Novel Validated Method for the Estimation of Capecitabine in Pharmaceutical Dosage forms by First Order Derivative Spectrophotometry
1Dept. of Pharmaceutical Quality Assurance, BLDEA’s SSM College of Pharmacy and Research Center, Vijayapur 586103.
2Dept. of Pharmaceutical Chemistry, BLDEA’s SSM College of Pharmacy and Research Center, Vijayapur 586103.
3Dept. of Pharmaceutics, BLDEA’s SSM College of Pharmacy and Research Center, Vijayapur 586103.
*Corresponding Author E-mail: santosh.karajgi@gmail.com
An easy, fast, precise and cost effective First derivative UV estimation method has been investigated for the estimation of Capecitabine in various pharmaceutical dosage forms. The novel method was developed using Shimadzu Spectronic 1800 model UV-Visible Spectrophotometer. The region on the UV-spectrum between 240nm to 400nm in distilled water as the solvent was found to be suitable for the first derivative peak measurement of Capecitabine with a Linearity range throughout the concentrations of 40 microgram/ml to 80 microgram/ml. The method was successfully employed for the routine estimation on tablet dosage forms. The method was validated according to ICH guidelines. No interference from additives was observed.
KEYWORDS: Capecitabine, Estimation, Analytical method, First derivative, UV method.
Capecitabine chemically is C15H22FN3O6 used for the treatment of Breast Cancer, Gastric Cancer and Colo- rectal Cancer. Inside the body, Capecitabine is converted into 5-fluorouracil, which further inhibits the synthesis of thymidine monophosphate, the active form of thymidine which is required for the de novo synthesis of DNA.1
Various authors have worked on UV-Visible spectrophotometric methods.2-12 As per the literature review; it is evident that only few methods are reported for the quantitative analysis of Capecitabine in pharmaceuticals. Seven methods for RP- HPLC determination,13-16 six methods for UV-Visible estimation17-22 and one method for Difference spectrophotometric methods23 are reported so far. No First derivative UV estimation method was reported, accordingly there was a scope for new method development.
Shimadzu 1800 spectronic model UV Spectrophotometer with 1cm matched quartz cells was used as the instrument for data collection and analysis. Distilled Water was used as the solvent. Tablet brands were obtained from the local market for assay and recovery studies.
Preparation of Standard Stock Solution:
An accurately weighed quantity of 25mg Capecitabine dissolved in little volume of about 10ml distilled water and transferred to 25ml volumetric flask; the volume was adjusted to 25ml using distilled water to get a concentration of 100microgram/ml concentration of Capecitabine.
A solution of 50microgram/ml concentration of Capecitabine was prepared by diluting appropriate volume of standard stock solution and scanned in the range of 240 to 400nm range of UV spectrum and the first derivative absorption maximum in distilled water found to be at 280nm with N=4. Therefore, the peak at 280nm was selected as analytical wavelength for first derivative UV method development.
Using the standard stock solution of Capecitabine, appropriate aliquots were prepared over the concentration range of 40 to 80microgram/ml for the purpose of observing linearity and to decide the range. The concentrations of 40microgram/ml, 50 microgram/ml, 60microgram/ml, 70microgram/ml and 80microgram/ml aliquots were subjected to the measurement at the peak 280nm with N=4 and a graph of area v/s concentrations was plotted. The data is provided in the table 1 and figures 1-3.
Table 1: Calibration data Table for Capecitabine
|
Sr. No. |
Concentration (Microgram/ml) |
Absorbance at 280nm First derivative UV Peak |
|
1 |
40 |
0.029 |
|
2 |
50 |
0.036 |
|
3 |
60 |
0.045 |
|
4 |
70 |
0.051 |
|
5 |
80 |
0.059 |
Calibration Graph of Area v/s Concentration for Capecitabine linearity
Figure 1: First derivative spectrum for 40 microgram/ml Concentration
Figure 2: First derivative spectrum for 60 microgram/ml Concentration
Figure 3: First derivative spectrum for 80 microgram/ml Concentration
Brand name- CAPGET® 500 Standard:
From the standard stock solution of Capecitabine, appropriate aliquots were pipette out into 25ml volumetric flask and dilutions were made with distilled water to obtain working standard solution of Capecitabine 50μg/ml. This concentration was scanned at the first derivative UV peak of 280nm with N=4.
