International Journal of ChemTech Research
CODEN (USA): IJCRGG, ISSN: 0974-4290, ISSN(Online):2455-9555
Vol.10 No.3, pp 666-670, 2017
Spectrophotometric determination of Furosimide in
pharmaceutical formulations by charge transfer complex
method
G. Dill Rani1A. Russia rani2and P. Venkateswarlu*
Dept of Chemistry, S.V.University, Tirupathi-517502, A.P, India
Abstract : The simple and sensitive spectrophotometric method for the determination of
furosimide reacts with 1ml of DDQ (2, 3 –dichloro -5, 6-dicylano-1, 4-benzoquinone) by
charge –transfer complex method. In this method the drug furosimide as n-electron donors
with acceptor 2, 3 dichloro-5, 6- dicyano 1,4- benzoquinone (DDQ) to form reddish pink color
charge-transfer complexes. This reaction is instantaneous and quantitative. The drug
maximum absorbance at 450 nm and Beer’s law limit was obeyed at 20-160 µg/ml.The optical
characteristics of the proposed method such as molar absorptivity, sandell’s sensitivity, slope
and intercept were 2.0847 L.mole-1cm-1 , 0.00208 µg.cm-2,0.0059 and 0.0061 for furosimide
respectively.The developed method was found to be simple, specific, robust, accurate and
precise for the determination of furosimide.
Key words : furosimide, chloroform, methanol, DDQ and UV-Spectrophotometric method.
1. Introduction
Furosimide, chemically known as 5-(aminosulfonyl-4-chloro-2-[(2-furanylmethyl) amino] benzoic acid,
is structurally a sulfonamide, an antibacterial agent. Furosimide is an anthracitic acid derivative extensively
used for its diuretic effect in the treatment of edema associated with pulmonary, cardiac, hepatic and renal
disease1, and of hypertension accompanied by fluid retention or impaired renal failure2. Because of its
predominant action on the loop of Hanley and the marked dieresis it can produce, this compound is often
designated as a loop diuretic and high ceiling diuretic. The adverse effects of furosimide are hypertension,
congestive heart failure3, 4, and cirrhosis of the liver.
Numerous methods have been reported for the determination of furosimide in pharmaceutical samples.
Most of the methods developed are based on different spectrophotometric methods 5-8,HPLC9,, rapid titrimetric
and spectrophotometric10,spectrophotometric – partial least squares (PLS-1)11, diffuse reflectance
spectroscopy12, developments in analytical method13, first digital derivative spectrophotometry14, ratio spectra
derivative spectrophotometry15. Second order derivative spectroscopy and area under curve (AUC) 16. These
methods are available for the determination of furosimide.
2. Experimental
2.1 Instrumentation
A Shimadzu UV-visible double beam spectrophotometer (model 2450) with 1 cm matched quartz cells
was used for the spectral measurements.
G. Dill Rani et al /International Journal of ChemTech Research, 2017,10(3): 666-670. 667
2.2 Chemicals and reagents
All the chemicals used were of analytical grade. Double distilled water was used for all the
experimental studies.
2.3 DDQ solution(1% w/v)
DDQ (2,3-dichloro5,6-dicyano-p-benzoqunone) (Loba Chem.,India) solution is prepared by dissolving
100 mg in 100 ml of distilled water.
2.4 Furosemide solution
An accurately weighed 50 mg of furosemide is dissolved in methanol and the volume was adjusted to
50 ml with methanol. Further dilution is made to obtain the working concentration of 100 g /ml.
2.5 Spectrum of furosimide treated with DDQ
1.0ml of furosimide standard solutions was taken into a standard flask. To this solution 1ml of DDQ
reagent is added to form a pale reddish pink colored solution. The final volume was brought to 10ml with
methanol. The solution was taken in 125 ml separating funnel and extracted with 5ml chloroform twice by
shaking for two minutes and allowed to stand for clear separation of two phases. The resultant solution is well
mixed and allowed to stand for 5 minutes for completion of the reaction. The absorbance of the reddish pink
colored solution is measured in the wavelength range of 400 to 700 nm against the reagent blank. The spectrum
is given in fig.1.from figure the drug treated with DDQ solution has maximum absorbance at 450 nm. Hence,
all further studies are made at 450 nm.
2.6 Assay procedures
Sample solutions ranging from 0.2-1.6 ml were transferred into a series of standard flaks and solution
of DDQ (1 ml) is added to produce a reddish pink color. The final volume is brought to 10 ml with methanol.
