Development And Validation of a LC-MS Compatible Method for Quantification of Degradation Impurities of Clofazimine Using UHPLC

Author(s): Nagulakonda Naga Veera Venkata Sri Surya Narayana Murty*, Akash Bhattacharjee*, Tatikonda Krishnamurthy, Muguda Ravi Prasada Rao, Gollapalli Nageswara Rao

Journal Name: Current Pharmaceutical Analysis

Volume 16 , Issue 7 , 2020

Become EABM
Become Reviewer

Graphical Abstract:


Abstract:

Background: Clofazimine has antibacterial and leprostatic properties, which has its use in Multidrug Therapy (MDT) of leprosy. As per the FDA guidance for industry, each NDA and ANDA must include the analytical procedures necessary to ensure the identity, strength, quality, purity, and potency of the drug substance and drug product. However, it was noticed that no stability indicating method is available in the literature for the estimation of degradation impurities of Clofazimine.

Objective: Objective of the proposed work is to develop and validate a rapid, specific, linear, robust, accurate and sensitive Ultra High-Performance Liquid Chromatography (UHPLC) method with LC-MS compatible mobile phase for the quantification of degradation impurities of Clofazimine in a pharmaceutical dosage form (topical gel 0.5% w/w).

Method: Ultra High-Performance Liquid Chromatography equipped with PDA and Tunable UV (TUV) detector at a wavelength of 284 nm, stationary phase with a fused core particle technology, LC-MS compatible mobile phase was employed in this study. Gradient elution was employed for ensuring the selectivity of degradation impurities and clofazimine. This method was validated in accordance with ICH Q2 guidelines. This is the first reported Ultra High-Performance Liquid Chromatography method for estimation of degradation impurities of clofazimine.

Results: The method showed good linearity over the range of 0.25 -1.5μg/ml of clofazimine. All the validation parameters were within the acceptance criteria. The product is found to degrade in the acid and peroxide degradation condition. The major degradant impurities are eluted at relative retention times of 0.35, 0.89 and 0.95. The developed method successfully separated the degradation products of clofazimine and able to quantitate accurately in its formulation.

Conclusion: To date, there is no UHPLC method for determination of degradation impurities of clofazimine. in pharmaceutical dosage forms. Being a specific, linear, accurate and robust method, this would help in determining the chemical stability of drug product during the product development as well as in the shelf life of the drug product.

Keywords: Clofazimine, leprosy, UHPLC, chromatography, degradation impurities, fused core particle.

