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Updated: Mar 29, 2026
Efficient Purification and LC-MS/MS-based Assay Development for Ten-Eleven Translocation-2 5-Methylcytosine Dioxygenase
Published on: October 15, 2018
Assay development for determination of tenofovir in human plasma by solid phase analytical derivatization and
Anthony T Podany1,2, Curtis Sheldon2, Daryl Grafelman2
1Antiviral Pharmacology Laboratory, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Background:
A novel method was developed and validated to measure tenofovir in human plasma. RESULTS/METHODOLOGY: This method employed solid phase analytical derivatization and analysis by LC-MS/MS. Stable-labeled internal standard was added to plasma samples followed by solid phase extraction. Retained analytes were derivatized on the solid phase extraction cartridges with a diazomethane solution to yield methyl-ester derivatives. Samples were analyzed using LC-MS/MS incorporating the use of a strong cation exchange column. The method was validated over a range of 5.00-750 ng/ml. The approach developed in this report for tenofovir could be applied to other analytes that share similar structural similarities.
Conclusion:
The tenofovir LC-MS/MS method was used to support a clinical study of over 400 samples with a 100% success rate.
