A validated RP-HPLC-UV/DAD method for simultaneous quantitative determination of rosmarinic acid and eugenol in Ocimum sanctum L.

摘要

Background: Ocimum sanctum L. is present as an ingredient in many herbal formulations and hencestandardization of its raw materials and extracts is significant. The study was aimed at developing andvalidating a RP-HPLC method for the standardization of O. sanctum L. raw material and extracts byselecting rosmarinic acid and eugenol as markers.Methods: The developed method used a Phenomenex Luna C18 column (250 4.6 mm; 5 mm) using agradient elution of 1 mM ammonium acetate buffer and acetonitrile as the mobile phase with a flow rateof 1.5 mL/min. The analytes were monitored at 278 nm. The method was validated according to the ICHguidelines.Results: The calibration curves showed good linear regression (r2 > 0.999) within test ranges. Specificityof the method was assessed using PDA profile and the peak purity of all the analytes was greater than99%. This method showed good reproducibility for the quantification of both the compounds in sampleswith an RSD of less than 2.5% respectively. Percentage recovery exceeded 90% for all analytes.Conclusion: The validated method was successfully applied to quantify rosmarinic acid and eugenol in O.sanctum raw material and extracts which provides a new basis for overall assessment on quality.