Controversies and conundrums in hydrogen sulfide biology

• Heightened interest in H2S is associated with some growing pains.
• Most reported plasma and tissue concentrations are unrealistic.
• High H2S volatility is rarely considered.
• Substances interfering with H2S measurement and H2S effects need further evaluation.
• Recent interest in using plasma H2S in clinical diagnoses requires proper statistical analysis.

Hydrogen sulfide (H2S) signaling has been implicated in physiological processes in practically all organ systems studied to date. At times the excitement of this new field has outpaced the technical expertise or practical knowledge with which to accurately assess these advancements. Recently, the myriad of proposed H2S actions has spawned interest in using indicators of H2S metabolism, especially plasma H2S concentrations, as a means of identifying a variety of pathophysiological conditions or to predict clinical outcomes. While this is a noteworthy endeavor, there are a number of contraindications to this practice at this time. First, there is little consensus regarding normal, i.e., “physiological” concentrations of H2S in either plasma or tissue. In fact, it has been shown that the methods most often employed for these measurements are associated with substantial artifact. Second, interactions, or presumed lack thereof, of H2S with other biomolecules (e.g., O2, H2O2, pH, etc.) or analytical reagents (e.g., reducing reagents, N-ethylmaleimide, phenylarsine, etc.) are often assumed but not evaluated. Third, the experimental design and/or statistical analyses may not be sufficient to justify using H2S concentration in tissue or blood as a predictive biomarker of pathophysiology. In this study, we first briefly review the problems associated with plasma and tissue H2S measurements and the associated errors and we provide some simple methods to evaluate whether the data obtained is physiologically relevant. Second we provide a brief analysis of H2S interactions with the above biomolecules. Third, we provide a statistical tool with which to determine the clinical applicability of H2S measurements. It is hoped that these points will provide a rational background for future work.