Vietnam Journal of Chemistry, International Edition, 55(6): 767-774, 2017

DOI: 10.15625/2525-2321.2017-00542

Extractive spectrophotometric methods for determination of

ciprofloxacin in pharmaceutical formulations using sulfonephthalein
acid dyes
Nguyen Trung Dung1*, Le Hoc Bau1, Le Quang Thao1, Nguyen Quang Dat1
Faculty of Physics and chemical Engineering, Le Quy Don Technical University, Ha Noi, Viet Nam
Received 17 August 2017; Accepted for publication 29 December 2017

Abstract
Three simple, rapid, sensitive and accurate extractive-spectrophotometric method for the determination of
ciprofloxacin in pharmaceutical preparation has been developed. These methods are based on the formation of yellow
ion-pair complexes between the examined drug and three sulfonephthalein acid dyes, namely; bromophenol blue
(BPB), bromocresol green (BCG), and bromothymol blue (BTB) in acidic medium. The formed complexes were
extracted with chloroform and measured at 420, the colored chromogen was stable for twenty four hours. The effect of
optimum conditions via pH, dye concentration, time and solvent are studied. Beer’s law is obeyed in the concentration
ranges 0.50-25.0 μg/mL with molar absorptivity of 1.46 104, 1.83 104 and 2.07 104 L. mol-1. cm-1 and limit of
detection (LOD) of 0.105, 0.101, 0.084 for BPB, BCG and BTB methods, respectively. No interference was observed
from common excipients present in pharmaceutical formulations. The proposed method has been applied successfully to
determine ciprofloxacin in pharmaceutical preparation (tablets, infusion and eye drops).
Keywords. Ciprofloxacin, extraction-spectrophotometry, ion pair complex; sulfonephthalein dyes.

1. INTRODUCTION ciprofloxacin include oxidative coupling with 3-

methyl-2-benzothiazolinonehydrazone
Ciprofloxacin (CPF), 1-cyclopropyl-6-fluoro-4-oxo- hydrochloride (MBTH) and cerium (IV) ammonium
7-(piperazin-1-yl)-quinoline-3-carboxylic acid sulfate, Fe(III)- MBTH, Fe(III)-1,10-phenanthroline,
(Fig.1), is a second generation fluoroquinolone Fe(III)-Bipyridil [5-6], charge-transfer complexation
antibacterial agent with a broad spectrum of activity with p-acceptors such as 2,3-dichloro-5,6-dicyano-q-
against a variety of gram positive and gram negative benzoquinone, 7,7,8,8-tetracyanoquinodimethane
bacteria. It is widely used in the treatment of acute (TCNQ), p-chloranil, p-nitrophenol and
sinusitis, lower respiratory tract infection, urinary tetracyanoethylene [7], ion-pair complex formation
tract infection, chronic bacterial prostatitis and non- with acid–dye reagents such as cobalt (II)
complicated intra abdominal infections caused by E. tetrathiocyanate, Bi (III) tetraiodide, sudan III,
coli, P. aeruginosa, Proteus mirabilis when used in methyl orange, supracene violet 3B, tropeolin 00,
combination with metronidazole[1]. bromophenol blue, bromothymol blue or
A number of analytical methods have been bromocresol purple [8-11].
reported for the determination of ciprofloxacin in However, the disadvantages of using these
pharmaceutical dosage forms and biological fluids methods are that the reaction is often narrow
including spectrofluorometric, micellar linearity range, requiring heating, long time for the
electrokinetic chromatography, differential pulse reaction to complete, low stability of the colored
voltammetry, flow injection analysis, high- product formed.
performance liquid chromatography, liquid Bromophenol blue (BPB), bromocresol green
chromatography tandem mass spectrometry and (BCG) and bromothymol blue (BTB) are known to
spectrophotometric [2-4]. Spectrophotometry is yield an ion-pair complex, which are applied in the
considered as the most convenient analytical determination of many pharmaceutical compounds
technique in pharmaceutical analysis because of its by extractive spectrophotometric[12-13]. The
inherent simplicity and availability in most quality methods based on ion pair complexes extractable
control and clinical laboratories. Spectrophotometric into a suitable organic solvent have been shown to
methods reported for the determination of be simple, sensitive, accurate and economical.

