title: A simultaneous determination of related substances by high performance liquid chromatography in a drug product using quality by design approach.
abstract: The combination of Abacavir, Lamivudine and Dolutegravir is an anti-retroviral formulation that displays high efficacy and superiority in comparison to other anti-retroviral combinations. Analysis of related substances in this combination drug product was very challenging due to the presence of nearly thirty peaks including the three active pharmaceutical ingredients (APIs), eleven known impurities and other pharmaceutical excipients. Objective of this study was to develop a single, selective, and robust high performance liquid chromatography method for the efficient separation of all peaks. Initially, one-factor-at-a-time (OFAT) approach was adopted to develop the method. But, it could not resolve all the critical peaks in such complex matrix. This led to the advent of two different HPLC methods for the determination of related substances, one for Abacavir and Lamivudine and the other for Dolutegravir. But, since analysis of a single sample using two methods instead of one is time and resource consuming and thus expensive, an attempt was made to develop a single and robust method by adopting quality by design (QbD) principles. Design of Experiments (DoE) was applied as a tool to achieve the optimum conditions through Response surface methodology with three method variables, pH, temperature, and mobile phase composition. As the study progressed, it was discovered that establishment of the design space was not viable due to the completely distant pH requirements of the two responses, i.e. (i) retention time for Lamivudine carboxylic acid and (ii) resolution between Abacavir impurity B and unknown impurity. Eventually, neglecting one of these two responses each time, two distinguished design spaces have been established and verified. Edge of failures at both design spaces indicate high probability of failure. It therefore, becomes very important to identify the most robust zone or normal operating range (NOR) within the design space with low risk of failure and high quality assurance. For NOR establishment, Monte Carlo simulation was performed on the basis of which process capability index (Cpk) was derived. Finally, the selectivity issue problem faced due to the pH dependency and the dissimilar pH needs of the two critical responses was resolved by introducing pH gradient into the program. This new ternary gradient program has provided a single robust method. Thus, two HPLC methods for the analysis of the combination drug product have been replaced with a selective, robust, and cost effective single method.
authors.affiliation: Department of Quality by Design, Cipla Ltd., Vikhroli (West), Mumbai 400083, India.
authors.forename: Trupti
authors.initials: T
authors.lastname: Tol
authors.affiliation: Formulation Analytical Development Laboratories, Cipla Ltd., Vikhroli (West), Mumbai 400083, India.
authors.forename: Nilesh
authors.initials: N
authors.lastname: Kadam
authors.affiliation: Formulation Analytical Development Laboratories, Cipla Ltd., Vikhroli (West), Mumbai 400083, India.
authors.forename: Nilesh
authors.initials: N
authors.lastname: Raotole
authors.affiliation: Formulation Analytical Development Laboratories, Cipla Ltd., Vikhroli (West), Mumbai 400083, India.
authors.forename: Anita
authors.initials: A
authors.lastname: Desai
authors.affiliation: Department of Quality by Design, Cipla Ltd., Vikhroli (West), Mumbai 400083, India. Electronic address: gautam.samanta@cipla.com.
authors.forename: Gautam
authors.initials: G
authors.lastname: Samanta
mesh_terms: D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015224:Dideoxynucleosides / Q000032:analysis; D006575:Heterocyclic Compounds, 3-Ring / Q000032:analysis; D019259:Lamivudine / Q000032:analysis; D009010:Monte Carlo Method; D010078:Oxazines; D004364:Pharmaceutical Preparations / Q000032:analysis*; D010879:Piperazines; D011728:Pyridones; D013696:Temperature; D000099227:Abacavir; D000099226:Dolutegravir
chemical_list: D015224:Dideoxynucleosides; D006575:Heterocyclic Compounds, 3-Ring; D019259:Lamivudine; D010078:Oxazines; D004364:Pharmaceutical Preparations; D010879:Piperazines; D011728:Pyridones; D000099227:Abacavir; D000099226:Dolutegravir
keywords: Abacavir; Antiretroviral; Dolutegravir; Lamivudine; Monte Carlo simulation; Normal operating range; Quality by design; Robustness