Haemolysis effect of oral suspension of Erythromycin Ethylsuccinate (Erythromycin 250mg/5 ml) in Bio analysis by liquid chromatography - tandem mass spectrometry

A rapid, simple and specific method for estimation of Erythromycin Ethylsuccinate in human plasmawas validated using Imipramine as internal standard. The analyte and internal standard wereextracted from plasma using simple solid phase extraction. The compound were separated on areverse-phase column with an isocratic mobile phase consisting of 0.1% formic acid in water andacetonitrile (20: 80 (v/v)) and detected by tandem mass spectrometry in positive ion mode. The iontransition recorded in multiple reaction monitoring mode were m/z 862.5 286.2 for ErythromycinEthylsuccinate and m/z 281.2 208.1 for internal standard Imipramine. Linearity in plasma wasobserved over the concentration range 10 – 1500 ng/mL for Erythromycin Ethylsuccinate. The meanglobal recovery for Erythromycin Ethylsuccinate was 62.72 % and 68.57% for Imipramine (IS). Thecoefficient of variation of the assay 6.09 % to 6.97% and accuracy was 95.18% to 97.08% at LLOQlevel. The validated method was applied to bioequivalence study of 250mg/5 ml ErythromycinEthylsuccinate suspension in 24 healthy human volunteers. Total 50 samples from individualvolunteers identified as Haemolyzed which were analyze initial and repeat again to cross check themethod’s reproducibity for Haeamolysis effect and compared which found acceptable range.