Research J. Pharm. and Tech.

11(1): January 2018

ISSN 0974-3618 (Print) www.rjptonline.org

0974-360X (Online)

RESEARCH ARTICLE

Quantification of Itopride Hydrochloride from tablet dosage form by

visible Spectroscopy
Salunke Poonam A.*, Kadam Lalit, Jaiswal Nikita, Jain Roshan, Patil Shital, Wagh Rupali,
Dr. Barhate S. D.
Shree Sureshdada Jain Institute of Pharmaceutical Education and Research, Jamner, Dist-Jalgaon.
*Corresponding Author E-mail: [email protected]

ABSTRACT:
The present study to describe the Quantification of Itopride Hydrochloride from tablet dosage form by visible
spectroscopy by applying principle of colorimetric study. It involves the measurement of the amount of
ultraviolet (190-380 nm) or visible (380-800nm) radiation absorbed by a substance in solution. The final
optimised method carried out on double beam UV-visible spectrophotometer (Shimadzu, model 1800) having
two matched quartz cells with 1 cm light path by using Bromothymol blue of concentration 0.5% w/v solution.
Itopride Hydrochloride detection wavelength was set at 479.40nm The linearity was found to be in the range of
10-50μg/ml. This method for the quantitative determination of the drug by validation in accordance with the
International Conference on Harmonization. The proposed method is economic, simple, sensitive, reproducible,
and accurate and no interference is observed due to common excipients of tablet. The proposed method was
successfully applied to determination of Itopride hydrochloride in pharmaceutical formulations.

KEYWORDS: Itopride Hydrochloride, UV-visible spectrophotometer, analytical method validation, acid-dye

method.

INTRODUCTION: Beer-Lambert law:2

The technique of ultraviolet-visible spectrophotometry is When a beam of light is passed through a transparent cell
one of the most frequently employed in pharmaceutical containing a solution of an absorbing substance,
analysis. It involves the measurement of the amount of reduction of the intensity of light may be occur.
ultraviolet (190-380 nm) or visible (380-800nm) A = log I0 /IT = abc
radiation absorbed by a substance in solution.
Instruments which measure the ratio or a function of the Chemical derivation:
ratio, of the intensity of two beams of light in the Indirect spectrophotometric assay are based on the
ultraviolet-visible region are called ultraviolet-visible conversion of the analyte by a chemical used, to ensure
spectrophotometers. Absorption of light in both the complete conversion, the absorbance of the derivative is
ultraviolet and visible regions of the electromagnetic usually, but not always, proportional to the concentration
spectrum occurs when the energy of light matches that of the analyte. The majority of indirect
required to induce in the molecules and electronic spectrophotometric procedures involve the conversion of
transition and its association vibrational and rotational the analyte to a derivative that has a longer. max
transitions (p.260). It is thus convenient to consider the and/or a higher absorptivity. Chemical derivatisation
technique of the ultraviolet spectrophotometry and procedures may be adopted any of several reason:
visible spectrophotometry together.1
a) If the analytical absorbs weakly in he ultraviolet
region, a more sensitive method of assay is obtained by
converting the substance to a derivative with a more
Received on 15.07.2017 Modified on 17.08.2017
intensely absorbing chromophore.
Accepted on 12.09.2017 © RJPT All right reserved
Research J. Pharm. and Tech. 2018; 11(1): 121-125
DOI: 10.5958/0974-360X.2018.00023.9

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 Research J. Pharm. and Tech. 11(1): January 2018

b) The interference from irrelevant absorption may be hydrochloride is chemically Hydrochloride salt of N-{[4-
avoided by converting the analyte to a derivative which (2- dimethylamino ethoxy) phenyl] methyl}-3, 4-
absorbs in the visible region, where irrelevant absorption
dimethoxy-benzamide is an anti-emetic drug and used in
is negligible the management of gastrointestinal symptoms like
nausea, vomiting, non ulcer dyspepsia, emesis and
c) Indirect spectrophotometric procedures are also used chronic gastritis5, 6, 7.
to improve the selectivity of an ultraviolet-absorbing
substance in a sample that contains other ultraviolet-
absorbing components.

