Talanta

ELSEVIER Talanta 42 (1995) 1849-1856

Fluorimetric determination of aminoglycoside antibiotics

using lanthanide probe ion spectroscopy
M. Rizk '~, Y. E1-Shabrawy a,., N.A. Zakhari a, S.S. T o u b a r '~, L.A. Carreira b
Anal Chem. Dept., Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt
Chemistry Dept., University of Georgia, Athens, GA, 30602, USA
Received 20 January 1995; revised 10 May 1995; accepted 12 May 1995

Abstract

The application of probe ion fluorimetry has succeeded in the microdetermination of six aminoglycoside
antibiotics: neomycin, streptomycin, gentamicin, tobramycin, amikacin and kanamycin as sulfate salts in
pure form and in some pharmaceutical preparations. The method is based on the reaction of Eu 3+ ions with
aminoglycosides through amino and hydroxy groups. Such interactions enhance the intensity of the 616 nm
fluorescence emission of the Eu 3+ ion. The fluorescence at 592 nm comes from a non-hypersensitive
transition and is not affected by the ligand which is bound to the probe ions. The intensity ratio R, defined
as I~92/16~6 was used to determine the amount of free and bound europium ions. A linear relationship
between bound europium ions and aminoglycoside was found within the concentration ranges 20-100 ppm
for neomycin, 5-60 ppm for streptomycin, and 10-70 ppm for gentamicin, tobramycin, amikacin, and
kanamycin as sulfate salts. The percentage recoveries ranged from 99.22 to 101.07, with standard deviations
ranging from + 1.5 to +4.38. The relative stability constants ranged from 5 x 103 to 2 x 104. The optimum
reaction conditions were studied and the results obtained compared favourably with the fluorimetric method
using fluorescmine reagent.

Keywords: Probe ion fluorimetry; Aminoglycoside antibiotics; Europium ions, Pharmaceutical analysis

1. Introduction Lanthanide ion probe spectrofluorometry

Numerous procedures are available for the
determination of aminoglycoside antibiotics
such as spectrophotometry [1-4], densitometry
[5], fluorimetry [6,7], polarography [8], gas
chromatography [9], radioimmunoassay [10],
H P L C [11,12], MS [13], and N M R [14]. Many
of these methods have been reviewed [15-19].
The official methods usually involve microbio-
logical procedures [20-22].
* Corresponding author. Present address: Chemistry
Dept., University of Georgia, Athens, Georgia, 30602,
USA. Fax: (706) 542-9454; e-mail: yaser@sunchem.
chem.uga.edu,
(LIPS) was introduced in 1979 by Horrocks
and Sundnick [23]. This technique employs two
emission lines of Eu 3+ located at 592 nm and
616 nm. The 616 nm emission line is produced
by a hypersensitive transition [24], ~Do ~ 7F2,
which is normally a forbidden transition, but
interaction with ligands makes the transition
allowed and enhances the intensity of its emis-
sion. The fluorescence at 592 nm comes from a
non-hypersensitive transition and is not affected
by the ligand which is bound to the probe ion
[25], Thus, the ratio of these intensities can be
used to determine free and ligated Eu 3+.
The applications of probe ion spectroscopy
in pharmaceutical analysis are limited in spite

