Quality by Design: Design of Experiments Approach Prior to the Validation of a Stability-Indicating HPLC Method for Montelukast

This paper describes a systematic design of experiments (DoE) approach by applying the principle of quality by design (QbD) to determine the design space for a stability-indicating HPLC method prior to validation. By employing DoE, a simultaneous multivariate approach was carried out for mobile phase pH, flow rate, percentage of organic content and column temperature. A two-level fractional factorial design (2⁴⁻¹ + 2 center points = 10 experiments) was employed and statistical analysis of the experimental data uncovered the significant influential chromatographic factors. The experimental data for USP tailing and resolution were analyzed statistically to screen the chromatographic factors. This approach determined the most influential chromatographic factors. During this process, inferences were evaluated from various data tables, for example, analysis of variance, summary of fit, lack of fit, and parameter estimates. The study also explained various plots such as actual vs. predicted plot, Pareto plot, and prediction profiler. The acceptable range of the chromatographic factors was displayed as a Contour plot defining the ‘design space’ of the method. The range of operating conditions that guarantee a satisfactory QbD was deduced to finalize the method prior to validation. The method is simple, rapid, and robust for the determination of montelukast in montelukast sodium oral granules dosage form. The method was validated according to ICH guidelines for accuracy, precision, linearity, range, specificity, ruggedness and robustness (one factor varied at a time). The method has been successfully transferred to the quality control department for quality analysis of manufactured batches and stability samples.

     

Application of Experimental Design Methodologies in the Enantioseparation of Pharmaceuticals by Capillary Electrophoresis: A Review

Chirality is one of the major issues in pharmaceutical research and industry. Capillary electrophoresis (CE) is an interesting alternative to the more frequently used chromatographic techniques in the enantioseparation of pharmaceuticals, and is used for the determination of enantiomeric ratio, enantiomeric purity, and in pharmacokinetic studies. Traditionally, optimization of CE methods is performed using a univariate one factor at a time (OFAT) approach; however, this strategy does not allow for the evaluation of interactions between experimental factors, which may result in ineffective method development and optimization. In the last two decades, Design of Experiments (DoE) has been frequently employed to better understand the multidimensional effects and interactions of the input factors on the output responses of analytical CE methods. DoE can be divided into two types: screening and optimization designs. Furthermore, using Quality by Design (QbD) methodology to develop CE-based enantioselective techniques is becoming increasingly popular. The review presents the current use of DoE methodologies in CE-based enantioresolution method development and provides an overview of DoE applications in the optimization and validation of CE enantioselective procedures in the last 25 years. Moreover, a critical perspective on how different DoE strategies can aid in the optimization of enantioseparation procedures is presented.     

Integrated quality by design (QbD) and design of experiments (DoE) approach for UFLC determination of telaprevir in rat serum

An ultrafast liquid chromatographic bioanalytical method was developed and validated for the determination of telaprevir in Wistar albino rat serum. Principles of quality by design (QbD) were implemented for enhancing the bioanalytical liquid–liquid extraction of telaprevir from rat serum. A Box–Behnken design was utilized in the studies by selecting extraction time, centrifugation speed, and vortex time as the critical method variables for evaluating their effect on the critical analytical attribute, i.e., %recovery of telaprevir. Chromatographic separation was achieved within a run time of 10?min using a C-18 column and mobile phase comprising of methanol:borate buffer of pH 9 (90:10 v/v) flowing at 1.2?mL/min. Photodiode array detection was performed at 270?nm. Results of validation studies were satisfactory. The method was linear over a concentration of 25–10,000?ng/mL. Limit of detection for the developed method was 10?ng/mL. Further, design of experiments (DoE) used for inter-day accuracy and precision study suggested superior method reliability. This integrated QbD- and DoE-based approach ensured the development of a validated and reliable analytical method for optimum bioanalysis of telaprevir in biological matrix.     

