Spectrophotometric analysis of some aminoglycosidic antibiotics

Introduction
Due to weak absorbances on the UV-Vis domain, direct spectrophotometric
determination of 2-
deoxystreptaminic aminoglycosides is not possible. UV-Vis absorbances of
aminoglycosides can be enhanced in the presence of metallic ions such as u2!.
"he aim of the present chapter of the thesis was to de#elop spectrophotometric
methods for the $uantitati#e determination of neomycin, kanamycin, gentamicin and
amikacin in the presence of u2! ions and to e#aluate the stability of the
aminoglycoside-u2! complexes.
%aterial and method
Determinations were conducted using an &gilent '()* spectrophotometer +&gilent
"echnologies, ,ermania-.
Data was processed using hemstation software. .pectra were recorded on the UV-Vis
2//-'// nm domain.
0inetics of the aminoglycoside-u2! complexes were studied using a .pectronic
,enesys ) +%ilton 1oy- spectrophotometer.
2ormation of aminoglycoside-u2! complex formation was studied using different
sol#ent mixtures +acetone, methanol, ethanol- at different p3 #alues. In the case of
neomycin optimum sol#ent pro#ed to be purified water4methanol (45 mixture, while in
the case of amikacin, kanamycin and gentamicin 5/ m% sodium hydroxide was used.
"o obtain an optimum u2! ion concentration, ul2.6327 was added to a /,5 mg8m9
concentration.
.pectra were recorded on the 2//-'// nm domain and processed both directly and #ia
the first deri#ati#e. 7n the non-deri#ati#e method +&- optimum wa#elengths were 26/
nm for neomycin, 2*/ nm for gentamicin, 2'/ nm for
kanamycin and *2) nm for amikacin. In the case of the deri#ati#e method +d&- results
were optimum at 2:: nm for neomycin, 2;5 nm for gentamicin, 2;/ nm for kanamycin
and 2'( nm for amikacin.
&t )-5) minutes after the preparation of the test solutions no significant modifications
were obser#ed
regarding optical stability, therefore the mentioned timeframe was considered suitable to
conduct all $uantitati#e determinations.
1esults
1egarding the non-deri#ati#e spectrophotometric methods, linearity was unsatisfactory,
the coefficients of correlation being below the r</,;;') in all cases. "hese methods
were not considered #alid.
.ince non-deri#ati#e spectrophotometry did not pro#ide #alid $uantitati#e determination
methods, $uantitation of aminoglycosides was performed using the first deri#ati#e of the
spectra. "he coefficient of correlation in this case was r=/,;;' for each studied
aminoglycosidic compound +amikacin, kanamycin, gentamicin and neomycin-. "he
e$uation of the calibration cur#e was calculated in each case. 1eco#eries were between
;;,';>-5//,'(> for neomycin, ;:,26>-5/5,55> for gentamicin, ;;,5:>-5/5,6(> for
kanamycin and ;',)6>-5/2,56> for amikacin. 1epeatability and intermediate precision
was assessed for each analyte. Intra-day 1.D> #alues were between 5,(/-2,*2> for
neomycin, 5,//-5,6*> for gentamicin, /,2:-/,''> for kanamycin and 5,56-5,);> for
amikacin. In case of intermediate precision 1.D> were 5,;/> for neomycin, 5,2:> for
gentamicin, 5,5'> for kanamycin, 5,2)> for amikacin.
9imits of detection and limits of $uantitation were e#aluated based on the signal-to-
noise ratio4
- neomycin4 97D</,/2) mg8m9, 97?</,/:) mg8m9@
- gentamicin4 97D</,//6 mg8m9, 97?</,/2/ mg8m9@
- kanamycin4 97D</,//; mg8m9, 97?</,/2; mg8m9@
- amikacin4 97D</,//( mg8m9, 97?</,/5* mg8m9.
"he de#eloped methods were used for the determination of the mentioned
aminoglycosides from acti#e pharmaceutical ingredients.