Evaluation of Lipopeptide (Daptomycin) Aggregation Using Fluorescence,Light Scattering,and Nuclear Magnetic Resonance Spectroscopy |
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| Affiliation: | 1. State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, China;2. The Department of Urology, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing210093, China;3. Medical School of Nanjing University, Nanjing 210093, China;4. Jiangsu Key Laboratory for Nano Technology, Nanjing 210093, China |
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| Abstract: | The aggregation behavior and critical aggregation concentration (CAC) values of daptomycin in aqueous solutions were evaluated under the external factors of pH, temperature, daptomycin concentration, and calcium ions concentration by using the complementary characterization techniques, fluorescence, dynamic and static light scattering, and nuclear magnetic resonance (NMR) spectroscopy. On the basis of the intrinsic fluorescence resonance energy transfer of daptomycin, the CAC values were identified by an upward inflection of the fluorescence emission from Kyn-13 at 460 nm. The pH-dependent CAC values were determined to be 0.14 mM at pH 3.0, 0.12 mM at pH 4.0, and 0.20 mM at pH 2.5 and 5.0. The CAC values obtained by fluorescence spectroscopy were confirmed by dynamic light scattering and NMR spectroscopy. |
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