QbD‐oriented development and validation of a bioanalytical method for nevirapine with enhanced liquid–liquid extraction and chromatographic separation

The present studies describe the systematic quality by design (QbD)‐oriented development and validation of a simple, rapid, sensitive and cost‐effective reversed‐phase HPLC bioanalytical method for nevirapine in rat plasma. Chromatographic separation was carried out on a C₁₈ column using isocratic 68:9:23% v/v elution of methanol, acetonitrile and water (pH 3, adjusted by orthophosphoric acid) at a flow rate of 1.0 mL/min using UV detection at 230 nm. A Box–Behnken design was applied for chromatographic method optimization taking mobile phase ratio, pH and flow rate as the critical method parameters (CMPs) from screening studies. Peak area, retention time, theoretical plates and peak tailing were measured as the critical analytical attributes (CAAs). Further, the bioanalytical liquid–liquid extraction process was optimized using an optimal design by selecting extraction time, centrifugation speed and temperature as the CMPs for percentage recovery of nevirapine as the CAA. The search for an optimum chromatographic solution was conducted through numerical desirability function. Validation studies performed as per the US Food and Drug Administration requirements revealed results within the acceptance limit. In a nutshell, the studies successfully demonstrate the utility of analytical QbD approach for the rational development of a bioanalytical method with enhanced chromatographic separation and recovery of nevirapine in rat plasma. Copyright © 2015 John Wiley & Sons, Ltd.     

Robust design space development for HPLC analysis of five chemical components in Panax notoginseng saponins

The present work describes the systematic development of a robust, precise, and rapid reversed-phase liquid chromatography method for the simultaneous analysis of five chemical components in Panax notoginseng saponins (PNS) using quality by design (QbD). The method was developed in two main phases: screening and optimization. During the screening phase, the most suitable stationary phase, column temperature, and flow rate were identified, while the secondary influential parameters, such as the gradient slope, the initial concentration of acetonitrile, and the initial isocratic hold of the gradient elution system were fine-tuned in the later optimization phase. In this phase, a 17-run experiment was used to examine multifactorial effects of these parameters on the critical resolution, analysis time, and peak symmetry. The Monte Carlo simulation method was successfully used to build the chromatographic design space and the process capability index C_p was introduced to evaluate the robustness of the design space. At last, the verification experiment was performed within the working design space by the accuracy profile methodology and model was found to be accurate. A robust combination of separation conditions predicted in the design space was obtained with the gradient slope of 1.6% · min^?1, the initial concentration of acetonitrile of 20%, and the initial isocratic hold of 20 min, and the total analysis time was no more than 40 min. The results demonstrated that rapid separation of five components of PNS herbal extract could be achieved on the chromatographic column packed with narrow size particles at backpressures available in ordinary high performance liquid chromatography (HPLC) instruments.     

Quality by Design Study of the Direct Analysis in Real Time Mass Spectrometry Response

A mass spectrometry method has been developed using the Quality by Design (QbD) principle. Direct analysis in real time mass spectrometry (DART-MS) was adopted to analyze a pharmaceutical preparation. A fishbone diagram for DART-MS and the Plackett-Burman design were utilized to evaluate the impact of a number of factors on the method performance. Multivariate regression and Pareto ranking analysis indicated that the temperature, determined distance, and sampler speed were statistically significant (P < 0.05). Furthermore, the Box-Behnken design combined with response surface analysis was then employed to study the relationships between these three factors and the quality of the DART-MS analysis. The analytical design space of DART-MS was thus constructed and its robustness was validated. In this presented approach, method performance was mathematically described as a composite desirability function of the critical quality attributes (CQAs). Two terms of method validation, including analytical repeatability and method robustness, were carried out at an operating work point. Finally, the validated method was successfully applied to the pharmaceutical quality assurance in different manufacturing batches. These results revealed that the QbD concept was practical in DART-MS method development. Meanwhile, the determined quality was controlled by the analytical design space. This presented strategy provided a tutorial to the development of a robust QbD-compliant mass spectrometry method for industrial quality control.