d) The adsorption of a visible spectrophotometric

procedure, instead of an ultraviolet procedure, may be Figure 1: Chemical structure of Itopride hydrochloride
based on cost considerations. In general, single-beam
manually adjusted visible spectrophotometers are much Need of work is that Literature survey reveals
cheaper than ultraviolet-visible spectrophotometers.3 spectrophotometric8, HPLC9,10 and HPTLC11 methods
several method of chemical derivation like as for the estimation of Itopride hydrochloride from
Diazotisation and coupling of primary aromatic amine, pharmaceutical formulation has been developed. One
Condensation reaction, Reduction of tetrazolium salt, colorimetric method was developed by using methyl red
The acid-dye method, Oxidation method, Metal-ligand but yet not developed colorimetric estimation of drug
complexation. based on the formation of coloured complex of drug with
Bromothymol blue. In proposed research work a
Acid-dye method4: colorimetric method has been developed and validate for
The addition of an amine in its ionised form to an estimation of Itopride hydrochloride from its tablet
ionised acidic dye, e.g. methyl orange, bromocresol formulations by colorimetric method.
purpul or bromocresol green, yields a salt (ion-pair) that
may be extracted in to an organic solvents such as MATERIAL AND EXPERIMENTAL WORK:
chloroform or dichloroformethane. The indicator dye is Material and Instrument:
added in excess and the pH of the aqueous solution is Bromothymol blue of concentration 0.5% w/v was
adjusted if necessary to a value where both the amine prepared in distilled water. Standard itopride
and dye are in the ionised forms. The ion-pair is hydrochloride drug and commercially available tablets of
separated from the excess indicator by extraction into the itopride hydrochloride were procured from local market.
organic solvent, and the absorbance is measured at the UV spectrophotometric method was performed on
max of the indicator in the solvents. Usually, the most double beam UV-visible spectrophotometer (Shimadzu,
intensely absorbing form of the indicator is measured, model 1800) having two matched quartz cells with 1 cm
with the addition, if required, of acidified or basified light path.
ethanol. Alternatively, the absorbance of the indicator
may be measured in aqueous solution after back
extraction from the organic solvent. The molar
absorptivities of ion-pairs formed between quaternary
ammonium compounds and methyl orange or
bromothymol blue are typically 2 × 104- 4 × 104. The
acid dye method therefore provides a more sensitive
technique for certain amines and quaternary ammonium
compounds that absorb weakly in the ultraviolet region,
e.g. hyoscine butylbromide (€257=202).

