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Heart homogenates were prepared from male rats in control conditions or after H<jats:sub>2<\/jats:sub>S infusion induced a transient cardiogenic shock (CS) or cardiac asystole (CA). Using a method of determination of gaseous H<jats:sub>2<\/jats:sub>S with a detection limit of 0.2 nmol, we found that the process of homogenization could lead to a total disappearance of free H<jats:sub>2<\/jats:sub>S unless performed in alkaline conditions. Yet, after restoration of neutral pH, free H<jats:sub>2<\/jats:sub>S concentration from samples processed in alkaline and nonalkaline milieus were similar and averaged \u223c0.2\u20130.4 nmol\/g in both control and CS homogenate hearts and up to 100 nmol\/g in the CA group. No additional H<jats:sub>2<\/jats:sub>S was released from control, CS, or CA hearts by using the reducing agent tris(2-carboxyethyl)phosphine or a strong acidic solution (pH &lt; 2) to \u201cfree\u201d H<jats:sub>2<\/jats:sub>S from combined pools. Of note, the reducing agent DTT produced a significant sulfide artifact and was not used. These data suggest that 1) free H<jats:sub>2<\/jats:sub>S found in heart homogenates is not a reflection of H<jats:sub>2<\/jats:sub>S present in a \u201cliving\u201d heart and 2) the pool of combined sulfides, released in a strong reducing or acidic milieu, does not increase in the heart in a measurable manner even after toxic exposure to sulfide.<\/jats:p>","DOI":"10.1152\/ajpheart.00464.2016","type":"journal-article","created":{"date-parts":[[2016,9,17]],"date-time":"2016-09-17T02:49:33Z","timestamp":1474080573000},"page":"H1445-H1458","source":"Crossref","is-referenced-by-count":14,"title":["H<sub>2<\/sub>S concentrations in the heart after acute H<sub>2<\/sub>S administration: methodological and physiological considerations"],"prefix":"10.1152","volume":"311","author":[{"given":"Takashi","family":"Sonobe","sequence":"first","affiliation":[{"name":"Division of Pulmonary and Critical Care Medicine, Department of Medicine, Pennsylvania State University, College of Medicine, Hershey, 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