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Method for detecting residual titer of erythromycin A in erythromycin fungi residues

A technology of erythromycin bacterial residue and determination method, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex bacteria residues, inability to accurately detect the content of erythromycin A, and many interfering substances.

Pending Publication Date: 2021-04-23
YILI CHUANNING BIOTECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there is no standard method for detecting the residual potency of erythromycin A in erythromycin residue at home and abroad, so the content of residual erythromycin A in erythromycin residue cannot be accurately detected, and its effect on the residue cannot be determined. The possible hazards and impacts on the environment have seriously restricted the resource reuse of erythromycin residues. Therefore, a method for the determination of erythromycin A residual potency in erythromycin residues has been established to clarify the judgment and release standards and exclude the residues of erythromycin. The possible negative impact of erythromycin A residue on the environment is very necessary and of great practical significance, providing a theoretical basis for the treatment and disposal of erythromycin residue
[0006] The residual potency of erythromycin A in the erythromycin residue is low, and the residue is complex and has many interfering substances. Therefore, it is necessary to effectively extract and separate the analyte to minimize the detection error and ensure the detection accuracy. accuracy of results

Method used

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  • Method for detecting residual titer of erythromycin A in erythromycin fungi residues
  • Method for detecting residual titer of erythromycin A in erythromycin fungi residues
  • Method for detecting residual titer of erythromycin A in erythromycin fungi residues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Determination of residual potency of erythromycin A in solid spray-dried inactivated erythromycin residue

[0049] 1) Preparation of mobile phase

[0050] Mobile phase A is 0.025mol / L phosphate buffer solution; mobile phase B is acetonitrile; mobile phase A: mobile phase B is 55:45.

[0051] 2) Extraction and separation of solid erythromycin residue

[0052] Take the spray-dried and inactivated solid erythromycin slag sample, grind it with a grinding bowl for more than 300 laps with medium force, and grind it to a fine and uniform powder, so that the powder particle size is D97=218.01μm; weigh the spray-inactivated erythromycin Put 5 g of the ground sample of the bacterial residue (accurate to 0.1 mg), into a 50 ml volumetric flask, add an appropriate amount of methanol, vortex for 1 min, ultrasonically assisted extraction for 10 min, centrifuge at 4500 rpm for 10 min, and take the supernatant.

[0053] 3) Detection of erythromycin residue titer

[0054] a. Chromatog...

Embodiment 2

[0075] Determination of residual potency of erythromycin A in liquid inactivated erythromycin residue

[0076] 1) Preparation of mobile phase

[0077] With step 1) among the embodiment 1.

[0078] 2) Extraction and separation of a batch of liquid erythromycin residue

[0079] Take the liquid inactivated erythromycin residue sample, weigh 5g of the liquid inactivated erythromycin residue sample (accurate to 0.1mg), add an appropriate amount of methanol to a 50ml volumetric flask, vortex for 1min, and ultrasonically assist extraction for 30min. Add methanol to dilute to the mark, centrifuge at 4500rpm for 10min, and take the supernatant.

[0080] 3) Detection of erythromycin residue titer

[0081] With step 3) in embodiment 1.

[0082] 4) Determination result data of erythromycin A residue in a batch of liquid inactivated erythromycin residue

[0083] For the results of the system suitability test and the determination of erythromycin residue A in a batch of liquid inactiva...

experiment example 3

[0088] Experimental Example 3 Repeatability Experiment

[0089] Accurately weigh two parts of about 40 mg of erythromycin reference substance, accurately weigh them, place them in two 10ml volumetric flasks respectively, dissolve them with methanol and dilute to the scale, shake well and repeat the measurement 6 times according to the chromatographic conditions of the analysis method, the results See Table 3.

[0090] Table 3 Repeatability experiment

[0091] serial number Erythromycin retention time (min) Erythromycin peak area (mAU*s) 1 15.469 1804.61218 2 15.459 1806.45020 3 15.443 1801.22144 4 15.430 1796.55554 5 15.417 1799.17029 6 15.408 1809.18335 RSD 0.036％ 0.33％

[0092] Conclusion: After 6 consecutive injections, the RSD of retention time was 0.036%, and the RSD of peak area was 0.33%, indicating good repeatability.

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Abstract

The invention provides a method for detecting residual titer of erythromycin A in erythromycin fungi residues. The method comprises the following steps: preparing a test solution; preparing a reference substance solution; determining the residual titer of erythromycin A: taking the test solution and the reference solution, and determining the peak area by using high performance liquid chromatography, wherein the chromatographic conditions of the high performance liquid chromatography are as follows: a mobile phase is composed of a mobile phase A and a mobile phase B, the mobile phase A is a phosphate buffer solution, the mobile phase B is acetonitrile, the volume ratio of the mobile phase A to the mobile phase B is 55: 45, a chromatographic column is an octadecylsilane chemically bonded silica filler chromatographic column, the flow velocity is 0.8-1.2 mL / min, the column temperature is 25-40 DEG C, the detection wavelength is 215 nm, and the sample introduction amount is 10 [mu]L; and calculating according to the peak areas of the reference substance and the test sample to obtain the erythromycin A residual titer. The detection method disclosed by the invention is high in sensitivity, high in separation efficiency, good in selectivity, good in repeatability and high in accuracy.

Description

technical field [0001] The invention belongs to the technical field of determining the residual potency of erythromycin residue, and relates to a method for determining the residual titer of erythromycin A in erythromycin residue. Background technique [0002] my country is a big producer of erythromycin. Erythromycin residue is the waste produced after the fermentation and extraction of erythromycin thiocyanate. The amount of production is huge. The residue of erythromycin A in the residue will cause livestock The drug resistance of poultry and ecological environment microorganisms will cause potential drug resistance risks to human health. Therefore, my country clearly stipulates that erythromycin residues cannot be directly used as feed and fertilizer and are listed as hazardous waste. [0003] The structure of erythromycin A has an endamide ring, the core is an amino erythromycin A alkanoic acid, and the molecular formula is C 37 h 67 NO 13 , the molecular weight is 73...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/30G01N30/32G01N30/34
Inventor 郑洁秦江涛李香霖王远莉陈楠
Owner YILI CHUANNING BIOTECH CO
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