www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1490
DEVELOPMENT AND VALIDATION OF
UV SPECTROPHOTOMETRIC METHOD FOR THE ESTIMATION OF PRAZIQUANTEL IN TABLET FORMULATION
Uma Parvathy S.*, Dhivya K. and Parvathi S.
K.K. College of Pharmacy, Gerugambakkam, Chennai – 600128.
ABSTRACT
Praziquantel is a drug which is used in the treatment of Helminthics caused by Tape worm & Roundworm. A simple, rapid, selective UV spectrophotometric method was developed for the estimation of praziquantel in tablet dosage form. The method was based on the measurement of maximum absorbance at about 261 nm wavelength of praziquantel using methanol as a solvent. The linearity studies indicated that estimation of praziquantel obeys beer’s law at the concentration of 250mcg to 750mcg/ml. This method was validated in terms of accuracy, precision and specificity as per ICH Q2 (R1)
guidelines. The percentage recovery range was found between 99.61-100%, that indicates the good index of precision and accuracy. Thus, this proposed method was found to be specific, precise, and cost-effective quality control tool for the routine analysis of praziquantel in tablet dosage form.
KEYWORDS: Praziquantel, UV spectrophotometric, validation, Precision.
INTRODUCTION
Spectroscopy is a tool for structure determination. Spectroscopy is defined as the study of the quantized interaction of electromagnetic radiations with matter. Electromagnetic radiations are produced by the oscillation of electric charge and magnetic field residing on the atom.
There are various forms of electromagnetic radiation, e.g. light (visible), ultraviolet, infrared, X-rays, microwaves, radio waves, cosmic rays etc.[1] UV spectroscopy is a type of absorption spectroscopy. In which light of ultraviolet region (200-400) is absorbed by the Molecule.
Absorption of the UV radiation results in the excitation of the electrons from the ground state
Volume 9, Issue 11, 1490-1498 Research Article ISSN 2278 – 4357
*Corresponding Author Uma Parvathy S.
K.K. College of Pharmacy, Gerugambakkam, Chennai – 600128.
Article Received on 28 August 2020,
Revised on 18 Sept. 2020, Accepted on 08 October 2020 DOI: 10.20959/wjpps202011-17571
www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1491 to higher energy state. The energy of the UV radiation that are absorbed is equal to the energy difference between the ground state and higher energy state.[2]
The principle of UV spectroscopy obeys the beer-lambert law, which states that: when a beam of monochromatic light is passed through a solution of an absorbing substance, the rate of decrease of intensity of radiation with thickness of the absorbing solution is proportional to the incident radiation as well as the concentration of the solution.[3-4]
A = log (Io/I) = abc Where, A = absorbance
Io = intensity of incident light c = molar concentration of solute b = pathlength
a = molar absorptivity
Steps Involved in UV spectrophotometer 1. Selection of solvent system.
2. Selection of Analytical wavelength.
3. Study of Beers- Lamberts law.
4. To perform Analysis of standard Laboratory mixture & Tablet formulations by proposed method.
5. To validate the developed method by using different validation parameters.
Analytical method validation
The process of method validation is used to establish the analytical procedure employed for a specific test is appropriate for its intended use. The result of the above used to judge the quality, reliability, and consistency of analytical results.[5]
According to ICH Q2 (R1) and US FDA Technical review guide, typical Analytical performance characteristics that should be considered in the validation of all types of methods are:
Accuracy
Precision:
a) Repeatability,
b) Intermediate precision (Ruggedness), c) Reproducibility.
www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1492
Specificity
Detection limit.
Quantitation limit.
Linearity
Range
Robustness
Stability
Praziquantel is chemically known as 2(cyclohexyl carbonyl)1,2,3,6,7,11b hexahydro -4H- pyrazino[2,1-a]iso quinoline- 4-one.
It is white crystalline powder. It is soluble in methanol, chloroform and slightly soluble in water. It is official in I.P & B.P
MATERIALS AND METHODS
Instrument: UV visible spectrophotometer with pair of 1cm Matched quartz cell, Software- UV probe 2.35 version, Model –UV 1800, Make- Shimadzu.
Reagents: Praziquantel working standard (potency-99.67%w/w) was collected from Medo pharm Pvt. ltd and AR-grade Methanol (stanchem chemicals) was used.