[1]
Suzuki, K.; Akama, T.; Kawashima, A.; Yoshihara, A.; Yotsu, R.R.; Ishii, N. Current status of leprosy: epidemiology, basic science and clinical perspectives. J. Dermatol., 2012, 39(2), 121-129.
[http://dx.doi.org/10.1111/j.1346-8138.2011.01370.x] [PMID: 21973237]
[2]
Venkatesan, K. Clinical pharmacokinetic considerations in the treatment of patients with leprosy. Clin. Pharmacokinet., 1989, 16(6), 365-386.
[http://dx.doi.org/10.2165/00003088-198916060-00003] [PMID: 2661102]
[3]
Patel, V.B.; Misra, A.N.; Marfatia, Y.S. A topical dosage form of liposomal clofazimine: research and clinical implications. Pharmazie, 1999, 54(6), 448-451.
[PMID: 10399190]
[4]
british pharmacopoeia commission. British Pharmacopoe-ia; British Pharmacopoeia Commission: Teddington; Mid-dlesex: UK, 2009, Vol. 3, pp. 2252-2254.
[5]
european pharmacopoeia commission and european directorate for the quality of medicines & healthcare. european pharmacopoeia 5.0 2005, 1.0, , 1324-1325.
[6]
Indian pharmacopoeia commission. Indian pharmacopoeia; indian Pharmacopoeia Commission, Ministry of Health & Family welfare; Government of India: Ghaziabad, India, 2014, Vol. 2, pp. 1428-1429.
[7]
Pharmacopeia, U.S. united states pharmacopeia and national formulary (usp 37–n32). 2014, 2, 2404-2406.
[8]
Siddiqui, M.R.; Othman, A.L. Z.A.; Rahman, N. Analytical techniques in pharmaceutical analysis: a review. Arab. J. Chem., 2017, 10, S1409-S1421.
[http://dx.doi.org/10.1016/j.arabjc.2013.04.016]
[9]
Kapoor, V.R. Shishu. A Novel Validation HPTLC method for the quantitative determination of clofazimine. Int. J. Pharma Bio Sci., 2013, 4(2), 819-828.
[10]
Saxena, S.; Singh, H.N.; Agarwal, V.K.; Rai, J.; Singh, S. Estimation of clofazimine in capsule dosage form by us-ing uv-vis spectroscopy. Int. J. Pharm. Pharm. Sci., 2013, 5(3), 635-638.
[11]
Krishnan, T.R.; Abraham, I. A rapid and sensitive high performance liquid chromatographic analysis of clofazimine in plasma. Int. J. Lepr. Other Mycobact. Dis., 1992, 60(4), 549-555.
[PMID: 1299710]
[12]
Queiroz, R.H.C.; Pereira, R.C.; Gotardo, M.A.; Cordeiro, D.S.; Melchior, E., Jr Determination of clofazimine in leprosy patients by high-performance liquid chromatography. J. Anal. Toxicol., 2003, 27(6), 377-380.
[http://dx.doi.org/10.1093/jat/27.6.377] [PMID: 14516492]
[13]
Shabada, A.; Badar, H.; Sultana, S.S.; Fatima, N. RP-HPLC Estimation of clofazimine in soft gelatine capsule: development, validation and stability indications. European J. Biomed. Pharmac. Sci., 2017, 4(12), 478-483.
[14]
Khedekar, G.; Mirgane, S. Development and validation of stability indicating HPLC assay method for Clofazimine capsules. Innovations in Pharmaceuticals and Pharma-cotherapy, 2017, 5(2), 112-120.
[15]
Lin, J.; Lin, Z.; Li, D.; Feng, M.; Yang, Q.; Hu, W.; Han, Y.; Zhu, W.; Li, M.; Yu, L. “Ghost peak” of clofazimine: A solution degradation product of clofazimine via nucleophilic substitution by nitrite leaching from certain glass HPLC vials. J. Pharm. Biomed. Anal., 2018, 150(052), 183-190.
[http://dx.doi.org/10.1016/j.jpba.2017.11.052] [PMID: 29245088]
[16]
Patel, H.A.; Patel, K.G.; Pathak, D.S.; Patel, D.J.; Meshram, D.B. Development and validation of stability indicating rp-hplc method for estimation of clofazimine in soft gelatine capsule. Int. J. Pharmaceut. Quality Ass., 2017, 8(1), 32-38.
[17]
Borner, K.; Hartwig, H.; Leitzke, S.; Hahn, H.; Müller, R.H.; Ehlers, S. HPLC determination of clofazimine in tissues and serum of mice after intravenous administration of nanocrystalline or liposomal formulations. Int. J. Antimicrob. Agents, 1999, 11(1), 75-79.
[http://dx.doi.org/10.1016/S0924-8579(98)00065-X] [PMID: 10075282]
[18]
O’Connor, R.; O’Sullivan, J.F.; O’Kennedy, R. Determination of serum and tissue levels of phenazines including clofazimine. J. Chromatogr. B Biomed. Appl., 1996, 681(2), 307-315.
[http://dx.doi.org/10.1016/0378-4347(96)00025-4] [PMID: 8811441]
[19]
Baijnath, S.; Naiker, S.; Shobo, A.; Moodley, C.; Adamson, J.; Ngcobo, B.; Bester, L.A.; Singh, S.; Kruger, H.G.; Naicker, T.; Govender, T. Evidence for the presence of clofazimine and its distribution in the healthy mouse brain. J. Mol. Histol., 2015, 46(4-5), 439-442.
[http://dx.doi.org/10.1007/s10735-015-9634-3] [PMID: 26208572]
[20]
Li, W.; Doherty, J.; Fu, Y.; Flarakos, J. Quantitative analysis of clofazimine (Lamprene®), an antileprosy agent, in human dried blood spots using liquid chromatography-tandem mass spectrometry. Biomed. Chromatogr., 2018, 32(2).,
[http://dx.doi.org/10.1002/bmc.4068] [PMID: 28833263]


Rights & PermissionsPrintExport Cite as

Article Details

VOLUME: 16
ISSUE: 7
Year: 2020
Published on: 17 August, 2020
Page: [856 - 866]
Pages: 11
DOI: 10.2174/1573412915666190304152329
Price: $65

Article Metrics

PDF: 24
HTML: 2