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📊 Separation & Purification · Method

Preparative HPLC

Gram-scale purification with far better resolution
🎯 Difficulty ★★★ Advanced Duration About 3–6 h 🎯 Use Purify milligram-to-gram samples by preparative HPLC to high purity.
📖 Principle
Pressure drives repeated adsorption/desorption so you can cut out pure peaks.
Below are 5 steps. Open each to see how to do it and add products to your shared list.

📋 Workflow

Full workflow, products add-to-cart directly
1

Develop the analytical method

1 h
Use an analytical column to fix mobile phase, gradient, and resolution.
💡 Tip Scale to prep only when R>1.5
⚠️ Caution Direct scale-up of small-column results overloads
🧪 Materials for this step (click to shop by spec/brand)
2

Prepare and dissolve sample

20 min
Dissolve the sample in a minimum of starting mobile phase; filter if needed.
💡 Tip Dissolving in starting mobile phase sharpens peaks
⚠️ Caution Large particles block the prep column
🧪 Materials for this step (click to shop by spec/brand)
3

Preparative injection and separation

1–3 h
Inject onto the prep column, run the developed method, collect the target peak by detector trigger.
💡 Tip Use DAD or MS to guide peak cutting
⚠️ Caution Overloading overlaps peaks
🧪 Materials for this step (click to shop by spec/brand)
4

Collect and concentrate fractions

40 min
Collect fractions by auto-collector, pool same-peak fractions, evaporate solvent.
💡 Tip Evaporate under vacuum to protect sensitive product
⚠️ Caution Over-vigorous evaporation loses sample
🧪 Materials for this step (click to shop by spec/brand)
5

Verify purity

20 min
Re-check purity on analytical HPLC; confirm by LC-MS if needed.
💡 Tip >95% is usually ready for downstream
⚠️ Caution UV-invisible impurities may still exist
🧪 Materials for this step (click to shop by spec/brand)

⚠️ 5 Common Beginner Mistakes

  • Not scaling flow with column diameter
  • Overload overlaps peaks
  • Cutting too wide pools impurity peaks
  • Unfiltered sample blocks the column
  • Solvent evaporation loses sample

❓ FAQ

+Yield vs resolution trade-off?
Increase load when R>1.5; otherwise reduce it.
+How to scale a small-column method?
Scale flow and load by the square of diameter ratio.
+Collected impurity?
Re-run that peak region or tweak the gradient.

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Data authenticity: Product categories from JW Supply Chain standard taxonomy; method→step→product connected. Last updated: 2026-08-23 · v3.0