or feces fermentation liquid using the most common HPLC-UV has not yet been reported

Method used

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  • Quantitative method of short-chain fatty acid
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  • Quantitative method of short-chain fatty acid

Examples

Experimental program
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Effect test

no. 1 example

[0043] The quantitative method of the short-chain fatty acid of the first embodiment of the present invention comprises the following steps:

[0044] (1) Sample processing

[0045] Collect 1 part of human feces samples from 6 volunteers, weigh 2 g of feces for each sample, and vortex with 30 wt% acetonitrile aqueous solution (add 7 parts of pure water first, then add 3 parts of acetonitrile) according to the weight ratio of 1:10 Extract for 2 minutes to obtain a suspension; centrifuge the suspension at 4000rpm / min for 10min at 10°C, take 400uL of the supernatant and place it in a 5mL centrifuge tube, add 200μL of 200mM 3NPH·HCl solution and 200μL of 120mM EDC·HCl and 6wt% pyridine mixed solution were reacted together at 40°C for 45min. After the reaction, add water to make it slightly more than 1 mL, centrifuge at 13,000 rpm at 4°C for 10 min, and take the supernatant to obtain a derivatized sample;

[0046] (2) SPE cartridge enrichment

[0047] Take 1mL of the derivatized ...

no. 2 example

[0057] Second embodiment (methodological verification of the method of the present invention)

[0058] (1) Determination of linear range and detection limit

[0059] Take mixed standard solutions of short-chain fatty acids with different concentrations, and measure them according to the method in the first embodiment "sample treatment, SPE small column enrichment and liquid chromatography separation". Determine the limit of detection and the limit of quantification with signal-to-noise ratios of 3 and 10 respectively, draw a curve with the peak area of ​​the characteristic absorption peaks of 10 short-chain fatty acids as the vertical coordinate y, and the concentration of short-chain fatty acids as the abscissa x, and obtain the detected short-chain fatty acids The linear range of , see Table 3.

[0060] Table 3 Linear regression equations and related parameters of 10 short-chain fatty acids

[0061]

[0062]

[0063] (2) Determination of stability, precision and accu...

no. 3 example

[0068] The third embodiment (determining the content of short-chain fatty acids in the human feces fermented liquid sample)

[0069] (1) Sample treatment: Collect human feces fermentation liquid samples, measure 700uL sample solution, add 300uL acetonitrile, and vortex for 2min.

[0070] (2) All the other processing steps and HPLC-UV analysis steps are the same as the first embodiment, such as Figure 4 and Figure 5 As shown, the measured content of short-chain fatty acids in the human feces fermentation broth is shown in Table 5.

[0071] Table 5 Results of quantitative detection of 10 kinds of short-chain fatty acids in human feces fermentation liquid samples

[0072]

[0073]

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Abstract

The invention discloses a quantification method of a short-chain fatty acid. 3-nitrophenylhydrazine is used as a derivatization reagent to carry out derivatization on the short-chain fatty acid in a biological excretion sample so as to generate a short-chain fatty acid derivatization product with a characteristic absorption peak in an ultraviolet region, and the short-chain fatty acid derivatization product is determined in the ultraviolet region by utilizing an HPLC-UV method based on a partition chromatography principle so that quantitative detection of the short-chain fatty acid is realized. The quantification method of the short-chain fatty acid is high in accuracy and precision and low in detection limit.

Description

technical field [0001] The present invention relates to the technical field of quantitative detection, in particular to a quantitative method for short-chain fatty acids, which specifically utilizes high performance liquid chromatography (HPLC-UV) to process and / or unprocessed biological excretion samples (specifically can be used Quantitative analysis of 10 kinds of short-chain fatty acids in human feces and / or feces fermentation broth) belongs to the detection technology method of endogenous substances. Background technique [0002] Short-chain fatty acids, also known as volatile fatty acids, are mostly organic fatty acids produced by anaerobic bacteria in the intestinal tract by fermenting carbohydrates in undigested and absorbed food residues, including acetic acid, propionic acid, butyric acid, isobutyric acid, and isovaleric acid and isocaproic acid etc. The type and content of short-chain fatty acids mainly depend on the type of intestinal flora, the fiber content in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8813
Inventor 詹华强董婷霞王怀友汪成
Owner HKUST SHENZHEN RES INST
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