Application of HPLC-MS analysis in the study of formylation 2-alkyl-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidine-7(4H)-ones
Abstract
Compounds based on [1,2,4]triazolo[1,5-a]pyrimidine have a wide range of biological activity. Despite considerable interest in [1,2,4]triazolo[1,5-a]pyrimidine derivatives, the issues of their synthesis and modification remain relevant. In particular, the formylation of these compounds is insufficiently covered in the literature. The purpose of this study was to study the Wilsmeyer-Haack reaction formylation of 2-alkyl-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidine-7(4H)-ones using high performance liquid chromatography in combination with mass spectrometry (HPLC-MS).
The initial 2-alkyl-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidine-7(4H)-ones were formylated using phosphorus chloride in anhydrous dimethylformamide medium.
A reaction mixture obtained as a result of the formylation of 2,5-dimethyl-[1,2,4]triazolo[1,5-a]pyrimidine-7(4H)-one is considered as a model system. When interpreting the results of the HPLC-MS analysis, it was found that in addition to the target 2,5-dimethyl-[1,2,4]triazolo[1,5-a]pyrimidine-6-carbaldehyde, the reaction mixture contains three products of unknown structure. The obtained mixture was separated using the method of preparative column chromatography. A silica gel fraction of 60-200 microns (Russia) was used as the stationary phase (NF), and a chloroform–methanol mixture (20:1) was used as the mobile phase (PF). Four fractions were obtained by elution of the resulting mixture. The first fraction contained a minor impurity, the second a major impurity, the third a target carbaldehyde, and the fourth a mixture of by–product and carbaldehyde.
The structure of the individually obtained compounds has been proven by 1H and 13C NMR and mass spectrometry. It was found that 2,5-dimethyl-[1,2,4]triazolo[1,5-a]pyrimidine-6-carbaldehyde is formed in the OH form, and the structure of 6-(dimethylamino)-2-methyl-4H-cyclopenta[d][1,2,4]triazolo[1,5-a]pyrimidine-7,8-diol. The analysis of substances corresponding to the minor impurity and by-product was not carried out, which is due to the difficulty of isolation in pure form and a yield value of less than 1%.
Similarly, 7-hydroxy-5-methyl-2-ethyl-[1,2,4]triazolo[1,5-a]pyrimidine-6-carbaldehyde and 6-(dimethylamino)-2-ethyl-4H-cyclopenta[d][1,2,4]triazolo[1,5-a]pyrimidine-7,8-diol were obtained.
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