A:
Prepare a test solution in water containing 10 mg of paromomycin per mL. Apply 25 µL of this solution and 25 µL of a Standard solution of
USP Paromomycin Sulfate RS containing 10 mg of paromomycin per mL to a suitable thin-layer chromatographic plate (see
Chromatography 621) coated with a 0.25-mm layer of chromatographic silica gel. Allow the spots to dry, place the plate in a developing chamber, and develop the chromatogram in a solvent system consisting of a mixture of freshly prepared ammonium acetate solution (4 in 100),
n-propyl alcohol, and ammonium hydroxide (30:10:6) until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow it to air-dry for 10 minutes. Heat the plate at 105
for 1 hour, allow to cool, and spray with a solution of ninhydrin in butanol (1 in 100). Heat the plate at 105
for 5 minutes: paromomycin appears as a red spot, and the
RF value of the principal spot obtained from the test solution corresponds to that obtained from the Standard solution.