ORIGINAL ARTICLE Chromatographic analysis of Hb S for the diagnosis of various sickle cell disorders in Pakistan Nazish Khalid Hashmi & Bushra Moiz & Maliha Nusrat & Mashhooda Rasool Hashmi Received: 24 December 2007 / Accepted: 2 April 2008 / Published online: 9 May 2008 # Springer-Verlag 2008 Abstract Sickle cell disease remains a relatively obscure theme in research on haemoglobinopathies in Pakistan. Limited data is available regarding its prevalence in the country. The objective of our study was not only to estimate the frequency of different sickle cell diseases but also to provide quantitative estimation of haemoglobin S and other haemoglobin variants using an automated high-performance liquid chromatography (HPLC) system. For this purpose, we retrospectively evaluated the results of HPLC performed on all patients with suspected haemoglobinopathies during the years 2005 and 2006. Information derived from various sources was used to identify a particular genotype by analysing each sample containing Hb S with respect to haemoglobin, red cell indices and levels of various associ- ated haemoglobin variants. Analysis of 15,699 samples identified 302 patients with Hb S (1.92%). The genotypes identified included Sβ 0 (46.7%), SS (19.2%), SA (11.6%), Sβ + (8.6%) and SD (2.3%). Thirty-five cases could not be categorised and were labelled ‘unclassified’. Majority of the patients (62.3%) were below the age of 18 years. Balochistan, which is the largest province based on the area, yielded the highest number of patients (n = 140). In the Sβ 0 group, the mean haemoglobin and Hb S were lower in children compared to adults (p value of 0.001 and 0.016, respectively). We conclude that sickle cell disorders are prevalent in Pakistan to a significant extent, being concen- trated in certain areas of the country. We present the first report of various haemoglobin S genotypes from our population. It is hoped that it will act as a database to characterise the same for our population. Keywords High-performance liquid chromatography . Sickle cell disorders . Hb S . Haemoglobinopathy Introduction In 1948, Linus Pauling and Harvey Itano employed the then new technique of haemoglobin electrophoresis to prove that haemoglobin molecules in patients suffering from sickle cell anaemia have a different electrical charge compared to that of normal individuals [1]. Eight years later, Vernon Ingram and J.A. Hunt first sequenced sickle haemoglobin (β s ) and discovered the single base substitution of glutamic acid by valine in the β globin gene [2]. These were the first instances of the biochemical diagnosis of sickle cell disease. Almost six decades after Pauling's initial discovery, sickle cell anaemia still remains a widely researched theme with upcoming novel approaches to diagnosis and management. The diagnosis of sickle cell anaemia is gaining impor- tance, as developing nations like Pakistan undergo a transition from an era of communicable to non-communicable disease [3]. Haemoglobinopathies are relatively common in Asian and African countries, probably due to their social customs and consanguineous marriages [4]. Indeed, the Hb S allele is highly prevalent in the Middle East, Africa and the Indian subcontinent [5]. Miller et al. conducted a survey of Ann Hematol (2008) 87:639–645 DOI 10.1007/s00277-008-0495-7 N. K. Hashmi (*) Department of Biological and Biomedical Sciences, Aga Khan University, Karachi, Pakistan e-mail: nazish.hashmi@aku.edu B. Moiz : M. R. Hashmi Department of Pathology and Microbiology, Aga Khan University, Karachi, Pakistan M. Nusrat Medical College, Aga Khan University, Karachi, Pakistan