Development and validation of a stability-indicating RP-HPLC method for assay of betamethasone and estimation of its related compounds.
Betamethasone (9alpha-fluoro-16beta-methylprednisolone) is one of the members of the corticosteriod family of active pharmaceutical ingredient (API), which is widely used as an anti-inflammatory agent and also as a starting material to manufacture various esters of betamethasone. A stability-indicating reverse-phase high performance liquid chromatography (RP-HPLC) method has been developed and validated which can separate and accurately quantitate low levels of 26 betamethasone related compounds. The stability-indicating capability of the method was demonstrated through adequate separation of all potential betamethasone related compounds from betamethasone and also from each other that are present in aged and stress degraded betamethasone stability samples. Chromatographic separation of betamethasone and its related compounds was achieved by using a gradient elution at a flow rate of 1.0mL/min on a ACE 3 C18 column (150mmx4.6mm, 3microm particle size, 100A pore size) at 40 degrees C. Mobile phase A of the gradient was 0.1% methanesulfonic acid in aqueous solution and mobile phase B was a mixture of tert-butanol and 1,4-dioxane (7:93, v/v). UV detection at 254nm was employed to monitor the analytes. For betamethasone 21-aldehyde, the QL and DL were 0.02% and 0.01% respectively. For betamethasone and the rest of the betamethasone related compounds, the QL and DL were 0.05% and 0.02%. The precision of betamethasone assay is 0.6% and the accuracy of betamethasone assay ranged from 98.1% to 99.9%.
Global Quality Services - Analytical Sciences, Schering-Plough Corporation, 1011 Morris Ave. Union, New Jersey 07083, USA. qiang.fu@spcorp.com
Qiang
Q
Fu
Minshan
M
Shou
Dwight
D
Chien
Robert
R
Markovich
Abu M
AM
Rustum
D001623:Betamethasone / Q000032:analysis* / Q000737:chemistry*; D002626:Chemistry, Pharmaceutical / Q000379:methods / Q000639:trends*; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000639:trends; D056148:Chromatography, Reverse-Phase / Q000379:methods / Q000639:trends
D001623:Betamethasone