INTRODUCTION:

Prucalopride succinate is a prokinetic agent. Prucalopride succinate is a serotonin-4 (5-HT₄) receptor agonist indicated for the treatment of chronic idiopathic constipation (CIC) in adults. Prucalopride succinate soluble in methanol and DMSO. Molecular Formula is C₂₂H₃₂ClN₃O_7.Molecular wt. is 485.96g/mole^1-2.

Structure of Prucalopride succinate

Literature survey reviews that there is LC-QTOF MS/MS³, method is available for Prucalopride succinate. Hence there is no any UV-Visible Spectrophotometric method is available for Prucalopride succinate. In the present study method have been developed for the quantitative estimation of Prucalopride succinate in bulk and its marketed formulations with simple, sensitive, precise, accurate and economical by UV-Visible Spectrophotometric method.

MATERIALS AND METHODS:

Chemicals:

Prucalopride succinate working standard was obtained from Macleod’s Pharmaceuticals, Mumbai, and Maharashtra. Methanol used was of analytical grade. Tablets (Pruvict-2, 2mg) were procured from local pharmacy.

Instruments:

JASCO model V-530 UV-Visible spectrophotometer with spectral bandwidth 2nm having wavelength accuracy 0.5nm and pair of 1cm quartz cells used to measure the absorbance of all the solutions. Spectra will automatically obtain using SPECTRA MANAGER software. Also precision analytical balance, ultrasonic bath used in the study.

Preparation of standard stock solution:⁴

10mg of Prucalopride is dissolved in 10ml of methanol to forms 1000ug/ml of solution. From this 1ml is solution is transferred to another volumetric flask and volume is made by using methanol to form 100ug/ml of solution. Again 1ml solution is transferred to another volumetric flask and volume is made by using methanol to form 10ug/ml of solution.

Preparation of sample solution:⁵

10 tablets were procured from local market and average weight was determined. The powder equivalent to 2mg of Prucalopride was weighed accurately and taken in separate 10ml volumetric flask, it was dissolved in 10 ml methanol by sonication, filtered through Whatman’s filter paper no. 41 and to give 1000μg/ml stock solution. From this solution, 1ml was withdrawn and diluted to 10 ml with methanol to get 100μg/ml solutions and from this further dilution was made similar to get 10ug/ml.

Absorption maxima method^4-5

Selection of maximum wavelength:

Proper wavelength selection depends upon nature of sample and solubility study.10mg of Prucalopride is dissolved in 10ml of methanol to forms 1000ug/ml of solution. From this 1ml is solution is transferred to another volumetric flask and volume is made by using methanol to form 100ug/ml of solution. Again 1ml solution is transferred to another volumetric flask and volume is made by using methanol to form 10ug/ml of solution. This solution is scanned against blank for UV range 200-400nm. Maximum wavelength is found to be 243nm.

Figure 1. λ-max of Prucalopride succinate

RESULT AND DISCUSSION:

Validation of method ^6-8

The analytical method was validated according to ICH validation parameters ⁹.

1. Linearity:

Fresh aliquots were prepared from standard stock solution ranging from 2-10μg/ml and the absorbance values of each concentration was recorded at 243nm for this method using methanol as blank. The drug shows linearity between 2-10μg/ml for this method.

Table 1: Linearity study

Sr. No.	Concentration (ug/ml)	Absorbance
1	2 ug/ml	0.3139
2	4 ug/ml	0.4382
3	6 ug/ml	0.5850
4	8 ug/ml	0.7340
5	10 ug/ml	0.9412

Figure 2: Calibration curve of Prucalopride succinate at 243 nm

2. Accuracy:

The accuracy for the analytical method for Prucalopride was determined at 80%, 100% and 120% levels of standard solution. Absorbance was measured at 243nm and results were expressed in terms of % recoveries.

Table 2. Accuracy study

Test sample	Accuracy Level	Amount of Std. Drug Added (μg/ml)	Mean Absorbance	% Recovery
	80%	8 ug/ml	0.7791	97.2%
10 ug/ml	100%	10 ug/ml	0.9613	98.2%
	120%	12 ug/ml	1.1461	97.1%

3. Precision:

Intra-day precision was resolute by examine the 6μg/mL of Prucalopride solution for three times in the similar day. Inter-day precision was estimated by 2, 6, 10μg/mL of Prucalopride for three days.

A) Intraday precision (repeatability study)

Table 3. Intraday Precision (Repeatability study)

Conc.	Absorbance (n=6)	Statistical analysis
6 ug/ml	0.6306	Mean=0.6289 S.D=0.0080 %RSD=1.27
6 ug/ml	0.6486
6 ug/ml	0.6287
6 ug/ml	0.6282
6 ug/ml	0.6289
6 ug/ml	0.6285

B) Intermediate precision (Inter-day precision)

Table 4. Intermediate Precision (Inter-day Precision)

Conc.	Absorbance (n=3)	Statistical analysis
2 ug/ml	0.3793	Mean = 0.3799 S.D = 0.00105 %RSD = 0.26
2 ug/ml	0.3792
2 ug/ml	0.3811
6 ug/ml	0.6212	Mean = 0.6265 S.D = 0.0056 %RSD = 0.81
6 ug/ml	0.6315
6 ug/ml	0.6270
10 ug/ml	1.0831	Mean = 1.0833 S.D = 0.0032 %RSD = 0.29
10 ug/ml	1.0833
10 ug/ml	1.0834

4. Robustness:

It is a study of small but deliberate variations in method parameters such as absorption maxima, pH and ratio of mobile phase solvents. In this present work the absorption maxima was decreased and increased 2nm and carried the process for 4μg/ml solution for 6 times. The % RSD was calculated.

