An effective ultra‐performance liquid chromatography and derivatization method for the quantification of potential genotoxic impurity Hydrazine in Gliclazide and its formulation – Robustness study by the design of experiments

Abstract

The current study presents an accurate, precise, and highly selective ultra‐performance liquid chromatography derivatization method for the quantification of Hydrazine in Gliclazide and its formulations. Gliclazide, derivatizing agent‐ 3,4,5‐trimethoxy benzaldehyde, and Hydrazine derivatives were separated on YMC Pack pro C18 column (50 × 3.0 mm id, 2 μm), column oven temperature maintained at 40°C, with flow rate 0.6 ml/min on ultra‐performance liquid chromatography system with gradient elution monitored at 333 nm. 0.1% trifluoroacetic acid and acetonitrile were used as mobile phase‐A and mobile phase‐B. Evaluated the robustness of the optimized method by employing a quality‐by‐design‐based design of experiments. The method was successfully validated as per the International Conference on Harmonization Q2 (R1) guideline and the United States Pharmacopeia < 1225 > general chapter. The United States Pharmacopeia resolution between the Gliclazide and Hydrazine derivative was 3.6. The limit of detection and limit of quantitation for Hydrazine derivative are 0.10 μg/ml and 0.30 μg/ml, respectively. The obtained regression coefficient (R2) was > 0.9998, and the %recoveries from the limit of quantitation to 150% level were 96.9%–100.8% from the linearity and accuracy studies, respectively. The method validation data and quality‐by‐design‐based robustness study indicate that the developed derivatization method is suitable for routine use in Quality Control laboratories.

A sensitive ultra‐performance liquid chromatography‐tandem mass spectrometry method for the simultaneous quantification of assay and trace‐level genotoxic tosylate analogs (methyl and ethyl) in empagliflozin and its tablet dosage forms

“…The current LC–MS/MS technique was successfully validated in accordance with ICH and regulatory guidelines (International Conference on Harmonisation, 2005) as well as reported papers in different journals (Al‐Sabti & Harbali, 2021; Konduru et al, 2018; Konduru, Gundla, et al, 2022; Konduru, Kethe, et al, 2022; Muchakayala, Katari, Saripella, Schaaf, Marisetti, Ettaboina, & Rekulapally, 2022; Muchakayala, Katari, Saripella, Schaaf, Marisetti, Kowtharapu, & Jonnalagadda, 2022; Nakka et al, 2022; Nakka, Katari, et al, 2023; Nakka, Muchakayala, et al, 2023). Specificity, limit of detection (LOD), limit of quantitation (LOQ), precision, linearity, accuracy, robustness, and solution stability were tested as part of method validation.…”

A sensitive UPLC-MS/MS method for the simultaneous assay and trace level genotoxic impurities quantification of SARS-CoV-2 inhibitor-Molnupiravir in its pure and formulation dosage forms using fractional factorial design

“…These methods, however, do not reveal the interaction effects of various factors on separation and elution. To conduct a minimum number of trials and quantitatively identify the effects of variable interactions, AQbD-established experiments are used [32] , [33] , [34] , [35] . A statistical quality-by-design is an effective technique for identifying probable risks and failures and understanding their interaction effect.…”