Simultaneous HPLC Analysis of Sucrose, Fosfomycin and Common Counterions on Amaze TH Mixed-Mode Column

Application description

Sucrose, fosfomycin, sodium and TRIS are part of the fosfomycin formulation. Sucrose, fosfomycin, sodium and tris are very polar in nature. Fosfomycin is a broad-spectrum antibiotic which contains an epoxy fragment as well as phosphonic acid. This makes this drug very polar and acidic. Sodium ion and tromethamine are used as counter-ions for this polar drug molecule. All four compounds are different in nature - sucrose is polar and neutral, sodium is polar and basic, TRIS is polar and basic, and fosfomycin is polar and acidic. It is hard to retain and separate these compounds in RP chromatography. HILIC can retain these polar compounds and mixed-mode HILIC on the Amaze TH column allows to introduce cation- and anion-exchange mechanisms for better retention and better retention control. The presence of ionic interaction, in addition to HILIC, allows better control of elution and retention. The retention time can be controlled by three parameters - amount of acetonitrile, amount of buffer and the buffer pH. This method can be used for the analysis of drug formulation and level of fosfomycin in biofluids (serum, blood, urine, etc). The method is robust, reproducible and compatible with all major detection techniques.

Conditions of Experiment
Column: Amaze TH
Separation Modes: HILIC, cation-exchange, anion-exchange
Column Dimenstions: 4.6x150 mm 5 um, 100A
Mobile Phase: ACN/water/Buffer
Detection: ELSD
Sample: 1 mg/ml
Injection: 10 uL
Flow rate: 1 ml/min
Class of compounds: Amines, Antibiotic, Drug, Inorganic cation, Sugar
Nature of compounds: Acidic, Basic, Hydrophilic, Neutral, Polar
Compounds: Fosfomycin, Sucrose, Sodium ion, Tromethamine