📋 Workflow
Full workflow, products add-to-cart directly1
Plate preparation
10 minChoose silica TLC plate and mark the spotting baseline
💡 Tip Mark with pencil to avoid dissolving
⚠️ Caution Do not touch the plate surface
🧪 Materials for this step (click to shop by spec/brand)
2
Spotting
15 minSpot samples on the baseline with capillary or spotting pipette
💡 Tip Small concentrated spots separate better
⚠️ Caution Too much sample causes tailing/overload
🧪 Materials for this step (click to shop by spec/brand)
3
Mobile phase & developing
30 minPlace mobile phase in the tank, insert the plate and develop away from light until the front
💡 Tip Saturate the tank before developing
⚠️ Caution Mobile phase surface below the spotting line
🧪 Materials for this step (click to shop by spec/brand)
4
Visualization & detection
15 minVisualize under UV lamp or spray with visualizing reagent
💡 Tip Check under UV 254 before spraying
⚠️ Caution Work under fume hood with spray reagents
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5
Rf calc & analysis
10 minMeasure spot migration distance and calculate/compar the Rf values
💡 Tip Run standards on the same plate for comparison
⚠️ Caution Rf affected by temperature and saturation
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Oversized spots causing tailing
- ② Unsaturated tank harming Rf reproducibility
- ③ Mobile phase above the spotting line
- ④ Hand contact contaminating the plate
- ⑤ Solvent evaporation altering mobile phase ratio
❓ FAQ
+What Rf is suitable?
Generally 0.2–0.5 gives good separation.
+Why broad streak?
Overloading or wrong mobile phase polarity.
+Can TLC be quantitative?
Semi-quantitative only; use HPLC for accurate quant.