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Pretreatment method for lincomycin fermentation liquor for HPLC (High Performance Liquid Chromatography) analysis

A technology of lincomycin and fermentation liquid, which is applied in the direction of analyzing materials, material separation, measuring devices, etc., can solve problems such as low efficiency, affecting accuracy, complicated process, etc., and achieve good results in analysis

Inactive Publication Date: 2012-07-18
ANHUI WANBEI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this lincomycin fermentation broth pretreatment method is that the process is complicated, multiple extraction is required, the extraction is incomplete, and only the content of lincomycin in the extraction phase can be measured, which affects the accuracy of the measurement and the efficiency is relatively low.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Take lincomycin production fermentation broth, centrifuge at 4000r / min for 10 minutes, remove mycelia and solid impurities, absorb the supernatant, add ethanol at a volume ratio of 1:2, mix well, and centrifuge at 6000r / min for 20 minutes minutes; then filter with a 0.22 μm metafluoride membrane; the filtered clear liquid is used for HPLC detection (HPLC detection conditions are the same as lincomycin hydrochloride, "Pharmacopoeia of the People's Republic of China" (2010) Part Two, page 727), the HPLC column After continuous analysis of 50 samples, the column pressure was 1298-1334psi, and the theoretical plate number of lincomycin peak was 6800-7500; The separation degree of chromatographic peaks was greater than 9. Before and after the analysis, the column pressure increased slightly. The analysis effect of HPLC was good, and the number of theoretical plates of the main peak did not change significantly.

Embodiment 2

[0020] Take lincomycin production fermentation liquid, centrifuge at 4000r / min for 15 minutes, remove mycelia and solid impurities, absorb the supernatant, add ethanol at a volume ratio of 1:3, mix well, and centrifuge at 6000r / min for 20 minutes minutes; then filter with a 0.22 μm metafluoride membrane; the filtered clear liquid is used for HPLC detection (HPLC detection conditions are the same as lincomycin hydrochloride, "Pharmacopoeia of the People's Republic of China" (2010) Part Two, page 727), the HPLC column After continuous analysis of 50 samples, the column pressure was 1316-1343psi, and the theoretical plate number of lincomycin peak was 6800-7500; The separation degree of chromatographic peaks is greater than 9. Before and after the analysis, the column pressure does not change significantly. The HPLC analysis effect is good, and the theoretical plate number of the main peak does not decrease.

Embodiment 3

[0022] Take lincomycin production fermentation broth, centrifuge at 6000r / min for 15 minutes, remove mycelia and solid impurities, absorb the supernatant, add ethanol at a volume ratio of 1:4, mix well, and centrifuge at 10000r / min for 15 minutes minutes; then filter with a 0.22 μm metafluoride membrane; the filtered clear liquid is used for HPLC detection (HPLC detection conditions are the same as lincomycin hydrochloride, "Pharmacopoeia of the People's Republic of China" (2010) Part Two, page 727), the HPLC column After continuous analysis of 50 samples, the column pressure was 1320-1337psi, and the theoretical plate number of lincomycin peak was 6800-7500; The separation degree of chromatographic peaks is greater than 9. Before and after the analysis, the column pressure does not change, the HPLC analysis effect is good, and the theoretical plate number of the main peak does not decrease.

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Abstract

The invention discloses a pretreatment method for lincomycin fermentation liquor for HPLC (High Performance Liquid Chromatography) analysis. The pretreatment method comprises the steps as follows: 1) centrifugally separating the lincomycin fermentation liquor to obtain supernatant; and 2) uniformly mixing the supernatant with ethanol by volume ratio of 1:2-4, and centrifugally separating out water-soluble protein to obtain clear liquid which then can be used for HPLC analysis. After the fermentation liquor is pretreated by the pretreatment method, the impurities in samples for the HPLC analysis are greatly reduced; when the pretreatment method is used for detection of the lincomycin fermentation liquor, both lincomycin component and lincomycin B component are effectively separated from an adjacent impurity peak, the analysis effect is good, the lincomycin B component can be visually represented with data and the analysis accuracy is high; and after hundreds of samples are analyzed continuously, column pressure is not obviously increased, and the chromatographic column theoretical plate number can meet detection requirements.

Description

technical field [0001] The invention relates to a pretreatment method for a fermented liquid to be detected by high performance liquid chromatography, in particular to a pretreatment method for a lincomycin fermented liquid for HPLC analysis. Background technique [0002] Lincomycin belongs to the lincomycin class of antibiotics. It is effective against most Gram-positive bacteria and some anaerobic Gram-negative bacteria. Its effect on positive bacteria is similar to that of erythromycin. It is mainly used for abdominal and gynecological infections caused by anaerobic bacteria, and is also commonly used for various serious infections caused by sensitive positive bacteria. Because it is concentrated in bone tissue, it is the first choice drug for aureus osteomyelitis. It is clinically used more extensive. The components of lincomycin include lincomycin and its component B, of which component B is an impurity component. At present, the most commonly used detection of lincomy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06
Inventor 姚健陈倩倩营丽李为全刘瑞华
Owner ANHUI WANBEI PHARMA
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