0039-9140/95/$09.50 1995 Elsevier Science B.V. All rights reserved

SSD I 0039-9140(95)01640-6
1850 M. Rick et al. / Talanta 42 (1995) 1849-1856
of the availability of the method for the deter-
mination of many pharmaceutical compounds.
Europium ions have been used for the fluori-
metric determination of tetracycline [26,27], di-
clofanec sodium [28] and theophylline [29] in
blood serum and in pharmaceutical prepara-
tions. In this article, we describe a spectro-
scopic method for the determination of
aminoglycoside antibiotics in pure form and in
pharmaceutical formulations using the LIPS
spectrofluorometric technique.
2. Experimental
2.1. Apparatus
A Lambda Physic E M G 102 XeCI excimer
laser and a FL 3002 tunable dye laser were
used in tandem to supply a high intensity tun-
able source. The average output of the XeC1
excimer was 1.6W ( 1 0 H z repetition rate) at
308 nm. A tunable ultraviolet emitting dye
laser was used to provide an excitation source
o f 394.3 nm, which corresponds to a resonant
absorption transition o f the Eu metal ions. The
Eu fluorescence was collected and frequency
analyzed with a Spex 1877 triple monochroma-
tor. The fluorescence was detected by a charge
coupled detector (cod 9000 Photometric Ltd.).
The recorded curves were fitted with Lab Calc
(Galactic Software) using a gaussian model.
The areas under the two peaks at 592 nm and
616 nm were integrated and the ratio of the
two peaks was taken as the R value of the
sample.
2.2. Materials
Neomycin sulfate, streptomycin sulfate, gen-
tamicin sulfate, tobramycin sulfate, amikacin
sulfate and kanamycin sulfate were obtained
from Sigma and the various dosage forms from
commercial sources. Europium(Ill) chloride
was from Aldrich. Hydrochloric acid, chloro-
form and ammonia solutions were Baker ana-
lyzed reagents.
2.3. Reagents
All chemicals were analytical reagent grade.
The following reagents were used:
(1) Stock solutions of aminogycosides were
freshly prepared as 0.5 mg ml -~ in deion-
ized water.
(2) I E - 4 M , I E - S M and 2.5E - S M Eu 3
ion solutions were prepared by dissolving
an appropriate amount of europium
chloride hexahydrate in 11 of deionized
water, and adjusting to pH 5.5 using hy-
drochloric acid solution.
(3) Aqueous solutions of pH 2, 5.5, 8, and
10 were prepared by serial dilution of
hydrochloric acid and sodium hydroxide
with deiionized water, and pH adjust-
ment hydrochloric acid solution or
sodium hydroxide solution using a
Fisher pH meter.
2.4. Procedure
A. Calibration lines
An aliquot volume of standard aminogly-
coside antibiotic solution equivalent to 0.2-
1.0 mg neomycin sulfate, 0.05-0.6 mg
streptomycin sulfate, and 0.1-0.7 mg gentam-
icin sulfate, tobramycin sulfate, amikacin sul-
fate and kanamycin sulfate was transferred into
a 10 ml calibrated flask. 1 ml of 2.5E -5 M Eu 3
solution was added to the neomycin sulfate and
streptomycin sulfate, 1 ml of 1E -a M Eu 3 so-
lution was added to the gentamicin sulfate and
tobramycin sulfate, and 1 ml of 1E - s M Eu 3
solution was added to the amikacin sulfate and
kanamycin sulfate. 2 ml o f a hydrochloric acid
solution of pH 5.5 was added and brought to
volume with deionized water. The fluorescence
was measured from 576 to 633 nm (excitation at
394.3 nm). The curves were integrated using a
gaussian model and the area under the two
peaks at 592 nm and 616 nm was taken as the
R value of the sample.
B. For injections
An accurate volume of the mixed contents o f
three vials, equivalent to 10 mg of drug, was
transferred into a CM (carboxylic acid) ion
exchange resin column [10,31] loaded with wa-
ter-hydrochloric acid solution pH 2, and then
eluted with 25 ml of water-hydrochloric acid
solution pH 2. The column was washed with
25 ml of pH 7 deionized water. The elution and
washing water were discarded. Aminoglycoside
was eluted using 30 ml of aqueous pH 11 solu-
tion and 10 ml of deionized water into a 50 ml
calibrated flask. The solution was mixed well
and its pH adjusted to about 5.5 using hy-
drochloric acid solution; it was brought to
volume with deionized water. 2 ml of this solu-
tion was assayed as described in the previous
section.
 M. Rizk et al. / Talanta 42 (1995) 1849-1856
i
.2 2
e-
m
1.5
<
E
tad
1
1
0.5
/~
Europium ion cone.,
Fig. I. Effect o f Eu 3 ions on the emission ratio of 1 x 10 - a M neomycin sulfate.
C. For tablets
Twenty tablets were weighed and powdered.
To the quantity of powder equivalent to
500mg was added 10ml of deionized water,