Standard preparation: 50mg of praziquantel working standard was taken in 100ml volumetric flask. Then 15ml of methanol was added, shaken well until it completely dissolved and made up to the volume with methanol (500 mcg/ ml).
Sample preparation: 20 tablets were weighed and powdered. Powdered tablet equivalent about 79mg was taken in 100ml volumetric flask and 20ml of methanol was added, shaken well for 15 minutes and then made up to the volume with water. The solution was filtered through 0.45µm membrane filter (500 mcg/ ml).
www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1493 Method validation: Specificity, Linearity, Precision, Accuracy, Robustness, Solution
RESULTS AND DISCUSSION
UV Method Development and Optimization: The wavelength selection of method based on solubility and nature of sample. our preliminary trials were by using different compositions of water, methanol, 0.1M HCl.by using methanol as a solvent gives best result.
Fig. 1: Spectrum of standard.
Fig. 2: Spectrum of sample.
www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1494 Fig. 3: Spectrum of blank.
Fig. 4: Spectrum of placebo.
Fig. 5: linearity curve for praziquantel.
www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1495 Table 1: Linearity data for praziquantel.
Linearity levels Concentration(μg/ml) Absorbance
Level I (50%) 250 0.274
Level II (80%) 400 0.431
Level III (100%) 500 o.537
Level IV (120%) 600 0.638
Level V (150%) 750 0.795
Correlation coefficient 1.0000
Intercept 0.0145
Slope 0.001 X
Table 2: Evaluation data for precision study.
Sample no. Assay in % SET Intra day Inter day
1 99.42% 100.50%
2 99.25% 99.78%
3 99.92% 99.38%
4 99.61% 99.86%
5 99.92% 100.04%
6 100.23% 99.51 %
Mean 99.56% 99.84%
% Rsd 0.40 0.40
Table 3: Evaluation data for accuracy study.
Level Ex.no Recovered in mg % Recovered
I (50%) 1 25.2 100.39%
2 25.05 99.40%
3 24.86 99.04%
Mean 25.06 99.61%
RSD 0.69%
II (100%) 1 50.38 100.5%
2 49.92 99.44%
3 50.01 100.02%
Mean 50.10 100%
RSD 0.56
III (150%) 1 74.79 99.58%
2 75.18 100.43%
3 74.86 99.57%
Mean 94.95 99.83%
RSD 0.4
www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1496 Table 4: Evaluation data for stability study.
Time % Assay % Rsd Absolute Difference
Initial 99.86% - -
2 hours 99.63% 0.31 0.03
4 Hours 99.48% 0.19 0.12
6 Hours 99.22% 0.25 0.2
Table 5: Evaluation data for robustness study (Change in wavelength for Praziquantel).
Conditions % mean assay % RSD
Actual conditions 99.56% -
Detector
Wavelength (261+2)
99.24% 0.42%
Detector
wavelength (261-2)
99.16% 0.38%
The amount of drug present in marketed formulation were analyzed and calculated. The amount of praziquantel was found in the range of 99.12 – 100.23%. The results show that there is no absorbance of blank and placebo at wavelength of 261nm. This parameter represents the specificity of the method. Linearity range was evaluated by plot of absorbance as a function of analyte concentration and the corresponding calibration curves. From the linearity studies, the specified concentration range was determined and the praziquantel was linear in the range of 250μg/ml to 750μg/ml. The linearity range was 50-150% with the correlation coefficient of 1.0000. The percentage recovery range was found between 99.61 – 100 % for praziquantel that indicates a good index of accuracy and precision of the method.
Study of ruggedness was carried out by two different analyst and the results were more precise, indicating the reproducibility of developed method.
CONCLUSION
The proposed method was found to be simple, accurate, precise and rapid. It could be used for routine analysis. The statistical parameters and recovery studies were carried out and the results were satisfactory.
ACKNOWLEDGEMENT
We are grateful to thank the principal prof. Dr. A. Meena, K.K college of Pharmacy, Chennai for her support.
www.wjpps.com │ Vol 9, Issue 11, 2020. │ ISO 9001:2015 Certified Journal │ 1497 REFERENCES
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