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A kind of method for the determination of sodium lauryl sarcosinate content by high performance liquid chromatography

A technology of sodium lauryl sarcosine and high performance liquid chromatography, which is applied in the field of detection, can solve problems such as inaccurate measurement results, and achieve the effects of wide application range, rapid separation, and accurate and reliable detection results

Active Publication Date: 2022-07-19
厦门德馨尚品医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, when determining the content of surfactants in biological products, the results are inaccurate due to the influence of the combination of proteins and surfactants

Method used

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  • A kind of method for the determination of sodium lauryl sarcosinate content by high performance liquid chromatography
  • A kind of method for the determination of sodium lauryl sarcosinate content by high performance liquid chromatography
  • A kind of method for the determination of sodium lauryl sarcosinate content by high performance liquid chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Preparation of Solutions

[0048] Precisely weigh 1.00 g of sodium lauryl sarcosinate reference substance, dissolve it in ultrapure water, and use a volumetric flask to dilute to 100 mL to obtain a 10 mg / mL sodium lauryl sarcosinate reference substance solution; Amount of bovine serum albumin sample containing lauryl sarcosine ammonia 5.00g, according to the estimated content of sodium lauryl sarcosinate, diluted with ultrapure water to the content of sodium lauryl sarcosinate in Between 0.05-2.00 mg / mL, a sample solution to be tested containing sodium lauryl sarcosinate was obtained; water for injection (WFI) was used as a blank control.

[0049] 2. Chromatographic conditions

[0050] Measure an appropriate amount of ultrapure water, add 1 mL of trifluoroacetic acid, add ultrapure water to 1000 mL, and filter with a 0.22 μm membrane to obtain a 0.1% trifluoroacetic acid-water solution as mobile phase A; weigh an appropriate amount of acetonitrile, add 0.85 mL The ...

Embodiment 2

[0057] The preparation of the solution, mobile phase A, mobile phase B, gradient elution procedure and chromatographic column are the same as in Example 1. The injection volume was 52 μL, the flow rate was 1.2 mL / min, and the detection wavelength was 218 nm. There are chromatographic peaks with the same retention time at the corresponding positions of the chromatograms of the reference solution and the sample solution to be tested.

Embodiment 3

[0059] 3.1 Solution preparation and chromatographic conditions

[0060] Precisely weigh 1.00 g of sodium lauryl sarcosinate, dissolve it in ultrapure water, and dilute the volume to 100 mL in a volumetric flask to obtain a reference solution of 10.00 mg / mL. When injecting, the mass concentration is 10.00 mg / mL 5 μL, 10 μL, 25 μL, 50 μL, 100 μL, 150 μL and 200 μL of the reference solution were injected respectively, that is, 0.05 mg / mL, 0.10 mg / mL, 0.25 mg / mL, 0.50 mg / mL, 1.00 mg / mL and 1.50 mg / mL were obtained respectively. mg / mL and 2.00 mg / mL reference solutions, labeled STD-0.05, STD-0.10, STD-0.25, STD-0.50, STD-1.00, STD-1.50, and STD-2.00.

[0061] According to the estimated content of sodium lauryl sarcosinate in a batch of ApoJ samples (referred to as sample 1 to be tested) and another batch of ApoJ samples (referred to as sample 2 to be tested), use ultrapure water Dilute until the sodium lauryl sarcosinate content is between 0.05-2.00 mg / mL.

[0062] The quality co...

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Abstract

The invention relates to a method for determining the content of sodium lauryl sarcosinate by high performance liquid chromatography. The chromatographic column is a C18-ST chromatographic column (4.6mm×250mm, 5μm,), the mobile phase A is a 0.1% trifluoroacetic acid-water solution, and the mobile phase B is a 0.085% trifluoroacetic acid-acetonitrile solution, using gradient elution. The content of sodium lauryl sarcosinate, especially in protein samples, can be quantitatively detected by external standard method. Add equal volume of acetonitrile to the blank control solution WFI, sodium lauryl sarcosinate reference solution and test sample solution containing sodium lauryl sarcosinate, mix well, centrifuge, and take the supernatant for testing. HPLC analysis. Sodium lauryl sarcosinate showed a good linear relationship in the mass concentration range of 0.05-2.00mg / mL, the regression equation was y=80.298x+1.835, R 2 = 0.9989, relative standard deviation RSD = 0.83, recovery 90-110%. The method for determining the content of sodium lauryl sarcosinate by the HPLC method of the invention has accurate and reliable results.

Description

technical field [0001] The invention belongs to the field of detection, and in particular relates to a method for determining the content of sodium lauryl sarcosinate by high performance liquid chromatography. Background technique [0002] When exogenous genes are expressed at high levels in E. coli, inactive protein aggregates or inclusion bodies are usually formed. Inclusion bodies are rich in expressed recombinant proteins. After separation, denaturation and dissolution, a suitable renaturation process is required to realize the refolding of the denatured proteins before obtaining biologically active proteins. In recent years, many strategies and methods have been developed to renature recombinant proteins from inclusion bodies. For example, adding a surfactant to the protein renaturation solution can effectively inhibit the hydrophobic interaction between protein molecules, reduce the formation of protein aggregates, and thus improve the renaturation rate. [0003] Hor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/86
CPCG01N30/02G01N30/06G01N30/34G01N30/8675G01N2030/047
Inventor 宇文镐杜佩云陈翔
Owner 厦门德馨尚品医疗科技有限公司
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