The tubes were taken thoroughly and placed in a boiling water bath for about 30 minutes. The reaction mixture
in each tube was called, transferred quantitatively into a 125ml calibrated flask and diluted to the mark with
distilled water. The absorbance of the reddish pink color solution was measured at 450 nm against the reagent
blank prepared in similar manner omitting drug solution. Calibration graph is obtained by plotting absorbance
values against the concentration of furosimide solution. The calibration curve is found to be linear over a
concentration range of 20 to 160g/ml of furosimide. The amount of furosimide drug present in the sample is
readfrom the calibration graph. The results are presented in fig 2.
2. 7 Effect of interferences
In order to apply the proposed method to the analysis of pharmaceutical formulations, the influence of
commonly used excipients starch, lactose, glucose, Dextrose sugar, and talc and additives was studied by
preparing solutions containing 2.0x10-3M furosimide and increasing concentrations of the potential interference
up to 1.0x10-3M.The results are shown in table .3
2.8 Assay in serum and urine samples
To apply the proposed method for biological samples, blood and urine samples were collected from
donors, and centrifuged at 3000 rpm for nearly 10 min. The resulted solutions were filtered and preserved in the
absence of light at a temperature of 4oC. From these solutions, various concentrations of the drug furosimide
were analyzed with the help of proposed analytical method and these results were recorded in table 4. Hence,
the proposed method can be successfully applied to recover furosimide in biological samples, viz. urine and
serum due to its high accuracy and good recoveries.
3.Results and discussion
The UV spectrum of furosimide is presented in fig 1. The absorption maximum was observed at 450nm
for furosimide abeyance to Beer’s law was confirmed by the linear of the calibration curve of furosimide, which
G. Dill Rani et al /International Journal of ChemTech Research, 2017,10(3): 666-670. 668
are presented in fig 2.The molar absorptivity and sandell’s sensitivity values show that method is sensitivity.
The regression analysis using method of least squares was made for the slope (b) intercept (a) and correlation
(r) obtained from different concentrations and results are summarized in the table 1. The value of correlation
coefficient (r) was 0.999 which indicated the good linearity of calibration lines.
Fig.1 Spectrum of furosimide react with DDQ
Fig.2 Calibration Curve of Furosimide
Table .1 Optical characteristics of the proposed methods
Parameters Proposed method
max(nm) 450
Beer’s law limit(g/ml) 20-160
Molar absorptivity (L.mole-1 cm-1) 2.0847
Sandal’s sensitivity (g.cm-2/0.001 A.U) 0.00208
Slope(b) 0.0059
Intercept(a) 0.00619
Correlation coefficient(r2) 0.9997
Relative standered deviation(RSD)% 0.20833
LOD (µg/ml) 0.50480
LOQ (µg/ml) 1.68101
Color Reddish pink
G. Dill Rani et al /International Journal of ChemTech Research, 2017,10(3): 666-670. 669
Table .2 Assay of furosimide in tablet formulations
Labelled Amount
Tablets amount found %Recovery SD % RSD
mg/ml mg/ml
Frusemene 250 249.53 99.81 0.3511 0.1407
Frusenes 250 249.53 99.81 0.3511 0.1407
Diucontin 250 249.43 99.77 0.8082 0.3240
*Average of five determinations
Table .3 Determination offurosimide in presence of excipients
Amount
*Found
Excipients taken Recovery % ±SD RSD %
mg/ml
mg/ml
Glucose 5 4.98 99.73 0.0152 0.3060
Sucrose 10 9.98 99.86 0.0152 0.1520
Lactose 15 14.96 99.77 0.0115 0.0770
Dextrose 20 19.97 99.88 0.0020 0.0458
Talc 30 29.96 99.87 0.0152 0.0501
Starch 20 19.97 99.89 0.0036 0.1018
Table .4 Method accuracy from recovery assay
Added *Found Recovery
Sample ±SD RSD%
mg/ml mg/ml %
0.5 0.49 99.66 0.001 0.2317
Serum 0.7 0.69 99.71 0.001 0.1432
samples 0.9 0.89 99.66 0.002 0.2229
1.1 1.09 99.75 0.002 0.1897
2 1.98 99.26 0.006 0.3238
Urine
2.2 2.19 99.81 0.001 0.0788
samples
2.4 2.39 99.88 0.002 0.0868
2.6 2.59 99.88 0.002 0.1018
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