Development and validation of a high performance liquid chromatographic method for quantitative determination of aporphine alkaloids from different samples of Cassytha filiformis.
A sensitive and accurate procedure based on an alkaloid extraction coupled to an HPLC-UV-MS determination has been developed for the separation and quantification of the major aporphines in Cassytha filiformis. The extraction step and the liquid chromatography conditions were optimized in order to improve the selectivity of the method. The HPLC mobile phase consisted of a mixture of water containing 10 mM ammonium acetate adjusted to pH 3 with acetic acid-acetonitrile (90: 10, v/v) (A) and acetonitrile (B) used in a gradient mode (0 to 40%). The stationary phase was an RP-select B (5 micro m) column. The method was completely validated using cassythine, one of the major aporphines in our samples, as reference standard and successfully applied to the determination of these pharmacologically interesting aporphines in seven different batches of C. filiformis. The detection and quantitation limits of cassythine were found to be 13 and 20 micrograms/mL, respectively. The results showed variations in the total alkaloid content in samples from 0.11 to 0.43%.
Stévigny
Caroline
C
Laboratoire de Pharmacognosie, Unité d'Analyse Chimique et Physico-Chimique des Médicaments, Université Catholique de Louvain, UCL 72.30-CHAM, Bruxelles, Belgium.
Wautier
Marie-Caroline
MC
Habib Jiwan
Jean-Louis
JL
Chiap
Patrice
P
Hubert
Philippe
P
Quetin-Leclercq
Joëlle
J
D002851:Chromatography, High Pressure Liquid / Q000592:standards*; D006801:Humans; D019770:Lauraceae; D008517:Phytotherapy; D035261:Plant Components, Aerial; D010936:Plant Extracts / Q000737:chemistry*; D015203:Reproducibility of Results
D010936:Plant Extracts