Application of experimental design for the optimization of forced degradation and development of a validated stability-indicating LC method for luliconazole in bulk and cream formulation

AbstractA stability-indicating LC method was developed and validated for the determination of luliconazole in bulk and cream formulation. Luliconazole was exposed to acid, alkali and water hydrolysis, oxidation effect by hydrogen peroxide, dry heat and photolytic conditions. Full factorial design was used during forced degradation experiments and the factors/combination of factors which were most likely to effect degradation of luliconazole under various conditions were identified and further were optimized using surface response curve. Drug was found to be stable under wet heat and dry heat conditions, but substantial degradation was observed under acid, alkali, oxidative and photolytic conditions. Drug and its degradation products were optimally resolved on HiQ sil C-18HS (250×4.6mm, 5μm) column with the mobile phase consisting of methanol and water (80:20, v/v) at a flow rate of 1mL/min, detection was performed at 296nm. The procedure was validated for specificity, linearity, accuracy and precision. There was no interference of excipients and degradation products in the determination of active pharmaceutical ingredient. The method was accurate and precise and the response was found to be linear in the range of 2–14μg/mL. The method was found to be simple and fast by making use of experimental design

Optimization of Forced Degradation Using Experimental Design and Development of a Stability-Indicating Liquid Chromatographic Assay Method for Rebamipide in Bulk and Tablet Dosage Form

A novel stability-indicating RP-HPLC assay method was developed and validated for quantitative determination of rebamipide in bulk and tablet dosage form. Rebamipide (drug and drug product) solutions were exposed to acid and alkali hydrolysis, thermal stress, oxidation by hydrogen peroxide and photodegradation. Experimental design has been used during forced degradation to determine significant factors responsible for degradation and to obtain optimal degradation conditions. In addition, acid and alkali hydrolysis was performed using a microwave oven. The chromatographic method employed the HiQ sil C-18HS (250 × 4.6 mm; 5 μm) column with mobile phase consisting of 0.02 M potassium phosphate (pH adjusted to 6.8) and methanol (40:60, v/v) and the detection was performed at 230 nm. The procedure was validated for specificity, linearity, accuracy, precision and robustness. There was no interference observed of excipients and degradation products in the determination of the active pharmaceutical ingredient. The method showed good accuracy and precision (intra and inter day) and the response was linear in a range from 0.5 to 5 μg mL−1. The method was found to be simple and fast with less trial and error experimentation by making use of experimental design. Also, it proved that microwave energy can be used to expedite hydrolysis of rebamipide

Implementing Quality by Design-A methodical approach in the RP-HPLC method development process

The concept of quality by design (QbD) has recently been adopted for the development of pharmaceutical processes to ensure a predefined product quality. Focus on applying the QbD concept to analytical methods has increased as it is fully integrated within pharmaceutical processes and especially in the process control strategy. Quality by design (QbD) refers to the achievement of certain predictable quality with desired and predetermined specifications. The QbD based method development helps in generating a design space and operating space with knowledge of all method performance characteristics and limitations and successful method robustness within the operating space. A very useful component of QbD is the understanding of factors and their interaction effects by a desired set of experiments. For the purpose of QbD for HPLC methods, robustness and ruggedness should be verified early in the method development stage to ensure method performance over the lifetime of the product. Quality-by-Design principles are applied to build in a more scientific and risk-based multi-factorial approach to the development and validation of analytical methods using HPL

Development and validation of RP-HPLC method for estimation of eplerenone in spiked human plasma

AbstractA rapid and simple high performance liquid chromatography (HPLC) method with a UV detection (241nm) was developed and validated for estimation of eplerenone from spiked human plasma. The analyte and the internal standard (valdecoxib) were extracted with a mixture of dichloromethane and diethyl ether. The chromatographic separation was performed on a HiQSil C-18HS column (250mm×4.6mm, 5μm) with a mobile phase consisting of acetonitrile:water (50:50, v/v) at flow rate of 1mL/min. The calibration curve was linear in the range 100–3200ng/mL and the heteroscedasticity was minimized by using weighted least squares regression with weighting factor 1/X