767
VJC, 55(6), 2017 Nguyen Trung Dung et al.

In this paper we report three simple, rapid and 2.2. Standard solutions
sensitive extractive spectrophotometric methods for
the determination of ciprofloxacin in pharmaceutical A stock solution of ciprofloxacin (1mg/mL) in
formulations. The methods are based on ion-pair double distilled water. The working standard
complexation between ciprofloxacin with anionic solution of ciprofloxacin containing 100 µg/mL was
dye namely bromophenol blue (BPB), bromocresol prepared by dilution.
green (BCG) and bromothymol blue (BTB) The dyestuffs were used as 0.025 % solutions in
subsequent extraction into chloroform and measure doubly distilled water.
the absorbance of color complex. The proposed
methods were applied to the determination of 2.3. Pharmaceutical preparations of ciprofloxacin
ciprofloxacin in pharmaceutical preparation.
. Tablets: Weigh and mix the contents of twenty
O O tablets (each one contains 500 mg ciprofloxacin), an
F accurately weighed amount of powder equivalent to
OH
0.1g of CPF transferred in to a 100 mL beaker.
N N Using a magnetic stirrer, the powder was completely
HN disintegrated in doubly distilled water, filtered
through a Whatman filter paper No 40 and diluted
up to 100 mL with doubly distilled water in a
. volumetric flask. The working solution of the drug
Figure 1: Chemical structure of ciprofloxacin containing 100 µg/mL was prepared by dilution and
the below procedure was followed.
2. EXPERIMENTAL Infusion solution (2 mg/mL) and eye drops (30
mg, 10 mL each ): a suitable volume was diluted to
2.1. Chemicals and equipment 100 µg/mL with double distilled water and the
below procedure was followed.
All chemicals used were of analytical grade and
double distilled water was used for dilution of 2.4. Procedure and calibration graph
reagents and samples. Ciprofloxacin hydrochloride
(Sigma-Aldrich, Germany, certified to be 99.0%), Into a series of 125 mL separating funnel, volumes
bromophenol blue (BPB), bromocresol green (BCG) of CPF working standard solution equivalent to 0.5-
and bromothymol blue (BTB) (Maya - R, China, 25 μg/mL were transferred. To each funnel, add 4.0
certified to be 99%) were used. The most common mL of 0.025% BPB, BCG, BTB, respectively and
solvents are chloroform, dichloromethane, carbon mixed well. Then 10 mL of chloroform was added to
tetrachloride, dichloroethane, benzene, toluene, n- each of the separating funnel. The contents were
hexane and other chemicals used were of analytical shaken for 2 min and allowed to separate the two
reagent grade. layers. The absorbance of the organic phase at 420
The following dosage forms containing nm was measured in each case against a reagent
ciprofloxacin were purchased from local pharmacy blank similarly prepared and and a calibration graph
market and employed in the study: 1 – Hasancip was constructed. The colored chromogen was stable
and Kacipro tablets equivalent to 500 mg for twenty four hours.
ciprofloxacin (Hasan-Dermapharm and Dong Nam
manufacturing – Trading pharmaceutical Co., Ltd, 2.5. Statistical analysis
Viet Nam), 2 – Ciprofloxacin infusion equivalent to
200 mg ciprofloxacin /100 mL solution for infusion Method was validated according to ICH
(Hebei Tiancheng Pharmaceutical Co., Ltd and Guidelines[14], in terms of linearity and range,
Shandong Hualu Pharmaceutical Co., Ltd, China) accuracy and precision, limit of detection (LOD),
and 3 – Ciprofloxacin 0,3% eye drops equivalent to limit of quantitation (LOQ).
30 mg ciprofloxacin/10 mL solution (Thanh Hoa Calculation and processing of data were done
pharmaceutical and mediacal supplies joint stock using the programs Origin Pro 8.0 (USA).
company, Viet Nam).
A Biochrom Model SP-60 double beam, UV- 3. RESULTS AND DISCUSSION
VIS spectrophotometer (Biochrom Ltd., UK) with
1.0 cm matched quartz cells was used for absorbance 3.1. Principles of the method
measurements.

768
VJC, 55(6), 2017 Extractive spectrophotometric methods for…

Ciprofloxacin contains a secondary amino two tautomers are present in equilibrium but due to
group, which is protonated in acid medium, while strong acidic nature of the sulphonic acid group, the
sulphonic acid group is present in BPB, BCG and quinoid body must predominate. Finally, the
BTB that is the only group undergoing dissociation protonated ciprofloxacin forms ion-pairs with
in the pH range 1-5. The colour of such dyes is due anionic dyes, which are quantitatively extracted into
to the opening of lactoid ring and subsequent chloroform. The possible reaction mechanisms are
formation of quinoid group. It is supposed that the proposed and given in figure 2.

