WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 7.421

Volume 8, Issue 5, XXX-XXX Research Article ISSN 2278 – 4357

SPECTROPHOTOMETRIC METHOD FOR ESTIMATION OF

TADALAFIL IN BULK AND TABLET DOSAGE FORM

Shrishail M. Ghurghure1, Priyanka R. Surwase1*, Mahewash A. Pathan1, Prasad M.

Gote1, Dr. Kirtee M. Gaikwad2

1
PG Pharmaceutical Department, DSTS Mandal’s College of Pharmacy, Solapur- 413004
Maharashtra, India.
2
Medical Officer at Municipal Hospital, Pimpari-Chinchwad Mahanagar Palica, Thergaon.

Article Received on
19 March 2019,
Revised on 09 April 2019,
Accepted on 30 April 2019,
DOI: 10.20959/wjpps20195-13847
ABSTRACT
UV spectroscopic method have been developed and validated for the
determination of Tadalafil in bulk and tablet formulation. The λmax was
found to be 284nm in methanol. Beer’s law was obeyed in the
concentration range of 2-10µg/ml. Good accuracy (105%), precision

*Corresponding Author (%RSD0.368), LOD (0.2012), LOQ (0.364) and linearity (0.999) were
Priyanka R. Surwase obtained. The result of analysis has been validated as per the ICH
PG Pharmaceutical guidelines. The developed method is simple, selective and reproducible
Department, DSTS
and can be used for routine of analysis of formulations containing
Mandal’s College of
Tadalafil.
Pharmacy, Solapur- 413004
Maharashtra, India.
KEYWORDS: Tadalafil, spectrophotometric validation, tablet dosage
priyankasurwase4595@gmail.com
form.

1. INTRODUCTION[1, 2]
Tadalafil, chemically pyrazino [1_,2_:1,6] pyrido [3,4-b] indole-1,4-dione,6-(1,3-
benzodioxol- 5-yl)-2,3,6,7,12,12 a-hexahydro-2-methyl-, (6R,12aR)-(Figure1), is an
impotence agent. It is indicated for the treatment of erectile dysfunction1-2. It is a selective
inhibitor of cyclic guanosine monophosphate (cGMP)-specific phosphodiesterase type 5
(PDE5)3. The aim of the present work was to develop simple, rapid, accurate and sensitive
UV spectrophotometric method for the determination of tadalafil in bulk and tablet
formulation. UV analysis of tadalafil was performed in methanol. The spectrum was recorded

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Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

from 200 nm to 400 nm. The quantitative analysis was carried out at 284 nm. The method
was validated and applied for the determination of tadalafil in tablet dosage form.

Figure 1: Chemical structure of tadalafil.

Several analytical methods that have been reported for the estimation of (TDF) in biological
fluids or pharmaceutical dosage forms are liquid chromatography and spectrophotometry.
The well-established spectrophotometric method employs direct UV spectrophotometric
estimation of (TDF). Other methods are based on ion-pair complex formation between basic
compounds and an anionic dye such as bromocresol purple (BCP) and methyl orange (MO),
At a specific pH, the ion-pair is extracted into an organic solvent, which is immiscible with
water, and the concentration of the resulting ion pair in the organic phase is determined
spectrophotometrically. In the present investigation, we report the development of accurate,
reproducible, less time consuming and adequately sensitive validated spectrophotometric
methods for the determination of (TDF).

MATERIALS AND METHOD

A double beam UV spectrophotometer (systronics, UV VIS 2201) was used with 1 cm
matched quartz cell. Tablet formulation [Glycomet-500, USV LIMITED regd.office: B.S.D.
Marg, Govandi Mumbai 400088] with labelled amount 500mg per tablet.

Solvent selection
Various solvents were selected for the solubility studies and it was found that Tadalafil was
soluble in the following solvents; methanol, water, ethanol, dichloromethane etc. in the
present investigation methanol was selected as solvent.

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Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

Preparation of stock solution

10mg of Tadalafil was weighed and transferred to a 10ml volumetric flask then dissolved in
the methanol. The volume was made up to the mark with same solvent to obtain
concentration of 1000µg/ml of Tadalafil. From the resulting solution 1 ml was diluted to
10ml with same solvent to obtain concentration of 10µg/ml of Tadalafil.

