• R. D. Godambe Tatyasaheb Kore College of Pharmacy Warananagar, Dist: Kolhapur (MS) India
  • J. I. Disouza Tatyasaheb Kore College of Pharmacy Warananagar, Dist: Kolhapur (MS) India
  • C. M. Jamkhandi Tatyasaheb Kore College of Pharmacy Warananagar, Dist: Kolhapur (MS) India
  • P. S. Kumbhar Tatyasaheb Kore College of Pharmacy Warananagar, Dist: Kolhapur (MS) India



Spectrophotometry, Fluorometry, Darunavir, Condensation reaction


Objective: The main objective of the present study is to develop newer simple, precise spectrophotometric and fluorometric methods of estimation for Darunavir using coupling agent O-pthaladehyde.

Methods: The experimental work was designed for both spectroscopic and fluorometric method development and validation. The method is based on formation complex of Darunavir with O-pthaladehyde. QbD approach was applied by varying different parameters. These parameters were designed into Ishikawa diagram.

Results: The complex Darunavir-Phthalaldehyde in methanol with 0.1 N HCl showed linearity for both spectrophotometric and fluorometric methods. The calibration curve by spectrophotometry is linear in concentration range of 2-22 µg/ml with regression coefficient (R2) = 0.998 at 355 nm and for fluorometry it is linear in concentration range of 0.5-5.0 ng/ml with regression coefficient (R2) = 0.999. This method was found to be rugged and robust in different testing criteria with % RSD less than 2. The limit of detection and limit of quantification was found to be 0.2 μg/ml and 0.8 μg/ml for a spectrophotometric method and 0.12 μg/ml and 0.43 μg/ml for fluorometric method respectively.

Conclusion: Both methods were found to be precise with % RSD of less than 2. The % recovery of the spectrophotometric and fluorometric methods was found to be 101.04 %, 98.15 % respectively. In this way, the results of all validation parameter were within the limits as per International Conference on Harmonization guideline.



Download data is not yet available.


Rodger D. MacArthur Darunavir: promising initial resultsâ€. Doi:10.1016/S0140-673660499-1.

Meyer S, Azijn H, Surleraux D, Jochmans D, Tahri A. TMC114, a novel human immunodeficiency virus type 1 protease inhibitor active against protease inhibitor-resistant viruses, including a broad range of clinical isolates. Antimicrob Agents Chemother 2005;49:2314–21.

Tie Y, Boross PI, Wang YF, Gaddis L, A Hussain K. High-resolution crystal structure of HIV-1 protease with a potent non-peptide inhibitor active against multi-drug resistant. J Mol Biol 2004;338:341–52.

King N, Jeyabalan M, Nalivaika E, PM Wigerinck, C Bethun. Schiffer, the structural and thermodynamic basis for the binding of TMC114, a next-generation human immunodeficiency virus type 1 protease inhibitor. J Virol 2004;78:12012–21.

Nishendu P, Nadpara, Rakshit V, Thumar, Vidhi N, Kalola, et al. International Journal of pharmaceutical sciences review and research. Rev Res 2012;17:20-8.

Ermer J. Quality by design in pharmaceutical analysis. Conference organized by European Compliance Academy, Vienna, Austria; 2013.

Warf SF. The state of QbD in the biopharmaceutical industry conference criticality assessment, design space implementation and control†in Proceedings of the Conference Sponsored by ISPE—Endorsed by CASSS with a Mutual Commitment to Quality Products, San Francisco, Calif, USA; 2013.

Lawrence XYu. Pharmaceutical quality by design: product and process development, understanding, and control. Pharma Res 2008;25:781-91.

Nethercote P, Borman P, Bennett T, Martin G, McGregor P. QbD for better method validation and transfer; 2010. Available from: http://www.pharmamanufacturing. com/articles/2010/ 060.html [Last accessed on 20 Oct 2017].

Elder P, Borman P. Improving analytical method reliability across the entire product lifecycle using QbD approaches. Pharm Outsourcing 2013;14:14–9.

Correa JC, D Arcy DM, Serra CH, Salgado HR. A critical review of properties of darunavir and analytical methods for its determination. Crit Rev Anal Chem 2014;44:16-22.

CM Jamakhandi, Chandrashekar Javali, Santosh Kumar, Sanjay Kumar DS, Dayanand Kadadevar. New fluorimetric method of development for lisinopril by a condensation reaction. Int J Pharm Pharm Sci 2010;2:209-11.

CM Jamakhandi, C Javali, JI Disouza, US Chougule, AK Mullani. Spectrophotometric determination of lisinopril dosage form by a condensation reaction. Int J Pharm Pharm Sci 2011;3:185-7.

International Conference on Harmonisation Tripartite Guideline Q9 Pharmaceutical Risk Management; 2005.

P Elliott, S Billingham, J Bi, Hu Zhang. Quality by design for biopharmaceuticals: a historical review and guide for implementation. Pharm Bioprocessing 2013;1:105-22.

P Borman. The application of quality by design to analytical methods. Pharma Tech 2007;31:142-52.

J Castillo, O Benavente, F Borrego, J Delri. Analysis of commercial hesperidin methyl chalcone by high-performance liquid chromatography. J Chromatography 1991;555:285-90.

Leposava M Jasmina. Development and validation of a fluorometric method for the determination of hesperidin in human plasma and pharmaceutical forms. J Serb Chem Soc 2012;77:1625-40.

International Conference on Harmonization (ICH) of Technical Requirements for Registration of Pharmaceuticals for Human Use, Topic Q9: Quality Risk Management, Geneva; 2005.

Y Li, GJ Terfloth, AS Kord. A systemic approach to RP-HPLC method development in a pharmaceutical QbD environment. Am-Pharma 2009;12:571-83.

FDA, Pharmaceutical GMPs for the 21st century–a risk-based approach (U. S Department of health and human services, Food and Drug Administration, U. S Government Printing Office, Washington DC; 2002.

MR Ghante, RS Shelar, SD Sawant, MM Kadam. Development and validation UV-spectrophotometric method for estimation of darunavir ethanolate in bulk and tablet dosage form. Int J Pharma Pharm Sci 2014;6:240-2.

K Balamuralikrishna. Development and validation of analytical procedure for the simultaneous estimation of efavirenz, lamivudine and zidovudine through new RP-HPLC method. J Pharm Res 2011;4:3766-8.



How to Cite

Godambe, R. D., J. I. Disouza, C. M. Jamkhandi, and P. S. Kumbhar. “DEVELOPMENT OF SPECTROPHOTOMETRIC AND FLUOROMETRIC METHODS FOR ESTIMATION OF DARUNAVIR USING QBD APPROACH”. International Journal of Current Pharmaceutical Research, vol. 10, no. 1, Jan. 2018, pp. 13-19, doi:10.22159/ijcpr.2018v10i1.24401.



Original Article(s)