METHOD FOR CHARACTERIZING SYNTHETIC OLIGONUCLEOTIDES OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH HIGH-RESOLUTION MASS SPECTROMETRIC DETECTION Russian patent published in 2025 - IPC G01N30/86 

FIELD: chemistry.

SUBSTANCE: invention relates to analytical chemistry, specifically to methods for assessing the quality of synthetic oligonucleotides for further use in various types of PCR and cell biology and can be used in medical, pharmacological, food, microbiological industry. Method of characterizing synthetic oligonucleotides using high-performance liquid chromatography with high-resolution mass spectrometric detection involves chromatographic separation of the analysed oligonucleotide on a chromatographic column with a reversed-phase C18 sorbent. Detection of the chromatographic flow is carried out using a high-resolution mass spectrometer equipped with an electrospray ionisation source (ESI) in the mode of negative ions. Multiply charged ions with charge numbers from 3 to 7 are registered, charge number of the most intense multiply charged ion is determined by the following formula: z=1/[(m/z)2-(m/z)1], where (m/z)1 is mass of the smaller neighbouring isotope peak, and (m/z)2 is the mass of the larger isotope peak. Experimental weight of the synthetic oligonucleotide is calculated by formula: M=z⋅(m/z+1.0073), where z is the charge number, m/z is the mass-to-charge ratio of the ion in the mass spectrum for the first isotope peak of the multicharged ion. Obtained experimental mass of oligonucleotide is compared with theoretical mass of said oligonucleotide sequence by first isotope peak, wherein compliance criterion is deviation of experimentally obtained value from theoretical value by not more than 5 ppm.

EFFECT: wider range of means of characterizing synthetic oligonucleotides.

2 cl, 2 dwg