Zhou Yuan-hu
2015
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Journal
Chinese Journal of Pharmaceutical Analysis
Abstract
Objective: To establish an HPLC method for determination of adenosine triphosphate disodium injection using adenosine as the reference. Methods: The method was established using the Elite Hypersil C8column( 4. 6 mm ×250 mm,5 μm) at the determination wavelength of 259 nm. The mobile phase was 0. 05 mol·L- 1 phosphate buffer( with 0. 161% tetrabutylammonium bromide,p H 7. 0). The flow rate was 1. 0 m L·min- 1and the column temperature was 35 ℃. The content was calculated by HPLC- external standard method using the relative molecular mass ratio( 2. 062) of adenosine triphosphate disodium( ATP- 2Na) to adenosine as the coefficient and adenosine as the reference. Results: The adenosine calibration curve was linear over the concentration range of 5. 056-404. 4 ng·m L- 1( linear regression equation was A = 3. 219 × 104 C + 31. 70,r = 0. 999 9) and the ATP- 2Na calibration curve was linear over the concentration range of 9. 939- 795. 1 ng·m L- 1( linear regression equation was A = 1. 567 × 104 C + 22. 16,r = 1. 000). The limit of quantification was 7. 6 × 10- 5mg·m L- 1for adenosine and was2. 6 × 10- 4mg·m L- 1for ATP- 2Na. RSDs of both repeatability and intermediate precision were 1. 0%. The recovery of ATP- 2Na was 101. 1%( RSD = 0. 9%,n = 9). The contents were 94. 2%- 109. 2%. Conclusion: The proposed HPLC method solved the issue of obtaining the ATP- 2Na reference with accurate and stable content,improving the specificity and accuracy of the method,which can be successfully applied in determination of the content of adenosine triphosphate disodium injection.