Optimisation of isolation procedure for pyrrolizidine alkaloids from Rindera umbellata Bunge

Procedure for isolation of pyrrolizidine alkaloids (PAs) from Rindera umbellata Bunge plant species was optimised. Different extraction media (methanol, ethanol and sulphuric acid), concentration and volume of sulphuric acid, pH of PA solution for alkaline extraction, extraction time and techniques (maceration, ultrasonic and overhead rotary mixer assisted extraction) were investigated. The yields of six PAs (7-angeloyl heliotridane, 7-angeloyl heliotridine, lindelofine, 7-angeloyl rinderine, punctanecine and heliosupine) were monitored by GC-MS/FID. The best results for the isolation all of six PAs were obtained when the extraction was performed with 1M sulphuric acid (30mL per 1.00g of dried sample) by overhead rotary mixer during three days. Optimal pH value for alkaline extraction of PAs with CH2Cl2 was 9, and the extraction should be performed with four portions of 30mL of CH2Cl2. This procedure could be also useful for a plant sample preparation for GC and LC analyses of PAs.