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Content detection method for lysine of compound ketoacid tablet

A detection method and lysine technology, applied in the field of lysine content detection, can solve the problems of significant double peaks of lysine and poor detection precision, and achieve the advantages of shortening detection time and improving sample injection precision. Effect

Inactive Publication Date: 2013-04-17
SHANGHAI JINGFENG PHARMA
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Problems solved by technology

[0007] When directly using the OPA reagent and amino acid detection method provided by Agilent to detect lysine samples, the double peak problem of lysine is obvious, and the detection precision is not good

Method used

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  • Content detection method for lysine of compound ketoacid tablet

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Effect test

Embodiment 2

[0045] Detection method of the present invention, self-prepared OPA reagent containing 3% (v / v) 3-mercaptopropionic acid:

[0046] (1) Preparation of 0.4mol / L boric acid buffer solution: Take 4.96g boric acid and 2.80g sodium hydroxide, add water to dissolve and dilute to 200ml. Adjust the pH to 10.20 with 2mol / L NaOH solution.

[0047] (2) Preparation of o-phthalaldehyde reaction solution: Weigh an appropriate amount of o-phthalaldehyde, add methanol or ethanol to dissolve to a constant volume, and prepare a 100 mg / ml o-phthalaldehyde stock solution; absorb an appropriate amount of o-phthalaldehyde stock solution, Add 3% (v / v) 3-mercaptopropionic acid, mix well, and then dilute with boric acid buffer to prepare a reaction solution with a concentration of phthalaldehyde of 10 mg / ml.

[0048] (3) Determination of sample solution: Under the high performance liquid chromatography that can use automatic pre-column derivatization, pre-column derivatization of lysine samples is car...

Embodiment 3

[0058] Detection method of the present invention, self-prepared OPA reagent containing 2% (v / v) 3-mercaptopropionic acid:

[0059] (1) Preparation of 0.4mol / L boric acid buffer solution: Take 4.96g boric acid and 2.80g sodium hydroxide, add water to dissolve and dilute to 200ml. Adjust the pH to 10.20 with 2mol / L NaOH solution.

[0060] (2) Preparation of o-phthalaldehyde reaction solution: Weigh an appropriate amount of o-phthalaldehyde, add methanol or ethanol to dissolve to a constant volume, and prepare a 100 mg / ml o-phthalaldehyde stock solution; absorb an appropriate amount of o-phthalaldehyde stock solution, Add 2% (v / v) 3-mercaptopropionic acid, mix well, and then dilute with boric acid buffer to prepare a reaction solution with an o-phthalaldehyde concentration of 10 mg / ml.

[0061] (3) Determination of sample solution: Under the high performance liquid chromatography that can use automatic pre-column derivatization, pre-column derivatization of lysine samples is car...

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Abstract

The invention relates to a content detection method for lysine of a compound ketoacid tablet, which comprises the following steps: preparing an o-phthalaldehyde reaction liquid (OPA reagent) with the concentration of 10 mg / ml, wherein 2-3 percent (v / v) of 3-thiohydracrylic acid is contained; afterwards, mixing lysine and the reaction liquid at ratio of 1:1(v / v) in a borate buffer system with a pH valve of 10.2 plus or minus 0.1 through an automatic pre-column derivation method, and then carrying out ambient temperature reaction; and finally, adopting a high performance liquid chromatography to measure the content of the lysine at 338 nm. The method provided by the invention can effectively solve the problem that bimodality of the lysine occurs in the current OPA pre-column derivation method, improves the sample introduction precision of the lysine in the detection through the automatic pre-column derivation method, greatly shortens the detection time, and is particularly suitable for detecting the content of the lysine of amino acid samples in a laboratory.

Description

Technical field: [0001] The invention relates to a method for detecting the content of lysine, in particular to a method for detecting the content of lysine in compound a-keto acid tablets. Background technique: [0002] At present, the most commonly used amino acid analysis techniques are amino acid analyzers and reversed-phase high-performance liquid chromatography combined with various pre-column or post-column derivatives. In recent years, great progress has been made in the determination of amino acids by reversed-phase high-performance liquid chromatography with pre-column derivatization. Compared with amino acid analyzers, the pre-column derivatization method has the advantages of rapidity, high sensitivity, and no need for special instruments. Commonly used pre-column derivatization reagents include o-phthalaldehyde (OPA), 9-fluorenylmethoxycarbonyl chloride (FMOC-Cl), phenylisothiocyanate (PITC) and dansyl chloride (Dansyl-Cl). Among them, OPA is the most widely u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 叶湘武付爱玲邱永锋
Owner SHANGHAI JINGFENG PHARMA
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