Estimation
of Ranitidine in Bulk and Formulation by First Order Derivative Area under
Curve UV-Spectrophotometry Methods
Audumbar
Digambar Mali *, Patil Manojkumar
Department of Pharmaceutics, Sahyadri College of Pharmacy, Methwade,
Sangola-413307, Solapur, Maharashtra, India
*Corresponding Author E-mail: maliaudu442@gmail.com
ABSTRACT:
Simple, fast and reliable spectrophotometric methods were
developed for determination of Ranitidine in bulk and pharmaceutical dosage
forms. The solutions of standard and the sample were prepared in Methanol. The
quantitative determination of the drug was carried out using the first order
Derivative Area under Curve method values measured at 329-335nm. Calibration
graphs constructed at their wavelengths of determination were linear in the
concentration range of Ranitidine using 2-10μg/ml (r²=0.999) for first order Derivative Area under
Curve spectrophotometric method. The proposed methods have been extensively
validated as per ICH guidelines. There was no significant difference between
the performance of the proposed methods regarding the mean values and standard
deviations. The developed methods were successfully applied to estimate the
amount of Ranitidine in pharmaceutical formulations.
KEYWORDS: Ranitidine, First order Derivative, Area
under Curve (AUC).
1. INTRODUCTION:
Ranitidine is
chemically Dimethyl[(5-{[(2-{[(E)-1-(methylamino)-2 nitroethenyl]amino} ethyl) sulfanyl]
methyl}furan-2-yl) methyl]amine. The H2 antagonists are competitive inhibitors
of histamine at the parietal cell H2 receptor. They suppress the normal secretion
of acid by parietal cells and the meal-stimulated secretion of acid.1,2 They accomplish this by
two mechanisms: histamine released by cells in the stomach is blocked from
binding on parietal cell H2 receptors which stimulate acid secretion and other
substances that promote acid secretion (such as gastrin
and acetylcholine) have a reduced effect on parietal cells when the H2
receptors are blocked.3-5
In our Literature survey reveals that for Ranitidine Spectrophotometric6 methods and HPLC7 methods have been reported
for its determination in commercial formulation.
To our notice, no UV- spectrophotometric method using First Order
Derivative Area under Curve has been reported for the determination of
Ranitidine in bulk and tablets. Hence an attempt has been made to develop new
First Order Derivative Area under Curve spectrophotometric method for
estimation of Ranitidine in bulk and pharmaceutical formulations with good
accuracy simplicity, precision and economy.
Fig.
1 Structure of Ranitidine
2. MATERIALS AND
METHODS:
2.1 Derivative
Spectrophotometric Methods:-
The first derivative spectrophotometry
was used in the wavelength ranges from 329 and 340 nm.
[dA/dλ=
f(λ ): first order
The first derivative spectrum of an absorption
band is characterized by a maximum, a minimum, and a cross-over point at the
λ max of the absorption band.
2.2 Area under curve (Area calculation):-
In this study area was integrated between
wavelength ranges from 329 and340 nm.
Area calculation:
(α+β) =
Where, α is area of portion bounded by curve data and a
straight line connecting the start and end
point, β is the area
of portion bounded
by a straight
line connecting the
start and end point on curve data and horizontal axis,
λ1 and λ2 are wavelength range
start and end point of curve region. 8
2.3 Apparatus and instrumentation:
A Shimadzu 1800 UV/VIS double beam spectrophotometer with 1cm
matched quartz cells was used for all spectral measurements. Single Pan
Electronic balance (CONTECH, CA 223, India) was used for weighing purpose.
Sonication of the solutions was carried out using an Ultrasonic Cleaning Bath
(Spectra lab UCB 40, India). Calibrated volumetric glassware (Borosil®) was used for the validation study.
2.4 Materials:
Reference standard of Ranitidine API was supplied as gift sample
by Cipla Pharmaceutical company,
Pune. Methanol was obtained from Research-Lab Fine Chem Industries, Islampur,
Mumbai, and Maharashtra. Tablet sample
with label claim 125 mg per Tablet were purchased from local market Mangalwedha, Solapur,
Maharashtra, India.
2.5 Method development:
2.5.1 Preparation of
Standard and Sample Solutions:-
Stock solution of 10μg/ml of Ranitidine was prepared in
Methanol, for First Order Derivative Area under Curve spectrophotometric
analysis. The standard solutions were prepared by dilution of the stock
solution with Methanol in a concentration range of 2, 4, 6, 8 and 10μg/ml with Methanol for First Order Derivative Area
under Curve spectrophotometric methods. Methanol was used as a blank solution.
Fig. 2 First order
derivative Area under Curve spectrum of Ranitidine in Methanol (10µg/ml).
Fig. 3 First order
derivative spectrum of Ranitidine in Methanol (10µg/ml).
Fig. 4 Linearity of Ranitidine.
2.5.2 Calibration curve for Ranitidine:
The dilutions were made from Standard Stock solution to get
concentration of 2, 4, 6, 8, and 10µg/ml respectively. These solutions were
scanned from 400 to 200 nm and First Order Derivative Area under Curve values
was integrated in the range of 329-340 nm. The calibration curve was plotted
between areas under curve values against concentration.
2.5.3 Assay of tablet
formulation:-
Twenty tablets each containing 125 mg of Ranitidine were weighed
crushed to powder and average weight was calculated. Powder equivalent to 10 mg
of Ranitidine was transferred in 100 ml of volumetric flask. A 50 ml of
Methanol was added and sonicated for 15 minutes. Then
solution was further diluted up to the mark with Methanol. The solution was
filtered using Whatmann filter paper no. 41, first 5 ml of filtrate was discarded. This solution was
further diluted to obtain 10µg/mL solution with
water, subjected for UV analysis using Methanol as blank. This procedure was
repeated three times.
