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Measuring method of CTAB content in meningococcal polysaccharide

A meningococcal and determination method technology, applied in the field of testing and analysis, can solve problems such as uncertainty, and achieve good repeatability

Active Publication Date: 2013-08-14
BEIJING MINHAI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no literature at home and abroad on the determination of CTAB residues in meningococcal polysaccharide stock solution and meningococcal polysaccharide vaccine, so how to detect CTAB in meningococcal polysaccharide vaccine is still uncertain

Method used

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  • Measuring method of CTAB content in meningococcal polysaccharide
  • Measuring method of CTAB content in meningococcal polysaccharide
  • Measuring method of CTAB content in meningococcal polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The investigation of embodiment 1 detection method

[0039] 1.1 Linear relationship inspection

[0040] Accurately weigh 10 mg of CTAB, dissolve it with 0.85% NaCl and dilute to 10 ml, as the reference substance stock solution (concentration is 1 mg / ml). Precisely draw 10ml of CTAB reference substance stock solution, and dilute to a 100ml volumetric flask as the reference substance working solution (concentration is 100μg / ml). Dilute the working solution of the reference substance to concentrations of 100 μg / ml, 50 μg / ml, 25 μg / ml, 12.5 μg / ml, and 6.25 μg / ml in sequence, absorb different concentrations of the reference substance solution, inject 20 μl sequentially, and record the chromatogram. Take the peak area (Y) as the ordinate and the reference substance solution X (μg / ml) as the abscissa to perform linear regression to obtain its regression equation. The results are shown in Table 1.

[0041] Table 1 Linear relationship test results

[0042] linear eq...

Embodiment 2A

[0057] Determination of CTAB content in group A meningococcus of embodiment 2

[0058] Precisely prepare 100μg / ml (C1) CTAB standard solution as sample 1;

[0059] Take the polysaccharide solution of group A meningococcus to be tested, spiked with CTAB at a concentration of 100 μg / ml, as sample 2;

[0060] Load sample 1 on a well-balanced high-efficiency gel chromatography column TSKgel G5000 chromatographic column, the sample volume is 20 μl, eluted with 0.9% NaCl solution, the elution flow rate is 0.5ml / min, and the chromatogram is recorded at a wavelength of 214nm: elution The corresponding time of peak start is 21.450min, and the corresponding time of peak end is 24.492min.

[0061] Integrate the chromatogram of sample 1, adjust the integration window from the corresponding time of peak start to 21.450min to the corresponding time of peak end of 24.492min, and obtain the peak area corresponding to sample 1 as 3289049;

[0062] Load sample 2 on a well-balanced high-effici...

Embodiment 3

[0067] Determination of CTAB content in group C meningococcus of embodiment 3

[0068] Precisely prepare 100μg / ml (C1) CTAB standard solution as sample 1;

[0069] Take the meningococcal group C polysaccharide solution to be tested as sample 2;

[0070] Load sample 1 on a well-balanced high-efficiency gel chromatographic column with a sample volume of 20 μl, eluted with 0.8% NaCl solution, and eluted at a flow rate of 0.4ml / min, and record the chromatogram at a wavelength of 214nm: the corresponding time of the elution peak It is 26.397min, and the corresponding time of peak end is 29.499min.

[0071] Integrate the chromatogram of sample 1, adjust the integration window from the corresponding time of 26.397 minutes from the peak start to the corresponding time of 29.499 minutes at the end of the peak, and obtain the peak area corresponding to sample 1 as 3240747;

[0072] Load sample 2 on a well-balanced high-efficiency gel chromatographic column, with a sample volume of 20 ...

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Abstract

The invention relates to a measuring method of the CTAB (Cetyltrimethyl Ammonium Bromide) content in meningococcal polysaccharide. The method comprises the following steps: taking CTAB standard solution and meningococcal polysaccharide solution samples, respectively placing the samples on a high-efficiency gel chromatographic column, and calculating the concentration of CTAB in the meningococcal polysaccharide solution sample according to a chromatogram. The measuring method of the CTAB content in the meningococcal polysaccharide, provided by the invention, is accurate, good in repeatability, quick and convenient.

Description

technical field [0001] The invention relates to the field of testing and analysis, in particular to a method for measuring CTAB content in meningococcal polysaccharides by using high-school liquid chromatography. Background technique [0002] CTAB, full name cetyltrimethylammonium bromide, has the property of precipitating nucleic acid and acidic polysaccharide from low ionic strength solution. During polysaccharide purification, CTAB acts as a chelating agent to precipitate complex sugars. However, CTAB is irritating to a certain extent and can cause harm to the human body through inhalation, ingestion or skin absorption. In recent years, during the declaration process of biological products, the state stipulates that the residues of organic substances such as CTAB should be specified to reduce the risk of products. Therefore, the accurate determination of CTAB residues in meningitis polysaccharide stock solution has an important impact on vaccine production and is of gre...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 王婷婷魏文进
Owner BEIJING MINHAI BIOTECH
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