Abstract: The method of determining ATP(adenosine-5'-triphosphate),ADP(adenosine-5'-dipphosphate),AMP(adenosine-5'-monophosphate),NAD~ (nicotinamide adenine dinucleotide) and NADH(nicotinamide adenine dinucleotide reduced) in vivo of pyruvate producing strain,Torulopsis glabrata 620 by high performance liquid chromatography(HPLC) was established and the profiles of both of energy charge and oxidationte-reduction state in vivo throughout the fermentation were studied.Energy charge in vivo plays a very important role in the biosynthesis of pyruvate.Pyruvate is accumulated intensively when the energy charge decreases to 0.1 and at the end of the production.It may represent a valuable tool to(improve) the production of pyruvate and to study the relationship between materials and energy in the cells.