Separation of Hydrophobic Compounds with HPLC & UHPLC
This AppNote shows separation of analytes within a range of hydrophobicity. A simple gradient is used to elute all the compounds and baseline separation is obtained for the critical pair (peaks 4 and 5) and the least hydrophobic compound is adequately retained.
A comparison is shown in the figure below with a 4μm Cogent Bidentate C18 Column and a similar 2.o (2.2µm) Column. The retention profiles are quite comparable, meaning Method Transfer from one Column to the other will be easy to achieve.
1. Acetanilide, 2. Benzonitrile, 3. Methyl Benzoate, 4. Ethyl Benzoate,
5. Phenyl Benzoate, 6. Benzyl Benzoate, 7. Bibenzyl 8. Triphenylamine
Columns: Cogent Bidentate C18 2.o™, 2.2μm, 120Å; Cogent Bidentate C18™, 4um, 100Å
Catalog Nos.: 40218-05P-2; 40018-05P-2
Dimensions: 2.1 x 50mm for both Columns
—A: DI Water / 0.1% Formic Acid (v/v)
—B: Acetonitrile / 0.1% Formic Acid (v/v)
Injection vol.: 1 µL
Flow rate: 0.3mL / minute
Detection: UV @ 254nm
Sample Preparation: Mixture of solutes in 80:20:0.1 Acetonitrile / DI Water / Formic Acid Diluent. Peak identities were confirmed with individual standards.
t0: 0.7 minutes
No 274 Transfer HPLC Method to UHPLC with Hydrophobic Compounds pdf 1 Mb Download File