Development of a validated HPLC method for the determination of sennoside A and B, two major constituents of Cassia obovata Coll.

Ghassemi-Dehkordi, N. and Ghanadian, M. and Arabha, S. (2014) Development of a validated HPLC method for the determination of sennoside A and B, two major constituents of Cassia obovata Coll. Journal of HerbMed Pharmacology, 3.

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Abstract

<p> <strong>Introduction: </strong><em>Cassia obovata</em> Coll is the only <em>Senna </em>species which grows wild in Iran. In the present study, an optimised reverse High Performance Liquid Chromatography (HPLC) validated method was established for quantification of sennosides A and B, the major constituents of <em>C. obovata</em> with a simple and accurate method. </p> <p><strong>Methods:</strong> HPLC analysis was done using Waters 515 pump on a Nova-Pak C18 (3.9 &times; 150 mm). Millennium software was used for the determination of the sennoside A and B in Cassia species and processing the information. The method was validated according to USP 32 requirements.</p> <p> <strong>Results:</strong> The solvent impact on the selectivity factor and partition coefficient parameters evaluated. Using a conventional RP-18 L1 column, 3.9 &times; 150 mm, the mobile phase was selected after several trials with different mixtures of water and acetonitrile. Sennosides A and B were determined using the external standard calibration method. Using USP 35-NF 30, the LOD and LOQ were calculated. The reliability of the HPLC-method for analysis of sennoside A + B was validated through its linearity, reproducibility, repeatability, and recovery. Fina1ly ethanol:water (1:1) extracts of <em>Cassia obovata</em> and <em>Cassia angustifolia </em>were standardized by assay of sennoside A and B through above HPLC validated method. </p> <p><strong>Conclusion:</strong> Through the above method, determination of sennosides in Cassia species are completely possible. Moreover, through comparing the results, even though sennosides are rich in <em>Cassia angustifolia</em> but, the results shows that <em>C. obovata</em> could be considered as an alternative source for sennosides A and B. </p>

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