A method for detecting betaine content in fermented liquid

A technology of betaine and fermentation liquid, which is applied in the field of chemical analysis, can solve problems such as the inability to accurately measure the betaine content in the fermentation liquid, the difficulty of effectively separating the matrix interference peak from the betaine peak, and the damage to the life of the alkyl silica gel chromatographic column. The detection method is simple and fast, the resolution is good, and the effect of eliminating the interference of complex matrix

Active Publication Date: 2020-06-30
河北华北制药华恒药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The all-aqueous mobile phase usually damages the life of the alkyl silica gel column, and the ion-pairing reagent mobile phase usually requires a long system equilibration time, and the post-cleaning of the system is also time-consuming
The samples in the examples are all high-purity, high-content betaine (50%). For the determination of low-content and complex matrix samples in the fermentation broth, there are also high background absorption, unstable baseline, and it is difficult to effectively separate the matrix interference peak and the betaine peak. Therefore, the patented method cannot accurately determine the content of betaine in the fermentation broth

Method used

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  • A method for detecting betaine content in fermented liquid
  • A method for detecting betaine content in fermented liquid
  • A method for detecting betaine content in fermented liquid

Examples

Experimental program
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Effect test

Embodiment 1

[0064] A method for detecting betaine content in fermented liquid, which utilizes high performance liquid chromatography to detect, and the specific detection steps are as follows:

[0065] A. Prepare betaine standard solutions of different concentrations, and use high performance liquid chromatography to establish a standard curve of betaine solution concentration and peak area.

[0066] A-1. Weigh betaine or betaine hydrochloride standard

[0067] Accurately weigh 0.6560g of the standard betaine hydrochloride (dried at 105°C) and dissolve it in a 100mL volumetric flask, dilute to the mark with water, and shake well to obtain a betaine 5.00mg / mL standard solution as a standard stock solution.

[0068] A-2. Betaine series standard solution preparation

[0069] Accurately draw 0.2mL, 0.4mL, 1.0mL, 2.0mL, 5.0mL, 10.0mL of betaine standard stock solution (in A-1) into 10mL volumetric flasks, dilute to volume with pure water, and mix well. Prepare standard series with mass conce...

Embodiment 2

[0091] A method for detecting betaine content in fermented liquid, which utilizes high performance liquid chromatography to detect, and the specific detection steps are as follows:

[0092] A. Prepare betaine standard solutions of different concentrations, and use high performance liquid chromatography to establish a standard curve of betaine solution concentration and peak area.

[0093]A-1. Weigh betaine or betaine hydrochloride standard

[0094] Accurately weigh 0.6560g of the standard product betaine hydrochloride (dried at 105°C) and dissolve it in a 100mL volumetric flask, dilute to the mark with acetonitrile-water mixed solution, wherein the volume ratio of acetonitrile and water is 55:45, shake well and Betaine 5.00mg / mL standard solution, as the standard stock solution.

[0095] A-2. Betaine series standard solution preparation

[0096] Accurately draw 0.2mL, 0.4mL, 1.0mL, 2.0mL, 5.0mL, 10.0mL of betaine standard stock solution (in A-1) into 10mL volumetric flasks, ...

Embodiment 3

[0116] A method for detecting betaine content in fermented liquid, which utilizes high performance liquid chromatography to detect, and the specific detection steps are as follows:

[0117] A. Prepare betaine standard solutions of different concentrations, and use high performance liquid chromatography to establish a standard curve of betaine solution concentration and peak area.

[0118] A-1. Weigh betaine or betaine hydrochloride standard

[0119] Precisely weigh 0.5768g of betaine monohydrate, dissolve it in a 100ml volumetric flask, dilute to the mark with acetonitrile-water mixed solution, wherein the volume ratio of acetonitrile and water is 45:55, and shake well to get betaine 5.00mg / mL standard solution, as a standard stock solution.

[0120] A-2. Betaine series standard solution preparation

[0121] Accurately draw 0.2mL, 0.4mL, 1.0mL, 2.0mL, 5.0mL, 10.0mL of betaine standard stock solution (in A-1) into 10mL volumetric flasks, dilute to the mark with acetonitrile-...

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Abstract

The invention discloses a method for detecting the content of betaine in fermented liquid and belongs to the technical field of chemical analysis. The method comprises the step of detecting by utilizing a high-performance liquid chromatography, wherein a chromatographic column in a high-performance liquid chromatographic condition is an inverse amino chromatographic column, a detector is an evaporation light scattering detector and a mobile phase is a gradient mobile phase formed by mixing acetonitrile and water. The detection method disclosed by the invention has a strong matrix interferenceresisting capability, the separation degree of the betaine and a matrix is high and interference of the complicated matrixes in the fermented liquid can be completely eliminated; the detection methodis simple and rapid and has good stability and an accurate and reliable result.

Description

technical field [0001] The invention relates to betaine, in particular to a method for detecting betaine content, which belongs to the technical field of chemical analysis. Background technique [0002] Vitamin B12 (Vitamin B12) is one of the B vitamins, also known as cobalamin, a complex organic compound containing cobalt. The molecular structure is a nucleotide composed of a cobalt ion-centered corrin ring and 5,6-dimethylbenzimidazole as a base. Vitamin B12 in nature is synthesized by microorganisms. Higher animals and plants cannot produce vitamin B12. Vitamin B12 is the only vitamin that requires the help of an intestinal secretion (endogenous factor) to be absorbed. The main physiological function of vitamin B12 is to participate in the manufacture of bone marrow red blood cells, prevent pernicious anemia, and prevent brain nerves from being damaged. [0003] In the process of producing vitamin B12 by Pseudomonas denitrification bacteria, betaine is the precursor of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8809
Inventor 李晓宇张现华谢广健王炜陈志彬刘欣章志峰宋海英
Owner 河北华北制药华恒药业有限公司
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