METHOD VALIDATION[1, 2, 3, 5]
Validation parameters
1. Linearity
2. Precision
3. Accuracy
4. Ruggedness
5. LOD
6. LOQ

METHODS
a) Description: colour and texture of tadalafil powder was compared with reported
characters.
b) Solubility: solubility of tadalafil was determined in various solvents like methanol, water,
ethanol, dichloromethane.
c) Melting point: capillary filled with tadalafil kept in melting point apparatus and
determined, the melting point was compared with the reference.

UV spectroscopic method for tadalafil

Spectrophotometric condition
 Mode: spectrum
 Measuring mode: absorbance
 Scanning speed: medium
 Wavelength range: 200-400nm
 Absorbance scale: 0.00 to 2Ao

Selection of solvent
Solution of Tadalafil (10µg/ml) was prepared in methanol. These solutions were scanned in
UV region (200-400nm) and maximum absorbance were determined for these solutions.

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Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

Preparation of solution
A primary stock solution of 10µg/ml of Tadalafil was prepared in methanol the standard
solution was prepared by dilution of the primary stock solution with methanol to obtain
working standard of concentration 100µg/ml.

Validation of UV spectrophotometer[4, 5, 6]
1. Linearity: From the standard stock (100µg/ml) solution, pipette out 1ml from stock and
dilute with methanol up to 10ml (10µg/ml).2, 4, 6, 8,10ml were transferred in a series of
10ml volumetric flask. The volume was made up to the mark with methanol to obtain the
concentration of 2, 4, 6, 8, 10µg/ml. The correlation coefficient (r2) of least square linear
regression of Tadalafil was calculated.

2. Range: The range of analytical method was decided from interval between upper and
lower level of calibration curve by plotting the curve.

3. Accuracy: Recovery study was carried out by standard edition method by adding the
known of Tadalafil (working standard) to the pre analysed sample at three different
concentration level that is 50%, 100%, 150% of assay concentration and percent recovery
were calculated.

From the 100µg/ml sample stock solution 1ml was transferred to 4 different 10ml volumetric
flask separately along with 1,2,4,6ml from the 10µg/ml solution the volume was made up to
the mark with methanol and respective absorbance were noted from the above data %
recovery were calculated.

4. Precision: The precision of the proposed method was ascertained by determination of six
replicates of same con centration of sample and standard for method precision and system
precision. Both intraday and inter precisions were carried out.

5. Limit of detection: detection limit was determined based on standard deviation of

absorbance of same concentration i.e. standard solution of Tadalafil (6µg/ml) prepared six
time and LOD calculated by
LOD=3.3(SD/S)
Where, SD-standard deviation; s-slope of curve.

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Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

6. Limit of Quantification: Quantification limit was determined based on the standard

deviation of peak area of same concentration that is standard solution of (6µg/ml) prepared
by six times and LOQ calculated by.
LOQ=10(SD/S)
Where, SD-standard deviation; s-slope of curve

7. Robustness: Robustness of the method was determined y carrying out the analysis at two
different temperature that is at room temperature 290C and 240C.respective absorbance were
noted and the result was indicated by % RSD.

RESULTS
Linearity
Table no.1 shows linearity results and figure no.1 shows linearity results.

Accuracy
Table no.2 shows accuracy results.

Precision
Intra day
Intra-day peecision results are shown in in table no.3.

Inter day
Inter-day precision results are shown in table no.4.

Detection Limit
LOQ (µg/ml) = 0.1394

Quantification Limit

LOQ (µg/ml) = 0.4225

Table no.1 Linearity results.

Concentration λmax Absorbance
2µg/ml 284.0 0.098nm
4µg/ml 284.0 0.2194nm
6µg/ml 284.0 0.276nm
8µg/ml 284.0 0.371nm
10µg/ml 284.0 0.457nm

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Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

Figure 1: Linearity graph of Tadalafil.

Table 2: Accuracy studies.

Amount Total
Level Amount
of standard Total amount amount
Sr.no Of % of sample %recovery
drug added added (µg/ml) found
recovery stock(ml)
(µg/ml) (µg/ml)
1 0 4 0 0 0 ---
2 50 4 2 6 5.99 99.25
3 100 4 4 8 7.99 98.61
4 150 4 6 10 9.8 98.66

Table 3: Precision studies: Intra day.

Sr.no. Concentration Absorbance %SD %RSD
1 6µg/ml 0.273
2 6µg/ml 0.274
3 6µg/ml 0.277 0.0018 0.6531
4 6µg/ml 0.278
5 6µg/ml 0.276
6 6µg/ml 0.276

Table 4: Precision studies: Inter day.

