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

1424109

USP

Methyl alcohol, United States Pharmacopeia (USP) Reference Standard

650188

Sigma-Aldrich

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

695017

Sigma-Aldrich

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

110086

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

79622

Sigma-Aldrich

Phosphoric acid, puriss. p.a., crystallized, ≥99.0% (T)

34888

Sigma-Aldrich

Acetonitrile, ≥99.9% (GC)

04107

Sigma-Aldrich

Phosphoric acid, puriss., meets analytical specification of Ph. Eur., BP, NF, FCC, 85.0-88.0%

34442

Sigma-Aldrich

Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)

60004

Sigma-Aldrich

Acetonitrile, ≥99.5% (GC)

494445

Sigma-Aldrich

Acetonitrile, biotech. grade, ≥99.93%

360457

Sigma-Aldrich

Acetonitrile, ACS reagent, ≥99.5%

438081

Sigma-Aldrich

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

733466

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

40430

Supelco

Potassium sorbate, certified reference material, TraceCERT^®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland

50462-U

Supelco

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

50461-U

Supelco

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

53793-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 15 cm × 10 mm

PHR1278

Supelco

Potassium sorbate, Pharmaceutical Secondary Standard; Certified Reference Material

P2650000

Potassium sorbate, European Pharmacopoeia (EP) Reference Standard

1601340

USP

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

1548407

USP

Potassium sorbate, United States Pharmacopeia (USP) Reference Standard

79617

Sigma-Aldrich

Phosphoric acid, BioUltra, ≥85% (T)

85520

Sigma-Aldrich

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

53802-U

Supelco

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

452289

Sigma-Aldrich

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

271004

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

45983

Supelco

Acetonitrile, analytical standard

53811-U

Supelco

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

53805-U

Supelco

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

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