and the mixture sonicated for 20 min. It was
then filtered into a 100 ml calibrated flask, the
powder washed three times each with 5 ml
water, and the combined filtrate mixed well
and brought to volume with deionized water.
2 ml o f the filtrate was transferred into a CM
ion exchange column and the procedure was
completed as described in the previous section.
D. For ointment
The contents of two bottles o f ointment were
weighed and mixed. A quantity o f ointment
equivalent to 20 mg of drug was transferred
into a separating funnel, and 50 ml of ether
was added. Gentamicin was extracted three
times with pH 8 water. The pH of the extract
was adjusted to pH 5.5 using hydrochloric acid
solution and brought to volume (100ml) with
deionized water. 2 ml of this solution was as-
sayed as described in Section 2.4 A.
3. Results and discussion
3.1. Lanthanide ion probe
In a study of any probe, the most fundamen-
tal and necessary characteristic is the ability to
determine quantitatively the concentration of
the free and bound metal ions simultaneously.
1851
I
c).&)()5 0.6Ol
The use of lanthanide ions as fluorescent
probes has been well documented [32]. The
europium metal ion has been the lanthanide of
choice in most applications and its spectral
features are widely documented [33]. This spe-
ciation information is uniquely encoded in the
experimental Eu 3+ profile. Fig. l shows a typi-
cal spectral titration plot o f Eu 3+ using
I E -4 M aminoglycoside (neomycin sulfate). In
the early stages of the titration process there is
only a small amount of fluorescent metal probe
ion compared to the available binding sites,
and hence the majority of metal ions are bound
to the ligand. Consequently, the hypersensitive
emission band at 616nm is larger than the
non-hypersensitive band at 592 nm, and the
intensity ratio R is small and close to the lower
limit value, Ls. As the titration progresses the
relative amount of total bound species gradu-
ally increases. Consequently, the intensity ratio,
R, gradually increases as more metal ions are
added to the system. At the end of titration the
ratio R is close to its upper limit value, La, and
the concentration of Eu 3 is much greater than
the concentration of ligand. The optimum con-
centration of Eu 3+ ions in a such study is that
required to produce a ratio value R between Ls
and La. A ratio value of around unity has been
shown to be the most sensitive for this type of
measurement [25,34]. All the aminoglycoside
antibiotics behave similarly to neomycin sul-
fate, except in the amount of Eu 3+ ions added
which is due to the differences in the molecular
weights.
1852
2000 -
1500
._=
o
,ooo
500
I~ ."",'!.4~f
I "<'~'-L,
. . . .
Fig. 2. Emission spectra of 1 ml of 2.5E-5 M of europium ions: (A) without neomycin sulfate; (B) in the presence of
600 lag neomycin sulfate; and (C) in the presence of 900 p.g neomycin sulfate, in a total volume of 10ml.
Suitable Eu 3 ion concentrations for such a
study were found to be 2.5E -5 M Eu(III) ions
for the determination of neomycin sulfate and
streptomycin sulfate, 1E - 5 M Eu 3 for the de-
termination of gentamicin sulfate and to-
bramycin sulfate, and I E - 4 M E u 3+ for the
determination of amikacin sulfate and
kanamycin sulfate. Fig. 2 shows emission spec-
tra of Eu 3 ions in the presence and absence of
neomycin sulfate.
3.2. Effect of buffer and pH
Fig. 3 shows that the complexes formed were
pH dependent and must be controlled between
pH 5 and 6. Acidic buffers such as acetate,
phosphate, MOPS, MES and T H A M buffer
systems were not suitable in such a study owing
to the formation of europium salts or com-
plexes which interfered with the measurement;
such observations have been made in our labo-
ratory [25]. At a pH below 5, the competition
between the proton and Eu 3+ will decrease the
formation of Eu-aminoglycoside complex, and
in neutral solution Eu(OH) 2+ is formed which
interferes with the measurement. Adjusting the
initial pH of the Eu 3+ solution to pH 5.5 and
using 2 ml of aqueous hydrochloric acid solu-
tion at pH 5.5 kept the pH of the mixtures
between 5 and 6.
3.3. Molar ratio
Eu is bound with aminoglycoside in a 1:1
molar ratio. Fig. 4 shows the spectroscopic
M. Rizk et al. / Talanta 42 (1995) 1849-1856
/f ,!!,,k i:, A \\
,,
~ ,L.-~
0,% ,'//
', ~ "-v~.~':L/
", ",,..\
':',
YL,
,
,
r ,,.,:
'! . . . . | . . . . i . . . . I . . . . i . . . . i
580 590 600 6'0 620 630
Wave length
titration of 1 ml of I x 1 0 - 4 M neomycin sul-
fate with 1 x 1 0 - 4 M Eu 3 ions.
