Ketoconazole Analyzed with HPLC - AppNote
October 9, 2012
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Date: 9-OCTOBER-2012 Last Updated: 24-AUGUST-2026
Lipophilic Ketoconazole Retained using HILIC HPLCKetoconazole is a significantly lipophilic molecule (log P ~ 3.8) and is not generally suitable to ordinary Methods based on Hydrophilic Retention (HILIC). However, in this Method it is well retained using Aqueous Normal Phase (Inverse Gradient), therefore it is an Orthogonal Assay Method. This can be used in addition to standard Reversed Phase approaches since demonstrating orthogonality is very beneficial in a pharmaceutical analytical Method.
The data presented illustrates how this Method can be used to retain Hydrophobic Compounds very well and with Precision and Robustness as shown by the two overlaid Chromatograms from two different Column Lots.
Peak:
Ketoconazole
Column: Cogent Diamond Hydride™, 4 μm, 100 Å
Catalog No.: 70000-7.5P
Dimensions: 4.6 x 75 mm
Mobile Phase:
-- A: DI Water with 0.1% Trifluoroacetic Acid TFA (v/v)
-- B: Acetonitrile with 0.1% Trifluoroacetic Acid TFA (v/v)
Gradient:
| Time (minutes) | %B |
|---|---|
| 0 | 95 |
| 1 | 95 |
| 6 | 50 |
| 7 | 95 |
Injection vol.: 2 μL
Flow rate: 1.0 mL / minute
Detection: UV @ 225 nm
Sample Preparation: 1 mg Ketoconazole USP Reference Standard was dissolved in 1 mL of 50:50 Solvent A / Solvent B. This stock Solution was diluted 1:10 for HPLC injections using the same diluent.
t 0 : 0.9 minute.
