Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for detecting protein content in 2-keto-L-gulonic acid

A technology of protein content and gulon acid, which is applied in the field of chemical analysis, can solve the problems of inability to detect protein content, and achieve the effects of low detection equipment requirements, good reproducibility, and good time stability

Active Publication Date: 2009-10-28
NORTHEAST PHARMA GRP
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The invention provides a detection method, the purpose of which is to solve the problem that the protein content in 2-keto-L-gulonic acid cannot be detected at present

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for detecting protein content in 2-keto-L-gulonic acid
  • Method for detecting protein content in 2-keto-L-gulonic acid
  • Method for detecting protein content in 2-keto-L-gulonic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Detection of protein content in 2-keto-L-sodium guronate fermentation broth;

[0055] Take the 2-keto-L-gulonic acid sodium fermentation liquid in production and centrifuge at 2000r / min for 20min, pipette 2ml of the supernatant into a test tube, then add 2ml of 40% KOH solution, boil in boiling water for 30min, wash it with deionized water into a 100ml volumetric flask, then add 4.5ml of 20% nitric acid solution and 10ml of a buffer solution with a pH of 7, and dilute to volume with deionized water.

[0056] Pipette one part of 1 mL of deionized water and two parts of the test solution into three test tubes, add 5 mL of Coomassie Brilliant Blue reagent to the first two test tubes, add 5 mL of deionized water to the third test tube, and detect the deionized water at 595nm and the absorbance of test tube 2 are 0.037 and 0.058L / (g.cm), adjust the absorbance of test tube 1 to 0, and detect the absorbance of test tube 3 within 30 to 60 minutes to be 0.162L / (g.cm),...

Embodiment 2

[0057] Example 2: Detection of protein content in 2-keto-L-sodium guronate ultrafiltrate

[0058] Take 20ml of post ultrafiltrate to a 50ml volumetric flask, then add 10ml of buffer solution with a pH of 7, and dilute to volume with deionized water.

[0059] Pipette one part of 1 mL of deionized water and two parts of the test solution into three test tubes, add 5 mL of Coomassie Brilliant Blue reagent to the first two test tubes, add 5 mL of deionized water to the third test tube, and detect the deionized water at 595nm and the absorbance of test tube 2 are 0.037 and 0.046L / (g.cm), adjust the absorbance of test tube 1 to 0, and detect the absorbance of test tube 3 within 30 to 60 minutes to be 0.205L / (g.cm). The protein content in the liquid is 0.089mg / ml.

Embodiment 3

[0060] Example 3: Detection of protein content in dry product of 2-keto-L-gulonic acid

[0061] Weigh 5g of the dry product of 2-keto-L-gulonic acid in production, dissolve it in deionized water and transfer it to a 100ml volumetric flask, then add 6.5ml of 20% KOH solution and 10ml of buffer solution with a pH of 7, and dilute to volume with deionized water .

[0062] Pipette one part of 1 mL of deionized water and two parts of the test solution into three test tubes, add 5 mL of Coomassie Brilliant Blue reagent to the first two test tubes, add 5 mL of deionized water to the third test tube, and detect the deionized water at 595nm The absorbance of test tube 2 and test tube 2 are respectively 0.037 and 0.039 L / (g.cm), adjust the absorbance of test tube 1 to 0, and detect the absorbance of test tube 3 within 30 to 60 minutes to be 0.115 L / (g.cm), which can be calculated by formula 1 The protein content in the dry product of 2-keto-L-gulonic acid is 0.036%.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Absorbanceaaaaaaaaaa
Absorbanceaaaaaaaaaa
Absorbanceaaaaaaaaaa
Login to View More

Abstract

The invention relates to a method for analyzing protein content in 2-keto-L-gulonic acid, which comprises the following steps: adopting an optimized Coomassie brilliant blue quantitative protein method (a Bradford method), preparing phosphate buffer solution, bovine serum protein standard solution and Coomassie brilliant blue G-250 staining agent, adding the Coomassie brilliant blue G-250 staining agent into the bovine serum protein standard solution, drawing a standard curve according to absorbency added values after the Coomassie brilliant blue is combined with the protein, then adding the 2-keto-L-gulonic acid into a Coomassie brilliant blue reagent, and calculating the protein content in the 2-keto-L-gulonic acid according to the absorbency added values and the slope coefficient of a protein concentration standard curve. The analyzing method is simple, convenient and sensitive, has the advantages of good stability, high accuracy, good repeatability and low requirement on detection equipment, is quite suitable for analyzing and detecting the protein content in the 2-keto-L-gulonic acid in the industrial production, and has important significance for ensuring and guiding normal production.

Description

1. Technical fields; [0001] The invention relates to a chemical analysis method, in particular to an analysis method for measuring the protein content in 2-keto-L-gulonic acid by Coomassie brilliant blue colorimetry. 2. Background technology; [0002] Vitamin C, also known as L-ascorbic acid, is an essential vitamin for human nutrition. It has a wide range of uses and is often used in food, feed and cosmetics. It is also widely used in medicine and clinical practice. It has long played a very important role in the international pharmaceutical trade. status. Its main production methods are the Reyes method and the two-step fermentation method. The two-step fermentation method is the first in my country. It replaces part of the chemical synthesis process of the Leyer route with a biological oxidation process, and then synthesizes vitamin C. [0003] The two-step fermentation method uses D-sorbitol as the raw material, and obtains the important intermediate of vitamin C, 2-ke...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N21/78G01N21/31
Inventor 于霞王雪李春杰陈宏权敖志刚董静董淑清符艳君王凤杰王彦春
Owner NORTHEAST PHARMA GRP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com