3.4. Effect of time and temperature
The complex formation between Eu 3 ions
and aminoglycoside was almost instantaneous,
as a stable ratio was produced within 5 rain
and the complexes were stable for at least 24 h.
Temperature had little effect on the complex
formation and its stability, so the reaction was
carried out at ambient temperature.
3.5. Effect of ionic strength
Ionic strength is a factor that sometimes
cannot be neglected. The effect of ionic strength
on the spectral titration plot was studied by
varying the concentration of the background
electrolyte, sodium chloride. The choice of
sodium chloride was based on the fact that the
Na ion is very weakly bound, if at all, to the
aminoglycoside binding sites, and CI - j is al-
ready in the system as a counter ion that comes
from dissolving EuCI3. Three titration plots
correspond to 0.0, 0.01 and 0.1 M sodium chlo-
ride added as background electrolyte. The addi-
tion of background electrolyte will decrease the
total concentration of bound species of the
probe ion. It should be emphasized here that
this decrease is not due to the competition effect
of sodium ion (similar to proton competition)
to occupy the same set of aminoglycoside sites,
but purely due to electrostatic forces as previ-
ously reported with humic acid substances [34].
 M. Rizk et al./ Talanta 42 (1995) 1849-1856
1.9
O
"~ 1.8
1.7
-~ 1.6
e-,
.o
.~ 1.5
~ 1.4
1.3
3.5 4 4.5
Fig. 3. Effect of pH on emission ratio of 50 lagml-J neomycin sulfate and 1.00ml of 2.5E-5 M EuJ+ ions.
4. Performance characteristics
The investigated aminoglycosides possess
amino and hydroxy groups which can form
complexes with metal ions. Evangelista and
Schapoval [1] have determined these com-
pounds by spectrophotometric determination
of their blue complexes with copper ions at
634 nm.
Aminoglycoside antibiotics complex Eu 3
ions, and such ligation leads to a hypersensi-
tive transition of Eu 3+ ions, and such ligation
leads to a hypersenstive transition o f Eu 3+
ions, 5Do--* 7F2 (616 nm). These hypersensitive
transitions are specific absorption or emission
transitions o f lanthanide ions that are ex-
tremely sensitive to ligation. The intensity o f
a hypersensitive transition is enhanced in
the metal-ligand complex relative to that
transition in aquo ions. Experimentally, this
is observed by making the concentration of
Eu 3 ions small relative to the aminoglycoside
concentration, so that the majority o f the
metal ions are complexed with aminogly-
cosides. Such effects are most obvious from
the observation o f several emissions of Eu 3
ions with neomycin sulfate concentrations
(Fig. 2). The absolute emission intensity is not
important here as we are concerned only with
the change in the integrated emission ratio R
(1592/16~6) due to the aminoglycoside antibi-
otics. These R values decrease upon the addi-
tion of antibiotics owing to the enhanced
emission at 616 nm relative to the emission at
592 nm. The 592 nm emission is not effected
by ligation.
1853
5.5 6 6.5
pH
Susetyo and co-workers [25,35] have devel-
oped a statistical model for the distribution of
binding sites in humic acid. This model was
applied to study the spectral titration plot gen-
erated by lanthanide ion probe spectroscopy.
They expressed the free and bound Eu 3+ ions
as a function of experimental parameters. The
free Eu 3+ ions could be expressed as
M = C M L a ( R -- Ls) (1)
R(LB Ls)
-
where M is the concentration of free metal
ions, CM is the total concentration of metal
added, LB is the ratio of the two Eu peaks at
592 nm and 616 nm when the concentration of
Eu 3+ was very large with respect to ligand
concentration, and Ls is the ratio when the
Eu 3+ concentration was very small with respect
to the ligand concentration. Table I shows LB
and Ls for each aminoglycoside. R is the ratio
1592/I616 of the free and bound metal species
which was quantitatively determined from the
experimentally observed parameters.
The bound Eu 3+ ion concentration could be
expressed as
ML = CM- M (2)
Calibration lines were calculated by plotting
M L against the concentration of aminogly-
coside. It was found that there was a linear
relation between the concentration of amino-
glycoside and the concentration of bound Eu 3
ions within the ranges shown in Table l, with
standard deviations from 4.338 to 1.50 ppm.
Table l also shows regression equations and
correlation coefficients o f aminoglycosides us-
1854 M. Rizk et al./ Talanta 42 (1995) 1849-1856

