Development of an Innovative Quality by Design (QbD) Based Stability-Indicating HPLC Method and its Validation for Clofazimine from its Bulk and Pharmaceutical Dosage Forms

  • Tulshidas S. Patil
  • Ashwini S. DeshpandeEmail author


The present research work discusses the systematic Quality by Design (QbD) enabled development of a simple, rapid, economical, and stability-indicating high-performance liquid chromatography (HPLC) method for effective analysis of clofazimine (CFZ). An Ishikawa fish-bone diagram was constructed for initial risk assessment. Analytical target profile (ATP) was defined and critical analytical attributes (CAAs) were assigned to meet these ATP requirements. Taguchi design was applied for screening critical material attributes (CMAs) and critical process parameters (CPPs) making an impact on the assigned CAAs. The major contributing factors were finely tuned using 33 Box-Behnken design with numerical and graphical optimization. Further, the method was validated as per the ICH guidelines and force degradation studies were carried out under different stress conditions. The optimum chromatographic separation was accomplished using 75:25% v/v ratio of methanol and ammonium acetate buffer (0.01 mol/L) as the mobile phase at flow rate 1.0 mL/min, and UV detection at 284 nm. The developed HPLC method was found highly sensitive, specific with linearity ranging between 2 and 10 µg/mL, and correlation coefficient (R2) 0.9995. It showed high accuracy with % recovery between 99.68 and 100.44%. It depicted detection limit and quantitation limit of 0.0066 µg/mL and 0.0199 µg/mL, respectively. In force degradation studies the drug was found to be highly susceptible in alkaline stress conditions. The results reveal successful applicability of the method for the estimation of CFZ from its marketed formulation which can be wisely extrapolated to assess the CFZ from its other formulation systems and different biological samples.


Clofazimine HPLC Quality by design Design space Validation Stress testing 



Authors are grateful to Chancellor, NMIMS (Deemed-to-be University) and management of SVKM’s NMIMS, School of Pharmacy and Technology Management, Shirpur for providing various reprographic sources for implementing this work.

Compliance with ethical standards

Conflict of interest

Authors declare no conflict of interest.

Ethical standards

The present work is in compliance with ethical standards.

Supplementary material

10337_2018_3660_MOESM1_ESM.pptx (71 kb)
Suppl. Figure 1 Ishikawa fish-bone diagram depicting the possible risk factors for the development of HPLC method (PPTX 71 KB)
10337_2018_3660_MOESM2_ESM.pptx (225 kb)
Suppl. Figure 2 (a-b) Chromatograms of CFZ standard solution with different solvents (Organic modifiers) (a) acetonitrile and (b) Methanol (PPTX 225 KB)
10337_2018_3660_MOESM3_ESM.pptx (143 kb)
Suppl. Figure 3 Overlay plot depicting the optimized conditions of the analytical method (PPTX 143 KB)
10337_2018_3660_MOESM4_ESM.pptx (142 kb)
Suppl. Figure 4 (a-b) Chromatograms of CFZ from (a) standard solution and (b) sample extracted from marketed soft gelatin capsule (PPTX 141 KB)
10337_2018_3660_MOESM5_ESM.docx (19 kb)
Supplementary material 1 (DOCX 18 KB)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.School of Pharmacy & Technology ManagementSVKM’s NMIMSShirpurIndia

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