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Kromasil CelluCoat

Kromasil CelluCoat

44 Scale up –

44 Scale up – With/without additive MWD1 D, Sig=290,10 Ref=360,100 (BKH\060512000001.D) MWD1 B, Sig=290,10 Ref=off (BKH\070116000021.D) mAU mAU 2000 2000 1750 1750 1500 1500 1250 1250 1000 1000 750 750 500 500 250 250 0 0 0 5 10 15 20 min 0 5 10 15 20 min Preparative sample: Metoprolol free base 40 mg/mL Mobile Phase: Heptane/2-propanol 90/10 Preparative load: 500 µl, 20 mg Column: Kromasil-10-CelluCoat 4.6 x 250 mm Mobile phase: Heptane/2-propanol 90/10, with and without 0.1% DEA as additive Flow rate: 1.0 mL/min Temperature: 25°C Preparative detection: UV @ 290nm

45 Scale up – With/without additive Elution profile reconstructed from fraction analysis data Metoprolol, 20 mg, 0.1% DEA in mobile phase Metoprolol, 20mg, no additive in mobile phase Peak area @ 270nm 16000,0 14000,0 12000,0 10000,0 8000,0 6000,0 4000,0 2000,0 0,0 0,0 5,0 10,0 15,0 20,0 Peak area @ 270nm 16000,0 14000,0 12000,0 10000,0 8000,0 6000,0 4000,0 2000,0 0,0 0,0 5,0 10,0 15,0 20,0 Time (min) Time (min) Pool Purity Recovery Pool Purity Recovery 1 100,00% 97,91% 1 100,00% 99,49% 2 99,73% 88,11% 2 99,48% 97,42%

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