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Adaptation of RPIP-HPLC method developed here to separate sulfonated products from reaction mixtures by gravity flow benchtop C18 resin. N-cbz O-sulfonated neomycin was eluted in 10 mM ammonium acetate, initially in 100% water, and then with addition of 30% ACN as indicated by the dashed arrow. 1 mL fractions were collected and analyzed for UV absorbance at 258 nm. UV-absorbing fractions were pooled, freeze-dried and injected on analytical HPLC, detected at 258 nm.