Figure 2: The possible reaction mechanism for the reaction between ciprofloxacin and bromocresol green,
bromothymol blue

3.2. Optimum reaction conditions for complex pH and effect of shaking time.
formation
3.1.2. Effect of pH
The optimization of the methods was carefully
studied to achieve complete reaction formation, The influence of pH on the ion-pair formations of
highest sensitivity and maximum absorbance. The ciprofloxacin with various dyes has been studied
following parameters were optimized such as dye using HCl 1 M and NaOH 1 M. It was noticed that
concentration, type of extracting solvent, effect of the highest absorbance value were observed at pH

769
VJC, 55(6), 2017 Nguyen Trung Dung et al.

3.3, 3.4 and 3.5 for BPB, BTB and BCG method, and stability of color of the formed ion-associates
respectively (Fig. 3). Thus, all the absorbance was chloroform for BCG, BPB and BTB (table 1).
measurements were made at pH 3.3, 3.4 and 3.5 with
BPB, BTB and BCG, respectively. Table 1: The effect solvent that required for ion-pair
complex formation ( max = 420 nm)
1.0
CPF- BCG
CPF- BPB
Absorbance (10 µg/mL of
0.8 CPF- BTB Organic solvent CPF)
BPB BCG BTB
0.6 Chloroform 0.452 0.565 0.695
Absorbance

0.4
Dichloromethane 0.436 0.553 0.663
Dichloroethane 0.182 0.325 0.140
0.2 Carbon
0.033 0.046 0.047
tetrachloride
0.0 Benzene 0.028 0.044 0.175
1 2 3 4 5 6 7
pH
Toluene 0.133 0.008 0.074
Figure 3: Effect of pH on the absorbance of 10 3.2.4. Effect of shaking time
g.mL-1 CPF acid-dye
The effect of shaking time on the formation and
3.2.2. Effect of dyestuff concentration stability of the ion-pair complex was studied by
measuring the absorbance of the extracted ion-
The effect of dyestuff concentrations was also associates at increasing time intervals (0-4.0 min), the
studied by adding different volumes of 0.025 % results showed that the ion-pair complex were
dyestuff (0.5-7.0 mL) to a constant amount of formed almost instantaneously in all cases at room
CPF(10 μg.mL-1). The results showed that the temperature with 2.0 min shaking time (Fig. 5). The
maximum color intensity of the complex was absorbances of the complexes were found to be
achieved with 4.0 mL of 0.025 % of each dye. Thus, stable for more than 24 h.
4 mL of each dyestuff was used for ion-pair
formation throughout the experiment (Fig. 4). 1.0
CPF-BPB
CPF-BCG
CPF-BTB
0.8

1.0
CPF-BPB
Absorbance

CPF-BCG 0.6
CPF-BTB
0.8
0.4
Absorbance

0.6
0.2

0.4
0.0
0 1 2 3 4
0.2
Shaking time (min)

0.0 Figure 5: Effect of shaking time on the ion

0 1 2 3 4 5 6 7 8
pair complexes
Volume of dye (mL)

3.2.5. Composition of ion-pair complexes

Figure 4: Effect of the volume of 0.025 % dyes with
10 g.mL-1 CPF Job’s method of continuous variation of equimolar
solutions was employed: a 3.0×10−4M standard
3.2.3. Effect of extracting solvent solution of ciprofloxacin and 3.0×10−4M solution of
BPB, BCG and BTB, respectively, were used. A
A number of organic solvents such as chloroform, series solutions was prepared in which the total
carbon tetrachloride, dichloromethane, benzene and volume of drug and reagent was kept at 10 mL for
toluene were examined for extraction of the ion-pair BPB, BCG and BTB, respectively. The absorbance
complex in order to provide an applicable extraction was measured at 420 nm for each dye. The molar
procedure. The most convenient solvent found to ratio of the reagents (drug:drug+dye) in the ion-pair
produce the highest absorbance, extraction power complexes was determined by the method

