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Method for detecting protein in sample

A detection method and protein technology, applied in the field of chemical detection, can solve the problems of inability to detect, high minimum limit concentration, inability to accurately detect the specific content of protein, etc., and achieve the effects of good stability and high sensitivity

Active Publication Date: 2021-06-15
山东尚诚质量检测有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through turbidity comparison, only the approximate range of protein concentration can be obtained, but the protein content cannot be accurately measured. When the protein concentration is low or similar, the resolution of the naked eye will not be able to determine whether there is a difference in turbidity or turbidity. Subtract interference from non-protein turbidity
Therefore, this method is still a qualitative inspection method and cannot accurately detect the specific content of protein.
[0007] Patent CN101865926A records that: the current pharmacopoeia (2010 edition) controls the protein content of traditional Chinese medicine injections by following the previous sulfosalicylic acid turbidimetric method, and the controlled protein limit is about 37-110 μg / ml. Protein pharmacopoeia methods are difficult to detect
Therefore, traditional Chinese medicine injections with a protein content of less than 37 μg / ml will be considered "qualified", which limits the scope of application of the sulfosalicylic acid method
This patent records the defects of the method of turbidimetric determination of protein, that is, the minimum limit concentration is too high, and the solution with protein content lower than 37 μg / ml cannot be detected by turbidimetric method

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Experimental program
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Effect test

preparation example Construction

[0117] (a) Preparation of calibration solution;

[0118] (b) preparation of the test solution;

[0119] (c) Preparation of standard solution;

[0120] (d) Assay.

[0121] The preparation method of the calibration solution is as follows: take the sample to be tested, add a solvent to dissolve it and prepare a solution with a suitable concentration, the purpose is to measure the turbidity of the sample itself, and the turbidity of the calibration solution is deducted from the turbidity of the test solution to obtain the test solution. The turbidity produced by the protein in the medium.

[0122] The solvent used for dissolving the calibration solution is water or an acid solution with a suitable concentration. The acid solution with suitable concentration is 0.1 mmol / L-10 mol / L hydrochloric acid solution, preferably 0.1 mol / L hydrochloric acid solution.

[0123] The preparation method of the test solution is as follows: take the sample to be tested and 5-sulfosalicylic acid,...

Embodiment 1

[0145] Example 1 The content detection method verification and content determination of protein in cefprozil

[0146] 1. Determination of the ratio between the concentration of 5-sulfosalicylic acid and the concentration of protein

[0147] 20% 5-sulfosalicylic acid solution: Weigh about 40g of 5-sulfosalicylic acid, put it in a 200ml measuring bottle, add water to dissolve and quantitatively dilute to the mark, shake well to get ready.

[0148] Stock solution: Take about 20.7mg of serum albumin (bovine) standard substance, weigh it accurately, dissolve it in water and quantitatively dilute to 250ml.

[0149] Proportion 1: Accurately measure stock solution 1, 2, 3, 4, 5ml and place in a 50ml measuring bottle, add 2ml of 20% 5-sulfosalicylic acid solution to obtain standard solution 2, 3, 4, 5, 6; Take another 2ml of 20% 5-sulfosalicylic acid solution, put it in a 50ml measuring bottle, quantitatively dilute to the mark with water, and shake well to obtain standard solution 1....

Embodiment 2

[0276] The content detection methodological verification of protein in the embodiment 2 glucose

[0277] 1. Linear relationship test

[0278] Stock solution: Take 20.51mg of serum albumin (bovine) standard substance, weigh it accurately, dissolve it in water and quantitatively dilute to 250ml.

[0279] Linear relationship test solution: Accurately measure stock solutions 1, 2, 3, 4, 5, 10, 25, 45ml and put them in a 50ml measuring bottle, add 5ml of 20% 5-sulfosalicylic acid solution, and dilute to the mark with water quantitatively, Shake well to get standard solution 2, 3, 4, 5, 6, 7, 8, 9; take 5ml of 20% 5-sulfosalicylic acid solution, put it in a 50ml measuring bottle, quantitatively dilute to the mark with water, shake well, Standard solution 1 was obtained.

[0280] Determination method: Take each standard solution, measure according to the law (Chinese Pharmacopoeia 2020 Edition General Rule 0902 Second Method), record the turbidity value, and calculate the linear re...

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Abstract

The invention belongs to the technical field of chemical detection, and particularly relates to a method for detecting protein in a sample. According to the detection method, a turbidity meter is adopted for determination. The sample can be dissolved in an acidic 5-sulfosalicylic acid aqueous solution. According to the method, the content of protein is measured by a turbidity meter method, the sensitivity of the method is high (the limit-of-detection concentration is 3.351 ng / ml, and the limit-of-quantitation concentration is 10.16 ng / ml), the stability is good (stability is achieved within 2-24 hours), the accuracy is high, and the content of protein in a sample can be accurately measured.

Description

technical field [0001] The application belongs to the technical field of chemical detection, and in particular relates to a method for detecting protein in a sample. Background technique [0002] Substances produced by enzymatic or fermentation methods may contain residues of enzymes or microbial tissues, and their main components are proteins. Exogenous proteins may cause adverse reactions such as human allergies. Therefore, in order to better control the quality of samples, it is necessary to The amount of protein is quantitatively detected and controlled to ensure the safety of the sample. [0003] Patent CN102000139A discloses the detection method of Aidi injection preparation, which involves protein inspection, the specific content is: protein: take 10-20 mg of freeze-dried powder for injection and add 1 ml of water for injection to dissolve, or take 4-10 ml of infusion and concentrate to 1 ml, Or take 1ml of water injection; add 1ml of newly prepared 30% sulfosalicyli...

Claims

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Application Information

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IPC IPC(8): G01N21/83G01N1/38
CPCG01N21/83G01N1/38
Inventor 曹悦兴张展任霞闫婷婷贾右刚
Owner 山东尚诚质量检测有限公司