GC–MS Lyophilized CCF was dissolved in GC grade dichloromethane (Merck) and derivatized with BSTFA (Sigma) following manufacturer’s protocol and run in an Agilent HP5 column (30 × 0.25x0.2) in temperature gradient of 70–260 °C with a ramping rate of 5 °C/min and helium flow of 1 ml/min in a GC/MS system (Agilent 7890A-5975C). Detection and identification of the isolated phytochemicals was done by NIST 2011 library.
LC–MS Lyophilized CCF was dissolved in HPLC grade methanol (Merck) and 3 µl of extract was injected in an Agilent G1316C column and run for 30 min with water and acetonitrile as mobile phase. The system is interfaced to a TOF/ Q-TOF mass spectrometer with an ion source analyzer- Dual AJSESI. The analysis was done from IIT, Mumbai.
Column chromatography A slurry was made from concentrated CCF and loaded in a glass column having silica gel (60–120). Elution was started with petroleum ether and then increasing concentrations of ethyl acetate and later with methanol. Sub-fractions were collected, concentrated and pooled. Pure fractions were tested for their cytotoxicity by MTT assay. Cytotoxic fractions were identified by NMR, ESI–MS studies.
LC–MS Lyophilized CCF was dissolved in HPLC grade methanol (Merck) and 3 µl of extract was injected in an Agilent G1316C column and run for 30 min with water and acetonitrile as mobile phase. The system is interfaced to a TOF/ Q-TOF mass spectrometer with an ion source analyzer- Dual AJSESI. The analysis was done from IIT, Mumbai.
Column chromatography A slurry was made from concentrated CCF and loaded in a glass column having silica gel (60–120). Elution was started with petroleum ether and then increasing concentrations of ethyl acetate and later with methanol. Sub-fractions were collected, concentrated and pooled. Pure fractions were tested for their cytotoxicity by MTT assay. Cytotoxic fractions were identified by NMR, ESI–MS studies.