Spectroscopic Estimation of Dapoxetine HCl using Multivariate Analysis

Krishnakumar P¹, Sundarrajan T¹, Kathiravan M K², Manikandan K³*

¹Department of Pharmaceutical Chemistry, SRM College of Pharmacy, SRM Institute of Science and Technology, Kattankulathur, Kancheepuram – 603203.

²209, Dr APJ Abdul Kalam Research Lab, Department of Pharmaceutical Chemistry, SRM College of Pharmacy, SRM Institute of Science and Technology, Kattankulathur, Kancheepuram – 603203.

³Department of Pharmaceutical Analysis, SRM College of Pharmacy, SRM Institute of Science and Technology, Kattankulathur, Kancheepuram – 603203.

*Corresponding Author E-mail: gurumani12@gmail.com

ABSTRACT:

A sensitive and exact UV spectroscopic technique with a multilinear regression technique for assurance for Dapoxetine HCL in the drug dosage form. This technique relies upon the utilization of the linear regression equations by utilizing the interconnection between absorbance and concentration at five selected wavelengths. The outcomes were dealt with numerically and were found highly precise, reproducible and accurate. This technique is precise, linear and accurate between the limits of 10-50μg/mL. The excipients do not have any conflicts. The numerical analysis with the multilinear regression method was utilized. The information procured for the evaluation of Dapoxetine HCL in bulk drug dosage form evidenced the high-level accuracy and precision after the multivariate analytical technique. Subsequently, this technique is extremely valuable with uncomplicated mathematical contents, is more dependable than the other spectroscopic techniques then it suggests an application in calibration models for routine analysis. It has been validated and developed for further studies and quantification of Dapoxetine HCL in Bulk and pharmaceutical formulation.

KEYWORDS: Dapoxetine Hcl, Multivariate calibration technique, Pharmaceutical formulations, ICH guidelines, Validation.

INTRODUCTION:

Dapoxetine Hcl is chemically as (S)-N,N-Dimethyl-α-(2-(naphthaleneyloxy)ethy) benzenemethamine having empirical formula of Dapoxetine is C₂₁H₂₃ON and the M.W of 305.413g¹. The Dapoxetine is primarily helpful in erectile dysfunction as selective serotonin reuptake inhibitor(SSRI)². SSRI’s are a class of drugs that are commonly used as antidepressant in the remedy of ailments like depression, anxiety disorders³. They are effective in the treatment of premature ejaculation, impotence and some cases of insomnia.

The mechanism of action of the drug is related to inhibiting the  neuronal reuptake of serotonin and consequent heightening of serotonin activity and increases the discharge time^4,5. The structure of Dapoxetine Hcl were revealed in (Figure 1).

Fig 1: Structure of Dapoxetine Hcl(DAP).

Multilinear Regression (MLR) relies upon statistical guidelines of multilinear measurements, which comprises examination and perception examination of more than one factual result variable at once^6,7,8. In structure and investigation, the strategy is utilized to execute exchange examines various concentrations while in consideration of the impacts of all factors on the reactions of premiums^9,10,11.

METHODOGY:

Chemicals:

· Dapoxetine Hcl,

· Methanol,

· The Marketed Tablet Formulation (label claim-30 mg)

Solubility:

Dapoxetine Hcl was dissolved in methanol.

Instrument Used:

The multilinear regression was executed in 1.0cm quartz cells using UV 7600 UV- Visible Spectrometer (UV Lamda., Eng.) with a fixed 1 nm spectral bandwidth and UV spectrum Rx v 1.04 software was utilize for all absorbance measurements.

Preparation of Standard Stock Solutions:

Standard stock solution for Dapoxetine was prepared by 25mg of Dapoxetine HCl were dilute in 100mL of methanol and the final volume makeup with methanol in 100mL Volumetric flask and 250µg/mL was determined as final concentration. The 30µg/mL was taken from the working standard solution were examined in the entire Ultraviolet range between 400-200nm to obtain overlay spectra and absorbance spectra¹².

Determination of λMax:

The standard stock solution of Dapoxetine Hcl was dilute with the solvent to get a concentration of 30 µg/mL. This scanned under the UV region from 400-200nm¹³. The maximum absorbance was found to be 292nm.

Preparation of Sample Solution:

For evaluation of marketed formulation, 250mg of the bulk drug weigh accurately and transfer into a 100mL volumetric flask containing the methanol. The above solutions were filtered through Watmann filter paper and the solution was transferred into a standard flask and makeup to the fixation with a solvent system to obtain a final concentration of 20µg/mL. All determinations were conducted with six variates. The solutions were sonicated for 15 minutes and finally makeup to the volume with solvent, filtered and diluted for further analysis.

Method of Validation:

This technique was evaluated as per the ICH Q2 R1 guidelines to examine validation parameters such as linearity, sensitivity, precision, and accuracy¹⁴.

Linearity:

The prepared standard stock solution 250µg/mL of Dapoxetine HCl was dilute with a solvent to get a concentration in the range 10-50µg/mL. To develop an interactive and underrate instrumental fluctuation absorbance of the solution was calibrated in certain wavelengths around the λmax of the drug 292nm i.e., 296, 294, 292, 290, 288nm. Then the overlay spectra show linearity of the absorbance were recorded at six various concentrations^15,16,17.

