UPLC-MS/MS测定姜黄素在MCF-7/Adr细胞中的亚细胞分布研究

o investigate distribution of Cur in subcellular compartments of resistant MCF-7/Adr cells, a specific and sensitive high-performance liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed and validated for the determination of curcumin (Cur) in intracellular compartments. Sequential extraction of four subcellular protein fractions (cytosolic, membrane/organelle, nucleic and cytoskeleton) was performed directly from MCF-7/Adr cells after incubation with 8 μg/mL Cur for 0.5, 1, 2 and 4 h, respectively. The calibration curves were linear over a concentration range of 5 - 2000 ng/mL for Cur （y = 0.00115x + 0.00517, r = 0.9910). The recoveries were 99.21%, 96.08% and 100.45% for three concentrations, respectively. The intra-day and inter-day precision was less than 15%. Cur was mainly distrubuted in nucleus and membrane (organelle) during 0.5 h and 2 h, seldom in cytosolic and cytoskeleton. In conclusion We quantitatively determin the location and mechanism of Cur in the subcellular compartment of resistant MCF-7/Adr cells. It will provide some experimental datas for the mechanism study of Cur in antitumor.