The correct choice of pH may permit the selective assay

of a mixture of an amine and a quaternary ammonium Figure 2. UV visible spectrophotometer
salt.
EXPERIMENTAL WORK:
The acid-dye technique is used for the assay of Preparation of standard stock solution (1000μg/ml)
formulation containing certain quaternary ammonium Accurately weighed 100 mg standard Itopride
salts or amines. hydrochloride transferred to 100 ml volumetric flask and
dissolved and diluted up to the mark with distilled water
Itopride (HCl) is a substituted benzamide, which is used to give a stock solution having strength 1000μg/ml. 100
for its prokinetic and antiemetic properties. Itopride μg/ml working standard solution was prepared by
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diluting 1 ml of stock solution to 10 ml with distilled different time points on different days and %RSD was
water with the same prepare different dilution in the calculated.
range of 10-50 μg/ml
Repeatability:
Preparation of calibration curve: Repeatability was carried out by using a minimum of 6
To 5 ml of each dilution taken in separating funnel, 5 ml determinations at one of the test concentration.
of bromothymol blue solution was added and shaken
vigorously. Then 2.5 ml of chloroform was added LOD AND LOQ:
reaction mixture was shaken vigorously and allowed to The LOD and LOQ were estimated from the set of6
stand so as to separate aqueous and chloroform layer. calibration curves used to determine method linearity.
The chloroform layer was separate out and transferred to LOD= 3.3*σ/S and LOQ= 10*σ/S
5 ml volumetric flask. Reaction mixture extracted further Where,
with 1.5 and 1 ml of fresh chloroform and added in σ = the standard deviation of y-intercepts of regression
previously extracted chloroform layer containing drug. lines
Scanned final solution wavelength was found 479.40nm. S = the slope of the calibration curve
Measure the absorbance of various dilutions at this
wavelength. Calibration curve was plotted between Ruggedness
concentration of drug and measured absorbance. The evaluation of Ruggedness should be considered
during the development phase and depends on the type
Analysis of tablet formulation: of procedure under study. It should show the reliability
Twenty tablets were accurately weighed and average of an analysis with respect to external factors in method
weight of tablet was determined. Tablets were to fine parameters such as instrument, analyst variation.
powder equivalent to 10 mg Itopride hydrochloride.
Dissolved weighed quantity of tablet powder in distilled RESULT AND DISCUSSION:
water. Sonicated for 15 min and filtered through
whatman filter paper. Filter paper was washed with
distilled water and made the volume to 100ml. From this
solution 3 ml was taken in 10 ml volumetric flask and
volume was made with distilled water to give
concentration of 30 µg/ml and followed same procedure
as per calibration curve. The absorbance of prepared
dilution was measured at 479.40 nm wavelength.
Concentration of drug in formulation was calculated
from calibration curve. Procedure repeated three times.
Figure 3: UV Spectrum of Itopride Hydrochloride
VALIDATION OF DEVELOPED METHOD12
Linearity: Wavelength final solution was found to be 479.40nm.
The linearity was determined by analyzing 6 Table 1 Linearity
independent levels of calibration curve in the range of Sr. No. Conc Absorbance
10-50μg/ml. Absorbance of each solution was recorded. 1 0 0
Absorbance vs. concentration was plotted and 2 10 0.0303
correlation co-efficient and regression line equation for 3 20 0.0608
4 30 0.0875
Itraconazole were determined. 5 40 0.1155
6 50 0.1389
Accuracy:
Accuracy was determined by performing recovery
studies within the analytical concentration range of the
proposed method at three different set at level of 80%,
100% and 120%. The amount of Itopride hydrochloride
and % Recoveries was calculated at each level.

Precision:
Inter-day precision was determined by analyzing Itopride
hydrochloride (10-50μg/ml) at three different time points
of the same day and Intra-day was determined by
analyzing Itopride hydrochloride (10-50 μg/ml) at three Figure 4 Calibration curve for Itopride Hydrochloride
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Calibration curve was plotted in the range of 10-50 30 µg/ml=0.52

