Introduction to the preparation method of 1,4-Dithiothreitol (DTT)|CAS:3483-12-3|

DTT is a small molecule organic reducing agent that is widely used in cell biology, biochemistry and biomedical applications. This article will introduce the method of subfamily preparing dithiothreitol, the specific steps are as follows:

(1) In the first solvent (not limited to dichloromethane), add 1,4-disulfonic acid-2-butene, control the reaction temperature at 30℃-35℃, add the oxidant in batches, and complete the addition within 3 hours. Control the reaction temperature not to be higher than 40°C, stir for 16 hours to obtain the first intermediate, the molar ratio of 1,4-disulfonic acid-2-butene to the oxidant is 1:8-13, and the oxidant is but not limited to tungstic acid/ One of hydrogen peroxide and peroxybenzoic acid, its reaction formula is:

(2)Hydrolyze the first intermediate obtained in step (1) in lye, the molar ratio of the first intermediate to lye is 0.1-0.2:1.5-3, stir and mix, stand for liquid separation, and divide into the first water phase and the first oil phase; then the remaining formula amount of lye is added dropwise to the first oil phase, stirred for liquid separation, divided into the second water phase and the second oil phase, Combine the first water phase and the second water phase to obtain the total water phase; heat the obtained total water phase to 60℃-70℃, react for 6 hours, and cool down; then add activated carbon, remove water, stir, filter, and then add sulfuric acid , Adjust the pH value to 4.0, stir and filter, then continue to add sulfuric acid to adjust the pH value to 1.0, and then filter to obtain the first solid, the color of the first solid is white; add the first solid to water and heat to 80℃, Stirred for 1h, then cooled to 5℃, precipitated crystals, filtered, and dried to obtain the second intermediate with a white color; the reaction formula is:

(3) The second intermediate is added to the second solvent (the second solvent is but not limited to tetrahydrofuran), the reaction temperature is controlled to 0℃, and the reducing agent is added in batches. The reducing agent is but not limited to lithium aluminum hydride and sodium borohydride The molar ratio of the second intermediate to the reducing agent is 1:6-8. After adding the reducing agent, continue to stir at 0℃ for 2h, then heat to 25℃, stir for 16h, and quench the reaction with dilute hydrochloric acid solution ; Then add water/ethyl acetate for extraction and remove the solvent to obtain the first organic layer and the first inorganic layer; then use ethyl acetate to extract the first inorganic layer to remove the solvent to obtain the second organic layer and in the second inorganic layer, the first organic layer and the second organic layer are combined to obtain a total organic layer; the obtained total organic layer is subjected to vacuum distillation to obtain 1,4-dithiothreitol.

The process of the preparation method is simple, the yield is high, and the yield is more than 77%

References

CN112028799A Method for preparing 1,4-dithiothreitol