Development And Validation Of A RP-HPLC Method To Determine Dehydrodiisoeugenol, Myristicin, And Safrole In Ethanol Extract Of Nutmeg (Myristica fragrans Houtt.)

A simple and validated analytical method to determine myristicin, safrole, and dehydrodiisoeugenol (DDIE) in nutmeg extract has been developed. High Performance Liquid Chromatography (HPLC) using C-18 LiChroCART 250-4, LiChrospher 100 RP 18e (5 μm) 250 mm column as stationary phase and methanol: water (73:27) as a mobile phase with the flow rate 1 mL/min was selected. Detection was done by using ultraviolet (UV) spectrophotometer at 282 nm. Retention time for myristicin, safrole, and DDIE were 8,260; 10,507; 13,900 minute. Limit of detection and limit of quantitation for myristicin, safrole, and DDIE were 0,991 μg/mL and 3,004 μg/mL; 0,668 μg/mL and 2,023 μg/mL; 0,981 μg/mL and 2,973 μg/mL, respectively. The recovery for myristicin, safrole, and DDIE were 99,754 ± 0,788 %; 101.421 ± 0,855 %; 100,242 ± 1,327 %, while the coefficient of variance for myristicin, safrole, and DDIE were 0,802 %; 0.838 %; 1,324 %. Mean concentration of MYR, SAF, and DDIE were 17,226%; 10,979%; 4,662%.