Table 5. Robustness Study

Concentration 4μg/ml	Absorbance At 241nm (-2nm)	Absorbance At 245nm (+2 nm)	%Mean RSD
Mean	Mean=0.2887	Mean=0.4830	1.25
SD	SD=0.00514	SD=0.0074
% RSD	%RSD=1.76	%RSD=0.74

Table 6: Analysis of tablet formulation

Sr. No.	Label claim	Amount taken	Amount found	% assay
1	2 mg	10 ug/ml	8.8 ug/ml	88 %

Table 7: Summary of validation parameters

Sr. No.	Parameters	Results
1	Absorption maxima (nm)	243nm
2	Beers range (μg/ml)	2-10ug/ml
3	Standard Regression Equation	Y=0.0775x+ 0.1373
4	Correlation Coefficient (R²)	0.990
5	%Assay	88%
6	Precision (%RSD) 1. Intra-day Precision 2. Inter-day Precision	1.27 0.45
7	Accuracy	97.1-98.3%
8	Robustness (%RSD) at 241 nm at 245 nm	1.76 0.74
9	LOD	0.81
10	LOQ	2.46

CONCLUSION:

A simple, selective, accurate, precise Spectrophotometric method for the estimation of Prucalopride succinate in bulk and pharmaceutical tablet dosage form has been developed and validated. All the procedure followed in the present study as per the ICH guidelines. Hence, this method can be easily used for routine quality control analysis of Prucalopride succinate. The wavelength 243 nm was selected for the estimation of Prucalopride succinate using methanol as a solvent. The linearity studies were performed by plotting different concentrations of standard solutions against their respective absorbance over the concentration range 2-10 μg/ml. The accuracy of the method was assessed by recovery studies by using standard addition method at concentration levels of 80%, 100% and 120% and recovery was found between 97.2- 98.3%.

The method was successfully applied for routine analysis of Prucalopride succinate in bulk and formulation. Hence, this method can be easily used for routine quality control analysis of Prucalopride succinate.

ACKNOWLEDGEMENT:

The authors would thankful to the principal and management of Rajgad Dnyanpeeth’s College of Pharmacy Bhor, for giving permission and providing required facilities to carry out my research work and also thankful to Macleod’s Pharmaceuticals, Mumbai for providing Gift sample Prucalopride succinate drug.

REFERENCES:

1. http:// www.drugbank.ca › salts › DBSALT002626

2. http://pubchem.ncbi.nlm.nih.gov › compound › Prucalopride-succinate

3. Mahamuni BS, Jajula A, Awasthi A, Kalariya P.D, Talluri MV. Selective separation and characterization of stress degradation products and process impurities of Prucalopride succinate by LC-QTOF MS/MS. J Pharm. Biomed. Anal. 2016; 125; 219-228.

4. Preetham DN, Sujana k, Sankar PD, Development and Validation of UV Spectrophotometric Method for the Estimation of Lisinopril in Bulk and Pharmaceutical Formulation. International Journal of Pharmaceutical Sciences Review and Research. 2014; 25(1); 257-259.

5. Sawant DK, Ige PP. Development and Validation of Simple UV-Spectrophotometric Method for Estimation Diclofenac Sodium in Bulk and Tablet Dosage Form. Inventi Rapid: Pharm Analysis & Quality Assurance. 2016; (4): 1-4.

6. Sharma K, Agrawal SS, Gupta M, Development and validation of UV Spectrophotometric method for the estimation of curcumin in bulk drug and pharmaceutical dosage forms. International Journal of Drug Development and Research. 2012; 4 (2): 375-380.

7. Elvis AM, Deepali MG, Development and Validation of UV Spectrophotometric Method for determination of Bisacodyl in Suppositories. International Journal of Pharm. Tech Research. 2011 ;3(1): 193-196.

8. Sethuraman S, Radhakrishnan K, Soloman T.A, Analytical Method Development and Validation of Caffeine in Tablet Dosage Form by Using UV- Spectroscopy. International Journal of Novel Trends in Pharmaceutical Sciences. 2013; 3 (4): 82-86.

9. PART 2. Validation of Analytical Procedure: Methodology Q2B, ICH Harmonized Tripartite Guidelines. 1996; 6-12.

Received on 01.07.2020 Modified on 18.08.2020

Research J. Pharm. and Tech. 2021; 14(8):4189-4191.

DOI: 10.52711/0974-360X.2021.00725