Precision:

Intra-day and Interday Precision was performed by examining the absorbance of the solution of concentration 30µg/ml at a selected wavelength. The concentration was scanned two times within a day (intraday precision) and for two days (interday precision). The absorbance value recorded at the selected wavelength for intraday and interday precision. The SD and %RSD values obtained at different wavelengths were calculated^18,19.

Assay:

The absorbance of the extracted sample solution was recorded at 292nm and the quantity of drug present in the formulation was determined^20,21.

Recovery Studies:

This method was executed to justify 0.5ml of standard formulation (100µg/mL) were taken in three 10ml standard flask and it 40%, 60%, 80% (i.e, 4mL, 6mL, 8mL) of working standard stock solution (100µg/ml) and make the volume up to the mark. The corresponding absorbance at 292nm was observed against the blank^22,23. The quantity of drug added concentration was determined from the obtained percentage recovery and absorbance values for each dosage form.

RESULTS AND DISCUSSION:

Dapoxetine HCl was predicted by the prospective multivariate UV spectroscopic method in tablets.  The bulk drug of Dapoxetine HCl was rapidly soluble in methanol then methanol was fixed as a solvent for Dapoxetine HCl to determine the UV spectrum in the range of 400-200nm. After the validation of the spectrum, the λmax of Dapoxetine present at 292nm were showed in (Figure 2).

The Overall Result of Multivariate Analysis of Dapoxetine HCl were shown in (Table 1). The technique was evaluated through ICH Q2 R1 guidelines for accuracy, linearity, precision, and sensitivity.

Linearity:

The concentration range 10-50µg/mL were scanned for the linearity at the selected wavelength of about 296, 294, 292, 290 and 288nm which was shown their calibration graphs in (Figure 3-7).

Fig 2:  λ max of Dapoxetine HCl

Table 1: Overall Result of Multivariate Analysis of Dapoxetine Hcl

Parameters	Wavelengths (nm)
	296	294	292	290	288
Linearity (µg/ml)	10-50	10-50	10-50	10-50	10-50
Correlation Coefficient R^2	0.9992	0.9994	0.9993	0.9992	0.9991
Slope(m)	0.0136	0.0144	0.0147	0.0147	0.0145
Intercept(c)	0.0720	0.0743	0.0728	0.0699	0.0682
LOD(µg/ml)	0.0034	0.0030	0.0032	0.0029	0.0038
LOQ(µg/ml)	0.0104	0.0090	0.0098	0.0088	0.0118
Precision Inter Day (%RSD)	0.0245	0.0272	0.0253	0.0292	0.0267
Precision Intra Day (%RSD)	0.0274	0.0236	0.0172	0.01748	0.0251
Assay (%w/w)	99.93-100.98	99.72-100.89	98.65-100.67	98.67-100.87	98.89-100.76
Percent Recovery(%w/w)	99.52-100.86	99.69-100.55	99.47-100.53	99.46-100.39	99.41-100.55

Precision:

Inter- day and Intra- day precision was done through scanning the absorbance of 30µg/mL solution at five selected wavelengths. The % RSD for Inter- day and Intra-day were found lie within range 0.0242 - 0.0295 and 0.0172 - 0.0274 and it was within well limited of less than 2% at all the wavelength. The Prepared solvent in 30µg/mL was scanned three times per day and for 3 days at the same intervals time for Intra-day and Inter-day precision. The least value of % Relative standard deviation shows that the destined technique was accurate and precise.

Assay:

The sample solutions were recorded for absorbance at 292nm and the amount of drug present in the dosage form was evaluated. The assay percentage of Dapoxetine HCL was found to be 98.65–100.67%  w/w.

Recovery:

The percentage recovery of the drug was calculated and was found to be within the range of 99.41 - 100.86 % w/w it shows a percentage recovery belongs to within the limits of 97 – 103 % w/w as per the ICH regulations.

Fig 3: Calibration graph at 296nm

Fig 4: Calibration graph at 294nm

Fig 5: Calibration graph at 292nm

Fig 6: Calibration graph at 290nm

Fig 7: Calibration graph at 288nm

CONCLUSION:

UV spectrophotometric of the Multivariate calibration technique was performed as per the ICH protocol shows that lie within the limit.  The method was developed in the present studies were establish to be accurate, precise, sensitive and reproducible for the assessment of Dapoxetine Hcl and its component formulation.  In this way, a basic and fast strategy utilizing scientific substance was created, which was discovered more unsurprising than the other spectroscopic strategies and is unequivocally suggested for the normal quality control analysis of Dapoxetine Hcl in pharmaceutical formulation.

List of Symbols/Abbreviations:

· nm = Nanometer

· µg/mL = Microgram per millilitre

· ICH = International Conference on Harmanonization

· UV = Ultraviolet

· g/mol = Gram per Mole

· USP = United States of Pharmacopoeia

· BP = British Pharmacopoeia

· HPLC = High Performance Liquid Chromatography

· HPTLC = High Performance Thin Layer Chromatography

CONFLICTS OF INTEREST:

Authors declare no conflicts of interest

ACKNOWLEDGEMENT:

The authors are thankful to rural pharmacy for providing the sample Dapoxetine HCl for our research work and very much thankful to Management & Dean, SRM College of Pharmacy, Kattankulathur, Kancheepuram district for providing all the facilities to carry out this work.

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Received on 21.09.2019            Modified on 06.11.2019

Accepted on 19.12.2019           © RJPT All right reserved

Research J. Pharm. and Tech 2020; 13(6): 2715-2719.

DOI: 10.5958/0974-360X.2020.00483.7