Finding a new Cyanide (CN) antidote is a great milestone in toxicology. Nithiodote™ and Cyanokit® are the two current CN antidotes used in the USA. Due to some of their limitations, there was a need to develop an effective intramuscular (IM) antidote to treat CN intoxication. The small lipophilic compound Dimethyl trisulfide (DMTS) has been proposed as a novel CN antidote. Protein binding and membrane binding are important aspects of drug development as they influence the bioavailability of a drug. The well-known Protein Equilibrium Dialysis (PED) method was used for the protein binding characterization for two molecules: the CN antidote candidate Poly-80 formulated Dimethyl trisulfide (DMTS),(Poly80-DMTS) and the antibiotic Cefuroxime. In this study, the Parallel Artificial Membrane Permeability Assay (PAMPA) method was employed for the protein binding and penetration studies. The moles in the donor cells and the acceptor cells for both drugs were determined using High Performance Liquid Chromatography (HPLC) method calibrated for the specific PED setup in PION PAMPA Buffers both in the presence and absence of Albumin. In PAMPA studies with Poly80-DMTS, the presence of Albumin decreased the membrane penetration by factor 1.27 and enhanced the membrane retention by factor 1.20. This suggests that DMTS has an affinity for binding to Albumin and the DMTS-Albumin complex is less capable of traveling through the artificial membrane. More experimental conditions are being investigated in the ongoing studies. In PED studies with Poly80-DMTS, the percentage of DMTS transferred to the acceptor plate in the absence of Albumin was about 1.5 times greater than the percentage transferred when Albumin was present. This also suggests that DMTS has significant affinity for binding to Albumin. In PED studies with Cefuroxime, the percentage of Cefuroxime B and Cefuroxime A transferred to the acceptor plate was lowered by a factor of 1.6, when Albumin was present in the donor plate, suggesting that Cefuroxime also has significant binding affinity to Albumin. DMTS shows better in vitro and in vivo antidotal character than any of the current CN antidotes. Presently DMTS is on the way to the FDA approval for being established as a therapeutic agent to treat CN intoxication. Clinical trials and metabolism studies are listed as to be done experiments.