HPLC Separation of Benzenesulfonamide and Sulfanilamide

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Condition 1

Column	Newcrom А, 3.2x50 mm, 3 µm, 100A
Mobile Phase	MeOH/H2O - 80/20%
Buffer	H2SO4 - 0.005%
Flow Rate	0.5 ml/min
Detection	275 nm

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Condition 2

Column	Primesep D, , Primesep 100 4.6x150 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O - 10/90%
Buffer	Formic Acid - 0.1%
Flow Rate	0.5 ml/min
Detection	UV 230 nm

Both Benzenesulfonamide and sulfanilamide are organic antibacterial compounds differing only in the addition of an extra amino group in sulfanilamide. The closely related compounds can be separated on a mixed-mode Primesep D column by hydrophobic and anion-exchange mechanisms or Primesep 100 column by hydrophobic and cation-exchange mechanisms with the elution order reversed using identical mobile phases. UV Detection at 230nm.

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Benzenesulfonamide
Sulfanilamide