Development and validation of a stability indicating RP-HPLC-DAD method for the determination of bromazepam

Prenylamine was initially used for the treatment of angina pectoris and later on withdrawn from the market in 1988 due to cardiac arrhythmias concern. The major phase I metabolite of prenylamine is p-hydroxy prenylamine that has a chiral center in the structure. Even though p-hydroxy prenylamine was synthesized earlier, it lacked complete analytical developments for chiral high-performance liquid chromatography (HPLC) separation. However, p-hydroxy prenylamine reference material is not commercially available. The innovation of this manuscript is the development and validation of a chiral HPLC separation method and more extensive characterization of the reference material than previously reported method. Therefore, it was hypothesized to develop and validate normal phase HPLC method for p-hydroxy prenylamine reference material. p-Hydroxy prenylamine was synthesized in two batches and characterized successfully using 13 C NMR, 1 H NMR, high-resolution mass spectrometry (HRMS), Fourier transform infrared spectroscopy (FT-IR), and thermogravimetric analysis (TGA). A normal phase chiral HPLC method was developed to analyze the p-hydroxy prenylamine purity. Separation of the p-hydroxy prenylamine enantiomers were achieved using ultra-high-performance liquid chromatography (UHPLC) on a ChiralCel ODH column at wavelength of 220 nm. The developed method was validated in terms of its linearity, accuracy, precision, and robustness for purification, purity assessment, and stability studies. Proton and carbon peaks were confirmed by nuclear magnetic resonance (NMR) analysis. Functional groups were confirmed by FT-IR. Loss on drying was 0.3% and 0.6% for Batches 1 and 2, respectively. The purity of the developed reference material for Batches 1 and 2 was found to be 99.59% and 100%, respectively. Therefore, the synthesized batches of p-hydroxy prenylamine can be used in dope testing as reference material.

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“…The developed method was validated using various parameters according to ICH Q2(R1) guidelines. [24][25][26]…”

Section: Methods Validationmentioning

confidence: 99%

Analytical developments of p‐hydroxy prenylamine reference material for dope control research: Characterization and purity assessment

Joseph

Kumar

Pawar

et al. 2021

Drug Testing and Analysis

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“…The system suitability was assessed by analyses of BEN (15 µg/ml) for six replicates. This validation test was conducted to determine the suitability of the chromatographic system for analysis and its reproducibility [16]. The method's suitability has been determined based on the theoretical plates, the relative standard deviation of the areas of each consecutive set of six standards (not more than 2%), and the acceptable chromatographic parameters of the produced peaks in terms of the tailing factor (acceptance limit is less than 2).…”

Section: System Suitabilitymentioning

confidence: 99%

Development and Validation of HPLC Method for Determination of Benidipine Hydrochloride in Lipid Vesicles Formulations

Syed¹,

Khater²,

Azmir³

et al. 2022

Int. J. Membrane Sci. Techno.

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In the current study, a novel high-performance liquid chromatographic method is being developed and validated to estimate benidipine hydrochloride in lipid-based pharmaceutical formulations including transfersome, ethosome, and transethosome. The chromatographic separation was accomplished on a ZORBAX Eclipse Plus C18 (4.6mm x 150mm) analytical column with a mobile phase consisting of a mixture of methanol and 50 mM phosphate buffer solution at a ratio of 70:30 (v/v). A standard calibration curve was used to quantitatively determine the medication at a UV wavelength of 237 nm. The limit of detection (LOD) and limit of quantitation (LOQ) were determined to be 0.005µg/ml and 0.015µg/ml, respectively. The relative standard deviation (%RSD) of the intra-day and inter-day studies for benidipine hydrochloride was less than 2%, and the percentage recovery of benidipine hydrochloride was found to be in the range of 98.57-100.27%. The method is specific, linear, accurate, precise, robust, and sensitive for its intended purpose, according to the results of the method validation. To determine the amount of benidipine hydrochloride and the effectiveness of drug entrapment in lipid-based formulated membrane like cell structures namely Transfersomes, Ethosomes, and Transethosomes the current method was successfully applied.

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“…Many analytical methods were reported as stability-indicating methods for BRZ. These methods applied spectroscopic [ 36 , 39 ] and liquid chromatographic [ 40 , 41 ] techniques for the selective determination of BRZ.…”

Section: Introductionmentioning

confidence: 99%

A Comparative Study between Screen-Printed and Solid-Contact Electrodes for the Stability-Indicating Determination of Bromazepam

Abdel-Gawad

Altharawi

2022

Molecules

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Stability-indicating methods are awesome tools to ensure the safety and efficacy of active pharmaceutical ingredients (APIs). An accurate comparative study involving the use of potentiometric sensors for the determination of bromazepam (BRZ) in the presence of its main product of degradation and impurity was performed by the fabrication of two membrane electrodes. A screen-printed electrode (SPE) and a solid-contact glassy carbon electrode (SCE) were fabricated and their performance optimized. The fabricated sensors showed a linear electrochemical response in the concentration range 1.0 × 10−6 M to 1.0 × 10−2 M. The electrodes exhibited Nernstian slopes of 59.70 mV/decade and 58.10 mV/decade for the BRZ-SPE and BRZ-SCE membrane electrodes, respectively. The electrochemical performance was greatly affected by the medium pH. They showed an almost ideal electrochemical performance between pH 3.0 and pH 6.0. The fabricated membranes were applied successfully for the quantification of BRZ in the presence of up to 90% of its degradation product. Moreover, a successful application of the fabricated electrodes was performed for the sensitive and selective quantification of BRZ in its tablet form without any pretreatment procedure.

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Development and validation of a stability indicating RP-HPLC-DAD method for the determination of bromazepam

Cited by 8 publications

References 40 publications

Analytical developments of p‐hydroxy prenylamine reference material for dope control research: Characterization and purity assessment

Analytical developments of p‐hydroxy prenylamine reference material for dope control research: Characterization and purity assessment

Development and Validation of HPLC Method for Determination of Benidipine Hydrochloride in Lipid Vesicles Formulations

A Comparative Study between Screen-Printed and Solid-Contact Electrodes for the Stability-Indicating Determination of Bromazepam

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