HPLC Analysis of 1,2,4-Triazole on Coresep 100 Mixed-Mode Column

Application description

Mannitol, ibandronic and citric acids are part of the formulation to reduce bone resorption, strengthen bones and treat Osteoporosis. Etidronic acid also prevents bone calcification. Citric acid is used as a preservative in the formulation. All three compounds are very polar in nature with citric and etidronic acids being acidic and mannitol being neutral. All three are not retained in RP chromatography. In HILIC, citric acid and etidronic acid can elute with a poor peak shape due to unspecified secondary interactions. We have developed a robust HILIC/anion-exclusion method for separation of these polar compounds. This method is compatible with major detection techniques for such compounds with no UV activity. (ELSD, CAD, RI and MS). The method can be used for analysis of polar acidic, basic and neutral compounds in HILIC, cation-exchange and anion-exclusion modes. The retention time is controlled by the amount of acetonitrile, buffer pH and buffer concentration. It is recommended to use salt buffer to achieve better retention and elution control.

Conditions of Experiment
Column: Coresep 100
Separation Modes: reversed-phase, cation-exchange
Column Dimenstions: 4.6x150 mm 5 um, 100A
Mobile Phase: ACN/water/TFA
Detection: UV 200 nm
Sample: 2 mg/ml
Injection: 5 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Amines, Aromatic base
Nature of compounds: Basic, Hydrophilic, Polar
Compounds: 1,2,4-Triazole