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Method for detecting zymoprotein residues in cefprozil prepared by enzymatic method

A technology for the preparation of cefprozil and enzymatic method, which is applied in the preparation of test samples and the measurement of color/spectral characteristics, etc., can solve the problems of patients' allergies, and achieve the effect of good reliability, high accuracy and simple operation

Pending Publication Date: 2020-04-03
广药白云山化学制药(珠海)有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If more remains in the finished product, it is an exogenous protein to the human body after taking the medicine, which may cause adverse reactions such as allergies in patients

Method used

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  • Method for detecting zymoprotein residues in cefprozil prepared by enzymatic method
  • Method for detecting zymoprotein residues in cefprozil prepared by enzymatic method
  • Method for detecting zymoprotein residues in cefprozil prepared by enzymatic method

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preparation example Construction

[0035] Preparation of the standard curve solution: take the standard protein stock solution and dilute it with sodium bicarbonate solutions of the same concentration and different volumes to obtain respective standard protein linear solutions; each take the same volume of the Coomassie Brilliant Blue G-250 dye solution, Add the above-mentioned standard protein linear solutions respectively, and mix well; after standing for reaction, the standard curve solution is obtained;

[0036] Preparation of the blank solution: Take the same volume of Coomassie Brilliant Blue G-250 staining solution as that used in the preparation of the standard curve solution, add the same concentration of sodium bicarbonate solution, mix well, leave it to react, and use it as a blank solution;

[0037] Preparation of the testing solution for the testing product: take an appropriate amount of cefprozil for the testing product, add the same concentration of sodium bicarbonate solution to dissolve and dilu...

Embodiment approach

[0040] As an embodiment, the preparation of Coomassie Brilliant Blue G-250 dye solution: take Coomassie Brilliant Blue G-250 and dissolve it in ethanol, then add an appropriate amount of phosphoric acid, and finally use purified water to make up the volume; preparation of standard protein stock solution: Weigh an appropriate amount of bovine serum albumin, dissolve and dilute it with purified water to obtain a standard protein stock solution.

[0041] As an embodiment, in the preparation process of the Coomassie Brilliant Blue G-250 dye solution, the ratio of the weight of Coomassie Brilliant Blue G-250, the volume of ethanol, the volume of phosphoric acid and the constant volume is 5g: (2~3)L : (5~6)L: 50L.

[0042] As an embodiment, the concentration of the standard protein stock solution is 0.1-5 mg / ml, and the concentration of the test solution is 5-100 mg / ml.

[0043] Further, the standing reaction time when the blank solution is prepared, the standing reaction time when...

Embodiment 1

[0056] The detection method of enzyme protein residue in the enzymatic preparation of cefprozil of the present embodiment comprises the following steps:

[0057] 1. Preparation of Assay Solution

[0058] (1) Preparation of Coomassie Brilliant Blue G-250 dye solution: Weigh 100 mg of Coomassie Brilliant Blue G-250, dissolve in 40-60 ml of ethanol, add 100-120 ml of phosphoric acid, and finally dilute to 1000 ml with purified water;

[0059] (2) Preparation of standard protein stock solution: 125 mg of bovine serum albumin was accurately weighed, dissolved in purified water to a volume of 50 ml, and used as a standard protein stock solution.

[0060] (3) Preparation of the standard curve solution: take the standard protein stock solution, dilute it with a sodium bicarbonate solution with a concentration of 10% g / ml as shown in Table 1, prepare 1 part of each concentration, and obtain 7 parts of the standard protein linear solution; Take 10ml of Coomassie Brilliant Blue G-250 dy...

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Abstract

The invention provides a method for detecting zymoprotein residues in cefprozil prepared by an enzymatic method. The method is characterized by comprising the following steps: preparing a Coomassie brilliant blue G-250 dye solution and a standard protein stock solution; preparing a standard curve solution by taking a sodium bicarbonate solution as a solvent; preparing a blank solution by taking asodium bicarbonate solution as a solvent; preparing a test sample determination solution by taking a sodium bicarbonate solution as a solvent; drawing an absorbance-protein concentration standard curve through blank solution correction by using an ultraviolet spectrophotometer; and calculating the residual content of zymoprotein contained in the test sample according to the absorbance value read by the test sample determination solution. Sodium bicarbonate is ingeniously selected as a solvent, cefprozil can be dissolved, zymoprotein can be fully dissolved, the residual quantity of zymoproteinin a cefprozil product prepared through an enzymatic method can be effectively measured, the medication safety of medicine is improved, and the detection method is easy to operate, high in accuracy and good in reproducibility and reliability.

Description

technical field [0001] The invention relates to the field of pharmacy, in particular to a method for detecting enzyme protein residues in the enzymatic preparation of cefprozil. Background technique [0002] The chemical name of Cefprozil is (6R,7R)-7-[(R)-2-amino-2-(p-hydroxy-phenyl)acetamido]-8-oxo-3-propene-5- Thia-1-azabicyclo[4,2,0]oct-2-ene-2-carboxylic acid-hydrate is a second-generation non-ester oral cephalosporin developed by Bristol-Myers Squibb It is a broad-spectrum antibacterial drug and the first oral cephalosporin antibiotic approved by the FDA for the treatment of otitis media and sinusitis in children. [0003] At present, the reported method of synthesizing cefprozil mainly adopts the chemical synthesis method, and a new green and environment-friendly enzymatic method for the preparation of cefprozil has also been reported in recent years. Compared with chemical synthesis, enzymatic synthesis uses less organic solvents and has simple routes. It can overc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/38G01N21/33
CPCG01N1/38G01N21/33
Inventor 李庆关晴
Owner 广药白云山化学制药(珠海)有限公司
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