A Validated LC Method for Determination of the Enantiomeric Purity of Montelukast Sodium in Bulk Drug Samples and Pharmaceutical Dosage Forms

A new and accurate chiral liquid chromatographic method has been developed for determination of the enantiomeric purity of montelukast sodium (R enantiomer) in bulk drugs and dosage forms. Normal phase chromatographic separation was performed on an immobilized amylose-based chiral stationary phase with n-hexane–ethanol–1,4-dioxane–trifluoroacetic acid–diethylamine 65:25:10:0.3:0.05 (v/v) as mobile phase at a flow rate of 1.0 mL min^?1. The elution time was approximately 15 min. The resolution (R _S) between the enantiomers was >3. The mobile phase additives trifluoroacetic acid and diethylamine played a key role in achieving chromatographic resolution between the enantiomers and also in enhancing chromatographic efficiency. Limits of detection and quantification for the S enantiomer were 0.07 and 0.2 μg, respectively, for a test concentration of montelukast sodium of 1,000 μg mL^?1 and 10 μL injection volume. The linearity of the method for the S enantiomer was excellent (R ² > 0.999) over the range from the LOQ to 0.3%. Recovery of the S enantiomer from bulk drug samples and dosage forms ranged from 97.0 to 103.0%, indicative of the high accuracy of the method. Robustness studies were also conducted. The sample solution stability of montelukast sodium was determined and the compound was found to be stable for a study period of 48 h.     

Rapid Determination of Montelukast in Human Plasma by LC-ESI-MS/MS and Its Application to a Bioequivalence Study

Abstract

A rapid liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS) method was developed for the determination of montelukast in human plasma. The extraction of montelukast from plasma (300 µL) involved protein precipitation. Quantitation was performed using LC-ESI-MS/MS, operating in the positive ion and selective reaction monitoring (SRM) mode. The total chromatographic run time for the analysis was 1.5 min. A linear dynamic range was established from 5 to 800 ng mL^?1 for montelukast. The method was fully validated especially with regard to real subject sample analysis. It was successfully applied to a bioequivalence study in 18 healthy human subjects under fed condition (human subjects were allowed to eat as per the prescribed diet for bioequivalence study).     

Applications of Monte-Carlo simulation and chemometric techniques for development of bioanalytical liquid chromatography method for estimation of rosuvastatin calcium

In the present work, we investigated the development of a bioanalytical HPLC method of rosuvastatin (RSV) calcium as per the Quality by Design (QbD)-based systematic chemometric tools. At first, the method objectives were framed and critical analytical attributes (CAAs) were chosen. Risk assessment and factor screening was performed using Hybrid Risk Matrix and Plackett–Burman design for identifying vital factors influencing the critical method parameters (CMPs). Monte-Carlo simulation analysis was conducted which confirmed excellent process robustness (Ppk >1.33) for the studied ranges of CMPs. Furthermore, systematic method development was carried out using custom experimental design, where mobile phase ratio, pH, and injection volume were taken as CMPs at three levels. The obtained trials were evaluated for peak area, retention time, theoretical plates, and peak tailing as CAAs. Mathematical response surface modeling was carried out and optimal chromatographic solution was identified using response optimizer plots. Method transfer was made to bioanalytical scale for estimation of the analyte in rat plasma samples. Extensive method validation was performed as per the ICH Q2 guideline, which indicated validation parameters within the acceptable limits. Overall, the studies construed successful development of QbD compliant HPLC method of rosuvastatin with potential utility bioanalytical testing.     

Application of quality by design to the development of analytical separation methods

Recent pharmaceutical regulatory documents have stressed the critical importance of applying quality by design (QbD) principles for in-depth process understanding to ensure that product quality is built in by design. This article outlines the application of QbD concepts to the development of analytical separation methods, for example chromatography and capillary electrophoresis. QbD tools, for example risk assessment and design of experiments, enable enhanced quality to be integrated into the analytical method, enabling earlier understanding and identification of variables affecting method performance. A QbD guide is described, from identification of quality target product profile to definition of control strategy, emphasizing the main differences from the traditional quality by testing (QbT) approach. The different ways several authors have treated single QbD steps of method development are reviewed and compared. In a final section on outlook, attention is focused on general issues which have arisen from the surveyed literature, and on the need to change the researcher’s mindset from the QbT to QbD approach as an important analytical trend for the near future.

