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Analysis of spectinomycin in fermentation broth by reversed-phase chromatography
Yan, Hong De, Xu, Pei Ya, Huang, Hai Chan and Qiu, Juan Ping Analysis of spectinomycin in fermentation broth by reversed-phase chromatography Chemical Papers, Vol.63, No. 6, 2009, 635-640
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Document type:
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Článok z časopisu / Journal Article |
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Collection:
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Chemical papers
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| Author(s) |
Yan, Hong De
Xu, Pei Ya
Huang, Hai Chan
Qiu, Juan Ping
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| Title |
Analysis of spectinomycin in fermentation broth by reversed-phase chromatography
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| Journal name |
Chemical Papers
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| Publication date |
2009
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| Year available |
2009
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| Volume number |
63
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| Issue number |
6
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| ISSN |
0366-6352
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| Start page |
635
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| End page |
640
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| Place of publication |
Poland
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| Publisher |
Versita
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| Collection year |
2009
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| Language |
english
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| Subject |
270000 Biological Sciences
270800 Biotechnology
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| Abstract/Summary |
A pre-column derivatized high-performance liquid chromatographic (HPLC) method with ultraviolet-visible detection was developed to measure the concentrations of spectinomycin in fermentation broth. Derivatization reagents, 2,4-dinitrophenylhydrazine in acetonitrile (5 mg mL−1) and trifluoroacetic acid in acetonitrile (0.8 mol L−1), were added to an aliquot of the fermentation broth, and the mixture was incubated for 60 min at 70°C. The resulting derivative was separated from other compounds by isocratic elution in a reversed-phase column Zorbax SB-C18 (250 mm × 4.6 mm, 5 µm). Mobile phase consisted of acetonitrile, tetrahydrofuran, and water (φ r = 40: 35: 25) and the flow rate was 1.0 mL min−1. The detection wavelength was 415 nm. The standard curve for spectinomycin sulfate was linear with correlation coefficients of 0.9997 in the range of 25 µg mL−1 to 600 µg mL−1. The relative standard deviation values ranged from 0.43 % to 2.18 % depending on the concentration of samples. The average recovery was 101.5 %. The limit of detection was 50 ng mL−1.
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