Ruckmani 2011

Original Article
A simple and rapid high-performance liquid

chromatographic method for determining
tobramycin in pharmaceutical formulations by
direct UV detection
Abs t r a c t
Background: Tobramycin, an aminoglycoside antibiotic, is a polar pharmaceutical

compound which lacks a UV absorbing chromophore. Due to the absence of a UV
absorbing chromophore and high polar nature of this antibiotic, the analysis of such
compounds becomes a major challenge. Objective: To overcome these problems, a novel
method for the determination aminoglycoside tobramycin was developed and validated
ȱȱȬȱȬȱȱ¢ȱǻȬ
Ǽȱ ȱȱ
detector. Materials and Methods:ȱȱȱȱȱȱȱěȱŖǯŖśȱȱ
diammonium hydrogen phosphate, pH adjusted to 10.0 using tetramethyl ammonium
hydroxide. Chromatography was carried out at 25°C on a Purosphere RP-8e, 250
mm x 4.6 mm, 5Pm. The detection was carried out using variable wavelength UV-Vis
ȱȱȱŘŗŖȱǯȱȱȱ ȱȱ¢ȱȱȱ¢ȱĚ ȱȱ
of 1.0 mL/min. Result and Discussion: Tobramycin retention time was about 9.0 min
with an asymmetry factor of 1.4. A logarithmic calibration curve was obtained from
ŖǯŚŝȱȱŖǯŝŗȱȦȱǻȱǁȱŖǯşşşŞǼǯȱȬ¢ȱƖȱ ȱŖǯŘşȱǻȱƽȱŜǰȱŖǯŜŖȱȦǼȱȱ
between-day %RSD was 0.54 ȱę¢Ȧȱ¢ȱ¡ȱȱȱȱ
K. Ruckmani, Saleem Z. ȱȱȱ¡ǰȱ¢ȱȱȱȱ ȱ ȱşşǯŖȮŗŖŖǯŖȱ
Shaikh1, Pavne Khalil1, percent. Conclusions: A HPLC method based on UV detection has been developed and
M. S. Muneera1 validated for determination of tobramycin from ophthalmic solution. The method is
ǰȱǰȱęǰȱȱǻȱŖǯŞŖƖǼǰȱȱǻȱ<ŘǯŖƖǼȱȱȱǻŘƽŖǯşşşŞǼǯȱ
Department of Pharmaceutical
The described method is suitable for routine analysis and quality control of ophthalmic
Technology, Anna University of
Technology, Thiruchirappalli, solution containing tobramycin.
Trichy, Tamilnadu, India, Key words: Amino glycoside, derivatization, isocratic, reverse phase chromatography,
1
Analytical Research and
tobramycin
Development, Jamjoom
Pharmaceuticals, Jeddah,
Saudi Arabia
Address for correspondence: INTRODUCTION

Dr. Saleem Z. Shaikh,
Analytical Research and
Tobramycin is an amino glycoside antibiotic used to treat various types of
Development, Jamjoom
Pharmaceuticals, P.O. Box- 6267, bacterial infections, particularly Gram-negative infections. It works by binding
Jeddah- 21 442, Saudi Arabia to a site on the bacterial 30S and 50S ribosome, preventing formation of the 70S
E-mail: salim2001p@hotmail.com complex. As a result, mRNA cannot be translated into protein and cell death
ensure. Tobramycin is preferred over gentamicin for Pseudomonas aeruginosa
Access this article online ȱȱȱĴȱȱȱȱȱ¢ǯ[1,2]
Website: www.phmethods.org
DOI: 10.4103/2229-4708.84455
Like all amino glycosides, tobramycin does not pass through the gastro-
intestinal tract, so for systemic use it can only be given intravenously or
Quick response code
intramuscularly.[1]ȱȱȱ¢ȱȱȱǻ¢ȬǼȱ ȱ
a tobramycin concentration of 0.3% is available in the market.
ȱȱȱŘŖŖŞȱǻȱřŗǼȱȱȱȱȱȱ
assay of raw material and tobramycin ophthalmic solution by high-performance
ȱ ¢ȱ ȱ £ȱ ȱ ŘǰŚȬĚ£ȱ
ȱȱǻ¢¡¢¢ȱǼȱǰȱ¡ȱȱěȱǻŘȱȱȱ
Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

