Informatics Publishing Limited

S. Kathirvel ¹, R. Raju ¹, B. Seethadevi ², A. Suneetha ³, K. Jyothisree ³

Affiliations
1 Department of Pharmaceutical Analysis, National College of Pharmacy, Manassery, Kozhikode, Kerala, 673602, IN
2 Department of Pharmacognosy, National College of Pharmacy, Manassery, Kozhikode, Kerala, 673602, IN
3 Department of Pharmaceutical Analysis, Hindu College of Pharmacy, Amaravathi road, Guntur, A.P, 522002, IN

Abstract

The study is a proposition of the application of high performance liquid chromatography (HPLC) with a spectrophotometric UV range detector to analyse the chemical purity and assay of tetrabenazine in bulk drug and tablet formulation and also to apply the proposed method to determine the content uniformity according to USP specification. During computer assisted literature search only a few publications were found about tetrabenazine, so the ultimate objective of the present investigation was aimed to develop stability - indicating LC method and the development of the newer analytical method was achieved under optimised chromatographic conditions by using mobile phase composed of potassium phosphate buffer: acetonitrile in the ratio (40:60v /v). The seperation was achieved by using isocratic elution mode with a flow rate of 1.0 mL/min. The effluent was monitored on a UV detector at 284 nm. The retention time of tetrabenazine was found to be at 5.05min and the standard calibration curve was linear over a concentration range of 6.25-37.5 μg/mL with r value of 0.999. The LOD and LOQ were found to be 0.562 and 1.704 μg/mL respectively. The recovery studies were performed and the percentage recoveries were found to be in the range of 98.64-98.88 %. The drug was also subjected to acidic, basic, oxidative and photolytic degradation as per the ICH guidelines. The developed method allows to separate all degradation from tetrabenazine and to quantitate the tetrabenazine amount. As there is no official monograph in the pharmacopeias about tetrabenazine, the performed full validation procedure makes the method ready to use in routine analysis.

Keywords

Content Uniformity, ICH Guidelines, Tetrabenazine And USP Specification.

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S. Kathirvel ¹, R. Raju ¹, B. Seethadevi ¹, A. Suneetha ², J. Pavani ²

Affiliations
1 Department of Pharmaceutical Analysis, National College of Pharmacy, Manassery, Mukkom Post, Kerala, IN
2 Department of Pharmaceutical Analysis, Hindu College of Pharmacy, Amaravathi Road, Guntur, A.P, IN

Abstract

The objective of the current study was to develop a validated, specific and stability-indicating reversed phase HPLC method for the quantitative determination of posaconazole and its related substances in API (Active Pharmaceutical Ingredient). The determination was done for active pharmaceutical ingredient in the presence of degradation products, and its process-related impurities. The chromatographic separation was achieved on a waters HPLC system with PDA detector and the column employed for the present investigation was inertsil ODS-3V C18 (150×4.6 mm with 5 μ particle size) and empower2 software provided by waters was used throughout the experiment. The method employed a linear gradient elution and the detection wavelength was set at 225 nm (for intermediate A impurity) and 260 nm (for intermediate B, diastereomer, formyl and benzyl posaconazole impurity). The drug was subjected to stress conditions of hydrolysis (acid and base), oxidation, photolysis and thermal degradation as per International Conference on Harmonization (ICH) prescribed stress conditions to show the stability-indicating power of the method. Significant degradation was observed during acid, oxidative, thermal and photo stress studies. In the developed HPLC method, the resolution between posaconazole and its process-related impurities was found to be greater than 2.0. Regression analysis shows an r value (correlation coefficient) of greater than 0.999 for posaconazole and it’s all the five impurities. The developed HPLC method was validated with respect to linearity, accuracy, precision and robustness.

Keywords

Degradation Products, Posaconazole, Process-Related Impurities, Regression Analysis.

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