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Method for detecting residual titer of cephalosporin C in cephalosporin residues

A method for measuring cephalosporins, which is applied in the field of determination of cephalosporin C residual potency in cephalosporin residues, can solve the problems of inaccurate detection of cephalosporin C content, inability to determine environmental hazards and impacts, and restrict bacteria residues Reuse of resources and other issues to achieve the effect of high accuracy, high sensitivity, and high separation efficiency

Inactive Publication Date: 2018-07-10
YILI CHUANNING BIOTECH CO
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

Poor stability
[0006] So far, there is no standard method for detecting the residual cephalosporin C potency in cephalosporin residues at home and abroad, so the content of residual cephalosporin C in cephalosporin residues cannot be accurately detected, and its possible harm to the environment cannot be determined Therefore, the method for determining the potency of cephalosporin C residues in cephalosporin residues is established, and the judgment and release standards are clearly defined, so as to exclude the possibility that the residues of cephalosporin C in cephalosporin residues may affect the environment. Negative impacts brought about by cephalosporins appear to be very necessary and of great practical significance, which can provide a theoretical basis for the treatment and disposal of cephalosporin residues

Method used

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  • Method for detecting residual titer of cephalosporin C in cephalosporin residues
  • Method for detecting residual titer of cephalosporin C in cephalosporin residues
  • Method for detecting residual titer of cephalosporin C in cephalosporin residues

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Example 1 Determination of residual potency of cephalosporin C in spray-dried inactivated cephalosporin residue

[0048] 1) Preparation of buffer

[0049] 0.1mol / L phosphate buffer (pH 5.0): Weigh 17.9g disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) (analytical pure) was dissolved in 500 ml of purified water, and the pH was adjusted to 5.0 with phosphoric acid (analytical pure).

[0050] 0.01mol / L sodium acetate buffer solution (pH 4.75): Weigh 0.4g sodium hydroxide (analytical grade), dissolve it with a small amount of purified water and transfer it to a 1000ml volumetric flask, then accurately measure 1ml of glacial acetic acid (analytical grade) and add Put it into a volumetric flask, dilute to the mark with purified water, shake well, and adjust the pH to 4.75 with glacial acetic acid (analytical grade).

[0051] 2) Extraction and separation of cephalosporin residue

[0052]Take the spray-dried and inactivated solid cephalosporin residue sample, grind it with...

Embodiment 2

[0083] Example 2 Determination of Cephalosporin C Residual Potency in Liquid Inactivated Cephalosporin Residue

[0084] 1) Preparation of buffer

[0085] With step 1) among the embodiment 1.

[0086] 2) Extraction and separation of cephalosporin residue

[0087] Take the liquid inactivated cephalosporin residue sample, and use a medium-strength grinding bowl for more than 300 laps to grind to a fine and uniform paste, so that the particle size is D97=33.20 μm; weigh 4 g of the liquid inactivated cephalosporin residue grinding sample, and put it in Into a 50ml polypropylene centrifuge tube, add 20ml of 0.1mol / L phosphate buffer (pH 5.0), vortex for 1min, ultrasonically assisted extraction for 30min, centrifuge at 4500rpm for 10min, and take the supernatant.

[0088] 3) Cephalosporin C residue titer detection

[0089] With step 3) in embodiment 1.

[0090] 4) Determination results of cephalosporin C residues in three batches of liquid inactivated cephalosporin residues

[0...

experiment example 1

[0096] Experimental example 1 Repeatability experiment

[0097] Weigh two parts of about 25 mg of cephalosporin C sodium salt reference substance, accurately weigh them, place them in two 100ml volumetric flasks respectively, dissolve and dilute to the mark with 0.01mol / L sodium acetate solution (pH4.75), weigh Spray inactivated cephalosporin residue grinding sample 2g, in a 50ml polypropylene centrifuge tube, add 20ml of 0.1mol / L phosphate buffer (pH 5.0), vortex for 1min, ultrasonically assisted extraction for 30min, centrifuge at 4500rpm for 10min, take supernatant. Take 0.5ml of the control solution and 1ml of the sample solution, mix and dilute to 10ml, repeat the measurement 5 times according to the chromatographic conditions of the analysis method, the results are shown in Table 3.

[0098] Table 3 Repeatability experiment

[0099] serial number

[0100] Conclusion: After 6 consecutive injections, the RSD of retention time was 0.05%, and the RSD of peak area...

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Abstract

The invention discloses a method for determining the residual titer of cephalosporin C in cephalosporin residues. The method comprises the following steps: (1) preparing a test solution: taking and grinding the cephalosporin residues, performing extraction by using a phosphate buffer solution, centrifuging the obtained solution, and taking the obtained supernatant to obtain the test solution; (2)preparing a reference solution: dissolving the cephalosporin C sodium salt in a mobile phase A to obtain the reference solution, wherein the mobile phase A is a sodium acetate buffer solution or an ammonium acetate buffer solution; and (3) detecting the residual tilter: taking the test solution, detecting the peak area by high performance liquid chromatography, and calculating the residual tilterof the cephalosporin C according to the peak areas of a reference substance and a sample, wherein the chromatographic conditions of the high performance liquid chromatography are as follows: the mobile phase is composed of the mobile phase A and a mobile phase B, the mobile phase B is acetonitrile, and a volume ratio of the mobile phase A to the mobile phase B is 97:3 to 99:1; and the chromatographic column is an octadecylsilane bonded silica gel column. The method has the advantages of high sensitivity, high separation efficiency, good selectivity and good repeatability.

Description

technical field [0001] The invention relates to a method for determining the residual titer of cephalosporin C in cephalosporin dregs, in particular to a method for determining the residual titer of cephalosporin C in cephalosporin dregs by high performance liquid chromatography. Background technique [0002] my country is a major producer of cephalosporins. Cephalosporin fermentation residues are wastes produced after the extraction of cephalosporin C. The amount of production is very large. The residues of cephalosporin C in the residues will cause livestock and poultry and ecological problems. The drug resistance of environmental microorganisms will cause potential drug resistance risks to human health. Therefore, my country clearly stipulates that cephalosporin residues cannot be directly used as feed and fertilizer and are listed as hazardous waste. [0003] The structure of cephalosporin C is similar to that of penicillin, it has an endamide ring, the nucleus is amino c...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 王继英邓仕宏王小莉王远莉李香霖徐文科梁金凤陈启军党建宁邓旭衡
Owner YILI CHUANNING BIOTECH CO
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