Development and validation of a novel stability-indicating HPLC method for the simultaneous assay of betamethasone-17-valerate, fusidic acid, potassium sorbate, methylparaben and propylparaben in a topical cream preparation.

Development and validation of a novel stability-indicating HPLC method for the simultaneous assay of betamethasone-17-valerate, fusidic acid, potassium sorbate, methylparaben and propylparaben in a topical cream preparation.

Journal of pharmaceutical and biomedical analysis (2014-04-16)

Jonathan Byrne, Trinidad Velasco-Torrijos, Robert Reinhardt

PMID24731970

ABSTRACT

A novel stability-indicating reversed phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous assay of betamethasone-17-valerate, fusidic acid and potassium sorbate as well as methyl- and propylparaben in a topical cream preparation has been developed. A 100mm×3.0mm ID. Ascentis Express C18 column maintained at 30°C and UV detection at 240nm were used. A gradient programme was employed at a flow-rate of 0.75ml/min. Mobile phase A comprised of an 83:17 (v/v) mixture of acetonitrile and methanol and mobile phase B of a 10g/l solution of 85% phosphoric acid in purified water. The method has been validated according to current International Conference on Harmonisation (ICH) guidelines and applied during formulation development and stability studies. The procedure has been shown to be stability-indicating for the topical cream.

MATERIALS

Product Number

Brand

Product Description

494437

Sigma-Aldrich

Methanol, BioReagent, ≥99.93%

320390

Sigma-Aldrich

Methanol, ACS reagent, ≥99.8%

646377

Sigma-Aldrich

Methanol, HPLC Plus, ≥99.9%

34885

Sigma-Aldrich

Methanol, suitable for HPLC, gradient grade, ≥99.9%

439193

Sigma-Aldrich

Methanol, suitable for HPLC, gradient grade, suitable as ACS-grade LC reagent, ≥99.9%

34860

Sigma-Aldrich

Methanol, suitable for HPLC, ≥99.9%

PHR1320

Supelco

Residual Solvent - Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material

685461

Sigma-Aldrich

Acetonitrile solution, contains 10.0% acetone, 0.05% formic acid, 40.0% 2-propanol

650609

Sigma-Aldrich

Methanol, HPLC Plus, ≥99.9%, poly-coated bottles

679186

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (w/v) ammonium formate, 5 % (v/v) water, 0.1 % (v/v) formic acid, suitable for HPLC

717975

Sigma-Aldrich

Methanol, NMR reference standard

698822

Sigma-Aldrich

Phosphoric acid solution, NMR reference standard, 85% in D₂O (99.9 atom % D), NMR tube size 5 mm × 8 in.

699608

Sigma-Aldrich

Phosphoric acid solution, NMR reference standard, 85% in D₂O (99.9 atom % D), NMR tube size 3 mm × 8 in.

50819-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 10 cm × 3 mm

50805-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 2 cm × 2.1 mm

50809-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 3 cm × 2.1 mm

50821-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 15 cm × 3 mm

50822-U

Supelco

Ascentis^® Express C18, 2 μm Guard Cartridge, 2 μm particle size, L × I.D. 5 mm × 2.1 mm, pkg of 3 ea

50814-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 15 cm × 2.1 mm

50812-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 7.5 cm × 2.1 mm

50811-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 5 cm × 2.1 mm

50813-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 10 cm × 2.1 mm

50816-U

Supelco

Ascentis^® Express C18, 2 μm UHPLC Column, 2 μm particle size, L × I.D. 5 cm × 3 mm

PHR1372

Supelco

Methanol, Pharmaceutical Secondary Standard; Certified Reference Material

574724

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid

574732

Sigma-Aldrich

Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC

50542-U

Supelco

Ascentis^® Express C18, 5 μm Guard Cartridge, 5 μm particle size, L × I.D. 5 mm × 4.6 mm, pkg of 3 ea

50518-U

Supelco

Ascentis^® Express C18, 5 μm HPLC Column, 5 μm particle size, L × I.D. 15 cm × 2.1 mm

50533-U

Supelco

Ascentis^® Express C18, 5 μm HPLC Column, 5 μm particle size, L × I.D. 7.5 cm × 4.6 mm

50527-U

Supelco

Ascentis^® Express C18, 5 μm HPLC Column, 5 μm particle size, L × I.D. 15 cm × 3 mm