To determine kinetic parameters of sulfate reduction by hydrocarbons (HC) without the initial presence of low valence sulfur, we carried out a series of isothermal gold-tube hydrous-pyrolysis experiments at 320, 340, and 360 °C under a constant confined pressure of 24.1 MPa. The reactants used consisted of saturated HC (sulfur-free) and CaSO₄ in an aqueous solution buffered to three different pH conditions without the addition of elemental sulfur (S₈) or H₂S as initiators. H₂S produced in the course of reaction was proportional to the extent of the reduction of CaSO₄ that was initially the only sulfur-containing reactant. Our results show that the in situ pH of the aqueous solution (herein, in situ pH refers to the calculated pH value of the aqueous solution at certain experimental conditions) can significantly affect the rate of the thermochemical sulfate reduction (TSR) reaction. A substantial increase in the TSR reaction rate was observed with a decrease in the in situ pH. Our experimental results show that uncatalyzed TSR is a first-order reaction. The temperature dependence of experimentally measured H₂S yields from sulfate reduction was fit with the Arrhenius equation. The determined activation energy for HC (sulfur-free) reacting with View the MathML sourceHSO₄⁻ in our experiments is 246.6 kJ/mol at pH values ranging from 3.0 to 3.5, which is slightly higher than the theoretical value of 227.0 kJ/mol using ab initio quantum chemical calculations on a similar reaction. Although the availability of reactive sulfate significantly affects the rate of reaction, a consistent rate constant was determined by accounting for the HSO₄⁻ ion concentration. Our experimental and theoretical approach to the determination of the kinetics of TSR is further validated by a reevaluation of several published experimental TSR datasets without the initial presence of native sulfur or H₂S. When the effect of reactive sulfate concentration is appropriately accounted for, the published experimental TSR data yield kinetic parameters that are consistent with our values. Assuming MgSO₄ contact-ion-pair ([MgSO₄]CIP) as the reactive form of sulfate in petroleum reservoir formation waters, a simple extrapolation of our experimentally derived HSO₄⁻ reduction kinetics as a proxy for [MgSO₄]CIP to geologically reasonable conditions predicts onset temperatures (130–140 °C) that are comparable to those observed in nature.