Introduction: Prolactin (PRL) exists in different forms in human serum. The predominant form is monomeric PRL (molecular mass 23 kDa) with smaller amounts of big PRL (molecular mass 50–60 kDa) and at times macroprolactin (molecular mass 150–170 kDa). Macroprolactin, generally considered to be biologically inactive, accounts for the major part of prolactin in some patients. Different immunoassays for prolactin differ in reactivity with this macromolecular complex. Aim: The present study was undertaken to assess the incidence of macroprolactinaemia in our cohort of hyperprolactinemic patients. Method: 204 samples with hyperprolactinemia were evaluated for macroprolactinemia by polyethylene glycol (PEG) precipitation and gel fi ltration chromatography (GFC). Recoveries <60% after PEG precipitation were considered to have macroprolactinaemia. Results: A total of 43 (21%) of these patients had less than 60% recovery after PEG precipitation. GFC confi rmed that in seven of these patients macroprolactin was the major part of the prolactin. Recoveries were < 40% PEG precipitation in these samples. Combined macro and hyperprolactinemia was observed in two samples and the recovery after PEG precipitation was >40% but 50%. The incidence of macroprolactinemia in our cohort of hyperprolactinaemic patients was noted to be 4.4%. Conclusion: Macroprolactin is a signifi cant cause of misdiagnosis, unnecessary investigation, and inappropriate treatment and hence it is useful to screen all patients with high PRL levels with PEG precipitation and to apply GFC to samples with recoveries <50%.