Much is still unknown concerning the mechanisms underlying the development of chronic muscle pain. The presence and magnitude of inflammatory substances and neurotransmitters in chronic painful conditions is not clear. The aims of the present study were to determine, with the use of microdialysis, the interstitial concentrations and the equilibration times for PGE2 and glutamate in the trapezius muscles of nine female subjects with chronic muscle pain, and nine pain-free age-matched controls. A microdialysis probe was implanted in the upper part of the trapezius muscle and perfused with Ringer-acetate solution at a flow rate of 0.3 microL/min. Samples were obtained every 30 min, during a 4-h rest period. At equilibration, the mean concentrations (+/-SE) of PGE2 were 0.71 (+/-0.11) ng/mL for the pain-group and 0.97 (+/-0.35) ng/mL for the controls. For glutamate the mean concentrations for the pain-group were 66.3 (+/-13.3) micromol/L and 60.6 (+/-22.9) micromol/L for the controls. For the pain group and the control group, respectively, equilibration for PGE2 was reached at 180 and 150 min, and for glutamate at 150 and 120 min. The present study showed no differences between groups in the concentrations of PGE2 and glutamate in the trapezius muscle. Further, it revealed that when using the slow-flow method, a period of at least 2.0-2.5 h is needed, after probe insertion, to reach steady state for glutamate and PGE2.