Separation of Anticholinergic Mydriatic Agents

In this Method, Tropicamide and Homatropine are separated with a simple Gradient. Homatropine is significantly hydrophobic yet still retains well.

Furthermore, the Separation between these two solutes is good, illustrating the Selectivity capabilities of this Method. Data is shown for two lots of Columns in order to demonstrate the robustness and precision of this Method.

1. Homatropine
2. Tropicamide

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

Time (minutes) %B
0 95
1 95
6 50
7 95

Injection vol.: 2μL
Flow rate: 1.0mL / minute
Detection: UV @ 220nm
Sample Preparation: 0.1mg / mL Homatropine and 0.01mg / mL Tropicamide reference standards in diluent of 50:50 Solvent A / Solvent B. Peak identities were confirmed with individual standards.
t0: 0.9 minutes

Note: Homatropine and Tropicamide are mydriatic (pupil-dilating) agents used in ophthalmoscopic examinations. Trade names include Homatropaire® for Homatropine and Mydral® for Tropicamide.


No 219 Tropicamide and Homatropine Analyzed with HPLC pdf  0.4 Mb  Download File