Int J Pharm Pharm Sci, Vol 8, Issue 2, 99-103Original Article


UV SPECTROPHOTOMETRIC ESTIMATION OF SUNITINIB MALATE IN PHARMACEUTICAL DOSAGE FORM

KAVITHA J.*, SAIDEVARAJ A. B., LAKSHMI K. S.

Department of Pharmaceutical Analysis, SRM College of Pharmacy, SRM University, Kattankulathur, Kancheepuram Dist 603203,
Tamil Nadu, India
Email: kavitha0208@gmail.com   

 Received: 21 Sep 2015 Revised and Accepted: 12 Dec 2015

ABSTRACT

Objective: To develop and validate simple zero (D0), first (D1) and second (D2) order derivative UV Spectrophotometric methods for the determination of Sunitinib malate in pharmaceutical dosage form.

Methods: Sunitinib malate was solubilised in distilled water and the resultant solution exhibits adsorption maximum (λmax) at 431, 457 and 489 nm in D0, D1 and D2 order derivative modes respectively. The developed method was validated as per ICH guidelines [1].

Results: Linearity was obtained over the concentration range of 2-12 µg/ml in all the derivative modes. Limit of detection (LOD) was found to be 0.291, 0.107, 0.327μg/ml and Limit of quantification (LOQ) was found to be 0.883, 0.324, 0.993μg/ml for D0, D1 and D2 order derivative modes respectively. The proposed method demonstrated an excellent intra-day precision and inter-day precision. Mean recovery was found within the range of 98.19-98.62% respectively, signifies the accuracy of the developed method.

Conclusion: The statistical results prove that the developed method can be effectively applied for the routine analysis of Sunitinib malate in industries and other analytical laboratories.

Keywords: Sunitinib malate, UV Spectroscopy, D0, D1 and D2 order derivative modes.

© 2016 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

INTRODUCTION

Sunitinib malate is described chemically as Butanedioic acid, hydroxy-(2S)-compound with N-[2-(diethylamino) ethyl]-5-[(Z)-(5-fluoro-1, 2-dihydro-2-oxo-3H-indol-3-ylidine) methyl]-2, 4-dimethyl-1H-pyrrole-3-carboxamide (fig. 1), is an orally administered small molecule that inhibits multiple receptor tyrosine kinases (RTKs). Sunitinib malate was approved by the US Food and Drug Administration (FDA) on January 26, 2006, for the treatment of metastatic renal cell carcinoma (mRCC) and gastrointestinal stromal tumor (GIST) in patients who have failed to respond to imatinib or were unable to tolerate it [2-3], with molecular formula C22H27FN4O2 • C4H6O5 and molecular weight 532.561g/mol. Sunitinib malate is an orange powder which is freely soluble in water.

3

Fig. 1: Chemical structure of Sunitinib malate

On surveying the literature, it was found that only a very few methods have been reported for the estimation of Sunitinib malate alone or in combination by employing RP-HPLC and UPLC [4-7]. Some of the RP-HPLC and UPLC method were not found to be done economical in terms of mobile phase employed and availability of columns. Hence there is a need for the development of newer method for the estimation of Sunitinib malate in pharmaceutical dosage form to overcome the above hurdles. At the same time, no UV Spectrophotometric method was reported for the estimation of Sunitinib malate. So it is felt worthwhile to develop a modest, rapid, accurate, precise and more economical UV Spectrophotometric method for the estimation of Sunitinib malate in pharmaceutical dosage form.

MATERIALS AND METHODS

Chemicals and reagents

The raw material of Sunitinib malate (99.78%w/w) was obtained a gift sample, which was used as reference material throughout the experiment without any prior treatment. Sunitinib malate tablets (SUTENT Capsule-25 mg, Pfizer Inc.) were purchased from a local market. All other reagents used were purchased from Merck laboratories and S. D. fine chemicals Ltd. Double distilled water was utilized throughout the process of analysis.

Instruments used

Solubility studies

The solubility studies of Sunitinib malate was carried out employing different solvent, and the results are as follows:

Selection of solvent

Sunitinib exists as a salt form of maleic acid and it is found that the drug is very freely soluble in water. Hence, water was chosen as the solvent to solubilise Sunitinib malate and to carry out further analysis.

Preparation of standard stock solution of sunitinib malate

The standard stock solution of Sunitinib malate was prepared by accurately weighing 25 mg of the drug and it was kept in a 25 ml standard flask. Half the volume of water was added. The solution was sonicated for 15 min and then the volume was made up to the mark with distilled water. The resultant solution was filtered and suitably diluting with distilled water to get the working standard solutions.