/ 0 ~ 0
.o_ /
w 0

.o_
~

1.5
2

J
i,i . . . . t i . . , ,. i 1 - - - i

0.5 1 1.5 2 2.5

Molar Ratio
Fig. 4. Spectroscopic titration of 1 ml of 1 10 -4 M neomycin sulfate with 1 x 10 -4 M Eu 3+ ions.

ing the proposed method. This has proved to Table 2 shows the assay results of aminogly-
be an accurate method for the determination of cosides by the proposed method and
aminoglycosides. As compared with conven- fluorescamine fluorimetric methods [6]. The
tional organic fluorescent probes [6,7], fluores- proposed method, when compared to the
cent lanthanide chelates can offer several official microbial assays, offers advantages with
advantages. They have narrow-banded emis- regards to rapidity, simplicity and precision, so
sion lines well distinguished from any dis- we compared these results to the fluorimetric
turbing background. Chelates have method using fluorescamine. The calculated
exceptionally wide Stokes' shifts, over 200 nm, student's t and variance ratio F tests were less
while with fluorescamine it is only 50-70 nm. than the respective tabulated one at P = 0.05,
In addition to spectral resolution, their emis- so the results obtained by the proposed method
sion can be distinguished from any organic have proved to be in good agreement with
background interference owing to the very long those obtained by the reference fluorimetric
fluorescence decay time ( ~ 110 las) associated method.
with Eu 3 fluorescence which makes them an Results of the determination of aminogly-
ideal choice for a fluorimetric system [36]. cosides in different dosage forms by the probe

Table I
Regression data, correlation coefficients, stability constants of Eu-aminoglycoside complexes, concentrations of Eu used
for calibration, and concentration ranges of aminoglycoside antibiotics determined by the proposed spectroscopic probe
ion method

Aminoglycoside Regression data a Cone. b La Ls

sulfate salt range
Co CI r k CM

Neomycin 3.59E - s 4.77E -9 0.99681 2.1E 4 2.5E -'~ 20-100 2.30 0.31
Streptomycin 1.08E -9 1.91E - s 0.99983 1.3E 4 2.5E --~ 5-60 2.30 0.31
Gentamicin 4.78E- 7 2.82E q - s 0.99086 3. I E 3 1.0E-4 10-70 2.55 0.61
Tobramyein 7.12E -7 4.29E - s 0.99892 2.5E ~ 1.0E -4 10-70 2.55 0.61
Amikaein 1.13E -7 2.48E -9 0.99857 3.5E 3 1.0E -5 10-70 2.17 0.47
Kanamyein I . I I E -7 3.52E -9 0.99935 5.7E ~ 1.0E -5 10-70 2.17 0.47

Co is the intercept, c~ is the slope, r is the correlation coefficient, K is the stability constant of the Eu-aminoglyoside
complex, CM is the molar concentration of Eu ~+ ions used for calibration, L a is taken to be the ratio of the two Eu ~+
peaks in the absence of aminoglycoside, Ls is taken to be the ratio when the Eu ~+ concentration was very small with
respect to a large ligand concentration.
"Average of at least seven triplicate determinations within the concentration range.
b Concentration ranges were calculated in ppm.
 M. Ri-.k et aL / Talanta 42 (1995) 1849-1856 1855

