The　progress　on　reversed-phase　high　performance　liquid　chromatography　(RP-HPLC)　techniques　for　analyzing　the　main　energy　substances　in　biological　tissues　and　cells　was　reviewed　in　this　paper.　From　the　pretreatment　of　samples,　mobile　phase　and　how　to　establish　the　chromatogram　conditions　were　discussed.　Many　energy　substances　were　detected　rapidly　and　exactly　by　reversed-phase　high　performance　liquid　chromatography.　The　technique　is　simple　and　convenient,　and　requests　for　this　test　technique　as　follows:　the　capability　of　each　components　of　the　apparatus　was　stable;　the　chromatographic　column　was　in　high　performance;　chromatographic　separation　was　good　and　elution　curve　was　clear;　the　selection　spots　were　fixed　to　insure　the　comparability;　samples　were　extracted　at　a　low　temperature　and　were　moved　into　perchloric　acid　solution　disrupted　completely　when　refining　in　order　to　extract　ATP　fully,　and　then　centrifuged　at　a　low　temperature　to　make　sure　of　the　accuracy　of　the　results.　As　an　efficient　separation　technology,　reversed-phase　high　performance　liquid　chromatography　would　be　used　in　the　area　of　separation　and　quantitative　analysis　of　complex　energy　substances.