Application of chemometric approach for development and validation of high performance liquid chromatography method for estimation of ropinirole hydrochloride

A systematic Quality by Design approach was employed for developing an isocratic reversed‐phase liquid chromatographic technique for the estimation of ropinirole hydrochloride in bulk drug and pharmaceutical formulations. LiChrospher RP 18‐5 Endcapped column (25 cm × 4.6 mm id) at ambient temperature (25 ± 2°C) was used for the chromatographic separation of the drug. The screening of factors influencing chromatographic separation of the active pharmaceutical ingredient was performed employing fractional factorial design to identify the influential factors. Optimization of the selected factors was carried out using central composite design for selecting the optimum chomatographic conditions. The mobile phase employed was constituted of Solvent A/Solvent B (65:35 v/v) (Solvent A [methanol/0.05 M ammonium acetate buffer, pH 7, 80:20 v/v] and Solvent B [high performance liquid chromatography grade water]) and used at 0.6 mL/min flow rate, while UV detection was performed at 250 nm. Linearity was achieved in the drug concentration range 5–100 µg/mL (R² = 0.9998) with limits of detection and quantification of 1.02 and 3.09 µg/mL, respectively. Method validation was performed as per ICH guidelines followed by forced degradation studies, which indicated good specificity of the developed method for detecting ropinirole hydrochloride and its possible degradation products in the bulk drug and pharmaceutical formulations.     

Orthogonal Design-Directed Optimization of an LC Method for Fingerprinting Mai-Luo-Ning Injection, and Validation of the Method

A novel hierarchical chromatographic response function (HCRF)-directed orthogonal design procedure has been used for optimization of an high-performance liquid chromatography method for fingerprinting Mai-Luo-Ning (MLN) injection. The method was then successfully validated. Five major controllable chromatographic conditions at four levels were included in the orthogonal design. A total of 16 chromatographic runs resulted in the optimum chromatographic conditions-a 250 × 4.6 mm i.d., 4-μm particle, C₁₈ column, a mixture of methanol and 0.025% aqueous formic acid in water as mobile phase, flow rate 0.8 mL min^?1, column temperature 35 °C, and detection wavelength 240 nm. The mobile phase gradient was then further optimized step by step by observation of the chromatographic profiles obtained. Fingerprints of MLN injection and its constituent single herb injections were separately acquired by use of the optimized method. Attribution of the 18 largest peaks observed in the MLN fingerprint indicated that Flos Lonicerae was the main ingredient. Validation of the method for precision, repeatability, and stability proved it was highly reproducible. This chromatography fingerprint method could be very useful for quality control of MLN injection. The original HCRF-directed orthogonal design approach proposed should be generally useful for developing chromatographic fingerprinting methods.     

A Scientific, General Approach to Robustness Validation for an Ion-Pair LC Assay Method Through a Case Study

This paper describes a scientifically sound, systematic approach to the robustness validation of an ion-pair LC assay method. Robustness validation was carried out on all chromatographic parameters, such as ion-pair reagent concentration, buffer concentration and pH, organic to buffer ratio, column oven temperature, and column age. Small, deliberate variations to each parameter were implemented according to reasonably expected laboratory variabilities. For instance, a typical measuring variability with a 2,000-mL graduated cylinder is 20 mL, and this translates into variations of ±1.3% for a volume of 1,560 mL of an aqueous phase. Details on the experimental design of robustness validation are described, such as the parameters considered, the variations chosen for each parameter along with rationales, the probes that were used to assess robustness, and corroboration of method robustness with intermediate precision validation. This robustness validation approach eliminates guess work, ensures scientifically appropriate and customized variations for each method parameter, and has won regulatory acceptance.     

