A novel method for rapid, room-temperature phosphorescence (RTP) chiral recognition of natural amino acids is presented. The method utilizes a single molecular-solid sensor, L-phenylalanine-derived benzamide, which can distinguish between natural (L) and unnatural (D) amino acids based on differences in RTP. The sensor is fabricated as nanocrystals through solvent evaporation, enabling efficient triplet-triplet energy transfer to chiral analytes. The results show that L-analytes induce strong RTP, while D-analytes produce minimal afterglow. This method expands the scope of luminescence chiral sensing by reducing the need for specific molecular structures. RTP from organic phosphors, particularly guest-host doped systems, has become a significant research area due to its long-lived lifetimes and flexibility. Guest-host RTP systems have been applied in various fields, including optoelectronics, bioimaging, and optical sensors. Recent studies have focused on the design and structure-property relationships of RTP systems with chiral moieties, such as circularly polarized luminescence. Understanding molecular chirality, excited state, and electron spin correlations can help elucidate fundamental physical principles and drive technological innovations. The study demonstrates that chiral-selective phosphorescence enhancement (CPE) in a donor-acceptor system is largely due to chirality-dependent energy transfer (CDET). The method allows for rapid chiral recognition using RTP by using a single energy donor to differentiate the chirality of various chiral structures attached to the same energy acceptor. The amino acid and naphthoyl chloride can be converted into a chiral system under ambient conditions. The L-phenylalanine derivative was selected as the universal triplet energy donor due to its ease of production and purification. The study shows that the enantioselective discrimination of amino acid-based systems can be quantified via RTP spectroscopy under rigorous test conditions. The host-to-guest energy transfer process is key to achieving enantioselective discrimination. The method was tested with 15 natural amino acids and their enantiomers, showing the best performance in both sensing time and substrate variety among all published studies. The method allows for rapid chiral recognition with sensing times as short as a few minutes. The study also explores the morphology and microstructure of solid-state samples, showing that different processing methods have minimal effects on the microscopic morphologies. The method is universal for RTP chiral sensing and can be applied to other naturally occurring amino compounds. The study also investigates the regulation of enantioselective differentiation and the mechanism of the process. The results show that the method can be used to optimize fine-tuning for the best recognition conditions. The study concludes that the method provides a universal design strategy for constructing an amino acid-based chiral guest-host RTP system with enantioselective discrimination photoluminescence performance in the solid state.A novel method for rapid, room-temperature phosphorescence (RTP) chiral recognition of natural amino acids is presented. The method utilizes a single molecular-solid sensor, L-phenylalanine-derived benzamide, which can distinguish between natural (L) and unnatural (D) amino acids based on differences in RTP. The sensor is fabricated as nanocrystals through solvent evaporation, enabling efficient triplet-triplet energy transfer to chiral analytes. The results show that L-analytes induce strong RTP, while D-analytes produce minimal afterglow. This method expands the scope of luminescence chiral sensing by reducing the need for specific molecular structures. RTP from organic phosphors, particularly guest-host doped systems, has become a significant research area due to its long-lived lifetimes and flexibility. Guest-host RTP systems have been applied in various fields, including optoelectronics, bioimaging, and optical sensors. Recent studies have focused on the design and structure-property relationships of RTP systems with chiral moieties, such as circularly polarized luminescence. Understanding molecular chirality, excited state, and electron spin correlations can help elucidate fundamental physical principles and drive technological innovations. The study demonstrates that chiral-selective phosphorescence enhancement (CPE) in a donor-acceptor system is largely due to chirality-dependent energy transfer (CDET). The method allows for rapid chiral recognition using RTP by using a single energy donor to differentiate the chirality of various chiral structures attached to the same energy acceptor. The amino acid and naphthoyl chloride can be converted into a chiral system under ambient conditions. The L-phenylalanine derivative was selected as the universal triplet energy donor due to its ease of production and purification. The study shows that the enantioselective discrimination of amino acid-based systems can be quantified via RTP spectroscopy under rigorous test conditions. The host-to-guest energy transfer process is key to achieving enantioselective discrimination. The method was tested with 15 natural amino acids and their enantiomers, showing the best performance in both sensing time and substrate variety among all published studies. The method allows for rapid chiral recognition with sensing times as short as a few minutes. The study also explores the morphology and microstructure of solid-state samples, showing that different processing methods have minimal effects on the microscopic morphologies. The method is universal for RTP chiral sensing and can be applied to other naturally occurring amino compounds. The study also investigates the regulation of enantioselective differentiation and the mechanism of the process. The results show that the method can be used to optimize fine-tuning for the best recognition conditions. The study concludes that the method provides a universal design strategy for constructing an amino acid-based chiral guest-host RTP system with enantioselective discrimination photoluminescence performance in the solid state.