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ORIGINAL ARTICLE
Year : 2015  |  Volume : 6  |  Issue : 1  |  Page : 1-4

Spectrophotometric method for simultaneous estimation of atazanavir sulfate and ritonavir in tablet dosage form


Departments of Quality Assurance, Shri Sarvajanik Pharmacy College, Mehsana, Gujarat, India

Date of Web Publication8-Jan-2015

Correspondence Address:
Disha A Patel
Department of Quality Assurance, Shri Sarvajanik Pharmacy College, Mehsana, Near Arvind Baugh, Mehsana, Gujarat
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2394-2002.148880

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  Abstract 

Background: Ritonavir (RTV) and atazanavir sulfate (ATV) are protease inhibitor and RTV mostly used as a booster for increasing the bioavailability of other protease inhibitors like ATV. Aims: Quality assessment of the new dosage form of RTV and ATV i.e., tablets is very essential and hence this work deals with to develop sensitive, simple and precise method for simultaneous estimation of ATV and RTV in tablet dosage form by absorbance correction method. Materials and Methods: The present work was carried out on Shimadzu Ultraviolate(UV)-1700 double beam spectrophotometer with 1 cm path length supported by S Shimadzu, model-1700(Japan), UV-Probe software, version 2.31 was used for spectral measurements with 10 mm matched quartz cells. Standard ATV and RTV were supplied by Cipla Pharmaceutical Ltd. Methanol was purchased from Finar Chemicals Pvt. Ltd. Results and Conclusion: The λmax or the absorption maxima for ATV and RTV were found to be 279 and 240 nm, respectively in methanol as solvent. The drugs follow Beer-Lambert's law in the concentration range 30-90 and 10-30 μg/mL for ATV and RTV, respectively. The percentage recovery was found to be 100-100.33% and 100-101.5% for ATV and RTV, respectively. The method was validated for different parameters as per the International Conference for Harmonization Guidelines.

Keywords: Absorbance correction method, atazanaviir sulfate, ritonavir, validation


How to cite this article:
Patel DA, Patel BN, Patel CN. Spectrophotometric method for simultaneous estimation of atazanavir sulfate and ritonavir in tablet dosage form. Drug Dev Ther 2015;6:1-4

How to cite this URL:
Patel DA, Patel BN, Patel CN. Spectrophotometric method for simultaneous estimation of atazanavir sulfate and ritonavir in tablet dosage form. Drug Dev Ther [serial online] 2015 [cited 2017 Dec 12];6:1-4. Available from: http://www.ddtjournal.org/text.asp?2015/6/1/1/148880


  Introduction Top


Atazanavir sulfate (ATV) is methyl N′-[(1S)-1-{N-[(2S,_3S)-2-hydroxyl-3-[-(2S)-2-[(methoxycarbonyl)_amino]-3,3-dimethylbutanamido]-4-phenylbutyl]-N′-{[4-(pyridin-2-yl)phenyl]methyl}hydrazine carbonyl}-2,2-dimethyl propyl]carbamate [Figure 1], an azapeptide is the 7 th protease inhibitor used in the treatment of human immunodeficiency virus type II infection. It is official in Indian Pharmacopeia(IP). [1] Ritonavir (RTV) chemically is (5S,_8S,_10S,_11S)-10-hydroxy-2-methyl-5-(1-methyl ethyl)-1-[2-(1-methyl ethyl)-4-thiazolyl]-3,6-dioxo-8,11-bis(phenylmethyl)-2, 4, 7, 12-tetraazatridecan-13-oic acid 5-thiazolyl methylester [Figure 2]. It is official in IP [2] , USP [3] , BP. [4] Both the drugs were used as antiretroviral agents. ATV is a known substrate for both hepatic metabolizing enzyme cytochrome P450 and intestinal drug efflux pump, P-glycoprotein so have low oral bioavailability. So co-administration of a small dose of RTV is recommended as booster.Literature survey revealed that ATV and RTV was quantitatively assayed in biological fluids either individually or in the presence of other retroviral drugs using the liquid chromatography, [5],[6],[7],[8] estimation of ATV and RTV alone or combination with other antiretro viral drug in bulk and pharmaceutical dosage forms by ultraviolate(UV)-visible spectrophotometric method, [9],[10],[11],[12],[13] reversed phase- high performance liquid chromatography method, [14],[15],[16],[17],[18],[19],[20] high performance thin layer chromatographic. [21],[22] The aim of this work is to develop a simple, accurate and precise absorbance correction method by using UV spectrophotometry for the simultaneous estimation of ATV and RTV in combined tablet dosage. The method was validated for different parameters like linearity, accuracy, precision, robustness, ruggedness, limit of detection (LOD) and limit of quantification (LOQ) according to International conference of Harmonazation Guideline. [23]
Figure 1: Chemical structure of atazanavir sulfate

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Figure 2: Chemical structure of chemical structure of ritonavir

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  Materials and Methods Top


Instrument

The present work was carried out on Shimadzu UV-1700 double beam spectrophotometer with 1 cm path length supported by [Shimadzu, model-1700(Japan), UV-Probe software, version 2.31] was used for spectral measurements with 10 mm matched quartz cells.

