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Esters of thiosulfoacids demonstrate a wide range of biological activity. One of their effects is the influence on the metabolism of proteins and lipids in the body. Therefore, the purpose of our experiment was to study the impact of synthesized thiosulfonates on the total content of proteins and phospholipids, as well as their spectrum in the blood, liver, and kidney of rats. For the experiment, allyl, ethyl, and methyl esters of thiosulfoacid were used. The protein profile of rat tissues was investigated by electrophoresis, and the ratio of different fractions of phospholipids by thin-layer chromatography. Our results have shown that short-term administration of thiosulfonates in a dose of 300 mg/kg of body weight did not cause significant changes in the content of total protein and its fractions in liver tissue, whereas the effect of allyl and ethyl esters of thiosulfoacid was accompanied by an increase in the total protein and albumin in the blood plasma. The decrease in total protein was found in the kidney tissue of rats injected with allyl- and methyl thiosulfonates. The newly synthesized compounds did not lead to significant changes in the total content of phospholipids in blood plasma and tissues of rats, except for methyl thiosulfonate, the effect of which was accompanied by an increase in the total phospholipids in the liver of rats. These data may indicate an adaptive reaction of the rat’s organism. Tissue-specific features of the phospholipid spectrum were detected in rats after short-term exposure to thiosulfonates. The most significant effect on the phospholipid profile in the blood was shown for allyl- and ethyl esters of thiosulfoacid. Their action was accompanied by a decrease in the phosphatidylserine and phosphatidylinositol fractions, while phosphatidylethanolamine and phosphatidylcholine increased, respectively. Esters of thiosulfoacid significantly influenced the ratio of different fractions of phospholipids in the liver and kidney tissues. The phospholipid composition of the liver was more influenced by the allyl and methyl esters of thiosulfoacid, whereas for the kidney tissue a greater effect was observed for ethyl and methyl esters. Thus, the action of allyl ester of thiosulfoacid caused a decrease in the asymmetry coefficient of hepatocyte membranes, indicating an elevation of the lipid bilayer saturation and the increase of membrane microviscosity. Similar changes were found in the kidney of rats treated with allyl- and ethyl thiosulfonates.