CN105510497A

CN105510497A - Quantitative detection method of L-alanyl-L-alanine

Info

Publication number: CN105510497A
Application number: CN201510851390.8A
Authority: CN
Inventors: 李斌水; 何连顺
Original assignee: JING JING PHARMACEUTICAL Co Ltd
Current assignee: JING JING PHARMACEUTICAL Co Ltd
Priority date: 2015-11-30
Filing date: 2015-11-30
Publication date: 2016-04-20

Abstract

The invention relates to the technical field of quantitative detection of L-alanyl-L-alanine and specifically discloses a quantitative detection method of L-alanyl-L-alanine. High performance liquid chromatography is used in the detection. A chromatographic column is an amino bonded silica gel column, and column temperature is 30 DEG C. An acetonitrile-0.05 mol/L monopotassium phosphate buffer solution (645:350) is used as a mobile phase, pH of monopotassium phosphate buffer salt is adjusted to 4.0 by the use of phosphoric acid, and flow velocity is 0.7 ml/min. detection wavelength is 215 nm. Through an external standard method, content of L-alanyl-L-alanine in a sample to be tested is measured. According to the method, content of L-alanyl-L-alanine in a chemical product can be measured simply, rapidly and accurately. The method can be used in detecting content of L-alanyl-L-alanine in chemical production and is applicable to industrial production.

Description

The quantitative detecting method of L-Ala-L-Ala

Technical field

The present invention relates to the quantitative measurement technology field of L-Ala-L-Ala.

Background technology

L-Ala-L-Ala is important biochemical reagents and chemical intermediate, is also a kind of amino acids drug developed simultaneously, in order to utilize L-alanylamino propionic acid, just needs to detect its content, thus judge product quality situation.But, so far, in prior art, also there is no a kind of method detecting L-Ala-L-Ala content.

Summary of the invention

The technical problem to be solved in the present invention is to provide a kind of quantitative detecting method of L-Ala-L-Ala, adopt high-performance liquid chromatography, the content of L-Ala-L-Ala in Chemical Manufacture product can be measured fast and accurately, be suitable for using in commercial production.

For solving the problems of the technologies described above, the technical solution used in the present invention is: the quantitative detecting method of L-Ala-L-Ala, and use high performance liquid chromatography to detect, chromatographic column used is amino bonded silicagel column, and column temperature is 30 DEG C; With acetonitrile-0.05mol/L potassium dihydrogen phosphate buffer solution 645:350 for mobile phase, its potassium dihydrogen phosphate buffer salt phosphoric acid adjusts PH=4.0, and flow velocity is 0.7ml/min; Determined wavelength is 215nm;

Detecting step is:

1) testing sample mobile phase dilution preparation is become sample liquid to be measured, L-Ala-L-Ala standard items being dissolved in mobile phase preparation becomes standard solution;

2) adopt identical testing conditions high performance liquid chromatograph to detect respectively liquid to be measured for sample, standard solution, obtain sample liquid chromatography figure to be measured, standard solution chromatogram, calculated the content of L-Ala-L-Ala in testing sample by external standard method.

In described external standard method measuring and calculating testing sample, the content method of L-Ala-L-Ala is:

X=C _sampleextension rate=(the C of × testing sample _right× A _sample/ A _rightthe extension rate of) × testing sample,

In formula: C _rightfor the concentration of L-Ala-L-Ala in standard solution, unit is mg/ml;

A _rightfor peak area or the peak height of standard solution chromatogram;

A _samplefor peak area or the peak height of sample liquid chromatography figure to be measured;

C _samplefor the concentration of solution to be measured in sample liquid to be measured, unit is mg/ml.

Further, the chromatographic column sample size of sample liquid to be measured and standard solution is 20 μ l.

Further, sample liquid to be measured and standard solution are all through organic membrane filtration of 0.45 μm

The beneficial effect that produces of technique scheme is adopted to be: the present invention can easy, the content that measures L-Ala-L-Ala in Chemical Manufacture product fast and accurately, the content of L-Ala-L-Ala in Chemical Manufacture can be detected, thus targetedly L-Ala-L-Ala is reclaimed, thus play the good effect improving income, there is significant social benefit, be suitable for using in commercial production.

