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Abstract: A rapid and reliable microwave extraction and the triple quadrupole-linear ion trap mass spectrometry method was developed and validated for the determination of eight alkaloids in Portulaca oleracea L. The optimal microwave extraction (MWE) condition was performed at 60°C for 12min with ethanol–water (70:30, v/v) as the extracting solvent, and the solvent to solid ratio was 30:1. The alkaloids were first detected simultaneously by electrospray ionization tandem mass spectrometry under positive–negative conversion multiple reaction monitor ((+/−)MRM) technique. With investigating three different columns, samples were separated in only 8min on a Waters ACQUITY UPLC HSS T3 (50×2.1mm2, 1.8μm) column using acetonitrile and formic acid–water solution as a mobile phase with a flow rate at 0.2mL/min. All calibration curves showed good linearity (r>0.999) within the test ranges. The method developed was validated with acceptable sensitivity, intra- and inter-day precision, reproducibility, and extraction recoveries. It was successfully applied to the determination of eight alkaloids in Portulaca oleracea L. from different sources and different harvest periods. The method also provide a reference for extraction and determination of alkaloids in other complex systems. [Copyright &y& Elsevier]