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

82762

Supelco

Methanol, analytical standard

W290017

Sigma-Aldrich

Phosphoric acid solution, 85 wt. % in H₂O, FCC, FG

322415

Sigma-Aldrich

Methanol, anhydrous, 99.8%

359769

Sigma-Aldrich

trans,trans-2,4-Hexadienoic acid potassium salt, ReagentPlus^®, 99%

PHR1372

Supelco

Methanol, Pharmaceutical Secondary Standard; Certified Reference Material

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

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

50814-U

Supelco

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

34885

Sigma-Aldrich

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

50537-U

Supelco

Ascentis^® Express C18, 5 μm HPLC Column, 5 μm particle size, L × I.D. 15 cm × 4.6 mm

50530-U

Supelco

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

50538-U

Supelco

Ascentis^® Express C18, 5 μm HPLC Column, 5 μm particle size, L × I.D. 25 cm × 4.6 mm

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

M1775

Sigma-Aldrich

Methanol, Absolute - Acetone free

676780

Sigma-Aldrich

Methanol, ACS reagent, ≥99.8%

320390

Sigma-Aldrich

Methanol, ACS reagent, ≥99.8%

32213

Sigma-Aldrich

Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)

494437

Sigma-Aldrich

Methanol, BioReagent, ≥99.93%

179337

Sigma-Aldrich

Methanol, ACS reagent, ≥99.8%

179957

Sigma-Aldrich

Methanol, Laboratory Reagent, ≥99.6%

PHR1320

Supelco

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

345245

Sigma-Aldrich

Phosphoric acid, 85 wt. % in H₂O, 99.99% trace metals basis

45983

Supelco

Acetonitrile, analytical standard

452289

Sigma-Aldrich

Phosphoric acid, ≥85 wt. % in H₂O, ≥99.999% trace metals basis

437557

Sigma-Aldrich

Acetonitrile, ≥99.5%, ACS reagent

271004

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

79617

Sigma-Aldrich

Phosphoric acid, BioUltra, ≥85% (T)

53816-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 15 cm × 3 mm

85520

Sigma-Aldrich

Potassium sorbate, purum p.a., ≥99.0% (NT)