Open AccessDesign of Experimental Approach to Analytical Robustness Study for UHPLC Method Developed for Separation and Quantification of Spironolactone and its Impurities in Drug Substances
Author(s) -
Y.V.S. Veerendra,
Pradeep Kumar Brahman,
Sharad D. Mankumare,
Jaya Raju,
Satish Jetta
Publication year - 2019
Publication title -
asian journal of chemistry/asian journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.145
H-Index - 34
eISSN - 0975-427X
pISSN - 0970-7077
DOI - 10.14233/ajchem.2020.22224
Subject(s) - robustness testing , robustness (evolution) , factorial experiment , chromatography , chemistry , design of experiments , elution , fractional factorial design , forced degradation , quality by design , impurity , methanol , central composite design , plackett–burman design , formic acid , analytical chemistry (journal) , response surface methodology , high performance liquid chromatography , computer science , reversed phase chromatography , mathematics , software , machine learning , particle size , statistics , biochemistry , organic chemistry , gene , programming language
The present work demonstrates the development of an optimal, robust, validated UHPLC method forquantification of related impurities and assay determination of spironolactone. Design of experimentprocedure, in combination with statistical evaluation of the data was used to test the robustness ofdeveloped method. A stability indicating method was established by forced degradation experiments.Analytical robustness was determined using design of experiment approach. The chromatographicseparation was achieved with Agilent SB-C18 RRHD column using gradient elution with mobilephase-A consists of a mixture of 0.1 % each formic acid and ammonia in water and methanol asmobile phase-B respectively. The developed method is exhaustively validated for parameters likeprecision, accuracy, linearity, LOD, LOQ, ruggedness and robustness. The stability tests were alsoperformed on drug substances as per ICH norms. Base line separation was achieved for all impurities,degradation products and the API. All impurities were eluted within 12 min, there was a remarkable3.5-fold decrease in runtime and a clear baseline separation between all peaks in comparison withPh.Eur monograph. A multi-dimensional design space was built to study the robustness of developedmethod using design expert software. Significant parameters such as effect of flow rate, buffer strengthand mobile phase compositions were optimized at three levels. Plackett-Burman design was appliedfor screening of chromatographic conditions and factorial design was applied for optimization ofessential factors in robustness studies.