Simultaneous assessment and validation of reverse phase-high performance liquid chromatography method for quercetin, eugenol, myristicin, and safrole from nutmeg, fruit and mace

Background: Nutmeg is the imperative spices having pharmacological importance. Objectives: The objective of this work was to standardize Nutmeg extract by RP-high performance liquid chromatography (HPLC) analysis. Settings and Design: An RP-HPLC method was developed for simultaneous quantification of quercetin (QUE), eugenol (EUG), myristicin (MYRS), and safrole (SAFR) from nutmeg fruit and mace extracts. Materials and Methods: RP-HPLC method was performed with Waters 2695 Alliance system using a 2996 photodiode array detector (PDA). QUE, EUG, MYRS, and SAFR were separated on a reverse-phase 250 × 4.6 mm, 5-m, Zorbax SB C18 column (Agilent). The mobile phase was prepared from 0.1% orthophosphoric acid in water of pH 2.5 (solvent-A) and acetonitrile (solvent-B). The gradient program was selected for separation. The PDA was set at 220 nm, which shows maximum response for all peaks. Statistical Analysis: Percent relative standard deviation (% RSD) and correlation coefficient (r 2 ) were calculated by standard formulas. Results: QUE, EUG, MYRS, and SAFR were satisfactorily resolved with retention time about 3, 7, 19 and 21 min. respectively. The method was validated and results obtained showed accepted values for correlation of coefficient and % RSD. Conclusions: The method was accurate and specific for analysis of nutmeg extract.