DEVELOPMENT AND VALIDATION OF A STABILITY INDICATING RP-HPLC METHOD FOR THE DETERMINATION OF POTENTIAL DEGRADATION PRODUCTS OF DIFLUPREDNATE IN OPHTHALMIC EMULSION

  • Murlidhar V. Zope Ramanbhai Patel College of Pharmacy, Charotar University of Science and Technology, Charusat Campus, Changa, 388421, Ta. Petlad, Dist.: Anand, Gujarat, India
  • Rahul M. Patel Sun Pharma Advanced Research Company Ltd., Tandalja, Vadodara, Gujarat, India
  • Ashwinikumari Patel Sun Pharma Advanced Research Company Ltd., Tandalja, Vadodara, Gujarat, India
  • Samir G. Patel Ramanbhai Patel College of Pharmacy, Charotar University of Science and Technology, Charusat Campus, Changa, 388421, Ta. Petlad, Dist.: Anand, Gujarat, India

Abstract

Objective: The objective of the current study was to develop and validate a simple, robust, precise and accurate RP-HPLC (reverse phase-high performance liquid chromatography) method for the quantitative determination of potential degradation products of Difluprednate (DIFL) in the ophthalmic emulsion.

Methods: Chromatographic separation was achieved on the YMC pack ODS-AQ (150× 4.6) mm, 3μm column with a mobile phase containing a gradient mixture of mobile phase A (0.02M Ammonium formate buffer pH 4.5 adjusted with formic acid) and Acetonitrile as mobile phase B, at flow rate of 1.5 ml/min and with UV detection at 240 nm.

Results: The peak retention time of DIFL was found at about 17.2 min, the RRT of degradation product-1 (DP-1), degradation product-2 (DP-2), and degradation product-3 (DP-3), were found to be about 0.49, 0.65 and 0.79 respectively (calculated with respect to Difluprednate). Stress testing was performed in accordance with an ICH (international council for harmonisation) guideline Q1A (R2) [1]. The method was validated as per ICH guideline Q2 (R1)[2]. The calibration curve was found to be linear in the concentration range of 0.1 to 0.75 µg/ml for Difluprednate, DP-1, DP-2 and DP-3. The LOD (Limit of detection) was found to be 0.1µg/ml and LOQ (Limit of quantification) of 0.15µg/ml for Difluprednate, DP-1, DP-2 and DP-3 respectively. The recovery from LOQ to 150% was within 90-110%. The forced degradation data confirms the stability indicating the nature of the method.

Conclusion: A simple, robust, precise and accurate RP-HPLC method for the quantitative determination of potential degradation products of Difluprednate in the ophthalmic emulsion was developed and validated. 

Keywords: Difluprednate, Stability indicating, RP-HPLC, Degradation products, Forced degradation

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References

1. ICH harmonized tripartite guideline, Q2A (R2) stability testing of new drug substances and product. Geneva: International Conference on Harmonization; 2003.
2. ICH harmonised tripartite guideline, Q2 (R1) validation of analytical procedures: text and methodology. Geneva: International Conference on Harmonization; 2005.
3. ICH harmonised tripartite guideline, Q1B stability testing: photostability testing of new drug substances and products. Geneva: International Conference on Harmonization; 2005.
4. Shin Ichi Y, Masako K, Akira O, Kazuaki K. Analysis of an anti-inflammatory steroidal drug, difluprednate, in aqueous humor by a combination of semi-micro HPLC and column switching method. J Pharm Biomed Anal 2003;30:1735-42.
5. Inoue J, Yamaguchi M, Sakaki H, Kida T, Tajika T. Preclinical pharmacokinetics of difluprednate emulsion. ‎Invest Ophthalmol Vis Sci 2007;48:2651.
6. Mangukiya RP, Patel RA, Shah PA, Patel KG. Chromatographic methods for simultaneous determination of moxifloxacin hydrochloride and difluprednate in the ophthalmic dosage form. Acta Chromatographica 2015;27:495–8.
7. Patel P, Patel D, Meshram D. Validated RP-HPLC method for simultaneous determination of difluprednate and gatifloxacin in opthalmic emulsion. Int J Adv Res 2014;2:484-92.
8. Pradhan PK, Patel M, Mavani D, Raiyani N, Upadhyay U. RP-HPLC method development and validation for simultaneous estimation of moxifloxacin and difluprednate in their combined dosage form. J Drug Delivery Thera 2014;4:60-5.
9. Nagaraju P, Siddartha V, Priyadarshini IG. New drug approval process in singapore-a detailed dissertation for registration of multi-source drug products. Int J Pharm Chem Sci 2015;4:61-6.
10. Patel P, Patel D, Desai S, Meshram D. Development and validation of analytical methods for simultaneous estimation of difluprednate and gatifloxacin in ophthalmic emulsion by uv-visible spectroscopy. Int J Pharm Sci Invent 2014;3:1-10.
11. ICH harmonised tripartite guideline, Q3B (R2) impurities in new drug products. Geneva: International Conference on Harmonization; 2006.
12. Mohammed IB, Prakash VK, Krishna MG. RP-HPLC method for simultaneous estimation of metformin and vildagliptin in bulk and its tablet formulation. J Global Trends Pharm Sci 2012;3:747-54.
13. Pratik Kumar Gupta, Vibha Chaturvedi. A study on forced degradation and validation of stability indicating RP-HPLC method for determination of benzalkonium chloride in azelastine hydrochloride pharmaceutical ophthalmic formulation. Asian J Pharm Clin Res 2017;10:374-82.
14. Babu C, Narasimha Rao KL, Devanna N, Suresh Reddy KVN. Development and validation of stability indicating reversed phase high performance liquid chromatographic method for the determination of related substances in fampridine drug substance and tablet dosage forms. Asian J Pharm Clin Res 2017;10:334-8.
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Zope, M. V., R. M. Patel, A. Patel, and S. G. Patel. “DEVELOPMENT AND VALIDATION OF A STABILITY INDICATING RP-HPLC METHOD FOR THE DETERMINATION OF POTENTIAL DEGRADATION PRODUCTS OF DIFLUPREDNATE IN OPHTHALMIC EMULSION”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 9, Sept. 2018, pp. 79-86, doi:10.22159/ijpps.2018v10i9.26342.
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