117
Ruckmani, et al: HPLC methods for determining tobramycin in pharmaceutical formulations
ȱ ǻ¢¡¢¢ȱ Ǽȱ ȱ ŘŖȱ ȱ ȱ MATERIALS AND METHODS
ŗȱȱǼȱȱȱȱȱȱŚŖǱŜŖȱ
Ȧȱ ȱ ȱ ȱ ȱ ȱ ȱ Ě ȱ ȱ ȱ ŗǯŘȱ Ȧ
Instrumentation
ǰȱȱȱȱǻřǯşȱƼȱřŖŖȱǼȱȱȱŗȱ
Integrated high-performance liquid chromatographic
packing, with detector wavelength set at 365 nm.[3]
systems LC-2010AHT from Shimadzu Corporation
ŘǰȱŚȬĚ£ȱȱȱǻ¢¡¢ȱ¢ȱ
(Chromatographic and Spectrophotometric Division,
Ǽȱȱȱȱȱśȱ¢ȱȱŚȱ
¢ǰȱǼȱȱȱȱ¢ȱȱ¢ǰȱ
hours respectively. Derivatization is carried out at
high-speed auto-sampler, column oven, and UV-
60°C constant temperature.
Vis detector. A Purospher RP-8e, 250 mm × 4.6 mm,
In comparison with derivatization, simple 5mȱ ¢ȱ ȱ ȱ ǰȱ ¢ǰȱ ȱ
reverse-phase chromatographic methods have the used as a stationary phase. Chromatograms were
advantages of reducing analysis time, enhancing recorded and integrated on PC installed with LC
sensitivity and flexibility, and lowering the cost ȱȱĞ ǰȱȱŗǯŘŘȱŗȱ
of the instruments and maintenance. The biggest ǻ£ǰȱ ¢ǰȱǼǯ
disadvantage of derivatization has been lack of
stability. The reaction products are not stable Reference substances, reagents, and chemicals
and have a short half-life possibly because of a Working standard of tobramycin was obtained from
spontaneous intermolecular rearrangement. Another Chongqing daxin pharmaceuticals laboratories Ltd.,
disadvantage of derivatization is that it reacts with China. Diammonium hydrogen phosphate and tetra
only few functional groups.[4] methyl ammonium hydroxide were purchased from
ȱ ǻǼȱ ǯȱ ȱ ȱ ȱ
The literature survey shows that several methods
ȱȱȱȬȱ¢ǰȱǰȱǰȱ
like HPLC with evaporative light scattering
ǰȱ ǯȱȱ ȱ ȱ ȱ ȱ ȱ ȱ
ǰȱȱǰȱȦǰȱ
ȱ
analytical or reagent grade. Reference standards
UV-Vis have been reported for the determination of
of tobramycin were obtained from United States
tobramycin with derivatization.[5-20] These reported
ȱ ǰȱ ǰȱ ǰȱ ǯȱ
methods and the USP method are not rapid for
Ophthalmic formulations containing tobramycin
assay of tobramycin in ophthalmic solutions. All
were developed and manufactured in our research
the reported and official methods are complex,
and development laboratory.
insensitive, and risky. However, as per bibliographical
revisions performed, no analytical method has
ȱ ȱ ȱ ȱ ǻ ȱ £Ǽȱ Chromatographic conditions
determination of tobramycin by high-performance ȱ ȱ ȱ ȱ ȱ ȱ ŖǯŖśȱ ȱ
liquid chromatography. diammonium hydrogen phosphate, pH adjusted to
ŗŖǯŖȱƹȱŖǯŖśȱȱ¢ȱȱ¢¡ȱ
The present study was aimed at developing a ǻŘśƖȱȱȱ Ǽǯȱȱȱȱ ȱęȱ
ǰȱęǰȱǰȱȱȱ
ȱȱ ȱȱȱȱȱęȱȱŖǯŚśȱΐȱ
for determination of tobramycin in commercially porosity under vacuum. A Purosphere RP-8e, 250
available pharmaceutical formulations such as mm × 4.6 mm, 5 mm analytical column was used as a
ophthalmic solution, injection, suspensions, and in- ¢ȱǯȱȱȱĚ ȱȱȱǯŖȱȦȱ
house prepared ophthalmic suspension, based on was employed throughout the analysis. A variable
ȱȬȱǰȱȱ ȱȱŗŖŖƖȱěȱ ȱȱ UV-Vis detector was set at 210 nm. All pertinent
as a mobile phase to determine the compounds for analyses were made at 25°C and volume of solution
use in routine quality control applications associated ȱ ȱ ȱ ȱ ȱ ȱ śŖȱ ΐǯȱ ȱ ȱ
with these ingredients. phase was used as diluent for standard and sample
preparations.
The proposed method for the determination of
tobramycin in pharmaceutical formulations by HPLC
ȱ ȱ ȱ ęȱ ȱ ȱ ȱ ȱ ȱ Samples
derivatization. This information could be very useful Test samples were ophthalmic solutions prepared
for many of the pharmaceutical industries for the in-house and purchased from the local market with
determination of this compound and for those who composition 3.0 mg/ml of tobramycin. Other test
do not have the costly instruments, selective detectors samples used were accelerated stability samples with
such as a refractive index detector. similar composition.

