📋 Workflow
Full workflow, products add-to-cart directly1
In vitro metabolism incubation
2–4 hIncubate drug with liver microsomes/hepatocytes and cofactor (NADPH) at multiple time points
💡 Tip Include no-NADPH and no-protein controls
⚠️ Caution Optimize protein and substrate concentrations
🧪 Materials for this step (click to shop by spec/brand)
2
Quench and sample preparation
1 hQuench with organic solvent to precipitate protein, centrifuge and collect supernatant
💡 Tip Include internal standard for precision
⚠️ Caution Handle on ice to prevent degradation
🧪 Materials for this step (click to shop by spec/brand)
3
LC-MS analysis
2–4 hUse LC-MS to separate and quantify parent drug and metabolites
💡 Tip MRM mode improves sensitivity
⚠️ Caution Validate method for reliable quantification
🧪 Materials for this step (click to shop by spec/brand)
4
Data analysis and evaluation
2 hCalculate metabolic half-life, clearance and intrinsic clearance to evaluate metabolic stability
💡 Tip Integrate with in vivo PK data
⚠️ Caution Assess cross-species predictability
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Wrong protein/cofactor bias metabolic rate
- ② Insufficient time points hamper half-life
- ③ Matrix effect disturbs MS quantification
- ④ Missing metabolite standards limit profiling
- ⑤ In vitro-in vivo extrapolation is challenging
❓ FAQ
+Microsomes vs hepatocytes?
Microsomes enriched in phase I are fast; hepatocytes retain phase I/II and transporters for holistic view.
+Poor metabolic stability?
Optimize structure to block soft spots, reduce lipophilicity or prodrug strategies.
+What is clearance?
Volume of drug cleared per unit time, governing dosing frequency and steady-state exposure.