Design of experiment techniques for the optimization of chromatographic analysis conditions: A review

Design of experiment (DoE) techniques have been widely used in the field of chromatographic parameters optimization as a valuable tool. A systematic literature review of the available DoE techniques applied to the development of a chromatographic analysis method is presented in this paper. First, the most common available designs and the implementation steps of DoE are comprehensively introduced. Then the studies in recent 10 years for the application of DoE techniques in various chromatographic techniques are discussed, such as capillary electrophoresis, liquid chromatography, gas chromatography, thin-layer chromatography, and high-speed countercurrent chromatography. Current problems and future outlooks are finally given to provide a certain inspiration of research in the application of DoE techniques to the different chromatographic techniques field. This review contributes to a better understanding of the DoE techniques for the efficient optimization of chromatographic analysis conditions, especially for the analysis of complex systems, such as multicomponent drugs and natural products.     

Validation of analytical methods involved in dissolution assays: Acceptance limits and decision methodologies

Dissolution tests are key elements to ensure continuing product quality and performance. The ultimate goal of these tests is to assure consistent product quality within a defined set of specification criteria. Validation of an analytical method aimed at assessing the dissolution profile of products or at verifying pharmacopoeias compliance should demonstrate that this analytical method is able to correctly declare two dissolution profiles as similar or drug products as compliant with respect to their specifications. It is essential to ensure that these analytical methods are fit for their purpose. Method validation is aimed at providing this guarantee. However, even in the ICHQ2 guideline there is no information explaining how to decide whether the method under validation is valid for its final purpose or not. Are the entire validation criterion needed to ensure that a Quality Control (QC) analytical method for dissolution test is valid? What acceptance limits should be set on these criteria? How to decide about method's validity? These are the questions that this work aims at answering. Focus is made to comply with the current implementation of the Quality by Design (QbD) principles in the pharmaceutical industry in order to allow to correctly defining the Analytical Target Profile (ATP) of analytical methods involved in dissolution tests. Analytical method validation is then the natural demonstration that the developed methods are fit for their intended purpose and is not any more the inconsiderate checklist validation approach still generally performed to complete the filing required to obtain product marketing authorization.     

Design of Experiment Approach for the Development and Validation of a Stability-Indicating High-Performance Thin-Layer Chromatography Method for the Estimation of Metformin Hydrochloride and Ursodeoxycholic Acid in Pharmaceutical Dosage Form

JPC – Journal of Planar Chromatography – Modern TLC - In the present study, a design of experiment (DoE) approach was used to optimize chromatographic conditions for the development of...     

Applying green analytical chemistry for development and validation of RP-HPLC stability indicating assay method for estimation of fenoverine in bulk and dosage form using quality by design approach

A green and robust reverse-phase liquid chromatographic method has been developed for the determination of fenoverine (FEN), by applying combined principles of green analytical chemistry and quality by design approaches on a Spherisorb C18 column (150?×?4.6?mm, 3?µm) with UV detection at 262?nm. A two level fractional factorial design (2^^7-3) Res IV was used for screening of influential chromatographic factors. The critical method parameters actively affecting critical quality attributes (CQAs) were identified and further optimized using Box–Behnken design. The predicted optimum assay conditions comprised of methanol and ammonium acetate buffer 20?mM, in an extent of 81:19% v/v individually having a flow rate of 1.0?mL/min with a column oven temperature of 33°C. The drug was stressed in hydrolytic, oxidative, reductive, thermal, and photolytic conditions. The developed method was validated successfully. The detector response was linear in the concentration of 0.5–160?µg/mL with a limit of detection (LOD) and limit of quantitation (LOQ) as 0.1 and 0.3?µg/mL, respectively. The % recovery was found to be 99.7%. The analytical method volume intensity value for developed method was 45?mL and the environment assessment tool (EAT) score was 41.07. The method is simple, environmentally benign, rapid, and robust for the determination of FEN in bulk and in its dosage form.     

