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Method for quantitatively determining 3-hydroxybutanone in fermentation liquid

A technology for the quantitative determination of hydroxybutanone, which is applied in the field of analysis and detection, to achieve the effects of reducing workload, fast detection speed and low cost

Active Publication Date: 2019-03-26
QILU UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Utilize 3,5-dinitrosalicylic acid and 3-hydroxybutanone azeotropic quantitative determination method of 3-hydroxybutanone at present, there is no report yet

Method used

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  • Method for quantitatively determining 3-hydroxybutanone in fermentation liquid
  • Method for quantitatively determining 3-hydroxybutanone in fermentation liquid
  • Method for quantitatively determining 3-hydroxybutanone in fermentation liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Draw 3-hydroxybutanone standard curve

[0039] ① Prepare 3-hydroxybutanone standard solution;

[0040] Accurately weigh 100 mg of 3-hydroxybutanone standard sample, 3-hydroxybutanone is pre-dried in an oven at 50°C to constant weight, placed in a small beaker, dissolved with a small amount of distilled water, quantitatively transferred to a 100mL volumetric flask, and Dilute to the mark with distilled water, shake well, and store in the refrigerator for later use.

[0041] ② Preparation of 3,5-dinitrosalicylic acid reagent;

[0042]Add 6.3g of 3,5-dinitrosalicylic acid and 262mL of 2mol / L NaOH solution to 500mL of hot water solution containing 185g of potassium sodium tartrate, add 5g of crystalline phenol and 5g of sodium sulfite, stir to dissolve; add distilled water after cooling Dilute to 1000mL to prepare 3,5-dinitrosalicylic acid reagent, store it in a brown bottle for later use;

[0043] ③Respectively take 0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2mL 3-hydroxybutanone st...

Embodiment 2

[0052] 3-Hydroxybutanone fermentation medium composition: glucose 50g / L, yeast extract 10.3 g / L, corn steep liquor 19.8g / L, manganese sulfate 0.193 g / L, ferrous sulfate 0.11 g / L, ammonium sulfate 7.0g / L, magnesium sulfate 0.2 g / L.

[0053] 3-Hydroxybutanone fermentation The fermentation conditions are: fermentation temperature 37°C, shaker rotation speed 180r / min, liquid seed inoculum amount 2-3%, adding 1% calcium carbonate to the medium to control the pH value.

[0054] When the shake flask was fermented for 72 hours, the glucose was exhausted, and the fermentation clear night was obtained by centrifugation.

[0055] After diluting the above fermented clear liquid with distilled water 100 times, add 3,5-dinitrosalicylic acid reagent, heat and boil for 5 minutes, and measure the absorbance of the solution after the reaction is completed;

[0056] Substitute the detected absorbance value into a linear equation to obtain the concentration of 3-hydroxybutanone in the fermentati...

Embodiment 3

[0062] 3-Hydroxybutanone fermentation medium composition: glucose 100g / L, yeast extract 11.5 g / L, corn steep liquor 20.5g / L, manganese sulfate 0.193 g / L, ferrous sulfate 0.11 g / L, ammonium sulfate 9.0g / L, magnesium sulfate 0.3 g / L.

[0063] 3-Hydroxybutanone fermentation The fermentation conditions are as follows: fermentation temperature 37°C, shaker rotation speed 180r / min, liquid seed inoculation amount 2-3%, adding 1.5% calcium carbonate to the medium to control the pH value.

[0064] When the shake flask was fermented for 80 hours, the glucose was exhausted, and the fermentation clear night was obtained by centrifugation.

[0065] After diluting the above fermented clear liquid with distilled water 100 times, add 3,5-dinitrosalicylic acid reagent, heat and boil for 5 minutes, and measure the absorbance of the solution after the reaction is completed;

[0066] Substitute the detected absorbance value into a linear equation to obtain the concentration of 3-hydroxybutanone ...

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Abstract

The invention discloses a method for quantitatively determining 3-hydroxybutanone in fermentation liquid and belongs to the field of analysis and detection. The method specifically comprises the following steps: after glucose is depleted by the 3-hydroxybutanone-containing fermentation liquid, adding a 3, 5-dinitrosalicylic acid reagent, azeotroping with the fermentation liquid and detecting the absorbance of the solution after the reaction is completed, wherein the absorbance of the solution and the concentration of the 3-hydroxybutanone in the fermentation liquid have a linear relation in acertain range; and substituting the detected absorbance value into a linear equation of a standard curve to obtain the concentration of the 3-hydroxybutanone in the fermentation liquid. The method fordetecting the content of the 3-hydroxybutanone in the fermentation liquid has the beneficial effects that the requirement on equipment is low; the cost is low; the detection speed is high; the methodis suitable for the detection of mass samples; and the workload of the product detection in selection and breeding processes of strains is significantly reduced.

Description

technical field [0001] The invention relates to the field of analysis and detection, in particular to a method for the quantitative determination of 3-hydroxybutanone in fermentation broth. Background technique [0002] 3-Hydroxybutanone, chemically named 3-hydroxy-2-butanone, is an alpha-hydroxy ketone with the molecular formula C 4 h 8 o 2 , molecular weight 88.12, 2 and 3 carbon atoms in the molecule are asymmetric carbon atoms, there are two chiral isomers, and its chemical structural formula is: [0003] [0004] 3-Hydroxybutanone monomer is a light yellow or colorless liquid with a creamy aroma. At low temperature, two molecules of 3-hydroxybutanone polymerize into a dimer, which is a white crystalline powder. 3-Hydroxybutanone has a boiling point of 148°C and a melting point of 15°C. It is spontaneously combustible, soluble in propylene glycol and ethanol, miscible in water, slightly soluble in ether, and insoluble in plants. [0005] 3-Hydroxybutanone quality ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N30/02
CPCG01N21/31G01N30/02
Inventor 徐慧李文婧徐贞刘建军田延军张俊娇
Owner QILU UNIV OF TECH
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