Selegiline HCl is an irreversible MAO-B inhibitor used to reduce symptoms in early-stage Parkinson's disease. It is used as an adjunct to drugs, such as L Dopamine (L-DOPA). The present study is designed to develop and validate a rapid, sensitive, and straightforward separation method with Electrospray ionization and triple quadrupole mass analyzer for the quantification of Selegiline HCl in bulk and pharmaceutical formulation. Zorbax C 18 column (50 mm × 4.6 mm i.d, 5 µ particle size) was used

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... for the separation of analyte and internal standard. The samples were eluted using 0.1% Formic acid in water and Methanol (40:60%v/v) which is delivered at 0.5 ml/minute flow rate, with a chromatographic runtime of 5 minutes. The eluents were monitored using a tandem mass spectrometer equipped with an electrospray ionization in positive mode and a triple quadrupole mass analyzer. The detection was carried out in multiple reaction-monitoring mode by quantifying the m/z 188.05→91.10 ion transition pair; with collision energy −29.0 V for Selegiline HCl. Linearity was achieved over the concentration range 3.5-6.5 ng/ml for the developed method. The limit of detection and limit of quantification were found to be 0.2 and 0.5 ng/ml, respectively. The correlation coefficient (r 2 ) value was ≥0.9985 for Selegiline HCl. This method offers a sensitive quantification of Selegiline HCl in the pharmaceutical formulation.