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Agilent Analysis of Peptides on a PLRP-S 100Å 10 m with ELS Detection Acetonitrile-free Eluents Manual
Due to global shortages, this document investigates alternative solvents for reversed phase HPLC peptide analysis. It details how switching from acetonitrile to methanol or ethanol can be achieved while maintaining separation efficiency using the Agilent PLRP-S 100Å column.
The primary focus is on utilizing the 385-LC ELS detector to overcome the high background absorbance and baseline drift caused by these alternative solvents in UV detection. The text provides specific gradient conditions, mobile phase compositions, and comparative data showing that the ELS detector produces flat baselines and sharp peaks comparable to acetonitrile-based runs.
Agilent PLRP-S 100A specifications
| Part Number | PL1512-3800 |
|---|---|
| Eluent | 0.1% TFA in 20% ACN / 80% water |
| Flow Rate | 1.0 mL/min |
| Detection | UV, 220 nm |
Source: Agilent Isocratic Purification of Synthetic Acyl Carrier Protein Fragment 65-74 Application Note
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How to use the product for one specific task, with worked examples.
Questions answered in these manuals
What alternative solvents are tested for acetonitrile-free peptide analysis?
The document tests methanol and ethanol as alternatives to acetonitrile.
How does the 385-LC ELS detector handle alternative solvents?
The 385-LC ELSD is capable of evaporating a wide range of different solvents, resulting in completely flat baselines and sharp peaks.
What media is used for the purification?
Macroporous, rigid polystyrene/DVB-based reversed phase media, PLRP-S.
From: Agilent Isocratic Purification of Synthetic Acyl Carrier Protein Fragment 65-74 Application Note
What is the maximum amount of crude synthetic ACP 65-74 chromatographed?
Approximately 1 mg of crude synthetic ACP 65-74 can be chromatographed isocratically.
From: Agilent Isocratic Purification of Synthetic Acyl Carrier Protein Fragment 65-74 Application Note
What column was used for the purification of leuprolide?
A PLRP-S 100A 10 um column was used for the purification.
From: Agilent PLRP-S 100A Leuprolide Purification Application Note
What is the amino acid sequence of leuprolide?
The sequence is p-Glu-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt.
From: Agilent PLRP-S 100A Leuprolide Purification Application Note