118
Solution preparation and detector wavelength at 205 nm resulted in no peak

Tobramycin standard solution ȱȱĞȱŜŖȱȱȱȱǯȱȱȱ
Tobramycin standard solutions were prepared by ȱ ȱ ȱ ȱ ŖǯŖŘȱ ȱ ȱ ȱ
transferring accurately about 66.0 mg of tobramycin ȱȱěȱȱȱȱȱ
working standard equivalent to 60.0 mg of anhydrous ratio 50:50, v/v, was tried in isocratic conditions on the
¢ȱȱȱŗŖŖȱȱȱĚǯȱȱŝŖȱȱ Spherisorb ODS-1, 250 mm × 4.6 mm, 5mm to obtain
of diluent was added and sonicated for few minutes symmetrical peak shapes and clear separation of the
to solubilize tobramycin. The solution was diluted signal peaks from the solvent front peaks.
to volume with the diluent and mixed. The solution
ȱęȱȱȱŖǯŚśȱΐȱȱęȱȱ Upon investigation of the two chromatographic systems
śŖȱΐȱ ȱǯ ȱŖǯŖśȱȱȱ¢ȱǰȱ
pH adjusted to 7.0 using potassium hydroxide, and
Estimation from formulations Ŗǯŗȱȱȱ¢ȱǰȱ
ȱȱ
ȱ Ĵȱ ȱ ȱ ǰȱ ȱ to 6.9 using a potassium hydroxide solution as the
tobramycin 3.0 mg/ml, were shaken gently and ȱ ȱ ȱ ȱ ȱ Ě ȱ ȱ ȱ ǯŖȱ Ȧǰȱ
transferred to a glass beaker and mixed. About BioSep SEC-S2000, 300 mm × 7.8 mm and a Purosphere
5.0 gm of ophthalmic suspension was weighed RP-8e, 250 mm × 4.6 mm, 5mm analytical column as
¢ȱȱȱŘśȱȱȱĚǰȱȱŗŖȱȱ a stationary phase and detector wavelength at 205
of diluent was added, shaken to disperse the sample, nm resulted in peak elution at 3.2 minutes and 9.7
and diluted to volume with diluent and mixed. ȱ ¢ǰȱ ȱ ęȱ ȱ ȱ
This solution theoretically contains 0.60 mg/ml of in tobramycin peak eluted very close to negative peak
¢ǯȱȱȱ ȱęȱȱȱŖǯŚśȱ ǻȱȱǼȱȱȱȱȱȱȱȱ
ΐȱȱęȱȱśŖȱΐȱ ȱȱ¢ȱ ȱȱȱǻǁřǼȱȱ ȱȱŗǯ
on to the column. ǰȱȱȱȱȱȱȱȱȱ
for the determination of tobramycin by UV detection,
