Expression Analysis of Pyocyanin by High Performance Liquid Chromatography
Objective A method for the determination of Pseudomonas aeruginosa metabolite was established by high performance liquid chromatography,and the expression of pyocyanin content was analyzed.Method Using Inertsil ODS-SP(4.6 mm×250 mm,5 μm)Chromatographic column separation,column temperature 40 ℃,flow rate 1.0 mL/min,sample injection amount 20 μL.The mobile phase was 0.01％formic acid water acetonitrile gradient elution with a detection wavelength of 268 nm.Results Pseudomonas aeruginosa had a detection limit of 0.10 μg and a quantitative limit of 0.20 μg under optimized conditions.The method's precision(RSD)was less than 0.80％in the addition range of 0.22-0.66 μg/mL and 1.10-3.30 μg/mL,and the target compound was linearly good in the linear range of 0.20-10.00 μg/mL.Preliminary tests using this method show that low quantities of levofloxacin promote Pseudomonas aeruginosa's secretion of pyocyanin,which may be connected to the bacterium's drug resistance mechanism.Conclusion The method is simple,accurate,with good precision and recovery,and can be used for the detection of pyocyanin.It has a certain guiding significance for further research on the relationship between the drug resistance mechanism of Pseudomonas aeruginosa and the secretion of pyocyanin.
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