Open AccessNovel, Rapid, Isocratic RP-HPLC Method for Simultaneous Estimation of Piperine and Embelin in Herbal Formulation
Author(s) -
Vandana Jain,
Revati So,
Leena Tandel
Publication year - 2021
Publication title -
international journal of pharmaceutical sciences review and research
Language(s) - English
Resource type - Journals
ISSN - 0976-044X
DOI - 10.47583/ijpsrr.2021.v70i02.001
Subject(s) - piperine , chromatography , chemistry , phosphoric acid , phosphate buffered saline , high performance liquid chromatography , methanol , retention time , correlation coefficient , relative standard deviation , detection limit , mathematics , statistics , organic chemistry
The objective of this paper was to develop and validate a novel, simple, rapid, precise and accurate, reverse-phase high-performanceliquid chromatographic (RP-HPLC) method for simultaneous quantitative estimation of piperine and embelin in the herbal formulationas per the International Conference on Harmonization guidelines (ICH). Chromatographic separation was achieved using a CosmosilC-18 (250*4.6mm) SH 5.0 µm column with a mobile phase consisting of methanol and 0.02 M phosphate buffer in ratio 98:2 v/v,(pH adjusted to 2.3 with ortho-phosphoric acid) at a flow rate of 1 mL/min and column temperature maintained at 28°C andultraviolet (UV) detection at 288 nm. The retention time of piperine and embelin was found to be 4.15 and 5.69 min respectively. Thelinearity of piperine and embelin was tested in the range of 5-40 ?g/mL. The correlation coefficient for piperine and embelin wasfound to be 0.997 and 0.995, respectively. The recovery values (98-102%) indicate a satisfactory accuracy. The method was found tobe precise as the percentage relative standard deviation was found to be <2 %. The proposed novel isocratic RP-HPLC method is rapid(short run time below 10 min), precise, accurate and sensitive. The method was successfully applied for the simultaneous analysis ofpiperine and embelin in herbal formulation.