Qualitative and quantitative evaluation of annonaceous acetogenins by high performance liquid chromatography

Abstract

Abstract A high performance liquid chromatographic (HPLC) method for the separation, identification and quantification of Annonaceous acetogenins under isocratic or gradient conditions, using ultraviolet, refractometric or evaporative light scattering (ELSD) detection is reported. The system, consisting of a μBondapak C 18 (10 μm) column and a combination of isocratic, step and linear gradient elutions with water and acetonitrile, allowed all six acetogenins of Annona muricata to be well resolved and separated. Well resolved and symetrical peaks were also obtained for other acetogenins with a variety of substitution patterns under isocratic conditions using methanol: water in the ratio of 90:10 to 80:20 or acetonitrile: water in the ratio of 90:10 to 70:30 as mobile phases. The effect of the number, position and type of substituents are discussed. HPLC using μBondapak C 18 (10 μm) column as stationary phase and acetonitrile: water as a mobile phase was successful for the separation of two epimeric acetogenins: rolliniastatin‐1 and rolliniastatin‐2. In order to demonstrate the possibility of quantifying the acetogenins in crude extracts, the quantitative evaluation of annonacin in the seeds of A. muricata was carried out. Semipreparative separations of some acetogenins by HPLC are also reported.