770
VJC, 55(6), 2017 Extractive spectrophotometric methods for…

continuous variations (Job’s method) (Fig. 6).The Harmonization (ICH) guidelines [14]. The limit of
results indicate that 1:1 (drug:dye) ion-pairs are detection (LOD) and quantification (LOQ) of the
formed through the electrostatic attraction between method are given by and respectively,
positive protonated CPF+ and negative BPB-, BCG-
relative standard deviation (RSD(%)= ;
and BTB-. The extraction equilibrium can be
represented as follows: where SD is the standard deviation of blank
CPF+(aq) + D-(aq) CPF+ D-(aq) CPF+ D-(org) absorbance values, b is the slope of the calibration
where CPF+ and D− represent the protonated curve equation, " is the average value of the
ciprofloxacin and the anion of the dye, respectively, measurement. Blank samples were prepared as
and the subcript (aq) and (org) refer to the aqueous described in section 2.4 but without CPF.
and organic phases, respectively. Under the described experimental conditions,
calibration curves for proposed methods were
1.0
CPF-BPB
constructed (Fig. 8). The linear regression equations,
CPF-BCG standard deviation, slopes and intercepts, correlation
CPF-BTB
0.8
coefficients, relative standard deviation of response
factors, and linearity ranges were given in (table 2)
Absorbance

0.6
for each proposed spectrophotometric method. The
0.4 molar absorptivities, Sandell’s sensitivity of each
methods was calculated and these values showed
0.2
that the molar absorptivity of BTB > BCG > BPB
0.0
ion-pair complexes.
0.00 0.25 0.50 0.75 1.00
[CPF]/[CPF+dye]

Figure 6: Job’s method of continuous variation

graph for the reaction of ciprofloxacin with acid-
dyes BPB BCG and BTB, [drug]=[dye] = 3×10-4 M

3.2.6. Spectral characteristics

The absorption spectra of the ion-pair complexes

extracted into chloroform are shown in Fig. 7. The
ion-pair complexes with BTB, BPB and BCG
absorbed maximally at 420 nm. The reagent blank
under similar conditions showed no absorption.

1.0
BPB
BCG
BTB
0.8
CPF-BPB
CPF-BCG
Figure 8: Standard curves of CPF ion pairs with
CPF-BTB BTB, BCG and BPB at max 420 nm
Absorbance

0.6

0.4 The accuracy and precision of the methods were

determined by preparing solutions of three different
0.2 concentrations of ciprofloxacin and analyzing them
in six replicates. Samples for analysis were prepared
0.0 as described in section 2.4. The precision of the
360 400 440 480 520
proposed methods was evaluated as percentage
Wavelength(nm)
relative standard deviation (RSD%) and accuracy as
Figure 7: Absorption spectra of ciprofloxacin percentage relative error (RE%). The percentage
(10µg.mL-1)-dye complex extracted into 10 mL relative error calculated using the following
chloroform equation: RE(%) =[(founded–added)/ added]x 100
The accuracy and precision results are shown in
3.3. Validation of the proposed method table 3.
The low values of the relative standard deviation
The proposed methods are successfully validated percentage and relative error percentage specify the
according to International Conference on high precision and the good accuracy of the method.

771
VJC, 55(6), 2017 Nguyen Trung Dung et al.

Table 2: Analytical characteristics of the proposed methods (n = 6)

Proposed methods
Parameters
BPB BCG BTB
Colour Yellow Yellow Yellow
Wavelengths λmax (nm) 420 420 420
pH 3.3 3.5 3.4
Stability (h) 24 24 24
Shaking time (min) 2 2 2
Stoichiometric ratio 1:1 1:1 1:1
Beer’s law range (µg.mL-1) 0.5-25 0.5-25 0.5-25
Limit of detection (µg.mL-1) 0.105 0.101 0.084
Limit of quantitation (µg.mL-1) 0.315 0.303 0.252
Molar absorbitivity (L.mol-1.cm-1) 1.46x104 1.83x104 2.07x104
Sandell’s sensitivity (µg/cm2) 0.0686 0.0562 0.0462
Regression equation (Y = bx +a)
Slope (b) 0.0441 0.0552 0.0625
Intercept (a) 0.0053 0.0287 0.0762
Correlation coefficient (R2) 0.996 0.998 0.997

Table 3: Evaluation of accuracy and precision of the proposed methods (n = 6)

Amount taken Amount found Recovery
Method RSD (%) RE (%)
( g.mL-1) ( g.mL-1) (%)
5.00 5.08 101.6 1.16 1.60
BCG 10.00 9.97 99.70 0.48 -0.30
15.00 14.84 98.93 0.75 -1.07
5.00 5.06 101.2 0.97 1.20
BPB 10.00 10.08 100.8 0.54 0.80
15.00 14.94 99.60 0.86 -0.4
5.00 4.94 98.80 1.02 -1.2
BTB 10.00 9.96 99.60 0.57 -0.4
15.00 15.18 101.2 0.94 1.2

