Sample descriptionPhyllanthus amarus
Metabolite extraction protocolThe samples were pulverized with liquid nitrogen. Around 100 mg of powder was weighed and mixed with 1 ml of ice-cold extraction solution (75% aqueous MeOH+0.1% FA). Sonicated the samples in water bath at room temperature with maximum frequency of 40 kHz for 15 min. Centrifugation was performed at 12,000 rpm for 15 min. The supernatant was filtered using sterile 0.22 µm filter and required quanity of filtrate to be used for subsequent mass spectrometry analysis
Instrument usedAgilent 6530 quadrupole time of flight LC/MS
Instrument typeHPLC-ESI-QTOF-MS and MS/MS
IonizationESI
Ionization modePositive
Column usedWaters Xbridge C18 column 5µm (4.6x250mm)
Run time60 min
Mobile phase(A) Deioinized water with 0.1% formic acid (B) Acetonitrile with 0.1% formic acid
Gradient program5-35% B at 0-45 min, 35-75% B at 45-47 min, 75% B at 47-52 min, 75-5% B at 52-54 min, 5% B at 54-60 min.
Flow rate used0.4 ml/min
Collision energyramp 10-60eV