Preparation of sample stock solution

Twenty capsules (STUTENT Capsule-25 mg, Pfizer Inc.) were accurately weighed and the sample stock solution was prepared by accurately weighing an amount of Sunitinib malate equivalent to 25 mg and transferred into a 25 ml standard flask. Half the volume of water was added. The solution was sonicated for 20 min and then the volume was made up to the mark with distilled water. The solution was filtered through Whatman filter paper. The aliquot portion of the filtrate was suitably diluting for further analysis.

Determination of λmax

The standard stock solution of Sunitinib malate was diluted suitably to get a concentration of 10µg/ml. The solution was scanned within the range of 200 nm-500 nm in D0, D1 and D2 order derivative modes respectively.

Method validation

Linearity

From the above prepared standard stock solution, 10 ml of the solution was diluted to 100 ml using water to get a concentration of 100µg/ml. From the above solution 2 ml, 4 ml, 6 ml, 8 ml, 10 ml and 12 ml of solutions were pipetted out into 6 different 100 ml volumetric flasks and the volume was made up to 100 ml using distilled water to get the final concentrations of 2µg/ml, 4µg/ml, 6µg/ml, 8µg/ml, 10µg/ml and 12µg/ml respectively. These aliquots were scanned in D0, D1 and D2 order derivative modes respectively.

LOD and LOQ

LOD and LOQ were calculated from the data obtained from the linearity studies. The slope of the linearity plot and standard deviation (SD) of the responses was determined. LOD and LOQ were calculated on the basis of SD and slope of the regression equation.

LOD = (3.3 x SD)/slope LOQ = (10 x SD)/slope

Precision

From the above prepared standard stock solution, 10 ml of the solution was diluted to 100 ml utilizing water to get a concentration of 100µg/ml. From the above solution 4 ml, 6 ml and 8 ml of solutions were pipetted out into 3 different 100 ml volumetric flasks and the volume was made with distilled water to get the final concentrations of 4µg/ml, 6µg/ml and 8µg/ml respectively. The intra-day and inter-day analysis was carried out to determine the precision of the developed method. These solutions were scanned in the three modes six times a day (inter-day precision). The solutions were scanned in the three modes for six days at the same time (intra-day precision). The calculated percentage relative standard deviation (% RSD) of the results was used to evaluate the method precision.

Recovery

The recovery study was carried out by the standard addition method. 10 ml of the standard stock solution of standard and sample was pipetted out separately to two 100 ml volumetric flasks and the volume was diluted to 100 ml with distilled water to get a concentration of 100µg/ml. From the above standard solution 4 ml was pipetted out into 4 different volumetric flasks and sample solutions of 0 ml, 2 ml, 4 ml and 6 ml were added to the above 4 volumetric flasks already containing standard solutions and the volume was made up to 100 ml with distilled water. The above solutions were scanned in the three modes. The percentage recovery was estimated.

Assay

The sample stock solution of Sunitinib malate was diluted suitably to get a concentration of 10µg/ml. The above solution was scanned in the three modes and the UV spectra’s were recorded and the percentage purity of Sunitinib malate in the pharmaceutical formulation was calculated.

RESULTS AND DISCUSSION

Selection of detection wavelength

The λmax was observed at 431, 457 and 489 nm in D0, D1 and D2 order derivative modes respectively and the UV spectra’s was shown in the fig. 2, 3 & 4 respectively.

4

Fig. 2: UV spectra showing maximum absorbance at D0 mode


5

Fig. 3: UV spectra showing maximum absorbance at D1 mode


6

Fig. 4: UV spectra showing maximum absorbance at D2 mode

Linearity Linearity of Sunitinib malate by the proposed method was made for different concentrations ranging from 2-12 µg/ml was recorded in all the three modes respectively. Linearity was found to obey Beer’s law in the concentration range employed, and the results were tabulated in table 1. The UV spectra slowing linearity and the calibration curves at D0, D1 and D2 modes were presented in fig. 5&6, 7&8, 9&10 respectively.

