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🧪 Separation & Purification · Method

Column Chromatography

The workhorse separation for pulling out product
🎯 Difficulty ★★☆ Medium Duration About 2–4 h 🎯 Use Purify crude synthetic products by silica column chromatography.
📖 Principle
Different components adsorb to silica differently, so solvent elutes them one by one.
Below are 4 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

Pack and settle the column

30 min
Pack silica by dry or wet method, tap until bubble-free.
💡 Tip Pre-wet and settle for even packing
⚠️ Caution Air bubbles cause uneven flow and tailing
🧪 Materials for this step (click to shop by spec/brand)
2

Load the sample

15 min
Dissolve crude product in a little solvent and load onto the top bed along the wall.
💡 Tip Dry-load on silica to save solvent
⚠️ Caution Overloading ruins the separation
🧪 Materials for this step (click to shop by spec/brand)
3

Gradient elution and collection

1–2 h
Elute with a polarity gradient, collecting fractions in tubes.
💡 Tip Fix the gradient from prior TLC first
⚠️ Caution Too fast a flow degrades resolution
🧪 Materials for this step (click to shop by spec/brand)
4

Combine and concentrate

30 min
Combine TLC-identical product fractions and concentrate by rotary evaporator.
💡 Tip Weigh combined product to get recovery
⚠️ Caution Don't pool edge fractions that still contain impurity
🧪 Materials for this step (click to shop by spec/brand)

⚠️ 5 Common Beginner Mistakes

  • Uneven packing causes edge flow
  • Overloading tails and fails to separate
  • Too steep a gradient overlaps impurities
  • Failing to check TLC pools impurity with product
  • Moisture in solvent worsens separation

❓ FAQ

+How to pick the gradient?
Use TLC to find an eluent giving Rf ~0.2–0.3.
+Can silica be reused?
Not recommended unless reactivated; residual impurities linger.
+Column blocked?
Often precipitate or bubbles; gentle pressure or tapping helps.

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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