title: Simultaneous determination of secondary metabolites from Vinca rosea plant extractives by reverse phase high performance liquid chromatography.
abstract: BACKGROUND
  Vinca rosea (Apocynaceae) is one of the most important and high value medicinal plants known for its anticancer alkaloids. It is the iota of the isolated secondary metabolites used in chemotherapy to treat diverse cancers. Several high performance liquid chromatography (HPLC) methods have been developed to quantify the active alkaloids in the plant. However, this method may serve the purpose in quantification of V. rosea plant extracts in totality.
  
  
  OBJECTIVE
  To develop and validate the reverse phase (RP)-HPLC method for simultaneous determination of secondary metabolites, namely alkaloids from V. rosea plant extracts.
  
  
  METHODS
  The quantitative determination was conducted by RP-HPLC equipped with ultraviolet detector. Optimal separation was achieved by isocratic elution with mobile phase consisting of methanol:acetonitrile:ammonium acetate buffer (25 mM) with 0.1% triethylamine (15:45:40 v/v) on a column (Zorbax Eclipse plus C(18), 250 mm % 4.6 mm; 5 μm). The standard markers (vindoline, vincristine, catharanthine, and vinblastine) were identified by retention time and co-injected with reference standard and quantified by external standard method at 297 nm.
  
  
  RESULTS
  The precision of the method was confirmed by the relative standard deviation (R.S.D.), which was lower than 2.68%. The recoveries were in the range of 98.09%-108%. The limits of detection (LOD) for each marker alkaloids were lower than 0.20 μg. Different parts of the V. rosea extracts shows different concentrations of markers, flower samples were high in vinblastine content, while methanol extract from the leaves contains all the four alkaloids in good yield, and there is no significant presence of markers in water extracts.
  
  
  CONCLUSIONS
  HPLC method established is appropriate for the standardization and quality assurance of V. rosea plant extracts.
authors.affiliation: School of Pharmaceutical Sciences, Universiti Sains Malaysia, P. Penang 11800, Malaysia.
authors.forename: Mohammad Jamshed Ahmad
authors.initials: MJ
authors.lastname: Siddiqui
authors.forename: Zhari
authors.initials: Z
authors.lastname: Ismail
authors.forename: Noor Hafizoh
authors.initials: NH
authors.lastname: Saidan