3.4. Effects of interference in its dosage forms.

The extent of interference by various excipients Table 4: Effect of foreign species on the
(magnesium stearate, glucose, lactose, starch and determination of 10 µg.mL-1 ciprofloxacin
sodium chloride) which often accompany the
Tolerance limit
pharmaceutical preparations was studied in a total Excipients
(μg. mL-1)
volume of 10 mL chloroform. The interference was
Magnesium stearate 500
determined by measuring the absorbance of a
solution containing 10 µg/mL of ciprofloxacin. This Glucose 250
study was carried out by following the proposed Lactose 500
procedures for a 10 mL sample system, by adding a Sodium chloride 500
known amount of foreign species to a ciprofloxacin Starch 250
solution of 10 µg/mL. The tolerance limits of
interfering species were established at those 3.5. Comparison with other spectrophotometric
concentrations that do not cause more than ±2.0% methods
error. The tolerance limits of excipients are listed in
table 4. The results indicated that there is no The proposed method compares favorably with other
interference from the degradation, indicating a high reported methods. As shown in table 5 the proposed
selectivity for determining the studied ciprofloxacin method is more high sensitivity than other methods,

772
VJC, 55(6), 2017 Extractive spectrophotometric methods for…

needs no heating, the product is stable for a longer common excipients.

time and and are free from interference with

Table 5: Comparison of VIS spectrophotometric methods for ciprofloxacin determination

Range of Molar
max\
No. Reagent determination absorbitivity Remarks Ref.
(nm)
( g. mL−1) (L.mol-1.cm-1)
Involves contact
1 Ce(IV)- MBTH 630 10-50 - [5]
time
Fe(III)-1,10- Involves contact
3 510 0.04-7.2 3.4 104
phenanthroline time and heating [6]
Involves contact
4 Fe(III)-Bipyridil 522 0.05-9 2.95 104
time and heating
Involves contact
5 CL 520 16-96 -
time and heating
[7]
Involves contact
6 TCNE 335 0.25-15 -
time and heating
Co(II)
7 623 20-240 8.38 102 Less sensitive [8]
tetrathiocyanate
Involves heating
8 Sudan III 566 0.4-10.4 2.38 104 [9]
/cooling samples
9 Eosin Y 547 2-8 3.56 104
[10]
Less stable colour
4
10 Merbromin 545 2-15 1.23 10
11 BPB 420 0.5-25.0 1.46×104 Short reaction
time, high This
12 BCG 420 0.5-25.0 1.83×104 sensitive and high work
colour stability
13 BTB 420 0.5-25.0 2.07×104
MBTH: 3-methyl-2-benzothioazolin-2-one-hydrazone; CL: p-chloranil; TCNE: Tetracyanoethylene; BPB:
Bromophenol blue, BCG: Bromocresol green and BTB: Bromothymol blue

3.6. Application of the proposed methods formulations. The relative standard deviation values
are below 2 % indicating the precision of the
The proposed method was successfully applied to method. The validations of the proposed methods
determine ciprofloxacin in different pharmaceutical were further confirmed by recovery studies. The %
preparations (tablets, capsules and eye drops). The recovery vary from 97.41 to 101.20 indicating high
results given in table 6 of the analysis showed that accuracy of methods.
the data are consistent with the label claim of the

Table 6: Results of ciprofloxacin determination in pharmaceutical preparations

Pharmaceutical Labeled amount Recovery (%) RSD (%)

preparation (mg/form) BCG BPB BTB BCG BPB BTB
Hasancip tablet 500 mg /tablet 99.75 99.36 98.89 0.59 0.28 0.16
Kacipro tablet 500 mg /tablet 98.36 98.57 101.20 0.74 0.45 0.24
Shandong
200 mg/100 mL 102.53 100.54 97.41 0.43 0.32 0.26
infusion
Hebei infusion 200 mg/100 mL 98.05 101.08 97.69 0.87 0.65 0.48
Eye drops 30mg/tube 100.24 99.02 98.53 0.78 0.51 0.39

