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Method for detecting nosiheptide contents in nosiheptide fermentation broth

A technology of Nosiheptide and fermentation liquid, which is applied in the field of detection of Nosiheptide content in Nosiheptide fermentation liquid, which can solve the problems of inability to monitor the quality of the fermentation production process, inability to accurately guide production regulation, and complicated preparation of mixed reagents. To achieve the effect of convenient filtering operation, saving time and simplifying the processing steps

Inactive Publication Date: 2018-12-21
HEBEI SHENGXUE DACHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The solvent DMF used in this detection method is highly toxic and used in large amounts, has a high viscosity and is difficult to filter, the preparation of mixed reagents is complicated, there are many operating steps, and the microbial detection cycle is long, so it cannot be used for process quality monitoring of fermentation production.
Although spectrophotometry is easy and fast to operate, it cannot accurately guide production regulation because of its poor specificity and is prone to errors caused by interference from impurities.

Method used

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  • Method for detecting nosiheptide contents in nosiheptide fermentation broth
  • Method for detecting nosiheptide contents in nosiheptide fermentation broth
  • Method for detecting nosiheptide contents in nosiheptide fermentation broth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] a. Preparation of Nosiheptide Reference Substance Solution: Accurately weigh 25 mg of Nosiheptide Reference Substance, dissolve it in DMF to a constant volume in a 25 mL brown volumetric flask, shake well, and use it as a stock solution. Accurately pipette 2.5mL stock solution into a 25mL brown volumetric flask, add methanol to dilute to volume, and use it as the reference solution;

[0059] b. Preparation of nosiheptide sample solution: draw 2.5mL uniform sample into a 25mL brown volumetric flask, add extraction solution (volume ratio: ethanol / water=90 / 10) to constant volume, shake well, ultrasonically extract for 25min and then shake well , let it stand for 15 minutes, take the supernatant and filter it with a 0.45 μm organic filter membrane to obtain the sample solution to be tested;

[0060] c. Sample measurement and result calculation:

[0061] Chromatographic conditions:

[0062] Liquid chromatograph: Waters e2695-2489;

[0063] Chromatographic column: C18 5.0μ...

Embodiment 2

[0077] a. Preparation of Nosiheptide Reference Substance Solution: Accurately weigh 50 mg of Nosiheptide Reference Substance, dissolve it in DMF to a constant volume in a 25 mL brown volumetric flask, shake well, and use it as a stock solution. Accurately pipette 1.0mL stock solution into a 25mL brown volumetric flask, add ethanol to dilute to volume, and use it as the reference solution;

[0078] b. Preparation of nosiheptide sample solution: draw 10.0mL uniform sample into a 50mL brown volumetric flask, add extraction solution (volume ratio: ethanol / water=75 / 25) to constant volume, shake well, ultrasonically extract for 30min and then shake well , let stand for 20 minutes, take the supernatant and filter it with a 0.45 μm organic filter membrane to obtain the sample solution to be tested;

[0079] c. Sample measurement and result calculation:

[0080] Chromatographic conditions:

[0081] Liquid chromatograph: Waters 1515-2489;

[0082] Chromatographic column: C18 5.0μm 4....

Embodiment 3

[0096] a. Preparation of Nosiheptide Reference Substance Solution: Accurately weigh 30 mg of Nosiheptide Reference Substance, dissolve it in DMF to a constant volume in a 25 mL brown volumetric flask, shake well, and use it as a stock solution. Accurately pipette 2.0mL stock solution into a 25mL brown volumetric flask, add acetonitrile to dilute to volume, and use it as the reference solution;

[0097] b. Preparation of nosiheptide sample solution: draw 2.5mL uniform sample into a 50mL brown volumetric flask, add extract solution (concentration 95% ethanol) to constant volume, shake well, shake well after ultrasonic leaching for 20min, let it stand for 10min, take The supernatant was filtered with a 0.45 μm organic filter to obtain the sample solution to be tested;

[0098] c. Sample measurement and result calculation:

[0099] Chromatographic conditions:

[0100] Liquid chromatograph: Diane U3000;

[0101] Chromatographic column: C18 5.0μm 4.6*150mm;

[0102] Column tempe...

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Abstract

The invention discloses a method for detecting nosiheptide contents in nosiheptide fermentation broth, relates to the technical field of detection, and relates to a method for detecting nosiheptide contents. The method comprises the following steps that: a: preparing the reference substance solution of the nosiheptide; b: taking a nosiheptide fermentation broth sample, determining the contents bytaking ethanol water mixed solvent as leach liquor, and then, carrying out ultrasonic extraction to obtain the sample solution of the nosiheptide; and c: injecting the reference substance solution andthe sample solution in sequence for measurement by HPLC (High Performance Liquid Chromatography), recording a chromatogram and a peak area, calculating a correction factor by the mass concentration and the peak area of the main ingredient nosiheptide A in the reference substance solution, utilizing the obtained correction factor to calculate the contents of the nosiheptide A component in the fermentation broth sample, and meanwhile, obtaining the contents of the nosiheptide B component. The method has the advantages of being simple, quick, accurate, efficient and low in toxin and cost, can beused as a fermentation process monitoring method of a nosiheptide production enterprise, and provides monitoring data for a production process in time.

Description

technical field [0001] The invention relates to the technical field of detection, and relates to a method for detecting the content of nosiheptide, in particular to a method for detecting the content of nosiheptide in a fermented liquid of nosiheptide. Background technique [0002] Nosiheptide is a sulfur-containing polypeptide antibiotic with the English name Nosiheptide. As a new type of feed additive, nosiheptide can not only promote the growth of livestock and poultry, improve feed efficiency, but also has the characteristics of not easy to produce drug resistance, no residue, little impact on the environment, and safe for humans and animals. Additive requirements; and it can be widely used not only in livestock and poultry industry but also in aquaculture, with certain social and economic benefits. Therefore, as a new type of feed additive, nosiheptide has a very broad application prospect. At present, it has been approved and widely used at home and abroad. [0003]...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 杜精精马盼盼曹建全
Owner HEBEI SHENGXUE DACHENG PHARMA
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