Sr.no. Concentration Absorbance %SD %RSD
1 6µg/ml 0.274
2 6µg/ml 0.277
3 6µg/ml 0.273 0.0018 0.6754
4 6µg/ml 0.276
5 6µg/ml 0.276
6 6µg/ml 0.278

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Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

Table 5: Results of validation parameters.

Sr.no Parameters Values
1 Beer’s law limit(µg/ml) 10-5
2 Regression equation(y=mx+c) y=0.0455x+0.0051
3 Correlation coefficient(r2) 0.9993
4 Slope(m) 0.0455
5 Intercept(c) 0.0051
6 Linearity 0.998
7 Limit of detection(µg/ml) 0.1394
8 Limit of quantification(µg/ml) 0.4225
9 Precision(%RSD)
Intraday 0.6531
Interday 0.6751
10 Accuracy(%recovery) 98.61%
11 % mean recovery for Tadalafil 98.33%

DESCUSSION
Preliminary analysis of Tadalafil
Tadalafil being UV absorbing has been successfully employed for its quantitative
determination by UV spectrophotometric method. Being freely soluble in methanol, stock
solutions and working standards were made in methanol. The λmax of the drug for the analysis
was determined by taking scan of the drug sample solution in the entire UV region (200-
400nm). The correlation of the standard curves for the drug was 0.998 (graph). The
commercial dosage form showed 100.4% recovery by this method which was within the
specified limits of content uniformity.

The proposed method showed absorption maxima at 284nm and Beer’s law in the
concentration range of 2-10µg/ml. The limit of detection (LOD) was found to be
0.1394µg/ml and limit of qualification (LOQ) to be 0.4225µg/ml. the percentage recovery
value of percentage relative standard deviation shows that the developed method was precise.
All statistical data prove validity of proposed method, which can be applied in industries for
routine analysis of metformin from tablet.

Assay of Marketed formulation

Amount of drug present in marketed formulation was calculated using equation.
y=0.0455x+0.0051. Amount of Tadalafil was found to be 98.33% of label claim. This method
can be employed for routine analysis of Tadalafil.

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Surwase et al. World Journal of Pharmacy and Pharmaceutical Sciences

A simple yet precise, rapid and accurate UV method for determination of Tadalafil from pure
and its tablet formulation has been developed and validated. Recoveries from formulation
were in good agreement with their respective label claim. The proposed methods can be used
for the routine determination of Tadalafil in bulk and pharmaceutical dosage forms. The
proposed UV spectrophotometric, method has been evaluate over the linearity, accuracy,
precision, LOD and LOQ. The proposed method was found to be convenient and effective for
the quality control.

Summary of UV spectrophotometric method of Tadalafil: It is shown in table no.5.

CONCLUSION
For the routine purpose it is always of interest to establish methods capable of analysing the
sample in a short period with due to accuracy and precision. The main purpose of this study
was to develop accurate, precise and economic method for the determination of Tadalafil.
The UV-Visible technique namely Zero-order Spectroscopic method, was applied without
using any prior cost effective fast and efficient. Finally, the proposed method could be useful
and suitable for determination of Tadalafil in bulk and dosage form.

REFERENCES
1. Yunoos M., Sanker G. UV spectrophotometric method for the estimation of Tadalafil in
bulk and tablet dosage form. E-Journal of chemistry, 2010; 17(3): 833-836.
2. Zamir K., Amod P., Atul S. Estimation of Tadalafil using derivative spectrophotometry in
bulk material and in. pharmaceutical formulation. International journal of spectroscopy,
2014; 14: 1-6.
3. Safwan F. Spectrophotometric methods for determination of Tadalafil in pharmaceutical
form. International journal of pharmacy and pharmaceutical sciences, 2014; 7(6):
443-445.
4. Ali K., Ayman G. Spectrophotometric determination of Tadalafil in pure and dosage
form. Chemical industry chemical engineering quarterly, 2011; 17(2): 125-132.
5. Gunjan A., Bhavin C., Ashutosh P. Development and validation dual wavelength UV
spectrophotometric method for simultaneous estimation of Tadalafil and Dapoxetin
hydrochloride in their combined tablet dosage forms. International journal of
pharmaceutical research bioscience, 2012; 1(2): 247-255.
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