Determination of the Content and Related Substances of Sodium Deoxyribonucleotide by HPLC

OBJECTIVE To establish a method for determining the content and related substances of sodium deoxyribonucleotide by HPLC. METHODS The Agilent Zorbax SAX(4.6 mm×250 mm, 5 μm) was used, the mobile phase was 0.05 mol·L^-1 potassium dihydrogen phosphate solution (adjust pH to 3.3 by phosphoric acid) at the flow rate of 0.5 mL·min^-1, the detective wavelength was 258 nm. RESULTS The calibration curves showed good linearity of 2’-deoxycytidine 5’-monophosphate (d-CMP), 2’-deoxyadenosine 5’-monophosphate (d-AMP) , 2’-deoxythymidine 5’-monophosphate (d-TMP), 2’-deoxyguanosine 5’-monophosphate (d-GMP) over the range of 0.017 6~3.382 7 μg, 0.019 6~3.758 0 μg, 0.014 8~2.847 7 μg and 0.018 0~3.452 8 μg, r=1.000. The average recovery rates of d-CMP, d-AMP, d-TMP and d-GMP were 96.73%, 97.51%, 97.98% and 98.39%, RSD were 0.17%, 0.39%, 1.32% and 0.32%(n=9). CONCLUSION The established method has excellent resolution for the related substances, and it is accurate and sensitive for testing the content and related substances of sodium deoxyribonucleotide which can be used for control of the production.