Does anybody know an HPLC method for the determination of alkaloids in the Casuarina equisetifolia extracts?
I can use both UV and RID detectors and maybe a ESI-MS (ion trap) instrument.
Can anybody suggest a supplier of the casuarina alkaloids standards?
Casuarina equisetifolia leaves extract contain alkaloids.The dried leaves of Casuarina equisetifolia (2 kg) are finely powdered and exhaustively extract them with 100% methanol, by maceration at room temperature. The crude methanolic extract should be evaporated to dryness under reduced pressure. The process of maceration and evaporation must repeated till exhaustion of the plants powder, and then the residues are combined and weighed. Separation and quantification of phenolic compounds can be conducted on Agilent Technologies 1200 Series Separations Module (GmbH, Germany) equipped with G1322A Vacuum degasser, G1311A Quaternary Pump, G1314B Variable Wavelength Detector (SL), G1328B Manual Injector and G1316A Thermostatted Column Compartment can used for HPLC analysis. The extract can be separated at 35°C on a reverse phase HPLC, ACE 5 µm C18 column with dimensions 250 × 4.6 mm, detection at 280 nm. The mobile phase will be gradient of A (CH3COOH 2.5%), B (CH3COOH 8%) and C (acetonitrile). The best separation can be obtained with the following gradient: at 0 min, 5% B; at 20 min, 10% B. The solvent flow rate should keep 1 m/min. The volume injected should 20 µl. Phenolic compounds can be quantified by using standard calibration for each compound and expressed as mg/100 g.
Separation and quantification of flavonoids
This was done using the above mentioned HPLC system and the same column with a mobile phase of methanol: water 1:1 (0–10 min) and 7:3 (10–20 min) at a flow-rate of 1 ml/min and detection at 339 nm. Each identified flavonoid was quantified by using standard calibration for each compound and expressed as mg %.
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