Quantitation ěȱȱȱȱȱ ȱ¢ȱ
Peak areas were recorded for all peaks. Peak areas [Table 1] to achieve the best signal response and
were taken into account to quantitate the label amount retention time.
in milligram per ml of ophthalmic suspension by using
the following formula: ¢ǰȱȱȱȱȱȱ ǱȱŖǯŖśȱȱ
diammonium hydrogen phosphate, pH adjusted to
Tobramycin mg/ml = Ru /Rs × C/100 × 25/W × 1/L × P × D 10.0 ± 0.05 using tetra methyl ammonium hydroxide
ǻŘśƖȱȱȱ ǼȱȱȱȱĚ ȱȱȱŗǯŖȱ
where Ru is the peak area obtained from tobramycin in ml/min and detector wavelength set at 210 nm, using
the investigation solution; Rs is the peak area obtained a Purosphere RP-8e, 250 mm × 4.6 mm, 5mm column,
from tobramycin in the standard solution; C is the was found to be appropriate, allowing well signal
ȱǻǼȱȱ¢ȱ ȱȱȱ response of tobramycin.
to prepare the standard solution; Wȱȱȱ ȱǻǼȱ
of the test sample; P is purity of tobramycin working
Optimization of HPLC
standard, L is the labeled amount of tobramycin in mg/
ȱ
ȱ ȱ ȱ ȱ ȱ ȱ ěȱ ȱ ¢ȱ
ml of ophthalmic suspension, and D is the density of
retention time as well as the detection sensitivity.
the ophthalmic suspension.
ȱ Řȱ ȱ ȱ ȱ ȱ ȱ ȱ
ǻȱ Ǽǰȱ Ĝ¢ȱ ǻ ȱ ȱ ȱ ȱ Ȧ
RESULTS AND DISCUSSION Ǽȱȱ¢ȱȱȱ¢ȱȱěȱ
pHs. The optimal pH 10.0 ± 0.05 was chosen for the
Chromatography determination of tobramycin.
Our method development started with the search for
the suitable column and mobile phase. ȱ ȱ ȱ ěȱ ȱ ȱ ȱ ȱ
ȱȱȱȬȱǯȱȱřȱ ȱȱ
ȱȱ¢ȱȱŖǯŖŘȱȱȱ ¢ȱ ȱ ȱ ȱ ȱ ǻȱ Ǽȱ
ǱȱǻśŖǱȱśŖȱȦǼǰȱȱȱȱȱȱȱ ȱ ȱ ȱ ȱ ȱ ěȱ ǯȱ ȱ
ȱĚ ȱȱȱǯŖȱȦǰȱȱǰȱŘśŖȱȱ ȱ ȱ ȱ ȱ ȱ ŖǯŖśȱ ȱ ȱ
× 4.0 mm, 5mm analytical column as a stationary phase, hydrogen phosphate was used. This may be due

