TITLE:
Optimization and Comparisons for Separation, Detection and Quantification of 12 Aminoglycosides Using 2 Chromatographic Conditions by LC-MS/MS
AUTHORS:
Samia Mokh, Farouk Jaber, Abir Kouzayha, Hélène Budzinski, Mohamad Al Iskandarani
KEYWORDS:
Aminoglycosides, Ion Pairing, ZIC-HILIC, LC-MS/MS
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.5 No.14,
October
29,
2014
ABSTRACT: Aminoglycosides are
a family of antibiotics with important applications in veterinary medicine.
Their ionic character, the similarity structures and the high polarity due to
the presence of two or more amino and hydroxyl groups cause a difficulty in
separation and make these compounds poorly retained on the reversed phase
column. An analytical method for the separation and detection of 12
aminoglycosides has been optimized using two kinds of chromatographic
conditions (HILIC, Ion
pairing). In Hydrophilic Interaction, ZIC_HILIC column was
used, by which the following parameters for the mobile phase were evaluated:
concentration of ammonium acetate buffer, percentage of formic acid and effect
of acid type. The maximum and adequate concentration of ammonium acetate for
the majority of analytes was set to 30 mM. The percentage 0.1% of formic acid
increases the response for the majority of analytes. On the other side, the use
of 0.1% of trifluoroacetic acid improves the response when compared with the response
obtained with 0.1% of formic acid except for Spectinomycin Dihydrostreptomycin
and Streptomycin. For ion pairing chromatography, the concentration of pentafluoropropionic
acid was tested and the greatest value appeared to be 9.2 mM. Therefore, the
comparison between the two separation methods shows that the response area of
the majority of analytes tested increases when using the ion pair mode. Also,
the high value of S/N and the lower detection limit (5 - 15 μg m·L﹣1 for most aminoglycosides studied make the ion pairing method more preferable than HILIC interaction.