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Theoretical analyses and Monte Carlo simulation are performed to investigate the detection limit of glucose concentration with near-infrared spectroscopy.The relation between detection limitation of glucose concentration and source-detector separation is derived.Monte Carlo simulation performed with a skin-layered model shows that the ratio of effective photons from the target layer could excess 50% by selecting proper source-detector separation,and that the detection limit of glucose concentration approaches to 0.28mM,which satisfies the requirement of food and drug administration for noninvasive glucose sensing.