119
a b
Figure 1: (C%JTQOCVQITCOQHVQDTCO[EKPUJQYKPIſTUVKPXGUVKICVKQP. Chromatographic column: BioSep SEC-S2000, 300 mm × 7.8 mm, mobile
RJCUG/RQVCUUKWOFKJ[FTQIGPRJQURJCVGR*CFLWUVGFVQWUKPIRQVCUUKWOJ[FTQZKFGƀQYTCVGONOKP&GVGEVQTYCXGNGPIVJPO
(b) Chromatogram of tobramycin showing second investigation. Chromatographic column: Purosphere RP-8e, 250 mm × 4.6 mm, 5Pm, mobile
RJCUG/RQVCUUKWOFKJ[FTQIGPRJQURJCVGR*CFLWUVGFVQWUKPIRQVCUUKWOJ[FTQZKFGƀQYTCVGONOKP&GVGEVQTYCXGNGPIVJPO
6CDNG'ORNQ[GFOQDKNGRJCUGUEQNWOPUCPFGNWVKQPVKOGFWTKPIVJGKPXGUVKICVKQPQHVQDTCO[EKP
Column Mobile phase Elution time
Silica column, 250 mm × 4.0 mm, 5 mm 0.02 M formic acid: acetonitrile (50: 50 v/v) *No peak eluted
Spherisorb ODS-1, 250 mm × 4.6 mm, 5 Pm 0.02 M ammonium phosphate: acetonitrile, (50: 50 v/v) *No peak eluted
BioSep SEC-S2000, 300 mm × 7.8 mm. 0.05 M potassium dihydrogen phosphate, pH adjusted to 7.0 **14 minutes
using potassium hydroxide.
Purosphere RP-8e, 250 mm × 4.6 mm, 5 mm 0.1 M potassium dihydrogen phosphate, pH adjusted to 6.9 ***4.3 minutes
using potassium hydroxide solution
BioSep SEC S2000, 300 mm × 7.8 mm 0.05 M sodium acetate: acetonitrile (50: 50 v/v) *No peak eluted
Waters, Spherisorb ODS-1, 250 mm × 4.6 mm, 5 Pm 0.05 M boric acid: acetonitrile (50: 50 v/v) *No peak eluted
Waters, Spherisorb ODS-1, 250 mm × 4.6 mm, 5 Pm 0.1% aqueous glacial acetic acid: acetonitrile, (50: 50 v/v) *No peak eluted
Alltech allsphere, ODS-2, 250 mm × 4.6 mm, 10 Pm 4.8% aqueous protriptylene hydrochloride: acetonitrile *No peak eluted
(50: 50 v/v)
BDS, Hypersil C18, 50 mm × 4.6 mm, 5 Pm 2.4% aqueous protriptylene hydrochloride: acetonitrile *No peak eluted
(50: 50 v/v)
Waters, Spherisorb ODS-1, 250 mm × 4.6 mm, 5 Pm 0.05 M aqueous diammonium hydrogen phosphate: acetonitrile, *No peak eluted
(50: 50 v/v)
Supelcosil LC-8, 250 mm × 4.6 mm, 5 Pm 0.01 M sodium dihydrogen phosphate: acetonitrile (950: 50 v/v) *No peak eluted
pH adjusted to 6.9 using potassium hydroxide
Purosphere RP-8e, 250 mm × 4.6 mm, 5 Pm 0.05 M diammonium hydrogen phosphate, pH adjusted to 10 8.9 minutes
using tetramethyl ammonium hydroxide
*No peak tobramycin eluted after 60 minutes., ** Tobramycin peak eluted very close to negative peak (peak from diluents), ***Peak shape was not acceptable, tailing
factor high.
to highly aqueous environment that is unfavorable 5RGEKſEKV[

ȱ ȱ ǯȱ ǰȱ
ȱ ŗŖǯŖȱ ȱ ŖǯŖśȱ ȱ No interferences were observed due to the obvious
diammonium hydrogen phosphate was chosen for presence of excipients and mobile phase.
estimation of tobramycin. Typical chromatogram of
ȱȱȱȱ ȱȱȱřǯ Linearity
Peak areas versus concentration in milligram per milliliter
ȱĴȱȱ¢ȱȱȱȱȱ
Method validation between 80% and 120% of the target level. Tobramycin
The test method for the determination of tobramycin showed linearity between 0.47 and 0.71 mg/ml with a
was validated to include the essential demands of ȱĜȱǻŘǼȱȱŖǯşşşŞǯ
ȱ ȱ ȱ
£ȱ ǻ
Ǽȱ
guidelines.[21]ȱ ȱ ȱ ę¢ǰȱ ¢ǰȱ Accuracy
accuracy, precision, range, robustness, and system Accuracy of the proposed HPLC determination was
suitability were examined. evaluated from the assay results of the components.