773
VJC, 55(6), 2017 Nguyen Trung Dung et al.

4. CONCLUSION determination of amoxicillin, ciprofloxacin and

piroxicam in pure and pharmaceutical formulations,
This article reports the use of BCG, BPB and BTB J. Pharm. Biomed. Anal, 29(5), 859-864 (2002).
as an anionic dyes for the extractive 7. Mostafa S, El-Sadek M, Alla E. A.
spectrophotometric determination of ciprofloxacin. Spectrophotometric determination of ciprofloxacin,
enrofloxacin and pefloxacin through charge transfer
The performance order of the proposed methods is complex formation, J. Pharm. Biomed. Anal, 27(1-2),
BTB > BCG > BPB. No interference from common 133-142 (2002).
excipients was encountered. The proposed methods 8. Amina Mohamed El-Brashy, M. E.-S. M., and Fawzi
are found to be simple, sensitive, selective, accurate, Abdallah El-Sepai. Spectrophotometric
precise, economical and can be used in the Determination of Some Fluoroquinolone
determination of ciprofloxacin in different Antibacterials through Charge-transfer and Ion-pair
pharmaceutical preparations (tablets, infucsions and Complexation Reactions. Bull. Korean Chem. Soc.,
eye drops). 25 (3), 365-372 (2004).
9. Alaa S. Amin. Quantitation of Some Recently
Introduced Antibacterial Drugs Using Sudan III as
REFERENCES Chromogenic Reagent, Microchimica Acta, 134(1),
89-94 (2000).
1. Sharma PC, Jain A, Jain S, Pahwa R, Yar MS. 10. El-Brashy A. M, Metwally M. E, El-Sepai F.A.
Ciprofloxacin: review on developments in synthetic, Spectrophotometric determination of some
analytical, and medicinal aspects, J. Enzyme Inhib fluoroquinolone antibacterials by binary complex
Med Chem, 25(4), 577-89 (2010). formation with xanthene dyes, II Farmaco, 59(10),
2. Janis Vella, Francesca Busuttil et al. A simple HPLC– 809-817(2004).
UV method for the determination of ciprofloxacin in
human plasma, Journal of Chromatography B, 11. Tadesse Haile Fereja, Muluneh Fromsa, Tizita Yirga
989(1), 80-85 (2015). Mola. UV-Visible Spectrophotometric Method
3. 3.O. Szerkus, J. Jacyna, A. Gibas, M. Sieczkowski, Development and Quantification of Ciprofloxaciline
D. Siluka, M. Matuszewski, R. Kaliszan, M.J. in Tablets Dosage Form, American Journal of
Markuszewski. Robust HPLC–MS/MS method for Pharmacy and Pharmacology, 2(1), 1-8(2015).
levofloxacin and ciprofloxacin determination in 12. Abdel-Ghani NT, Rizk MS, Mostafa M. Extractive
human prostate tissue, Journal of Pharmaceutical and determination of ephedrine hydrochloride and
Biomedical Analysis, 132(5), 173-183 (2017). bromhexine hydrochloride in pure solutions,
4. Kuldeep Kaur, Ashwini Kumar, Ashok Kumar Malik, pharmaceutical dosage form and urine samples,
Baldev Singh, and A. L. J. Rao. Spectrophotometric Spectrochimica Acta Part A: Molecular and
Methods for the Determination of Fluoroquinolones: Biomolecular Spectroscopy, 111, 131-41 (2013).
A Review, Critical Reviews in Analytical Chemistry, 13. Ali E. A, Adawy A. M., El-Shahat M. F., Amin A. S.
38, 2-18 (2008). Simple spectrophotometric methods for
5. M. Rizk, F. Belal, F. Ibrahim, S. M. Ahmed, and N. determination of fluoxetine and clomipramine
M. El-Enany. A simple kinetic spectrophotometric hydrochlorides in dosage forms and in some post-
method for the determination of certain 4-quinolones mortem biological fluids samples, Egyptian Journal
in drug formulations, Scientia Pharmaceutica, 68(1), of Forensic Sciences, 6(4), 370-380 (2016).
173-188 (2000). 14. ICH Topic Q2 (R1). Validation of analytical
6. Nagaralli, B.S, Seetharamappa, J, Melwanki, M. B. procedures: text and methodology
Sensitive spectrophotometric methods for the (CPMP/ICH/281/95); accessed June 30 (2010).

Corresponding author: Nguyen Trung Dung

Faculty of Physics an chemical Engineering, Le Quy Don Technical University
236, Hoang Quoc Viet road, Cau Giay district, Hanoi, Viet Nam
E-mail: nguyentrungdung1980@gmail.com.

774