High-efficiency liquid-phase chromatography detection method of L-arginine-alpha-ketoglutarate

A high-performance liquid chromatography and ketoglutaric acid technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor precision, low accuracy, and decreased column efficiency of spectrophotometry, and achieve good precision and Accuracy, labor saving, quality controllable effect

CN104483406AActive Publication Date: 2015-04-01JING JING PHARMA
1 Cites 9 Cited by

Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2015-04-01

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

Abstract

The invention discloses a high-efficiency liquid-phase chromatography detection method of L-arginine-alpha-ketoglutarate, and relates to the technical field of column chromatography. According to chromatographic condition, a chromatographic column adopts a C18 chromatographic column; a mobile phase adopts a 0.02mol / L to 0.08mol / L phosphate buffer solution with pH value of 3.0 to 4.0; the flow rate of the mobile phase is 0.5 to 1.3 ml / min; the detection wavelength is 200nm to 210nm; the temperature of the chromatographic column is 15 to 30 DEG C, and the content of L-arginine and the content of alpha-ketoglutarate in L-arginine-alpha-ketoglutarate or preparations of the L-arginine-alpha-ketoglutarate are determined by carrying out the high-efficiency liquid-phase chromatographic analysis. The content of L-arginine and the content of alpha-ketoglutarate in L-arginine-alpha-ketoglutarate or preparations of the L-arginine-alpha-ketoglutarate can be accurately determined by virtue of one-step high-efficiency liquid-phase chromatographic detection, and the two components can be well separated; moreover, the balance time of the C18 chromatographic column is short and is about 30 to 40 minutes, and the working efficiency can be improved.
Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] The invention relates to the technical field of column chromatography. Background technique

[0002] L-arginine-α-ketoglutaric acid is an amino acid salt health product with increasing sales volume in the international market. It is mainly used to improve human protein and enhance human physique. There are two specifications of 1:1 and 2:1 (the molar ratio of arginine and ketoglutaric acid). The liver-protecting drugs developed by pharmaceutical companies can directly and quickly provide nutrition for liver dysfunction, provide energy for liver cells, and quickly maintain the normal function and health of the liver.

[0003] In the prior art, Zhongjing Hongwu et al. have disclosed the use of spectrophotometry to measure α-ketoglutarate (simultaneous determination of α-ketoglutarate and pyruvic acid by spectrophotometry, Journal of Tongji University, 1986, 14 (4): 531- 536), but the spectrophotometric method has poor precision and low accuracy, and ca...

Examples

Embodiment 1

[0038] Chromatographic conditions

[0039] Mobile phase: 0.05mol / L sodium dihydrogen phosphate buffer (use 0.05mol / L phosphoric acid to adjust the pH value to 3.4)

[0040] Mobile phase flow rate: 1.0ml / min

[0041] Detection wavelength: 205nm

[0042] Column temperature: 25°C

[0043] Experimental steps:

[0044] (1) Prepare mobile phase, adjust pH value, filter and degas;

[0045] (2) Adjust the conditions of the high-performance liquid chromatograph, and start to balance the chromatographic column;

[0046] (3) Configuration of reference substance and test solution:

[0047] Reference substance solution: Weigh 34.0 mg of L-arginine reference substance and 29.0 mg of α-ketoglutarate reference substance into a 100ml measuring bottle, add mobile phase to dilute to the mark, and shake well to obtain reference substance solution.

[0048]Test solution: Weigh 64 mg of L-arginine-α-ketoglutarate (the molar ratio of L-arginine to α-ketoglutarate is 1:1) and place it in a 100m...

Embodiment 2

[0054] Chromatographic conditions

[0055] Mobile phase: 0.05mol / L sodium dihydrogen phosphate buffer (use 0.05mol / L phosphoric acid to adjust the pH value to 3.4)

[0056] Mobile phase flow rate: 1.0ml / min

[0057] Detection wavelength: 205nm

[0058] Column temperature: 25°C

[0059] Experimental steps:

[0060] (1) Prepare mobile phase, adjust pH value, filter and degas;

[0061] (2) Adjust the conditions of the high-performance liquid chromatograph, and start to balance the chromatographic column;

[0062] (3) Configuration of reference substance and test solution:

[0063] Reference substance solution: Weigh 68.0 mg of L-arginine reference substance and 29.0 mg of α-ketoglutarate reference substance into a 100ml measuring bottle, add mobile phase to dilute to the mark, and shake well to obtain reference substance solution.

[0064] The test solution: Weigh 96 mg of L-arginine-α-ketoglutarate (the molar ratio of L-arginine to α-ketoglutarate is 2:1) and place it in a...

Embodiment 3

[0088] The difference with Example 1 is that the chromatographic conditions:

[0089] Mobile phase: 0.05mol / L potassium dihydrogen phosphate buffer (use 0.05mol / L phosphoric acid to adjust the pH value to 3.4);

[0090] Mobile phase flow rate: 0.5ml / min;

[0091] Detection wavelength: 200nm;

[0092] Column temperature: 15°C.

[0093] Configuration of reference substance and test solution:

[0094] Reference substance solution: Weigh 16 mg of L-arginine reference substance and 14 mg of α-ketoglutarate reference substance in a 100ml measuring bottle, add mobile phase to dilute to the mark, and shake well to obtain reference substance solution.

[0095] For the test solution, weigh 30 mg of L-arginine-α-ketoglutarate (the molar ratio of L-arginine to α-ketoglutarate is 1:1) and place it in a 100ml measuring bottle, add flow Phase dissolved and diluted to the mark, shake well, that is.

[0096] Perform HPLC analysis to determine the content of L-arginine and α-ketoglutarate ...