1

Fig. 5: UV spectra showing linearity at D0 mode


2

Fig. 6: Calibration graph of D0 mode


3

Fig. 7: UV spectra showing linearity at D1 mode


4

Fig. 8: Calibration graph of D1 mode


5

Fig. 9: UV spectra showing linearity at D2 mode


6

Fig. 10: Calibration graph of D2 mode


Table 1: Linearity data at D0, D1 and D2 modes

Parameters

Results

D0

D1

D2

Linearity range (µg/ml)

2-12 µg/ml

Regression equation

y=0.048x-0.025

y=0.178x-0.063

y=0.003x-0.001

Correlation coefficient (r2)

0.999

0.999

0.999

Slope

0.048

0.178

0.003

Y-Intercept

0.025

0.063

0.001

SD

0.0075

0.0058

0.0005

Table 2: LOD and LOQ values at D0, D1 and D2 modes

Parameters

D0 Mode

D1 Mode

D2 Mode

LOD (µg/ml)

0.291

0.107

0.327

LOQ (µg/ml)

0.883

0.324

0.993

LOD and LOQ

The low SD value of responses, LOD and LOQ indicates that the developed method is precise and sensitive. The calculated results are listed in table 2.

Precision

The intra-day and inter-day precision studies (intermediate precision) were performed. The %RSD was considered to be within 0.470-1.870% (intra-day) and 0.870-1.830% (inter-day) which was well within the acceptance criteria, i.e., less than 2% and the results were tabulated in table 3&4.

Recovery

The percentage recovery of the drug was in the range of 97.20-99.36% w/w, which was well within the acceptance limit of 97-103% w/w as per ICH guidelines and the results were tabulated in table 5.

Assay

The mean percentage assay was found to be 97.32-102.28% w/w.

The UV spectra’s were shown in fig. 11, 12 & 13 and the results were listed in table 6.

Table 3: Intra-day precision at D0, D1 and D2 modes

Mode

D0

D1

D2

Concentration

(µg/ml)

4

6

8

4

6

8

4

6

8

Absorbance

0.1659

0.2573

0.3714

0.5831

0.9342

1.3384

0.0100

0.0155

0.0219

0.1639

0.2548

0.3753

0.5820

0.9332

1.3392

0.0096

0.0156

0.0226

0.1626

0.2541

0.3672

0.5813

0.9350

1.3389

0.0098

0.0158

0.0215

0.1632

0.2557

0.3702

0.5792

0.9338

1.3402

0.0099

0.0159

0.0223

0.1645

0.2568

0.3692

0.5840

0.9351

1.3368

0.0099

0.0152

0.0225

0.1632

0.2555

0.3720

0.5754

0.9202

1.3372

0.0096

0.0158

0.022

Mean (±SD)*

0.1638±0.0011

0.2557±0.0011

0.3708±0.0027

0.5808±0.0031

0.9319±0.0057

1.3384±0.0012

0.0098±0.0001

0.0156±0.0002

0.0221±0.0004

%R.S.D

0.720

0.470

0.740

0.540

0.620

0.095

1.700

1.650

1.870

mean±SD* = average of six determinations


Table 4: Inter-day precision at D0, D1 and D2 modes

Mode

D0

D1

D2

Concentration

(µg/ml)

4

6

8

4

6

8

4

6

8

Absorbance

0.1659

0.2573

0.3714

0.5831

0.9342

1.3384

0.0100

0.0155

0.0219

0.1644

0.2610

0.372

0.5734

0.9430

1.3440

0.0099

0.0158

0.0223

0.1660

0.2582

0.3623

0.5728

0.9229

1.3560

0.0099

0.0162

0.0215

0.1643

0.2600

0.3719

0.5801

0.9448

1.3652

0.0102

0.0160

0.0223

0.1672

0.2632

0.3682

0.5790

0.9280

1.3620

0.0101

0.0163

0.0225

0.1679

0.2560

0.3721

0.5629

0.9450

1.3760

0.0098

0.0161

0.0220

Mean (±SD)*

0.1659±0.0014

0.2592±0.0026

0.3696±0.0038

0.5752±0.0072

0.9363±0.0094

1.3569±0.0139

0.0099±0.0001

0.0159±0.0002

0.0220±0.0003

%R. S. D

0.870

1.016

1.053

1.260

1.010

1.030

1.470

1.830

1.630

mean±SD* = average of six determinations


Table 5: Recovery studies at D0, D1 and D2 modes

Mode

Amount present (µg/ml)

Amount added (µg/ml)

Absorbance

Amount recovered (µg/ml)

