OraChrom, Inc.

FYI NOTE

FYI041698

Resolving capabilities of a 25 cm column without the flow and pH restrictions of Silica.

Human Angiotensins with one to three amino acid variances can be readily separated with high resolutions even at very high speed by altering their retentions through pH adjustments.

Angiotensin I: Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu

Angiotensin II: Asp-Arg-Val-Tyr-Ile-His-Pro-Phe

Angiotensin III: Arg-Val-Tyr-Ile-His-Pro-Phe

Such separation is not possible with Silica under normal operating conditions:

The same separation could be achieved significantly faster by using the same polymeric column at higher flow rates:

Operating parameters:

HPLC System.	HP 1100, standard cell.
Column	As indicated on each chromatogram.
Mobile Phase
pH = 2	A: 0.1% TFA in H2O B: 0.1% TFA in Acetonitrile/H2O (95/5)(v/v)
pH = 11.5	A:7 mM PO4Na3 in H2O B: 7 mM PO4Na3 in ACN:H2O (50:50)
Gradient	Optimized in each case
Temperature	20°C.
Detection	214 nm

Polymeric Stationary Phases, Setting new standards in High Speed, High Performance,

Liquid Chromatography.

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