Additional purification was carried out using a SPE C18 cartridge, previously activated with 3 ml methanol and 20 ml water, as flavourings and fat could not be separated by carrez clarification. 2 ml of the carrez clarified filtrate was added to the activated cartridge and eluted with 10 ml of mobile phase A, followed by 10 ml of mobile phase B. The isolate obtained was collected in a 25 ml volumetric flask and finally the volume was made up to the mark with a mixture of mobile phases A and B (1:1). A blank experiment was also performed simultaneously. A 20 ll aliquot of the filtrate
was injected into the HPLC.