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A Rapid Method for Analyzing Amino Acid Mixtures

A rapid analysis, amino acid technology, applied to the analysis of materials, material separation, measurement devices, etc., can solve the problems of short service life and high cost of chromatographic columns, and achieve the effect of reducing analysis cost, low price and good separation

Active Publication Date: 2016-05-04
BENGBU BBCA MEDICINE SCI DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the existing methods for detecting amino acids use amino-bonded silica gel columns, which have the disadvantages of being unable to achieve effective separation of multiple amino acids at the same time, short service life of the chromatographic column, and high cost.

Method used

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  • A Rapid Method for Analyzing Amino Acid Mixtures
  • A Rapid Method for Analyzing Amino Acid Mixtures
  • A Rapid Method for Analyzing Amino Acid Mixtures

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1HPLC (high performance liquid chromatography) measures arginine quantitative limit, detection limit

[0039] Chromatographic column: amide-bonded silica gel column (WATERS 3.5 μm 4.6×150 mm).

[0040] Mobile phase: Acetonitrile: Phosphate buffer = 60:40; Phosphate buffer: Take 3.2664g of potassium dihydrogen phosphate, add 240mL of water to dissolve, add 5mL of ammonia water, and adjust the pH to 4.5±0.1 with phosphoric acid.

[0041] Detection wavelength: 205nm, flow rate: 1.0mL / min, column temperature: 25°C.

[0042] Take an appropriate amount of arginine, dissolve it with 65% acetonitrile, inject 20 μL of the solution into the liquid chromatograph, and record the liquid chromatogram (such as figure 1 shown). The test product was diluted step by step and detected by liquid chromatography. When the signal-to-noise ratio was 10:1, the quantification limit of arginine was established, and the quantification limit was obtained: 1.5×10 -4 mg / mL, dilute the t...

Embodiment 2

[0043] The ratio of aspartic acid and ornithine in the determination of gate ornithine by HPLC in embodiment 2

[0044] Chromatographic column: amide-bonded silica gel column (WATERS 3.5 μm 4.6×150 mm).

[0045] Mobile phase: Acetonitrile: Phosphate buffer = 60:40; Phosphate buffer: Take 13.61g of potassium dihydrogen phosphate, add 500mL of water to dissolve, add 5mL of 25-28% ammonia water, add water to dilute to 1000mL, mix with phosphoric acid Adjust the pH to 4.2 ± 0.1.

[0046] Detection wavelength: 205nm, flow rate: 1.0mL / min, column temperature: 25°C.

[0047] The sample composition of this embodiment is: aspartic acid and ornithine.

[0048] Take an appropriate amount of ornithine, add 65% acetonitrile to dissolve and dilute to make a solution of about 4 μg / mL as the test solution; accurately measure 20 μL, inject it into a liquid chromatograph, and record the chromatogram (such as image 3 shown). After comparing with the reference substance solution, the results...

Embodiment 3

[0049] Embodiment 3HPLC measures glycine content

[0050] Chromatographic column: amide-bonded silica gel column (WATERS 3.5 μm 4.6×150 mm).

[0051]Mobile phase: Acetonitrile: Phosphate buffer = 60:40; Phosphate buffer: Take 13.61g of potassium dihydrogen phosphate, add 500mL of water to dissolve, add 5mL of 25-28% ammonia water, add water to dilute to 1000mL, adjust with phosphoric acid after mixing pH to 4.2 ± 0.1.

[0052] Take an appropriate amount of glycine, weigh it accurately, add 65% acetonitrile to dilute to make each 1mL solution contain about 2μg of glycine, the detection wavelength is 205nm, the flow rate is 1.0mL / min, and the column temperature is 25°C. Inject 20 μL into the liquid chromatograph, record the liquid chromatogram, see Figure 4 , about 2.7 minutes in the figure is the peak of glycine. Under this measurement condition, the peak shape of glycine is good. The main component self-control method is used for 6 measurements, and the average value of the...

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Abstract

The invention relates to the technical field of pharmaceutical analysis, and provides a method for rapid analysis of an amino acid mixture. The amino acid mixture to be measured is diluted into a test solution by a mobile phase, and then the amino acid mixture is analyzed by high performance liquid chromatography, wherein chromatographic column is an amide bonded silica gel column. The method has the following advantages: 1, the method can conveniently detect a plurality of amino acids at the same time achieve good separation; 2, the method has high analysis sensitivity, accurate and reliable results, is simple and quick, and can be used to analysis detection of amino acids or drugs containing amino acid impurities; 3, amide bonded silica gel column has lower background noise than amino bonded silica gel, is conducive to the protection of the chromatographic column, and has good stability; 4, compared with amino bonded silica gel column, the amide bonded silica gel column has low price and reduces analysis cost; and 5, the peak type obtained from amide column detection is better than that obtained from the existing amino chromatographic column.

Description

technical field [0001] The invention relates to the technical field of drug analysis, in particular to a method for rapidly analyzing amino acid mixtures using high-performance liquid chromatography. Background technique [0002] Amino acids have been widely used in food and medicine. It has been found through research that some medicines often contain a variety of amino acids, and the related substances in various amino acids are mainly other amino acids. Most of the methods currently used are thin-layer chromatography, which is qualitative and quantitative. Difficulties, especially other amino acids, when the content is around 0.1%, no impurity spots can be seen on the TLC plate. The proportion of amino acids directly affects the quality of the preparation, thereby affecting the efficacy of the drug. During the chromatographic separation process, the chromatographic column, column temperature, flow rate, and pH of the mobile phase will all affect the chromatographic separ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 李国艳孙建华李园刘铜梅熊媛媛
Owner BENGBU BBCA MEDICINE SCI DEV