Chiral chemicals have attracted significant interest in the pharmaceutical industry, yet the separation methods to get pure enantiomers from racemic mixture are still challenging. To date, the separation of enantiomers still mainly depends on chromatography using high-cost chiral stationary phases. Herein, wood channels were used as the handheld integrated device, and enantiomer separation was simultaneously detected using an electrochemical detector. In this method, a chiral UIO-66 (L-UIO-66) modified enantiomer separation zone and carbonized wood based online detection zone are integrated along a single wood column. Based on the in situ separation results from the chronoamperometry data, the wood device shows excellent separation ability for a wide range of electrochemically active enantiomers, including 3,4-dihydroxyphenylalanine, amino acids, ascorbic acid, carnitine, and penicillamine with high chirality purity. The unbiased molecular dynamic simulations indicate that the excellent chiral recognition and separation are attributed to the different barriers from the bound states to the dissociated state of the enantiomers in the homochiral microenvironment of the framework. This integrated enantiomer separation-electrochemical detection device provides a novel, easy, and low-cost platform for the separation of pure enantiomer from racemic mixture.