An Application of Quality by Design and Analytical Greenness Assessment Approach for the Development of Erlotinib Stability Indicating Method

Pharmaceutical regulators are worried about medication quality and stability since drug degradation may result in harmful chemicals. Erlotinib (ERL) is a tyrosine kinase inhibitor associated with the epidermal growth factor receptor (EGFR) containing susceptible functional groups such as quinazoline and amine ketone, methoxy, and ethoxy leads to a reduction in pharmaceutical quality. According to the ICH-Q1A (R2) guideline, the goal of ERL stability studies is to establish its susceptibility to degradation under various environmental conditions. A novel isocratic stability–indicating liquid chromatography method has been developed using systemic quality by design (QbD) approach. The QbD strategy includes screening and optimization as phases. Placket Burman was used for primary parameters screening, and critical factors were optimized with response surface design. The prepared degradation samples (acid, base, neutral hydrolysis, oxidative, photolytic, and thermal) were separated using a Shimadzu GIST C18 column (250 mm?×?4.6 mm, 5 µm) with 15 mM ammonium formate: ACN (58:42% v/v) as mobile phase, 0.9 mL/min flow rate, and 246 nm wavelength, which was found to be LC–MS compatible. A total of six degradation products (DPs) were identified with the optimized chromatography method. The drug was sensitive toward acidic and basic hydrolysis, but it remained stable under neutral, oxidative, thermal, and photolytic stress conditions. The optimized method was sensitive, specific, and robust, with linearity ranging from 10 to 35 µg/mL, with a correlation coefficient (R²?=?0.9997). The analytical method greenness score was calculated and observed that the developed method is green.

     

Quality by Design applications in biosimilar pharmaceutical products

A process is well understood when all critical sources of variability are identified and explained, variability is managed by the process design and monitoring, and product quality attributes are accurately and reliably predicted over the design space. Quality by Design (QbD) is a systematic approach to product development and process control that begins with predefined objectives, emphasizes product and process understanding, and sets up process control based on sound science and quality risk management. The Food and Drug Administration (FDA) and the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) have recently started promoting QbD in an attempt to curb rising development costs and regulatory barriers to innovation and creativity. QbD is partially based on the application of multivariate statistical methods and a statistical Design of Experiments strategy to the development of both analytical methods and pharmaceutical formulations. In this paper, we review the basics of QbD and their impact on the innovative, generic, and biosimilar pharmaceutical industry. In particular, we consider the challenge of mapping the control space in biotechnological processes and how advances in statistical methods can contribute to QbD.     

Innovative high-performance liquid chromatography method development for the screening of 19 antimalarial drugs based on a generic approach, using design of experiments, independent component analysis and design space

An innovative methodology based on design of experiments (DoE), independent component analysis (ICA) and design space (DS) was developed in previous works and was tested out with a mixture of 19 antimalarial drugs. This global LC method development methodology (i.e. DoE-ICA-DS) was used to optimize the separation of 19 antimalarial drugs to obtain a screening method. DoE-ICA-DS methodology is fully compliant with the current trend of quality by design. DoE was used to define the set of experiments to model the retention times at the beginning, the apex and the end of each peak. Furthermore, ICA was used to numerically separate coeluting peaks and estimate their unbiased retention times. Gradient time, temperature and pH were selected as the factors of a full factorial design. These retention times were modelled by stepwise multiple linear regressions. A recently introduced critical quality attribute, namely the separation criterion (S), was also used to assess the quality of separations rather than using the resolution. Furthermore, the resulting mathematical models were also studied from a chromatographic point of view to understand and investigate the chromatographic behaviour of each compound. Good adequacies were found between the mathematical models and the expected chromatographic behaviours predicted by chromatographic theory. Finally, focusing at quality risk management, the DS was computed as the multidimensional subspace where the probability for the separation criterion to lie in acceptance limits was higher than a defined quality level. The DS was computed propagating the prediction error from the modelled responses to the quality criterion using Monte Carlo simulations. DoE-ICA-DS allowed encountering optimal operating conditions to obtain a robust screening method for the 19 considered antimalarial drugs in the framework of the fight against counterfeit medicines. Moreover and only on the basis of the same data set, a dedicated method for the determination of three antimalarial compounds in a pharmaceutical formulation was optimized to demonstrate both the efficiency and flexibility of the methodology proposed in the present study.