Chemical and solvent

Standard of ATV and RTV were procured by Cipla Pharmaceutical Ltd. Methanol was purchased from Finar Chemicals Pvt. Ltd.

Method: Absorbance correction method

In this method, [24] absorbance of interfering component, i.e., absorbance of RTV was corrected from the total absorbance by the following formula:

Cx = A1/ax1 (1)

Cy = [A2-(ax2*Cx)]/ay2 (2)

where Cx and Cy are the concentrations of ATV and RTV in g/100 mL in the sample, respectively, A1 and A2 are the absorbance of the sample at λ1 (279 nm) and λ2 (240 nm), respectively, ax1 and ax2 are the specific absorptivities of ATV at λ1 (279 nm) and λ2 (240 nm), respectively, ay2 is the specific absorptivities of RTV at λ2 (240 nm).

Procedure

Solubility test


Solubility test for the drug was performed by using various solvents. The solvents include water, methanol, acetonitrile, 0.1 N HCl, and 0.1 N NaOH. However, methanol was chosen as a solvent for developing the method.

Preparation of standard stock solutions and calibration curve

Standard stock solutions of pure drug containing 1000 μg/mL of ATV and RTV were prepared separately in methanol. Standard stock solutions were further diluted with methanol to get working standard solutions of analytes in the concentration range of 30-90 and 10-30 μg/mL of ATV and RTV, respectively and scanned in the range of 200-400 nm.

Preparation of sample solution and formulation analysis

Twenty tablets were weighed accurately and a quantity of tablet powder equivalent to 30 mg of ATV (10 mg RTV) was weighed and dissolved in the 30 mL of methanol with the aid of ultrasonication for 10 min and the solution was filtered through Whatman paper no. 41 into a 100 mL volumetric flask. Filter paper was washed with solvent, adding washings to the volumetric flask; volume was made up to the mark with methanol. The solution was suitably diluted with methanol to get 30 μg/mL ATV and 10 μg/mL of RTV. Measure the absorbance at 240 and 279 nm [Figure 3] and [Figure 4].
Figure 3: Overlay spectra of atazanavir sulfate

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Figure 4: Overlay spectra of ritonavir

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  Results and Discussion Top


The proposed method was found to be simple, accurate, and economic for routine simultaneous estimation of these two drugs in combined dosage form. The values of relative standard deviation (RSD) are satisfactory low and recovery was closed to 100%, indicating reproducibility and accuracy of the method. This method gave excellent result and can be employed for routine analysis.

Assay of formulation

Absorbance correction method was found to be suitable to quantify ATV and RTV in tablet dosage form. Assay of ATV and RTV were found to be within IP limit [Table 1].
Table 1: Assay of formulations


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Method validation

Linearity


The calibration curves were constructed at optimum experimental conditions using absorbance values versus concentration in the range of 30-90 and 10-30 μg/mL for ATV and RTV, respectively. The results are reported in [Table 2].
Table 2: Optical characteristic of the method


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Recovery study

To check the accuracy of the method, recovery studies were carried out by the addition of standard drug solution to pre-analyzed sample solution at three different levels 80%, 100% and 120% [Table 3].
Table 3: Result of accuracy study


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Precision

Intra-day precision and inter-day precision were determined by analyzing ATV (30, 60 and 90 μg/mL) and RTV (10, 20 and 30 μg/mL) for 3 times on the same day and on different days. The results were reported in terms of %RSD [Table 4].
Table 4: Precision


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LOD and LOQ

The LOD and LOQ were calculated using following formula:

Where,

σ = standard deviation of response.

S = slope of the calibration curve.

Specificity

The formulation was assayed in the presence of the excipients by the proposed methods. It was found that there is no interference of the excipients which justifies the specificity of the drug for the proposed method.


  Conclusion Top


The proposed method is simple, specific, accurate and precise and hence can be used in routine for estimation of ATV and RTV in tablet dosage form. The percentage RSD for all parameters was found to be <2, which indicates the validity of the method and assay results obtained by this method are in fair agreement.

 
  References Top

1.
Government of India Mimistry of Health and Family Welfar. Indian Pharmacopoeia. Vol. 2. New Delhi: The Indian Pharmacopoeia Commission; 2010. P. 845-7.  Back to cited text no. 1
    
2.
Government of India Mimistry of Health and Family Welfar. Indian Pharmacopoeia. Vol. 2. New Delhi: The Indian Pharmacopoeia Commission; 2010. P. 845-7.  Back to cited text no. 2
    
3.
United State Pharmacopiea. Rockville: United State Pharmacopiea Convention. Inc.; 2004. p.3143.  Back to cited text no. 3
    
4.
British Pharmacopoeia. Norwich: Licensing Division HMSO; 2009. P.5227.  Back to cited text no. 4
    
5.
Dickinson L, Robinson L, Tlia J, Khoo S, Back D. Simultaneous determination of HIV protease inhibitors amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir in human plasma by high-performance liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2005;829:82-90.  Back to cited text no. 5
    
