Analysis of adenylosuccinate in biological matrices using hydrophilic interaction ultra-performance liquid chromatography-tandem mass spectrometry

Authors

• MM Hall
• AQ Wang
• X Xu

Abstract

Adenylosuccinate (ASA) is an important endogenous molecule of the purine nucleotide cycle (PNC), as well as for maintaining energy levels within the body, among other functions. ASA has been tested in clinical trials for diseases such as Duchenne’s muscular dystrophy (DMD). Although clinical development efforts on ASA for the treatment of DMD were abandoned, ASA still holds potential as a treatment for metabolic diseases involving the PNC such as Adenylosuccinate synthase-like 1 myopathy (ADSSL-1). Currently, there are few quantification methods which are crucial to the drug development process in the literature.
We developed an accurate and sensitive hydrophilic interaction ultra-performance liquid chromatography-tandem mass spectrometry (HILIC UPLC-MS/MS) quantitation method for ASA analysis in plasma and other biological matrices. Calibration standards ranging from 10 ng/mL to 100,000 ng/mL were prepared in plasma. Samples were extracted using protein precipitation with TFA and a stable isotope labeled internal standard in acetonitrile. The method utilizes a HILIC column with a 2-minute UPLC gradient which is paired with a triple quadrupole mass spectrometer to analyze ASA using positive ion electrospray. Due to the simple sample preparation procedures and short UPLC-MS/MS analysis time, the developed method will allow for the quantification of ASA in pharmacokinetic studies in order to develop ASA for the treatment of metabolic diseases such as ADDSL-1.

Scientific Focus Area: Molecular Pharmacology

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