Fig. 5 First order
derivative Area under Curve spectrum of Ranitidine of dosage form in Methanol
(10µg/ml).
Fig. 6 First order derivative spectrum of
Ranitidine of dosage form in Methanol (10 µg/ml).
Fig. 7 First order
derivative overlay of Ranitidine
at diff. Concentration.
Table 1: Assay of tablet
dosage form:-
|
Sr.No. |
Sample Solution Concentration (µg/ml) |
Amount found (%) |
Mean % found* |
%RSD* |
|
1 |
10 |
98.16 |
|
|
|
2 |
10 |
102.80 |
100.29 |
0.0237 |
|
3 |
10 |
99.91 |
|
|
*n=3, % RSD = % Relative Standard Deviation.
3. METHOD VALIDATION: 9
The above method
was validated for
various parameters such as
Accuracy, Linearity, Precision,
Limit of detection
(LOD) and Limit of Quantitation (LOQ) according to ICH guideline.
3.1 Accuracy:
The accuracy for
the analytical method
was evaluated at
80%, 100% and
120% levels of 10 µg/ml Sample solution. First Order
Derivative Area under curve (AUC) was measured in wavelength range 329-340 nm
and results were obtained in terms of percent recovery. Three determinations at
each level were performed and % RSD was calculated for each level.
Table 2: Accuracy results for Ranitidine :
|
Accuracy level |
Sample Conc. (µg/ml) |
Std. Conc. |
Total amount. Added (µg/ml) |
% Recovery |
Mean % Recovery |
% RSD |
|
80 |
10 |
12 |
8 |
99.39 |
|
|
|
100 |
10 |
15 |
10 |
102.78 |
100.96 |
0.0473 |
|
120 |
10 |
18 |
12 |
100.71 |
|
|
3.2 Precision:
The precision of an analytical procedure expresses the closeness
of an agreement (degree of scatter) between a series of measurements obtained
from multiple sampling of the same homogeneous sample under the prescribed
conditions intraday precision was studied by integrating area of standard
solution of 10 µg/ml concentration at six independent series in the same day.
Interday precision studies were performed by integrating area of standard
solution of 10µg/ml concentration on three consequent days. The % RSD Was
calculated.
Table 3: Precision Study:-
|
Parameter |
Intra day |
Inter-day |
|
Sample sol conc.
µg/ml |
10 |
10 |
|
AUC (mean) |
0.0057 |
0.0062 |
|
%RSD |
0.7752 |
0.8116 |
3.3 Limit of Detection and
Limit of Quantification:
The Limit of Detection (LOD) is the smallest concentration of the analyte that gives the measurable response. LOD was
calculated using the following formula
LOD = 3.3 σ /S
The Limit of Quantification (LOQ) is the smallest concentration of
the analyte, which gives response that can be
accurately quantified. LOQ was calculated using the following formula
LOQ = 10 σ/S
Where, σ is standard deviation of the response and
S is the slope
of the calibration curve.
LOD and LOQ of Ranitidine was found to be
0.62 µg/ml and1.67µg/ml respectively.
Table 4: Summary of
validation parameters:-
|
Parameter |
Result |
|
λ range |
329-340 |
|
Regression
Equation (y=mx+c) |
Y=0.059x + 0.01 |
|
Linearity range |
2-10µg/ml |
|
Slope |
0.059 |
|
Intercept |
0.01 |
|
Correlation
coefficient (R2) |
0.999 |
|
Limit of
Detection (LOD) µg/ml |
0.62 |
|
Limit of Quantitation (LOQ) µg/ml |
1.67 |
|
Accuracy (Mean %
Recovery) |
100.96 |
|
Precision (%RSD) |
0.0473 |
4.
RESULTS AND DISCUSSION:
The UV visible spectroscopic method for the Ranitidine by First
order derivative Area under Curve was found to be simple, accurate, economical
and reproducible. The drug concentrations were
found to be linear
in the range
of 2-10 µg/ml and the correlation
coefficient value of 0.999 indicates that
developed method was
linear. For Precision
the percent relative
standard deviation (% RSD) was found to be 0.0473 while, intra-day and
inter-day precision results in
terms of percent relative standard deviation
values were found to be 0.7752 and 0.8116 respectively thus the method
is observed as precise. The accuracy of the method was assessed by recovery
studies at three different levels i.e. 80%, 100%, 120%. The values of standard
deviation were satisfactory and the recovery studies were close to 100%. The %
RSD value is ≤ 2 indicates the accuracy of the method. The
Limit of Detection
and Limit of Quantitation
values were found to
be 0.62µg/ml and 1.67µg/ml
respectively. The result
of the analysis
for pharmaceutical
formulation by the
developed method was
consistent with the
label claim, highly reproducible
and reliable. The method can be
used for routine
quality control analysis
of Ranitidine in
bulk and pharmaceutical
formulations.
5.
CONCLUSION:
The UV spectroscopic AUC method for the analysis of Ranitidine by
First order derivative Area under Curve was found to be simple, precise, and
accurate; can be used for assay of bulk drug and pharmaceutical dosage
formulations.
6. ACKNOWLEDGEMENT:
The authors are highly thankful to the Sahyadri
College of Pharmacy, Methwade, Sangola,
Solapur, Maharashtra, India
for proving all the facilities to carry out the research work.
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Received on 05.06.2015 Accepted on 25.06.2015
© Asian Pharma
Press All Right Reserved
Asian
J. Pharm. Ana. 5(2): April-June 2015; Page 61-66
DOI: 10.5958/2231-5675.2015.00010.1