Embodiment

Do not have one fast to solve in prior art Chemical Manufacture, the method of accurate detection L-Ala-L-Ala content, causing can not to effective detection and control of L-Ala-L-Ala content in chemical products or intermediate product, the invention provides a kind of quantitative detecting method of L-Ala-L-Ala, adopt high-performance liquid chromatography, can be quick, measure the content of L-Ala-L-Ala in Chemical Manufacture product accurately, be suitable for using in commercial production, be convenient to monitor L-Ala-L-Ala in chemical products or intermediate product, thus adjustment production technology, improve productivity effect.

The present invention is further detailed explanation below.

A detection method for L-Ala-L-Ala, its step is as follows:

1) preparation of standard solution: take in mark product L-Ala-L-Ala solid 25mg to 25ml volumetric flask, use mobile phase constant volume; By the organic membrane filtration of solution with 0.45 μm, get in 1ml filtrate to 100ml volumetric flask and also use mobile phase constant volume, obtain standard solution.

2) preparation of sample solution to be measured: by the organic membrane filtration of liquid phase testing sample with 0.45 μm, then diluting with mobile phase is sample solution to be measured.

3) standard solution and sample solution to be measured are detected by high performance liquid chromatography respectively, testing conditions is:

Be filling agent with amino bonded silicagel column, with acetonitrile-0.05mol/L potassium dihydrogen phosphate (645:350) for mobile phase, determined wavelength is 215nm, and its potassium dihydrogen phosphate buffer salt phosphoric acid adjusts PH=4.0, and column temperature is 30 DEG C, and flow velocity is 0.7ml/min;

4) cubage: in analyte sample fluid, the content of L-Ala-L-Ala is:

A _rightfor peak area or the peak height of standard solution chromatogram;

Embodiment 1-3

Embodiment 1-3 gets different L-Ala-L-Ala samples respectively, and measure as stated above, measurement data sees the following form.

Claims

The quantitative detecting method of 1.L-alanyl-ALANINE, is characterized in that: use high performance liquid chromatography to detect.
2. the quantitative detecting method of L-Ala-L-Ala according to claim 1, is characterized in that, chromatographic column used is amino bonded silicagel column, and column temperature is 30 DEG C; With acetonitrile-0.05mol/L potassium dihydrogen phosphate buffer solution 645:350 for mobile phase, its potassium dihydrogen phosphate buffer salt phosphoric acid adjusts PH=4.0, and flow velocity is 0.7ml/min; Determined wavelength is 215nm;

Detecting step is:

1) testing sample mobile phase dilution preparation is become sample liquid to be measured, L-Ala-L-Ala standard items being dissolved in mobile phase preparation becomes standard solution;

2) adopt identical testing conditions high performance liquid chromatograph to detect respectively liquid to be measured for sample, standard solution, obtain sample liquid chromatography figure to be measured, standard solution chromatogram, calculated the content of L-Ala-L-Ala in testing sample by external standard method.
3. the quantitative detecting method of L-Ala-L-Ala according to claim 2, is characterized in that the content method of L-Ala-L-Ala in described external standard method measuring and calculating testing sample is:

X=C _sampleextension rate=(the C of × testing sample _right× A _sample/ A _rightthe extension rate of) × testing sample

In formula: C _rightfor the concentration of L-Ala-L-Ala in standard solution, unit is mg/ml;

A _rightfor peak area or the peak height of standard solution chromatogram;

A _samplefor peak area or the peak height of sample liquid chromatography figure to be measured;

C _samplefor the concentration of solution to be measured in sample liquid to be measured, unit is mg/ml.
4. the quantitative detecting method of L-Ala-L-Ala according to claim 2, is characterized in that the chromatographic column sample size of sample liquid to be measured and standard solution is 20 μ l.
5. the quantitative detecting method of L-Ala-L-Ala according to claim 2, is characterized in that sample liquid to be measured and standard solution all through organic membrane filtration of 0.45 μm.