R. D. Godambe, J. I. Disouza, C. M. Jamkhandi, P. S. Kumbhar


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.



Spectrophotometry, Fluorometry, Darunavir, Condensation reaction

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Rodger D. MacArthur “Darunavir: promising initial results”. 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.

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Spectrophotometry, Fluorometry, Darunavir, Condensation reaction





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International Journal of Current Pharmaceutical Research
Vol 10, Issue 1 (Jan-Feb), 2018 Page: 13-19

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Authors & Affiliations

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


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