---
title: "Method development for peptide purification: How to create a step gradient"
description: Guidelines for creating a step gradient for peptide purification, ultimately enabling complete separation of amino acid peptide from an impurity.
---

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# [Method development for peptide purification: How to create a step gradient](https://www.biotage.com/literature/technical-note/method-development-peptide-purification-create-step-gradient)

 Written by [Biotage](https://www.biotage.com/literature/author/biotage) | Dec 6, 2025 9:54:57 AM

Synthetic peptide purification represents the most significant bottleneck of the entire peptide workflow. Reversed-phase flash chromatography is gaining traction as a strategy to reduce this bottleneck given the significant increase in loading capacity and short purification times. Presented herein are guidelines for creating a step gradient for peptide purification, ultimately enabling complete separation of a 20 amino acid peptide from a 19 amino acid peptide impurity.

Literature number: UI467

[View full post](https://www.biotage.com/literature/technical-note/method-development-peptide-purification-create-step-gradient)

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