📋 Workflow
Full workflow, products add-to-cart directly1
Mobile phase prep
20 minMix ultrapure water with organic solvent at the target ratio, degas by sonication and filter
💡 Tip Use degassed mobile phase to avoid baseline noise
⚠️ Caution Keep organic phase ratio consistent
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2
Standard & sample prep
30 minDissolve standards and samples in mobile phase to suitable concentration and filter
💡 Tip Prepare external standard calibration curve
⚠️ Caution Filter samples before injection to remove particulates
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3
Column equilibration
30 minFlush the column with mobile phase until baseline stabilizes
💡 Tip Equilibrate for at least 10 column volumes
⚠️ Caution Purge lines when changing mobile phase
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4
Method setup & injection
20 minSet flow rate, column temperature and detection wavelength; inject blank, standards and samples
💡 Tip Run in isocratic or gradient mode
⚠️ Caution Avoid overpressure that damages the column
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5
Data processing & quant
30 minIntegrate peak areas and calculate sample concentration using the calibration curve
💡 Tip Use at least 5-6 calibration points
⚠️ Caution Check retention time drift
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Non-degassed mobile phase causing baseline spikes
- ② Unfiltered samples clogging the column
- ③ Too few calibration points causing quant bias
- ④ Not addressing abnormal pressure increase
- ⑤ Wrong wavelength reducing sensitivity
❓ FAQ
+Why HPLC-grade solvents?
Low impurities avoid ghost peaks interfering with quantitation.
+What if pressure rises?
Back-flush or replace the inlet frit; replace the column if needed.
+Isocratic or gradient?
Use isocratic for few components; gradient for wide polarity differences.