Isolation of Benzylideneacetophenone from a Crude Reaction Mixture
The Application Notebook
Benzylideneacetophenone forms the central core for a variety of important biological compounds, known collectively as chalcones. Many of these compounds have significant medicinal value as they demonstrate antibacterial, antifungal, antitumor, and anti-inflammatory properties.
Benzylideneacetophenone forms the central core for a variety of important biological compounds, known collectively as chalcones. Many of these compounds have significant medicinal value as they demonstrate antibacterial, antifungal, antitumor, and anti-inflammatory properties. They are synthesized by an aldol condensation between a benzaldehyde and an acetophenone in the presence of NaOH as catalyst. In this application note, benzylideneacetophenone is isolated from a crude reaction mixture using flash chromatographic technique, including consideration for scale-up.
Figure 1: TLC of crude mixture.
TLC of the Crude Reaction Mixture
TLC on silica gel Si60F254, developed in n-hexane/ethyl acetate 19:1, detection UV 254nm.
Sepacore Configuration
Cartridge 12 × 150 mm, prepacked with silica gel 60, 40–63 µm
2 Pump modules C-605
Fraction collector C-660
Control-Unit C-620 with SepacoreControl software
UV Photometer C-635
Figure 2: Separation 1.
Separation Conditions
Eluent: see below
Flow rate: 10 mL/min
100 mg crude mixture, dissolved in n-hexane and some drops of toluene (solubility of the sample in pure n-hexane too low)
Loading: approx. 0.4 mL
1. Test runs
Separation 1
Eluent: n-hexane with 2% ethyl acetate, isocratic
Figure 3: Separation 2.
Separation 2
Eluent: n-hexane with 1%, 2% and 3% ethyl acetate, step gradient. Each step was initiated after a component was completely eluted (at the end of a peak).
TLC Check
TLC on silica gel Si60F254, developed in n-hexane/ethyl acetate 19:1, detection: UV 254 nm
1 = Benzaldehyde
2 = Benzylideneacetophenone
3= Benzophenone
Figure 4: TLC check of Separation 2.
Recovery
Fraction 7: 65 mg
2. Scale-up
Calculation of the scale-up factor
Cartridge 12 × 150 mm: cross sectional area = 1.13 cm2
Cartridge 40 × 150 mm: cross sectional area = 12.56 cm2
Scale-up-factor = 12.56 cm2 / 1.13 cm2 = 11.1 ≈ 10
Figure 5: Scale-up separation.
Separation conditions
Eluent: n-hexane with 1%, 2% and 3% ethyl acetate, step gradient
Sample: 1 g crude mixture, dissolved in n-hexane/toluene 7:3 (solubility of the sample in n-hexane too low, toluene is eluted as front peak)
Injection volume: 2 mL
Figure 6: TLC check of scale-up.
TLC-Check
TLC on silica gel Si60F254, developed in n-hexane/ethyl acetate 19:1, detection: UV 254 nm
Recovery
Fraction 13–16: 512 mg crystalline product, mp 57 °C
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