Table 2 sUch as additives, stabilizers a n d a n t i o x i d a n t s

Assay results of aminoglycoside antibiotics in bulk using
o f tablets, a m p o u l e s a n d o i n t m e n t s , are re-
the proposed method and the reference method [6]
m o v e d b y this p r o c e d u r e a n d d o n o t interfere
Aminoglycoside % Recovery ~, 4-SD with the r e c o m m e n d e d m e t h o d . T h e y were sep-
sulfate salt a r a t e d from a m i n o g l y c o s i d e s in an ion ex-
Proposed method Reference method b c h a n g e resin ( c a r b o x y l i c acid resin) by a q u e o u s
buffer at p H 2 [30,31]. T h e p r o p o s e d p r o c e d u r e
Neomycin 100.16 + 1.05 100.04 4- 1.48
t=0.175 (2.179) F=1.987 (4.28) is as sensitive as the fluorescamine fluorimetric
m e t h o d [6], a n d gives g o o d a g r e e m e n t with it.
Streptomycin 98.61 + 3.57 99.25 4- 1.86
t=0.421 (2.179) c F=3.684(4.28) T h e p r o p o s e d m e t h o d is simpler a n d m o r e
rapid, a n d the reaction p r o d u c t s are m o r e
Gentamicin 100.82 + 2.30 99.00 4- 1.82 stable t h a n for c o n v e n t i o n a l f l u o r o m e t r i c meth-
t = 1.660 (2.179)c F = 1.689 (4.28)c
ods. M o r e o v e r , the d e t e r m i n a t i o n b o u n d Eu 3
Tobramycin 100.26 + 2.23 99.68 + 2.28 using the relative intensity r a t i o R as a function
t=0.481 (2.179) c F = 1.045 (4.28)
o f the c o n c e n t r a t i o n o f a m i n o g l y c o s i d e antibi-
Amikacin 98.81 4- 4.52 97.82 + 2.66 otics is m o r e reliable, a n d has less e x p e r i m e n t a l
t=0.500 (2.179) c F=2.875 (4.28) c e r r o r ( a b o u t + 5 % ) , t h a n that using the a b s o -
Kanamycin 100.19 + 1.50 99.14 4- 2.49 lute emission intensity in c o n v e n t i o n a l o r g a n i c
t = 0.956 (2.179) c F = 2.756 (4.28) ~ fluorescent m e t h o d s . In c o n v e n t i o n a l o r g a n i c
f l u o r o m e t r i c m e t h o d s the a b s o l u t e emission
a Average of at least seven triplicate determinations.
b Fluorometric determination of antibiotics using value is s t r o n g l y effected b y m a n y systemic
fluorescamine [6]. errors d u e to b l a n k a n d a n a l y t e interferences.
Values in parentheses are theoretical values at P = 0.05. M o r e o v e r , the a b s o l u t e emission intensity is
highly d e p e n d e n t o n the intensity o f the excita-
ion m e t h o d a n d the fluorescamine fluorimetric tion source, the q u a n t u m yield o f the c o m -
m e t h o d are p r e s e n t e d in T a b l e 3. T h e p r o p o s e d plexes which is low c o m p a r e d to the
p r o c e d u r e was selectivity e n h a n c e d by an ion E u 3 - c o m p l e x (-~0.18) [37], a n d the d e t e c t o r
e x c h a n g e resin r e s o l u t i o n o f the interferences, used for the m e a s u r e m e n t r a t h e r t h a n the
since it is well k n o w n that Eu 3+ ions are fluorescence o f the a n a l y t e [30]. Thus, the p r o -
s t r o n g l y b o u n d by c h a r g e d oxygen d o n o r p o s e d p r o c e d u r e can be used for r o u t i n e a n a l y -
a t o m s such as c a r b o x y l i c o r p h e n o l i c g r o u p s sis a n d quality c o n t r o l o f a m i n o g l y c o s i d e s in
[29]. I n g r e d i e n t s o t h e r t h a n a m i n o g l y c o s i d e s , pharmaceutical preparations.

Table 3
Assay results of aminoglycoside antibiotics in some pharmaceutical preparations by the proposed method and the
reference method

Drug % Recovery, +SD ~

Proposed method Reference method b

Neomycin Tablets 500 mg 100.99 + 0.36 98.98 + 2.16

(Memphis Co., Egypt)
Gentamicin Oint., 0.1% 104.74 4- 1.6 97.54 4- 1.98
(Memphis Co., Egypt)
Gentamicin Amp. 20 mg 101.22 4- 2.39 98.23 + 1.71
(Memphis Co., Egypt)
Gentamicin Amp. 80 mg 102.53 4- 1.03 98.93 4- 1.98
(Memphis Co., Egypt)
Nebcin Amp. 20 mg Tobramycin 103.49 4- 0.64 99.68 + 2.28
(Lily France S.A. France)
Amikin Amp. I g Amikacin 101.20 4- 1.66 99.68 4- 0.87
(Briston Co., USA)
Kanamycin vial, 1 g per vial 99.27 + 0.41 99.52 _+ 1.62

"Average of at least three triplicate determinations, calculated relative to nominal content.

b Fluorometric determination of aminoglycoside antibiotics using fluorescamine [6].

TAL ~ IZ,~
1856 M. Rizk et al. / Talanta 42 (1995) 1849-1856
5. Conclusion
From the foregoing discussion, the Eu 3
probe ion fluorimetric technique has been ap-
plied to the determination of six aminogly-
cosides in pure form and in pharmaceutical
preparations. Ion exchange was carried out for
resolution of aminoglycoside antibiotics from
other additives such as EDTA, sodium citrate,
phenol, and other injection and tablet additives.
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