The change of intracellular adenosines concentration is the sensor of cellular energy metabolism changes, and it will contribute to monitor the effect of drugs on cellular energy metabolism with establishment of high performance liquid chromatography-tandem mass spectrometry to detect intracellular adenosines concentration. The adenosines of cells were extracted by ultrasonic extraction with perchloric acid containing Na-EDTA. The chromatographic separation was achieved with a UPLC HSS T3 column and waters containing 8 mmol/L N, N-dimethylhexylamine (DMHA) and acetonitrile as mobile phases at 0.25 ml/min flow rate. The identification and quantification were achieved by using ESI-MS/MS in positive ion and with multiple reactions monitoring (MRM) mode. With linear ranges of AMP ((0.1814～14.5164) μmol/L), ADP ((0.2342～18.7354) μmol/L) and ATP ((0.2003～16.0260) μmol/L), the external calibration method was well used for the quantitation, and the correlation coefficient of AMP, ADP and ATP were 0.9984, 0.9964 and 0.9990 respectively. The limit of detections (LOD, S/N > 3) of AMP, ADP and ATP were 1.9291, 1.8794 and 166.5 nmol/L, and their limit of quantifications (LOQ, S/N > 10) were 1.9632, 1.9672 and 185.6 nmol/L, respectively. The recoveries of the spiked standards varied from 81.8% to 107.8% with relative standard deviations (RSDs) less than 7.55%. The intraday precisions of AMP, ADP and ATP were 6.16%, 5.13% and 7.66%, and their interday precisions were 6.36%, 2.74% and 6.77%, respectively. The method was fast, simple, sensitive and suitable for the quantitative analysis of AMP, ADP and ATP in cell extracts. Analysis of AMP, ADP and ATP in human lung cancer A549 cells treated with different concentration volatile oils of Alpinia officinarum Hance, the results showed that the total level of cellular adenylate energy and ECP were decline with concentration dependence. The levels of ATP/TAN were decreased significantly when the concentration was 500 mg/L but the level of AMP/ATP in A549 cells were increased with concentration dependent. It was suggested that the volatile oils of Alpinia officinarum Hance inhibited cell proliferation by affecting the cellular energy metabolism.