Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - Monitoring the mode of action of antibiotics using Raman spectroscopy
T2 - investigating subinhibitory effects of amikacin on Pseudomonas aeruginosa
AU - López-Diez, E. Consuelo
AU - Winder, Catherine L.
AU - Ashton, Lorna
AU - Currie, Felicity
AU - Goodacre, Royston
PY - 2005/5/1
Y1 - 2005/5/1
N2 - During the last 20 years the rate at which new antimicrobial agents are produced has decreased dramatically, with concomitant increase in the number of pathogens that are becoming multidrug resistant. Together these have created a patient healthcare risk and this is of great concern. A crucial aspect for the discovery of new antibiotics is the development of new techniques that allow rapid and accurate characterization of the mode of action of the pharmacophore. In this work UV resonance Raman (UVRR) spectroscopy has been developed to monitor the concentration effect of antibiotics on bacterial cells. UVRR was conducted at 244 nm and spectra were collected in typically 60 s. Supervised multivariate analysis and 2D correlation spectroscopy were used to evaluate whether the UVRR spectra contained valuable information that could be used to study the mode of action of antibiotics. The clustering pattern in the discriminant factors space correlated directly to the concentration of amikacin, and partial least squares (PLS) regression analysis of the UVRR spectra was able to predict the concentration of amikacin to which bacterial cells had been exposed. 2D correlation spectroscopy contour maps indicated that spectral changes due to the presence of amikacin in the growth media occur according to the known mode of action of the studied antibiotic. Therefore, we conclude that UVRR spectroscopy, when coupled with chemometrics and 2D correlation spectroscopy, constitutes a powerful approach for the development and screening of new antibiotics.
AB - During the last 20 years the rate at which new antimicrobial agents are produced has decreased dramatically, with concomitant increase in the number of pathogens that are becoming multidrug resistant. Together these have created a patient healthcare risk and this is of great concern. A crucial aspect for the discovery of new antibiotics is the development of new techniques that allow rapid and accurate characterization of the mode of action of the pharmacophore. In this work UV resonance Raman (UVRR) spectroscopy has been developed to monitor the concentration effect of antibiotics on bacterial cells. UVRR was conducted at 244 nm and spectra were collected in typically 60 s. Supervised multivariate analysis and 2D correlation spectroscopy were used to evaluate whether the UVRR spectra contained valuable information that could be used to study the mode of action of antibiotics. The clustering pattern in the discriminant factors space correlated directly to the concentration of amikacin, and partial least squares (PLS) regression analysis of the UVRR spectra was able to predict the concentration of amikacin to which bacterial cells had been exposed. 2D correlation spectroscopy contour maps indicated that spectral changes due to the presence of amikacin in the growth media occur according to the known mode of action of the studied antibiotic. Therefore, we conclude that UVRR spectroscopy, when coupled with chemometrics and 2D correlation spectroscopy, constitutes a powerful approach for the development and screening of new antibiotics.
KW - UV RESONANCE RAMAN
KW - 2-DIMENSIONAL CORRELATION SPECTROSCOPY
KW - DEPENDENT SPECTRAL VARIATIONS
KW - INFRARED CORRELATION SPECTROSCOPY
KW - PURE LIQUID-STATE
KW - RAPID IDENTIFICATION
KW - MASS-SPECTROMETRY
KW - N-METHYLACETAMIDE
KW - E. COLI
KW - BACTERIA
U2 - 10.1021/ac048147m
DO - 10.1021/ac048147m
M3 - Journal article
VL - 77
SP - 2901
EP - 2906
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 9
ER -