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Method of detecting trihydroxymethyl aminomethane (Tris) in bioproduct by means of ion chromatography

A technology of tris(hydroxymethyl)aminomethane and ion chromatography, applied in the field of analysis and detection, can solve the problems of inability to quantitatively detect Tris, high limit of quantification, interference, etc., and achieve the effect of low detection limit, low quantitative limit and good selectivity.

Inactive Publication Date: 2018-03-09
SHANGHAI RAAS BLOOD PRODUCTS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the quantitative detection of Tris mainly adopts the method of acid-base titration, which has a high limit of quantification and is easily interfered by various acid-base groups.
However, due to the large amount of carboxyl groups, amino groups, and anions and cations contained in biological products, Tris in the products cannot be quantitatively detected by titration.

Method used

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  • Method of detecting trihydroxymethyl aminomethane (Tris) in bioproduct by means of ion chromatography
  • Method of detecting trihydroxymethyl aminomethane (Tris) in bioproduct by means of ion chromatography
  • Method of detecting trihydroxymethyl aminomethane (Tris) in bioproduct by means of ion chromatography

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The test sample is freeze-dried human thrombin produced by Shanghai RAAS Blood Products Co., Ltd., reconstituted with water for injection according to the regulations, diluted 500 times, and injected for detection. The instrument used in the experiment was ICS 5000 ion chromatograph from Thermo Fisher Company, and the chromatographic conditions were: Dianne Ionpac CS16 cation exchange column, including analytical column (250mm×5mm) and guard column (50mm×5mm), and the column temperature was 30°C; The post-column rehydration derivation method is adopted, the post-column rehydration is 500mmol / L sodium hydroxide solution; the mobile phase is methanesulfonic acid solution, and the flow rate of the mobile phase is 1.0mL / min; the detector is a pulsed amperometric detector with Au as the electrode , the Ag-AgCl electrode is the reference electrode, and the waveform is Carbohydrates; the Tris ion chromatogram is shown in figure 1 .

[0024] Standard solutions of different con...

Embodiment 2

[0026] The test sample is a 1% glycine solution, which is diluted 500 times and injected for detection. The instrument used in the experiment is the ICS 5000 ion chromatograph of ThermoFisher Company, and the chromatographic conditions are: Dianne Ionpac CS16 cation exchange column, including the analytical column (250mm × 5mm) and the guard column (50mm × 5mm), and the column temperature is 30°C; Post-column replenishment derivation method, the post-column replenishment is 500mmol / L sodium hydroxide solution; the mobile phase is methanesulfonic acid solution, and the flow rate of the mobile phase is 1.0mL / min; the detector is a pulsed amperometric detector with Au as the electrode, The Ag-AgCl electrode is the reference electrode, and the waveform is Carbohydrates; the Tris ion chromatogram is shown in figure 2 .

[0027] Because the pulsed amperometric detector can detect small molecular compounds containing hydroxyl and amino groups, it can be seen from the figure that th...

Embodiment 3

[0029] The test sample is freeze-dried human blood coagulation factor FVIII (AHF) produced by Shanghai Raas Blood Products Co., Ltd., reconstituted with water for injection according to the regulations, diluted 500 times, and injected for detection. The instrument used in the experiment is the ICS5000 ion chromatograph of Thermo Fisher Company, and the chromatographic conditions are: Dianne Ionpac CS16 cation exchange column, including the analytical column (250mm × 5mm) and the guard column (50mm × 5mm), and the column temperature is 30°C; Post-column replenishment derivation method, the post-column replenishment is 500mmol / L sodium hydroxide solution; the mobile phase is methanesulfonic acid solution, and the flow rate of the mobile phase is 1.0mL / min; the detector is a pulsed amperometric detector with Au as the electrode, The Ag-AgCl electrode is the reference electrode, and the waveform is Carbohydrates; the Tris ion chromatogram is shown in image 3 .

[0030] No Tris w...

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Abstract

The invention discloses a method of detecting trihydroxymethyl aminomethane (Tris) in a bioproduct by means of ion chromatography. The method comprises the following steps: drawing a concentration-peak area standard curve of Tris by means of an external standard method by adopting a pulse ampere detector and a post-column liquid supplementation deviating method through using CS16 cation-exchange chromatography to obtain a regression equation of the standard curve, wherein the mobile phase is a methanesulfonic acid solution and the post-column supplemented liquid is an alkaline solution; and calculating the content of Tris by means of the regression equation. According to the method, the lowest detection limit of Tris is 3.14<-5>mmol / L, the lowest limit of quantitation is 1.05<-4>mmol / L, the recovery rate is 93.8-100.1%, and the repeatability range is 0.10-0.52% (RSD, n=5). Without deviation and pre-treatment of a complicated sample, the method is neither interfered by acid-base groupscarried on proteins in the bioproduct and nor interfered by anions and cations in the sample, is high in detection sensitivity and high in analytical speed, and can be applied to content detection andquality judgment of Tris in the bioproducts in related industries.

Description

technical field [0001] The invention relates to a method for detecting trishydroxymethylaminomethane (Tris) in biological products by ion chromatography, belonging to the field of analysis and detection. Background technique [0002] Tris (Tris) is also known as tromethamine; tromethamine; 2-amino-2-(hydroxymethyl)-1,3-propanediol. Tris is a weak base that is often used as a biological buffer system in the production of biological products. Tris can cause hypoglycemia, hypotension, nausea, vomiting, and can also inhibit breathing or even stop breathing. Excessive intake or renal insufficiency can cause alkalemia. [0003] At present, the quantitative detection of Tris mainly adopts the method of acid-base titration, which has a high limit of quantification and is easily interfered by various acid-base groups. However, since biological products contain a large number of carboxyl groups, amino groups, and anions and cations, Tris in the products cannot be quantitatively det...

Claims

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Application Information

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IPC IPC(8): G01N30/64G01N30/96
CPCG01N30/96G01N30/64
Inventor 尉小平余进
Owner SHANGHAI RAAS BLOOD PRODUCTS CO LTD
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