Method for analysizing amino acid

An analysis method and amino acid technology, applied in the field of amino acid analysis, can solve problems affecting the chromatographic separation results of DNFB-AA derivatives, etc., and achieve the effect of improving reproducibility and expanding the application range

Active Publication Date: 2006-03-22
TIANJIN PHARMA GROUP CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mobile phases prepared on different dates and different brands, or even columns of the same brand with different batch numbers may affect the chromatographic separation results of DNFB-AA derivatives

Method used

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  • Method for analysizing amino acid
  • Method for analysizing amino acid
  • Method for analysizing amino acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1 kidney disease injection (Amino acid for renal insufficiency, Hoechst Marion Produced by Roussel ) and liver disease injection (Amino acid preparation for hepatic insufficiency, Manufactured by Hoechst Marion Roussel Analysis of AA in )

[0048] 1. Reagents

[0049] 1.1.0.1M HCl 4mL hydrochloric acid mixed with 480mL water.

[0050] 1.2 1% DNFB 1g DNFB was dissolved in 100mL acetonitrile.

[0051] 1.3.0.5M NaHCO 3 2.1g NaHCO 3 Dissolve in 50mL water.

[0052] 1.4.0.1M phosphate buffer, pH6.5 Weighs 1.74g K 2 HPO 4 Dissolve in 100mL water to get 0.1MK 2 HPO 4 . Weigh 1.36g KH 2 PO 4 Dissolve in 100mL water to get 0.1M KH 2 PO 4 . To 0.1M KH 2 PO 4 Add 0.1M K 2 HPO 4 to pH=6.5.

[0053] 1.5.36mM triethylamine phosphate (TEAP), pH2.75 Take ~950mL water in a beaker, and add 5mL triethylamine to it with a pipette. After mixing, adjust the pH to 2.75 with phosphoric acid. Transfer the solution to a 1000mL volumetric flask and flush to t...

Embodiment 2

[0093] Example 2 Determination of content of main component propylglutamine in the bulk drug (produced by Tianjin Tiancheng Pharmaceutical Co., Ltd.) of propylglutamine (L-alanyl-L-glutamine)

[0094] 1. Reagents

[0095] 1.1. See Example 1.

[0096] 1. 2.6N HCl Concentrated hydrochloric acid and water are mixed in a ratio of 1:1 (V / V).

[0097] 2. Preparation of standard solution Take out the proglutamine reference substance from the desiccator, accurately weigh about 15mg into a polyethylene tube with a cover (1), add 1mL 6N HCl with a pipette, cover tightly, and shake well for later use.

[0098] 3. Preparation of sample solution Accurately weigh about 15 mg of proglutamine sample into a polyethylene tube with a cover (2), add 1 mL of 6N HCl with a pipette, cover tightly, and shake well for later use.

[0099] 4. Acid hydrolysis of standard solution and sample solution Heat polyethylene tubes (1) and (2) at 105°C for 6 hours. Let cool, shake well and set aside.

[0100]...

Embodiment 3

[0127] Determination of Proglutamine Content in Example 3 Proglutamine Injection (Sichuan Kelun Pharmaceutical Co., Ltd. Production)

[0128] 1. Reagents

[0129] 1.1. See Example 1.

[0130] 1.2.6.7N HCl 167mL concentrated hydrochloric acid diluted with water to 300mL.

[0131] 2. Preparation of standard solution Accurately weigh about 20mg of proglutamine or 13.5mg of glutamic acid reference substance into a polyethylene tube with a cover (1), add 100μL of water and 900μL of 6.7N HCl, cover tightly, and shake well for later use.

[0132] 3. Preparation of the sample solution Take 100 μL of the sample solution and 900 μL of 6.7N HCl in a polyethylene tube with a cover (2), cover it tightly, and shake well for later use.

[0133] 4. Acid hydrolysis Put the polyethylene tubes (1) and (2) in a heating block, heat at 105°C for 6 hours, take it out to cool, shake well and set aside.

[0134] 5. Derivatization reaction Use a pipette to draw 20mL each of the acid hydrolysis stand...

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Abstract

The amino acid analyzing method includes high efficiency liquid phase chromatographic separation of the 2, 4-dinitro fluoro benzene derivative of amino acid in C18 column with pH 2-3 or pH 6-7 TEAP/ACN as flowing phase; and 360nm ultraviolet detection at column temperature 35deg c. The present invention has raised separation result reproducibility, optional water phase buffering area and trimmed pH value for analysis of chromatographic column and sample, and accurate analysis result. The present invention is suitable for common HPLC analysis lab and the application range includes various amino acid analysis, indirect analysis of other matter capable of being converted into amino acid, and other matter capable of taking quantitative deriving reaction with DNFB.

Description

technical field [0001] The present invention relates to an analysis method for amino acids, based on the separation of amino acid-2,4-dinitrofluorobenzene derivatives using triethylamine phosphate / acetonitrile as the mobile reverse phase, using common high-performance liquid chromatography and chromatographic conditions, no longer need Other instrument configurations can complete the analysis of various amino acid samples and give accurate results. Background technique [0002] Amino acid (amino acid, AA) samples are relatively complex, and there are more than 20 common AAs. The chromatographic analysis method must have sufficient selectivity to distinguish the AA to be determined in the sample. Most AA has no obvious absorption to ultraviolet light. For detection, the most common method is to react the amino group in the AA molecule with a derivatizing agent to generate derivatives with ultraviolet absorption or fluorescence properties. [0003] AA samples are generally an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/88
Inventor 朱彭龄朱润之
Owner TIANJIN PHARMA GROUP CORP
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