Fluconazole Tablet Analyzed with HPLC – AppNote – MicroSolv Technology Corporation

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June 17, 2013
Bill Ciccone

AppNotes ANP/HILIC Methods / AppNotes Cogent Diamond Hydride / AppNotes Library / AppNotes Pharmaceuticals / ARTICLES, APPNOTES, FAQ

Assay Method for Amine Containing API

This Application Note shows a simple Gradient Method for Assay of Fluconazole Tablets. In Reversed Phase HPLC, notable tailing was observed due to the two heterocyclic amine groups. Use of Aqueous Normal Phase (ANP) HPLC produced a sharp Peak for this API. Data from three different Column lots is shown in order to demonstrate the Method Reproducibility and Robustness.

Peak:
Fluconazole

Method Conditions
Column: Cogent Diamond Hydride™, 4μm, 100Å
Catalog No.: 70000-7.5P
Dimensions: 4.6 x 75mm
Mobile Phase:
—A: DI Water with 0.1% Formic Acid (v/v)
—B: Acetonitrile with 0.1% Formic Acid (v/v)
Gradient:

Time (minutes)	%B
0	95
1	95
6	40
7	40

Post Time: 3 minutes
Injection vol.: 1μL
Flow rate: 1.0mL / minute
Detection: UV @ 260nm
Sample Preparation: 150mg strength Fluconazole Tablet was ground and added to 50mL volumetric flask containing 25mL 50:50 Solvent A / Solvent B diluent. The solution was sonicated 10 minutes, diluted to mark, and mixed. A portion was filtered through a 0.45μm Nylon Syringe Filter (MicroSolv Tech Corp.).
t0: 0.9 minutes

Note: Fluconazole is a triazole antifungal drug. Its mode of action is inhibition of fungal Cytochrome P450 enzyme 14a-demethylase. It is available under the trade name Diflucan®.

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