% Recovery

Mean (±SD)*

D0

4

0

0.1715

3.9745

99.36

98.620±0.9654

4

2

0.2685

5.8881

97.20

4

4

0.3705

7.9592

98.98

4

6

0.4934

9.9576

98.94

D1

4

0

0.6591

3.9663

99.16

98.375±0.6456

4

2

1.0293

5.952

97.60

4

4

1.3896

7.9298

98.24

4

6

1.8408

9.9100

98.50

D2

4

0

0.0098

3.9596

98.99

98.1925±0.5901

4

2

0.0162

5.9632

98.16

4

4

0.0203

7.9219

98.05

4

6

0.0286

9.7945

97.57

 mean±SD* = average of four determinations


Table 6: Assay at D0, D1 and D2 modes

Mode

Theoretical yield (mg)

Practical yield (mg)

Percentage yield (%w/w)

D0

24.33

25

97.32

D1

24.925

25

99.70

D2

25.57

25

102.28

7

Fig. 11: UV spectra showing assay at D0 mode


8

Fig. 12: UV spectra showing assay at D1 mode


9

Fig. 13: UV spectra showing assay at D2 mode


CONCLUSION

Sunitinib malate is an oral, small-molecule, multi-targeted receptor tyrosine kinase (RTK) inhibitor that was approved by the FDA for the treatment of renal cell carcinoma (RCC) and imatinib-resistant gastrointestinal stromal tumor (GIST).

It was the first cancer drug that was approved. The development of a simple, rapid, sensitive and precise Spectrophotometric method for the routine quantitative determination of samples will definitely reduce unnecessary tedious sample preparations and the cost of materials and labour. Sunitinib malate is a UV-absorbing molecule with specific chromophores in the structure that absorbs at a particular wavelength and this fact was successfully employed for their quantitative determinations using the UV Spectroscopic method and validating the same. The λmax was observed at 431, 457 and 489 nm in D0, D1 and D2 order derivative modes respectively.

This needed to be done as a first step to quantifying the amount of drug present in pharmaceutical dosage form. Various validation parameters as discussed above were analyzed and they were found to comply with the guidelines laid by ICH. Therefore, it was concluded that the proposed UV Spectroscopic method was novel, simple, accurate, precise, reproducible, economical and sensitive which would be used for the estimation of Sunitinib malate in their pharmaceutical dosage form in routine analysis.

ACKNOWLEDGEMENT

Authors are highly thankful and we acknowledge the management of SRM University, for providing the facilities required for completing the work successfully.

CONFLICT OF INTERESTS

Declare none

REFERENCES

  1. International Conference on Harmonization of Technical Requirements for Registration of Pharmaceutical for Human Use ICH Guideline: Validation of Analytical procedures text and methodology Q2(R1); 1994.
  2. Edwin P Rock, Vicki goodman, Janet X Jiang, Kooros Mahjoob, S Leigh Verbois, David Morse, et al. Food and drug administration drug approval summary: sunitinib malate for the treatment of gastrointestinal stromal tumor and advanced renal cell carcinoma. Oncologist 2007;12:107-13.
  3. Val R Adams, Markos Leggas. Sunitinib malate for the treatment of metastatic renal cell carcinoma and gastrointestinal stromal tumors. Clin Ther 2007;29:1338-53.
  4. andhya B, Chinnari Harika V, Bikshal Babu Kasimalla, Reshma Syed, Kalpana Pammi. RP-HPLC method development and validation for the analyisis of Sunitinib in pharmaceutical dosage forms. Int J Sci Innovations Discoveries 2011;1:441-50.
  5. De Bruijn P, Sleijfer S, Lam MH, Mathijssen RH, Wiemer EA, Loos WJ. Bioanalytical method for the quantification of Sunitinib and its n-desethyl metabolite SU12662 in human plasma by Ultra performance liquid chromatography/tandem triple quadrupole Mass spectrometry. J Pharm Biomed Anal 2010;51:934-41.
  6. Marie Christine Etienne-Grimaldi, Nicole Renée, Hassan Izzedine, Gérard Milano. A routine feasible HPLC analysis for the anti-angiogenic tyrosine kinase inhibitor, Sunitinib, and its main metabolite, SU12662, in plasma.J Chromatogr 2009;877:3757-61.
  7. Benoit Blanchet, Carole Saboureau, Anne-Sophie Benichou, Bertrand Billemont, Fabrice Taieb, Stanislas Ropert, et al. Development and validation of an HPLC-UV-visible method for sunitinib quantification in human plasma. Clin Chim Acta 2009;404:134-9.