◈ Quick Reference

Chemicals

MeOHMethanol

CHCl₃Chloroform

H₂ODistilled Water

NH₃Ammonia (basify)

HClHydrochloric acid

Hexanen-Hexane

Key Volumes — M1 JT Baker

Condition3 mL MeOH

Equilibrate3 mL H₂O

Sample20 mL

Wash1 mL 5% MeOH

Elute3 mL × 2

Key Volumes — M2 Florisil

Condition3 mL MeOH

Equilibrate3 mL CHCl₃

Sample20–50 mL

Wash3 mL CHCl₃

Elute6 mL CHCl₃ (×2)

pH Reference

Alkaline target~8.7

Filter target6–7

Dry temp40–60 °C

Sample weight0.50–1.00 g

⚠ Safety Notes

CHCl₃ — Toxic & volatile. Use fume cupboard at all times.

Hexane — Flammable. No open flames near workspace.

Dry bath max 60 °C. Do not overheat eluent.

Laboratory Procedure Reference

Solid-Phase
Extraction

Three-Method Guide — SPE Protocol

3Methods

C18 · Florisil · SoftgelCartridges

Syarief AzmanPrepared by

Overall Progress 0%

Completed: 0

Remaining: 24

Total Steps: 24

OverviewJT Baker 500 mg / 6 mL — Non-Polar Extraction

Powder · Tablet · Capsule Solvent: MeOH / H₂O SPE Busc Config

🔬CiQ samples: 10 mL sample + 10 mL H₂O → sonicate → measure pH → filter. Uses vacuum manifold with back-vessel insert.

Step ASample Preparation — tap each step to check off

Weigh

Weigh sample 0.50–1.00 g in beaker

Dissolve

Add 20 mL water

Sonicate

Sonicate for 5–10 minutes

⏱ 10 min

pH Check

Measure the solution with pH indicator

Adjust pH / Basify

Neutralize / Basify / Acidify using NH₃ or HCl. To basify → use NH₃ (target pH > 7). To acidify → use HCl (target pH < 7).

⚠ pH Critical

Filter

Filter solution before SPE loading

Step BSPE Extraction Procedure

Condition

Add & draw each cartridge 3 mL MeOH

Draw

Equilibrate

Add & draw each cartridge 3 mL H₂O

Draw

Load Sample

Add 20 mL sample & draw

Draw

Release Vacuum

Discard waste fluids — insert back vessel

Wash

Add 1 mL 5% MeOH in water (v/v) & draw

Draw

Release Vacuum

Discard waste fluids — insert back vessel

Elute

Add 3 mL × 2 MeOH — release vacuum, sampai habis dan collect

★ Collect

Evaporate

Place eluent in evaporating dish

Dry

Dry sample residue on water bath at 40–60 °C

⏱ 30 min

OverviewFlorisil 500 mg / 6 mL — Polar Extraction

Oil & Cream-Based Solvent: CHCl₃ / Hexane SPE Chloroform Method

☠️CHCl₃ is toxic & volatile. All steps must be performed in or near the fume cupboard. Drying must be done under fume cupboard ventilation.

Step ASample Preparation

Dissolve

Dissolve 0.50–1.00 g in Chloroform or Hexane (~20–50 mL)

pH Adjust

Adjust pH before filtration as necessary

Filter

Filter solution before loading onto SPE cartridge

Step BSPE Procedure — Florisil Column

Condition

Add each cartridge 3 mL MeOH & draw

Draw

Equilibrate

Add each cartridge 3 mL CHCl₃ & draw

Draw

Load Sample

Add 20–50 mL sample — do not draw away

★ Collect

Wash

Add 3 mL CHCl₃ — do not draw

★ Collect

Elute

Add 3 mL × 2 = 6 mL CHCl₃ & collect eluent

★ Collect

Dry

Place eluent in evaporating dish — dry under fume cupboard

☠ Fume Hood

OverviewSoftgel Column — Methanol-Dissolved Samples

Softgel / Gel-Based Sample in Methanol Non-Polar Extraction

🧬Jika sampel dilarutkan dengan Methanol, jalankan SPE seperti dibawah. Combine both load-through AND elute fractions before evaporating.

⚠️Kira kuantiti: if sample quantity after filter > 0 mL (e.g. 0.5 mL), account for this in final volume calculation.

Step ASPE Procedure — Vacuum Pump Control Critical

Condition

Add & draw each cartridge 3 mL MeOH — Vacuum pump ON

Pump ON

Equilibrate

Add & draw each cartridge 3 mL H₂O

Draw

Release Vacuum

Discard waste fluids — insert back vessel — Pump OFF

Pump OFF

Load Sample

Add 20 mL sample & collect the solution — Pump OFF

★ Collect

Wash

Add 1 mL 5% MeOH in water (v/v) & draw — Pump OFF

Draw

Release Vacuum

Discard waste fluids — insert back vessel — Pump OFF

Elute

Add 3 mL (×2) MeOH & collect eluent

★ Collect

Combine & Evaporate

Combine loading fraction + elute fraction → evaporating dish

★ Combine Both

Dry

Dry sample residue on water bath at 40–60 °C

⏱ 30 min

Solid-PhaseExtraction

Solid-Phase
Extraction