Table 2: Assay for Capecitabine in Tablet Form
|
Brand Name |
Label Claim (mg/tablet) |
Amount Found (mg/tablet) |
% Of Label Claim |
Mean |
SD |
CV |
|
CAPGET® 500 |
500 |
500.99 |
99.45 |
99.88 |
0.2358 |
0.0022 |
|
500 |
500.12 |
100.03 |
||||
|
500 |
500.98 |
99.97 |
||||
|
500 |
501.14 |
100.01 |
||||
|
500 |
500.97 |
99.91 |
Table 3: Result for accuracy parameters of Capecitabine (Brand Name: CAPGET® 500)
|
Label % recovery |
Amount present (mg/tablet) |
Amount Of Standard added (mg/tablet) |
Amount Recovered (mg/tablet) |
Total % recovery |
% mean recovery |
SD |
CV |
|
80 |
500 |
400 |
899.99 |
99.94 |
100.07 |
0.1257 |
0.0013 |
|
80 |
500 |
400 |
900.02 |
100.2 |
|||
|
80 |
500 |
400 |
900.04 |
100.2 |
|||
|
100 |
500 |
500 |
899.98 |
99.91 |
100.04 |
0.2353 |
0.0021 |
|
100 |
500 |
500 |
901.01 |
100.4 |
|||
|
100 |
500 |
500 |
901.02 |
99.86 |
|||
|
120 |
500 |
600 |
899.98 |
99.96 |
99.56 |
0.6611 |
0.0065 |
|
120 |
500 |
600 |
899.98 |
98.81 |
|||
|
120 |
500 |
600 |
899.99 |
99.94 |
Ten tablets of the branded tablets CAPGET® 500 containing 25mg of Capecitabine weighed, and finally powered using a mortar. Each uncoated tablet contains 25mg of Capecitabine. A quantity of powder segment corresponding to 25mg of Capecitabine was taken into volumetric flask. And dilution was made to get concentration of 50μg/ml respectively. These all concentrations were scanned at the first derivative UV peak of 280nm with N=4. (Table 2)
Recovery experiments are used for the study of accuracy method. This study was carried out by adding known amount bulk sample to the tablet and recovery was performed at three levels, 80, 100 and 120% of Capecitabine standard concentration. Samples for recovery studies were prepared according to before mentioned procedure. Three samples were prepared for each recovery level. The solutions of sample were analyzed and % recoveries were calculated by using following formula.
Observed amount of compound in sample
% Recovery = –––––––––––––––––––––––––––––––––––––––– × 100
Amount of all compound present in sample
The recovery values are summarized in following table 3.
The precision (inter–day) was carried out by using four independent sample of Capecitabine. The intermediate precision (inter-day precision) of the method was evaluated using four different analysts in the same laboratory. (Table 4)
Table 4: Determination of Precision of Capecitabine
|
Sample Number |
Assay of Capecitabine as % of Labelled amount (inter – day precision) |
|||
|
Analyst 1 |
Analyst 2 |
Analyst 3 |
Analyst 4 |
|
|
1 |
99.36 |
99.54 |
99.75 |
99.56 |
|
2 |
100.10 |
99.44 |
99.35 |
99.59 |
|
3 |
99.35 |
99.39 |
100.71 |
100.88 |
|
4 |
100.89 |
100.09 |
99.74 |
99.88 |
|
Mean |
99.91 |
99.60 |
99.89 |
99.96 |
|
SD |
0.7295 |
0.3312 |
0.57684 |
0.6247 |
|
CV |
0.0072 |
0.0033 |
0.0057 |
0.0062 |
The standard solutions of Capecitabine prepared in distilled water subjected to a scan in the area between 240nm to 400nm for area under curve method using Shimadzu UV-Visible spectrophotometer 1800 spectronic model because the first derivative UV absorption maximum was found to be located at 280nm. The calibration graph of Capecitabine was linear at a concentration range over 40 to 80μg/ml at the first derivative peak of 280nm.
As a result, it was noticeable that Capecitabine can be estimated in the presence of distilled water as the only reagent with no interruption of any extraneous substance in pharmaceutical products. With the intention of determining the practicability of the developed technique for the evaluation of commercially available (CAPGET® 500) identified formulations, the method is by the side, primarily applied on bulk drugs in their bulk sample, then concentrations in formulations were subjected to evaluation. Subsequently this new method was applied to the determination of formulation obtained from the market and satisfactory results were confirmed within the set limits as per the of the label content claimed for Capecitabine.
Validation of presently designed method was carried out as per the internationally accepted parameters and guidelines. This area under curve method for the quantitative investigation of Capecitabine was tried on exposing to various parameters of validation guidelines like specificity and selectivity in presence of substances in need to prepare dosage formulations, deliberated for linearity (y = 0.00075x-0.001) and range at distorted levels of concentrations and calibration standards where the determination range was established, accuracy was affirmed by recovery studies at special levels of concentrations, precision was confirmed through inter- day and intraday precision studies, in which the samples were exposed to altered conditions other than optimized parameters.
This newly developed first derivative UV estimation method is accurate and precise because the standard deviation of various parameters was close to 1 and co efficient of variation was close to zero. The method is fast because the procedure can be completed within 15- 20 minutes. The method is economical because distilled water is the only reagent used for the method development. Moreover, the method can be applied to dosage formulations for their routine analysis successfully in a simple manner because the same zero order spectra can be processed into first order UV derivative spectrum within minutes by using options available in the instrument itself.
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Received on 05.09.2020 Modified on 17.10.2020
Accepted on 10.11.2020 © RJPT All right reserved
Research J. Pharm. and Tech. 2021; 14(10):5238-5241.
DOI: 10.52711/0974-360X.2021.00912