120
Accuracy was done by performing the assay of involving different weighing. Each solution was
samples and calculated the peak area responses of injected in triplicate under same conditions and the
ěȱȱ¢ȱȱȱ¢ȱǯ mean value of peak area response for each solution was
taken. Corrections in area were made for each weight
Stock solution that had been taken to prepare six sample solutions
A Stock solution was prepared by dissolving accurately and relative standard deviation of the contents of
weighed portions of about 666 mg of tobramycin in tobramycin the six sample solutions was calculated.
portions of the mobile phase and diluted to produce Relative standard deviation was 0.29 for tobramycin.
100 ml solution.
Intermediate precision was performed by analyzing the
An appropriate portion of the stock solution was ȱ¢ȱ ȱěȱ¢ȱ¢ȱěȱ
spiked into a blank placebo matrix to produce ǯȱ ȱ ȱ ȱ ȱ ¡ȱ ěȱ
concentrations of 80, 100, and 120 of the target level. samples at 100% target level were prepared by each
ȱ ¢ȱ ȱ ȱ ȱ ȱ şşǯŚśȱ Ɩȱ ȱ analyst. The relative standard deviation obtained from
tobramycin [Table 2]. 12 assay results by two analysts was 0.42 for tobramycin.
Precision Robustness
Instrumental precision was determined by six Robustness of the proposed method was performed
replicate determinations of standard solution and the
by keeping chromatographic conditions constant with
relative standard deviation was 0.29 for tobramycin.
the following deliberate variations:
ȱ ȱ ȱ Ȭ¢ȱ ȱ ȱ ǻǼȱȱȱȱȱȱ
performed by preparing six different samples ǻǼȱȱȱĚ ȱȱȱŗǯŖȱȦȱȱŗǯŘȱȦǯ
Figure 2: 'HHGEV QH R* QP

RGCM CTGC CPF GHſEKGPE[
UJQYP CU Figure 3: A typical chromatogram of test sample by proposed
plate number N/column) and capacity factor of tobramycin column: methods column: Purosphere RP-8e, 250 × 4.6 mm, 5Pm, mobile
Purosphere RP-8e, 250 × 4.6 mm, 5Pm, mobile phase: 0.05 M phase: 0.05 M diammonium hydrogen phosphate, pH adjusted to
diammonium hydrogen phosphate, pH adjusted to 10 using tetramethyl WUKPI VGVTCOGVJ[N COOQPKWO J[FTQZKFG ƀQY TCVG ONOKP
ammonium hydroxide Oset at 210 nm
6CDNG#EEWTCE[FCVC
CPCN[VGTGEQXGT[6QDTCO[EKP
Amount std. added Amount std. recovered Determined Recovered (%) Bias (%)
(mg/ml) (mg/ml) (% of target level)
1 0.4727 0.4687 79.32 99.15 -0.85
0.4729 0.4696 79.44 99.29 -0.75
0.4726 0.4678 79.19 98.98 -1.02
2 0.5909 0.5903 99.91 99.91 -0.09
0.5911 0.5903 99.87 99.87 -0.13
0.5908 0.5888 99.68 99.68 -0.32
3 0.7090 0.7054 119.39 99.49 -0.51
0.7093 0.7051 119.28 99.40 -0.60
0.7089 0.7042 119.20 99.30 -0.70