6.
Ehrhardt M, Mock M, Haefeli WE, Mikus G, Burhenne J, Monitoring of lopinavir and ritonavir in peripheral blood mononuclear cells, plasma, and ultrafiltrate using a selective and highly sensitive LC/MS/MS assay. J Chromatogr B Analyt Technol Biomed Life Sci 2007;850:249-58.  Back to cited text no. 6
    
7.
Weller DR, Brundage RC, Balflour HH Jr, Vezina HE. An isocratic liquid chromatography method for determining HIV non-nucleoside reverse transcriptase inhibitor and protease inhibitor concentration in human plasma. J chromatogr B Analyt Technol Biomed Life Sci 2007;848:369-73.  Back to cited text no. 7
    
8.
Turner ML, Reed-Walker K, King JR, Acosta EP. simultaneous determination of nine antiretroviral compounds in human plasma using liquid chromatography. J Chromatogr B Analyt Technol Biomed Life Sci 2003;784:331-41.  Back to cited text no. 8
    
9.
Khanage SG, Deshmukh VK, Mohite PB. Development of derivative spectrophotometric estimation of atazanavir sulfate in bulk drug and pharmaceutical dosage forms. Int J Pharm Health Sci 2010;3:149-54.  Back to cited text no. 9
    
10.
Chiranjeevi K, Channabasavaraj KP, Reddy PB. Development and validation of spectrophotometric method for quantitative estimation of ritonavir in bulk and pharmaceutical dosage forms. Int J of Chem Technol Res 2011;3:58-62.  Back to cited text no. 10
    
11.
Dey S, Reddy Y, Reddy T. Method development and validation for the estimation of atazanavir and dosage forms and its stress degradation studies using Uv-vis spectrophotometric method. Int J Pharm Bio Sci 2010;1:47-52.  Back to cited text no. 11
    
12.
Bari NA, Kela SP, Sharma SN. Spectrophotometric simultaneous determination of atazanavir and ritonavir in combined tablet dosage form by spectroscopic method. Der Pharm chem 2012;4:208-13.  Back to cited text no. 12
    
13.
Dinakaran SK, Machana S,Avasarala H, Desari AN. Spectrophotometric method development and validation for atazanavir sulphate and ritonavir in bulk and tablet dosage form using absorption ratio method. Am J Pharmtech Res 2013;3:645-54.   Back to cited text no. 13
    
14.
Konidala SK, Rani P. New validated RP-HPLC method for the determination of atazanavir sulphate in bulk and dosage form. Der Pharm Chem 2012;4:1305-10.  Back to cited text no. 14
    
15.
Suneethaa A, Kathirvela S, Ramachandrikaa G. A validated RP-HPLC method for simultaneous estimation of lopinavir and ritonavir in combined dosage form. Int J Pham Pharma Sci 2011;3:49-51.  Back to cited text no. 15
    
16.
Reddish CV, Devi PR, Makkanti K. Simultaneous estimation of atazanavirsulfate and ritonavir by RP-HPLC method in combined tablet dosage forms and its in vitro dissolution assessment. Novus Int J Anal Innov. 2012;1:5-14.  Back to cited text no. 16
    
17.
Ravindra RY, Swetha MA. Method development and validation of atazanavir and ritonavir in a combined dosage form by RP-HPLC method. Int J Pharm Technol 2011;3:3316-34.  Back to cited text no. 17
    
18.
Tiyyaguru A, Gunda A, Chitturi AR. Method development andvalidation for the simultaneous estimation of atazanavir and ritonavir in pharmaceutical dosage form by RP-HPLC. Int J Pharm Chem Bio Sci 2011;3:44-54.  Back to cited text no. 18
    
19.
Gadhvi MP, Bhandari A, Suhagia BN, Desai UH. Development and validation of RP-HPLC method for simultaneous estimation of atazanavir and ritonavir in their combined dosage form. Res J Pharm Technol 2013;6:200-3.  Back to cited text no. 19
    
20.
Rao RN, Ramachandra B, Vali RM, Raju SS. LC-MS/MS studies of ritonavir and its forced degradation products. J Pharm Biomed Anal 2010;53:833-42.  Back to cited text no. 20
    
21.
Nanda RK, Yadav PB. Stability-Indicating validated HPTLC method for Simultaneous estimation of atazanavir sulfate and ritonavir in Pharmaceutical dosage form. Asian J Res Chem 2011;4:1381.  Back to cited text no. 21
    
22.
Gadhvi MP, Bhandari A, Suhagia BN, Desai UH. Simultaneousdetermination of ritonavir and atazanavir in combined tablet dosage form by HPTLC. Asian J of Biomed Pharm Sci 2012;2:15-9.  Back to cited text no. 22
    
23.
The European Agancy for the Evaluation of Medical Products. ICH Topic Q2B, Guideline on Validation of Analytical Procedures: Methodology, GPMP/ICH/254/1-18;1996.  Back to cited text no. 23
    
24.
Beckett AH, Stanlake JB. Practical Pharmaceutical Chemistry. 3 rd ed. London The Athrone Press of University;1976; p. 249-50.  Back to cited text no. 24
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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