Amikacin

For determination of amikacin (Figure 1, structure 1), the serum sample is loaded onto the silica gel column, followed by addition of o-phthalaldehyde (a derivatizing reagent). The column is eluted with 95% ethanol (pH 10) and the eluent is heated at 50°C. After cooling, the mixture is injected onto an ODS

ch,oh ch,oh

nh2 (1)

ch(r,)nhr2

2 ch3 ch3

4 ch3 h

Figure 1 Chemical structures of amikacin (1) and gentamycins C (2), C1a (3) and C2 (4).

column. Mobile phase is methanol-water-aceto-nitrile (65 : 30 : 5) containing 0.2% tripotassium ethylenediamine tetraacetic acid (EDTA). Amikacin is detected fluorometrically at 350 nm for excitation and 420 nm for emission. The detection limit is 1 |gmL~\ In order to resolve amikacin from its three isomers, a similar method is used except that the mobile phase is methanol-water (7:3). Tobramycin may be analysed with the same method.

Pre-column derivatization of amikacin is also conducted by addition of 1-fluoro-2,4-dinitrobenzene to plasma or urine, leading to formation of a stable chromophore, which can be detected at 360 nm. An ODS column is used with a mobile phase consisting of acetonitrile-water (68 : 32). The detection limit is 1 |gmL~\ Amikacin may be extracted from serum using a cation exchange solid-phase extraction column prior to derivatization.

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Solar Panel Basics

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