µg/ml. Linearity was found R2=0.998
% RSD of Intraday precision was found to be
Table 2 Assay of tablet formulation 10 µg/ml=0.80
Formulation Labeled Amount %drug %RSD
amount(mg) recovered recoverd
20 µg/ml=1.01
Tablet 50 49.12±0.0347 99.812 0.834 30 µg/ml=0.98
% drug recovered was found to be 99.812
Table 5 LOD AND LOQ
Table 3 Accuracy Sr Conc. Abs I II III Mean SD %
80% No. RSD
Sr Conc. Amt Abs Amt Amt % rcvd 1 10 0.0303 0.0305 0.0306 0.03 0.000153 0.50
no. adde found rcvd 2 20 0.0608 0.0605 0.0627 0.06 0.001193 1.95
d 3 30 0.0875 0.087 0.0866 0.09 0.000451 0.52
1 30 24 0.0485 23.75 24.42 101.74 4 40 0.1155 0.117 0.1156 0.12 0.066399 0.72
2 30 24 0.0486 23.80 24.47 101.94 5 50 0.1389 0.1366 0.1367 0.14 0.0013 0.95
3 30 24 0.0485 23.75 24.42 101.74 Avera 0.013899
Mean 23.77 24.43 101.81 ge SD
SD 0.03 0.03 0.12
%RSD 0.12 0.12 0.12 Sr.no. Parameter Result
1 LOD 9.173
100%
2 LOQ 27.79
Sr Conc. Amt Abs Amt Amt % Recv
no. adde found recvd
Table 6 Ruggedness
d
ANALYST I ANALYST II
1 30 30 0.0657 32.35 33.02 110.06
Sr No. Conc. Abs Conc. Abs
2 30 30 0.0655 32.25 32.92 109.72
1 30 0.0863 30 0.0875
3 30 30 0.0676 33.30 33.97 113.22
2 30 0.0872 30 0.0878
Mean 32.63 33.30 111.00
3 30 0.0864 30 0.0874
SD 0.58 0.58 1.93
4 30 0.0878 30 0.0879
%RSD 1.78 1.74 1.74
5 30 0.0879 30 0.0868
120% 6 30 0.0871 30 0.0906
Sr Conc. Amt Abs Amt Amt % Mean 0.09 Mean 0.09
no. added found rcvd Rcvd %RSD 0.77 %RSD 1.51
1 30 36 0.0832 41.10 41.77 116.02
2 30 36 0.0839 41.45 42.12 116.99 % RSD for Ruggedness was found to be
3 30 36 0.0842 41.60 42.27 117.41 Analyst I=0.77
Mean 41.38 42.05 116.81 Analyst II=1.51
SD 0.26 0.26 0.71
%RSD 0.62 0.61 0.61 Table 7 Repeatability
Sr. No. Conc. Absorbance Amt Found %Amt Found
%RSD for recovery study of itopride hydrochloride was 1 20 0.0606 19.85 99.75
found to be for 2 20 0.0608 20.00 100.00
3 20 0.0609 20.65 100.83
80%=0.12
4 20 0.0608 20.00 100.00
100%=1.74 5 20 0.0608 20.00 100.00
120%=0.61 6 20 0.0608 20.00 100.00
Mean 20.78 100.58
Precision SD 0.44 1.46
Table 4 1) Interday Precision %RSD 1.30 1.30
Sr Conc. Abs II III Mean %RSD %RSD for Repeatabiliy was found to be 1.30
No.
1 10 0.0303 0.0305 0.0306 0.03 0.50 Table 8 Result summary
2 20 0.0608 0.0605 0.0627 0.06 1.95 Sr. Validation Parameter Result
3 30 0.0875 0.087 0.0866 0.09 0.52 No.
1 Solubility Chloroform and
2) Intraday Precision bromothymol
Sr Conc Abs II III Mean % RSD green solution
No. 2 UV detection Maximum wavelength 479.40 nm
1 10 0.0312 0.0315 0.0317 0.03 0.80 3 Linearity range 10-50 µg/ml
2 20 0.0618 0.0615 0.0627 0.06 1.01 4 Standard regression equation y = 0.002x + 0.002
3 30 0.0865 0.0862 0.0878 0.09 0.98 5 Coefficient of regression (R2) 0.998
6 % Recovery (n=3)%RSD
% RSD of Interday precision was found to be 80% 0.12
100% 1.74
10 µg/ml=0.50 120% 0.61
20 µg/ml=1.95 7 Precision (%RSD)

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Interday (n=3)
10µg/ml 0.50
20 µg/ml 1.95
30 µg/ml 0.52
Intraday(n=3)
10 µg/ml 0.80
20 µg/ml 1.01
30 µg/ml 0.98
8 Repeatability(n=6)%RSD 1.30
9 LOD 9.173
LOQ 27.79
10 Ruggedness (n=6)%RSD
Analyst I 0.77
Analyst II 1.51

CONCLUSION:
The proposed method was economic, simple, sensitive,
reproducible, and accurate and no interference was
observed due to common excipients of tablet. Statistical
analysis of the result has been carried out revealing high
accuracy and good precision. The proposed method was
successfully applied to the determination of Itopride
hydrochloride in pharmaceutical formulations. Thus the
proposed method can be used for the routine analysis of
Itopride hydrochloride in bulk as well as in its
pharmaceutical preparations.

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