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HPLC (high performance liquid chromatography) detection method of arginine ketoglutarate

A technology of ketoglutaric acid and arginine salt, which is applied in the field of biomedicine, can solve the problems of control reporting, low accuracy, and poor precision of spectrophotometry, and achieve the effect of solving separation problems and ensuring quality

Inactive Publication Date: 2013-01-30
BEIJING JIASHILIANBO PHARM SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

α-Ketoglutarate-L-arginine salt is easy to dissociate, so it is difficult to analyze its content, purity and related substances
In the prior art, there are only reports on the determination of α-ketoglutaric acid or L-arginine monomer content or related substances. Jing Hongwu et al. Simultaneous determination of diacid and pyruvic acid by spectrophotometry. Tongji University Journal, 1986, 14(4):531-536), but the accuracy of spectrophotometry is poor, and it can only measure α-ketoglutarate Content
Mao Ying et al disclosed the content of ketoglutaric acid determined by high performance liquid chromatography, which used Agilent ODS C 18 column, with 0.5% (NH 4 ) 2 HPO 4 -H 3 PO 4 The buffer solution is the mobile phase, the flow rate is 110ml / min, and the ultraviolet detection wavelength is 233nm (the content of ketoglutaric acid is determined by HPLC. China Pharmaceutical Affairs, 2008, 22(10): 598-599), but this method can only It is used to determine the content of ketoglutaric acid, and the content detection and inspection of related substances cannot be carried out for α-ketoglutarate-L-arginine salt
Until recently, there was no report on the quality control of α-ketoglutarate-L-arginine salt in the prior art

Method used

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  • HPLC (high performance liquid chromatography) detection method of arginine ketoglutarate
  • HPLC (high performance liquid chromatography) detection method of arginine ketoglutarate
  • HPLC (high performance liquid chromatography) detection method of arginine ketoglutarate

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Chromatographic conditions

[0046] Mobile phase: acetonitrile-phosphate buffered saline solution (0.02mol / L sodium dihydrogen phosphate solution, adjust the pH value to 5.2 with 0.02mol / L disodium hydrogen phosphate solution) = 60:40;

[0047] Flow rate: 1.0ml / min;

[0048] Detection wavelength: 205nm

[0049] Column temperature: room temperature

[0050] Experimental steps:

[0051] The test solution: Weigh 10 mg of α-ketoglutarate-L-arginine salt (1:2) into a 10ml measuring bottle, add mobile phase to dissolve and dilute to the mark, shake well, and you will get it.

[0052] Precisely draw 20 μl of the test solution, inject it into a high-performance liquid chromatograph, and record the chromatogram. The results are shown in the attached figure 1 . Depend on figure 1 It can be seen that peaks 1-3 are impurity peaks, peak 4 is L-arginine peak, and peak 5 is α-ketoglutarate peak. The separation between peaks 1-3 and the main peak was greater than 2.5. α-Ketoglu...

Embodiment 2

[0054] Chromatographic conditions

[0055] Mobile phase: acetonitrile-phosphate buffered saline solution (0.02mol / L sodium dihydrogen phosphate solution, add 0.2% triethylamine, adjust the pH value to 5.2 with dilute phosphoric acid) = 60:40;

[0056] Flow rate: 1.0ml / min;

[0057] Detection wavelength: 205nm

[0058] Column temperature: room temperature

[0059] Experimental steps:

[0060] The test solution: Weigh 10 mg of α-ketoglutarate-L-arginine salt (1:2) into a 10ml measuring bottle, add mobile phase to dissolve and dilute to the mark, shake well, and you will get it.

[0061] Precisely draw 20 μl of the test solution, inject it into a high-performance liquid chromatograph, and record the chromatogram. The results are shown in the attached figure 2 . Depend on figure 2 It can be seen that the main peak and the impurity peaks in the sample are well separated. Peak 1-2 and peak 4-5 are impurity peaks, peak 3 is L-arginine peak, and peak 6 is α-ketoglutarate peak...

Embodiment 3

[0063] Chromatographic conditions

[0064] Mobile phase: acetonitrile-phosphate buffered saline solution (0.02mol / L sodium dihydrogen phosphate solution, adding 0.2% triethylamine, adjusting the pH value to 5.2 with oxalic acid) = 60:40;

[0065] Flow rate: 1.0ml / min;

[0066] Detection wavelength: 205nm

[0067] Column temperature: room temperature

[0068] Experimental steps:

[0069] The test solution: Weigh 10 mg of α-ketoglutarate-L-arginine salt (1:2) into a 10ml measuring bottle, add mobile phase to dissolve and dilute to the mark, shake well, and you will get it.

[0070] Precisely draw 20 μl of the test solution, inject it into a high-performance liquid chromatograph, and record the chromatogram. The results are shown in the attached image 3 . Depend on image 3 It can be seen that the main peak and the impurity peaks in the sample are well separated. Peaks 1-3 are impurity peaks, peak 4 is L-arginine peak, and peak 5 is α-ketoglutarate peak. The separation b...

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Abstract

A method for detecting L-arginine-alpha-ketoglutarate using high performance liquid chromatography(HPLC). Chromatographic conditions include: chromatographic column is amino column; mobile phase is methanol-acetonitrile-phosphate buffer in volume ratio of 0-20:30-70:20-50 and detection wavelength is 200-220nm.

Description

technical field [0001] The invention relates to a method for measuring α-ketoglutarate-L-arginine salt in the field of biomedicine, in particular to a method for measuring α-ketoglutarate-L-arginine salt and The content of its preparation and the method of related substances. Background technique [0002] α-ketoglutaric acid-L-arginine salt (1:2) and its preparations are liver-protecting drugs developed by French Ruifalan Pharmaceutical Factory. It targets liver dysfunction, can directly and quickly provide nutrients, provide energy to liver cells, and quickly maintain the normal function and health of the liver. [0003] In the process of new drug research, accurate determination and analysis of drug content and impurities are important conditions to ensure its quality. α-Ketoglutarate-L-arginine salt is easy to dissociate, so it is difficult to analyze its content, purity and related substances. In the prior art, there are only reports on the determination of α-ketoglut...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/60
CPCG01N30/88G01N2030/8813G01N30/74G01N30/34
Inventor 温守明常龙陈小平
Owner BEIJING JIASHILIANBO PHARM SCI & TECH
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