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

322415

Sigma-Aldrich

Methanol, anhydrous, 99.8%

W290017

Sigma-Aldrich

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

612162

Sigma-Aldrich

Phosphoric acid solution, NMR reference standard, 85% in D₂O (99.9 atom % D), NMR tube size 4.2 mm × 8 in. , WGS-5BL Coaxial NMR tube

82762

Supelco

Methanol, analytical standard

717975

Sigma-Aldrich

Methanol, NMR reference standard

466123

Sigma-Aldrich

Phosphoric acid, crystalline, ≥99.999% trace metals basis

359769

Sigma-Aldrich

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

50816-U

Supelco

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

53992-U

Supelco

Ascentis^® Express C18, 2.7 Micron Capillary HPLC Column, 2.7 μm particle size, L × I.D. 5 cm × 300 μm

53998-U

Supelco

Ascentis^® Express C18, 2.7 Micron Capillary HPLC Column, 2.7 μm particle size, L × I.D. 5 cm × 500 μm

53501-U

Supelco

Ascentis^® Express C18, 2.7 Micron Guard Cartridge, 2.7 μm particle size, L × I.D. 5 mm × 2.1 mm, pkg of 3 ea

53508-U

Supelco

Ascentis^® Express C18, 2.7 Micron Guard Cartridge, 2.7 μm particle size, L × I.D. 5 mm × 4.6 mm, pkg of 3 ea

54271-U

Supelco

Ascentis^® Express C18, 2.7 Micron Capillary HPLC Column, 2.7 μm particle size, L × I.D. 15 cm × 300 μm

53504-U

Supelco

Ascentis^® Express C18, 2.7 Micron Guard Cartridge, 2.7 μm particle size, L × I.D. 5 mm × 3 mm, pkg of 3 ea

721956

Sigma-Aldrich

Methanol solution, NMR reference standard, 4% in methanol-d₄ (99.8 atom % D), NMR tube size 3 mm × 8 in.

650188

Sigma-Aldrich

Methanol solution, (Methanol:Dimethyl sulfoxide 1:1 (v/v))

54273-U

Supelco

Ascentis^® Express C18, 2.7 Micron Capillary HPLC Column, 2.7 μm particle size, L × I.D. 15 cm × 500 μm

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

50811-U

Supelco

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

PHR1372

Supelco

Methanol, Pharmaceutical Secondary Standard; Certified Reference Material

50813-U

Supelco

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

50805-U

Supelco

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

50809-U

Supelco

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

50819-U

Supelco

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

699608

Sigma-Aldrich

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

698822

Sigma-Aldrich

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

50536-U

Supelco

Ascentis^® Express C18, 5 μm HPLC Column, 5 μm particle size, L × I.D. 10 cm × 4.6 mm