Introduction: HPLC (High Performance Liquid Chromatography) is the commonest method endorsed for screening of Thalassemia and other hemoglobinopathies in our country. Universal screening of antenatal mothers in the early first trimester has been employed under the Thalassemia control programme launched as a part of National Health Mission. Estimated prevalence of thalassemia in India is as high as 10000-15000 per year and varies from state to state, and even district to district owing to the multi-ethnic population of India and the scattered tribal belts. Case Reports: Here, our discussion has highlighted four scenarios which are reasonably common in Indian subcontinent, but problematic none-the-less due to various reasons. The differential diagnoses, decision making and strategies for further choice of investigations has been elaborated in details. Conclusion: As a tool of screening, the outcome based on haemoglobin HPLC greatly relies on the successful interpretation of the patterns of the graphs and thorough understanding of the gray zones and pit falls inherent to the procedure employed.