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

345245

Sigma-Aldrich

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

53805-U

Supelco

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

53811-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 5 cm × 3 mm

45983

Supelco

Acetonitrile, analytical standard

452289

Sigma-Aldrich

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

53802-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 3 cm × 2.1 mm

271004

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

79617

Sigma-Aldrich

Phosphoric acid, BioUltra, ≥85% (T)

53804-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 7.5 cm × 2.1 mm

53816-U

Supelco

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

54273-U

Supelco

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

85520

Sigma-Aldrich

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

P5811

Sigma-Aldrich

Phosphoric acid, BioReagent, suitable for insect cell culture, 85%

53812-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 7.5 cm × 3 mm

53818-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 3 cm × 4.6 mm

47848

Supelco

Potassium Sorbate, analytical standard

466123

Sigma-Aldrich

Phosphoric acid, crystalline, ≥99.999% trace metals basis

609781

Sigma-Aldrich

Phosphoric acid-¹⁶O₄ solution, 70 wt. % in D₂O, 99.9 atom % ¹⁶O

695017

Sigma-Aldrich

Phosphoric acid, ACS reagent, ≥85 wt. % in H₂O

110086

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

L010300

Sigma-Aldrich

Ultrapure Acetonitrile

PHR1278

Supelco

Potassium sorbate, Pharmaceutical Secondary Standard; Certified Reference Material

1601340

USP

Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard

1424109

USP

Methyl alcohol, United States Pharmacopeia (USP) Reference Standard

PHR1551

Supelco

Acetonitrile, Pharmaceutical Secondary Standard; Certified Reference Material

P2650000

Potassium sorbate, European Pharmacopoeia (EP) Reference Standard

50461-U

Supelco

Ascentis^® Express C18 PCP HPLC Columns, 2.7 μm particle size, L × I.D. 10 cm × 4.6 mm

50462-U

Supelco

Ascentis^® Express C18 PCP HPLC Columns, 2.7 μm particle size, L × I.D. 15 cm × 4.6 mm

494445

Sigma-Aldrich

Acetonitrile, biotech. grade, ≥99.93%

360457

Sigma-Aldrich

Acetonitrile, ACS reagent, ≥99.5%