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

50462-U

Supelco

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

34442

Sigma-Aldrich

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

53819-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 7.5 cm × 4.6 mm

79622

Sigma-Aldrich

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

53822-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 5 cm × 2.1 mm

53814-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 10 cm × 3 mm

733466

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

34998

Sigma-Aldrich

Acetonitrile, HPLC Plus, ≥99.9%

30417

Sigma-Aldrich

Phosphoric acid, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥85%

01-0622

Sigma-Aldrich

Acetonitrile, ≥99.8%, JIS 1000

01-0623

Sigma-Aldrich

Acetonitrile, JIS 5000

01-0620

Sigma-Aldrich

Acetonitrile, ≥99.8%, JIS 300

721956

Sigma-Aldrich

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

53793-U

Supelco

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

01-0600

Sigma-Aldrich

Acetonitrile, SAJ first grade, ≥99.0%

53816-U

Supelco

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

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

01-0590

Sigma-Aldrich

Acetonitrile, JIS special grade, ≥99.5%

632546

Supelco

Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%

01-0645

Sigma-Aldrich

Acetonitrile

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

54271-U

Supelco

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

675415

Sigma-Aldrich

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

PHR1278

Supelco

Potassium sorbate, Pharmaceutical Secondary Standard; Certified Reference Material

1424109

USP

Methyl alcohol, United States Pharmacopeia (USP) Reference Standard

P2650000

Potassium sorbate, European Pharmacopoeia (EP) Reference Standard

695017

Sigma-Aldrich

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

110086

Sigma-Aldrich

Acetonitrile, ReagentPlus^®, 99%

53799-U

Supelco

Ascentis^® Express C18, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 2 cm × 2.1 mm