Development and validation of analytical methods for estimation of Levosimendan in injectable dosage form

A simple, rapid, precise and accurate gradient reversed phase stability indicating HPLC method was developed and validated for the determination of levosimendan in injectable dosage form. The chromatographic separation was achieved on phenomenex Gemini C18 (250×4.6 mm, 5 μm) column using a mobile phase consisting of Acetonitrile, water and 0.1% TFA in the ratio of 40:10:50 v/v/v having pH 3.0 at a flow rate of 1.0 ml/min and UV detection at 375 nm. The linearity of the proposed method was investigated in the range of 0.1–15 μg/ml (r 2 =0.9967) for levosimendan. Degradation products produced as a result of stress studies did not interfere with the detection of levosimendan and the assay can thus be considered stability-indicating.