Key facts
- Brand
- Agilent
- Model
- 10-C18
- Industry
- Industrial
- Documents
- 2
- Languages
- EN
- Last added
- 2012-06-11
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Agilent SepTech ST150 10-C18 Peptide Resolution Application Note
Agilent Technologies investigated alternative solvents for reversed phase HPLC analysis of peptides due to global shortages in acetonitrile. This document evaluates ethanol as a viable substitute, comparing its performance against acetonitrile and three leading competitor columns.
The study utilizes the SepTech ST150 10-C18 column with a mixture of four peptides (Oxytocin, Angiotensin II, Angiotensin I, and Insulin). Results indicate that while ethanol provides similar solvent strength to acetonitrile, it alters selectivity, causing partial co-elution between insulin and angiotensin I. Additionally, ethanol exhibits higher UV absorbance at 220 nm compared to acetonitrile.
Agilent 10-C18 specifications
| Column Type | SepTech ST150 10-C18 |
|---|---|
| Flow Rate | 1 mL/min |
| Detection Wavelength (Acetonitrile) | 220 nm |
| Detection Wavelength (Methanol) | 280 nm |
All Agilent 10-C18 documents by type
How to use the product for one specific task, with worked examples.
Questions answered in these manuals
What is the stated particle size of the SepTech ST150 10-C18 column?
The column has a particle size of 10 micrometers.
From: Agilent SepTech ST150 10-C18 Peptide Resolution Application Note
What is the recommended flow rate for the peptide separation described?
The procedure specifies a flow rate of 1.0 mL/min.
From: Agilent SepTech ST150 10-C18 Peptide Resolution Application Note
Which alternative solvent is best for peptide analysis on the ST150 10-C18 column?
Methanol was found to be the best alternative, providing separation similar to acetonitrile with greater spacing between peptides.
Why must the detection wavelength be changed when using methanol?
Methanol has a high UV response at 220 nm, so the separation must be run at 280 nm to avoid background absorbance issues.