121
6CDNG%QORCTKUQPQHU[UVGOUWKVCDKNKV[RCTCOGVGTU6QDTCO[EKP
Parameters Proposed method Change in buffer composition -5% &KDQJHLQÀRZUDWH
Retention time 8.93 8.88 8.50
Peak asymmetry 1.43 1.32 1.36
Theoretical plates 6566.38 6490.63 6023.84
Capacity factor 1.98 1.88 1.92
6CDNG#RRNKECVKQPQHVJGFGXGNQRGF*2.%OGVJQFVQVJGFGVGTOKPCVKQPQHVQDTCO[EKPKPFQUCIGHQTOU
Test sample Concentration Assayb %
Ophthalmic solution 0.3 % 99.87
Injection 40 mg/ml after reconstitution 99.68
Suspension Tobramycin 0.3% and dexamethasone 0.1% 100.02
In-housea (suspension) Tobramycin 0.3% and dexamethasone 0.1% 101.23
Stress samplea (suspension) Tobramycin 0.3% and dexamethasone 0.1% 100.05
a
Formulation was prepared in the research and development lab. bEach value is the average of two determination
The standard solution was injected six times in CONCLUSIONS

replicate for each minor change. System suitability
parameters like peak asymmetry, theoretical plates, A HPLC method based on UV detection has been
capacity factor, and relative standard deviation developed and validated for determination of
were recorded for each peak and found to be within tobramycin from an ophthalmic solution. The
acceptable limits. method is simple, rapid, specific, accurate (error
ŖǯŞŖƖǼǰȱȱǻȱ<ŘǯŖƖǼǰȱȱȱǻŘƽŖǯşşşŞǼǯȱ
Six test samples at the target concentration level The described method is suitable for routine analysis
were prepared and analyzed for each change. The and quality control of the ophthalmic solution and
ȱěȱȱ¢ȱȱȱȱ injection containing tobramycin.
deviations was calculated during each change and
found to be ± 0.5% and less than 1.0, respectively.
It was noted that slight addition of solvents in the
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2.
Ĵȱ ǯȱ ¢Ǳȱ ¡¢ȱ ȱ ǯȱ ȱ ȱ ȱ
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ǰȱ ȱ ǰȱ ȱ
ǰȱ ȱ
ǯȱ ȱ ¢ȱ ȱ ¢ȱ
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ǻŚŖķȦŘśƖ
Ǽȱ ȱȱȱ¢ȱȱ¢ǯȱ 1984;305:345-52.
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ěȱ ǯȱ ȱ
determination of amino glycoside antibiotics in serum: a comparison
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ěȱ ǯȱ Ȭȱ ȱ
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ȱ
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using high-performance anion exchange chromatography with
How to cite this article: Ruckmani K, Shaikh SZ, Khalil P, Muneera
ȱ ȱ ȱ ǯȱ ȱ ȱ ȱ ȱ
MS. A simple and rapid high-performance liquid chromatographic
2006;40:1006-12.
method for determining tobramycin in pharmaceutical formulations
17. ȱ ǰȱ ȱ ǯȱ ȱ ȱ ȱ
by direct UV detection. Pharm Methods 2011;2:117-23.
of a novel HPLC/ELSD method for the direct determination of
Source of Support: Nil, &RQÀLFWRI,QWHUHVW None declared.
tobramycin in pharmaceuticals, plasma, and urine. Anal Bioanal
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Original Article

A simple and rapid high-performance liquid

Background: Tobramycin, an aminoglycoside antibiotic, is a polar pharmaceutical

Address for correspondence: INTRODUCTION

Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

Solution preparation and detector wavelength at 205 nm resulted in no peak

Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

to highly aqueous environment that is unfavorable 5RGEKſEKV[

Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

Figure 2: 'HHGEV QH R* QP

Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

The standard solution was injected six times in CONCLUSIONS

Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

1978;24:1940-7. Chem 2005;382:290-6.

Pharmaceutical Methods | April-June 2011 | Vol 2 | Issue 2

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