{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,2]],"date-time":"2026-11-02T11:46:07Z","timestamp":1793619967787,"version":"4.5.0"},"reference-count":31,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"3","license":[{"start":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T00:00:00Z","timestamp":1751328000000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-sa\/4.0"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:sec>\n                    <jats:title>Introduction:<\/jats:title>\n                    <jats:p>Cation exchange high-performance liquid chromatography (HPLC) serves as a rapid, reproducible, and accurate method for diagnosing hemoglobinopathies. This study outlines the diagnostic approach and the challenges faced in the routine diagnosis of hemoglobinopathies through HPLC, particularly in laboratories with limited resources.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Aims and Objectives:<\/jats:title>\n                    <jats:p>The aim of the study was to identify the challenges encountered in identifying abnormal hemoglobin (Hb) variants and to determine the significance of HbA2 and\/or fetal Hb (HbF) analysis in the HPLC methodology for hemoglobinopathies.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Materials and Methods:<\/jats:title>\n                    <jats:p>A total of 1900 samples were analyzed using the ARKRAY ADAMS HA-8180T HPLC automated analyzer for the purpose of hemoglobinopathy testing. The samples were classified into normal or abnormal hemoglobin variants based on the percentage levels of HbA2, HbF, HbA, and the identification of any abnormal peaks. Among these, 113 cases were diagnosed to have thalassemia or hemoglobinopathy. The clinical presentations and red blood cell (RBC) indices were compared with the HPLC findings for each case, thereby contributing to the accuracy of the diagnosis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>The study examined the distribution of Hb variants, revealing that \u03b2-thalassemia trait was the most prevalent at 44.2%, followed by sickle cell trait at 13.3% and HbD Punjab trait at 10.6%. There were many challenging cases with elevated HbA2, like HbE thalassemia and Hb Lepore. Furthermore, there was identification of some abnormal peaks which were not exactly in the instrument\u2019s predetermined HbA2, HbF, HbA, or sickle windows, like HbJ Meerut. There were a few cases with abnormally elevated HbF, which can be seen in homozygous \u03b2-thalassemia, sickle cell disease, compound double heterozygous sickle cell \u03b2-thalassemia, \u03b4\u03b2-thalassemia, and hereditary persistence of HbF. Carriers of \u03b2-thalassemia were generally identified by an HbA2 level of 4% or higher; however, there were nine cases which exhibited borderline HbA2 levels ranging from 3.5% to 3.9%, which might turn out to be \u03b2-thalassemia trait, especially in high-prevalence areas like Gujarat.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion:<\/jats:title>\n                    <jats:p>Any case scenario with abnormally elevated HbA2 is not always \u03b2-thalassemia trait. Nor abnormally elevated HbF may always indicate \u03b2-thalassemia major. Furthermore, some clinico-pathologically relevant hemoglobinopathies might show an abnormal peak on HPLC at any retention time, which may not be necessarily determined by the machine to be in some specific window. We need to correlate the clinical context, RBC indices, HPLC findings, and family studies to effectively detect most Hb variants.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.4103\/ijabmr.ijabmr_70_25","type":"journal-article","created":{"date-parts":[[2025,8,20]],"date-time":"2025-08-20T20:01:28Z","timestamp":1755720088000},"page":"197-205","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":2,"title":["Challenges Associated with the Identification of Abnormal Hemoglobin Variants Utilizing the High-performance Liquid Chromatograph Technique: A Prospective Study in a Hospital Setting in Gujarat"],"prefix":"10.4103","volume":"15","author":[{"given":"Garima","family":"Anandani","sequence":"first","affiliation":[{"name":"Department of Pathology, AIIMS, Rajkot, Gujarat, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anita","family":"Motiani","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, AIIMS, Rajkot, Gujarat, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parth","family":"Goswami","sequence":"additional","affiliation":[{"name":"Department of Pathology, AIIMS, Rajkot, Gujarat, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amit","family":"Sonagra","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, AIIMS, Rajkot, Gujarat, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","published-online":{"date-parts":[[2025,8,20]]},"reference":[{"key":"R1-20250821","first-page":"112","article-title":"Prevalence of common hemoglobinopathies in Gujarat: An analysis of a large population screening program","volume":"3","author":"Patel","year":"2012","journal-title":"Natl J Community Med"},{"key":"R2-20250821","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/S0009-8981(01)00672-6","article-title":"Diagnosis of hemoglobinopathies: Electrophoresis versus HPLC","volume":"313","author":"Ou","year":"2001","journal-title":"Clin Chim Acta"},{"key":"R3-20250821","first-page":"257805","article-title":"Detection of Abnormal hemoglobin variants by HPLC method: Common problems with suggested solutions","volume":"2014","author":"Pant","year":"2014","journal-title":"Int Sch Res Notices"},{"key":"R4-20250821","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1007\/s12288-014-0409-x","article-title":"HPLC in characterization of hemoglobin profile in thalassemia syndromes and hemoglobinopathies: A clinicohematological correlation","volume":"31","author":"Khera","year":"2015","journal-title":"Indian J Hematol Blood Transfus"},{"key":"R5-20250821","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1007\/s12687-012-0114-0","article-title":"Prevalence of \u03b2-thalassemia and other haemoglobinopathies in six cities in India: A multicentre study","volume":"4","author":"Mohanty","year":"2013","journal-title":"J Community Genet"},{"key":"R6-20250821","first-page":"PO70","article-title":"Spectrum of hemoglobinopathies and thalassemias diagnosed on HPLC in a tertiary teaching hospital of Northern India","volume":"5","author":"Singh","year":"2016","journal-title":"Natl Lab Med"},{"key":"R7-20250821","first-page":"133","article-title":"Spectrum of haemoglobinopathies in sikh population of Bangalore: A pilot study","volume":"3","author":"Kaler","year":"2018","journal-title":"IP J Diagn Pathol Oncol"},{"key":"R8-20250821","doi-asserted-by":"crossref","first-page":"5269","DOI":"10.18203\/2320-6012.ijrms20164193","article-title":"Spectrum of hemoglobinopathies by high performance liquid chromatography with special reference to role of HbA2 levels at tertiary care centre","volume":"4","author":"Bhokare","year":"2016","journal-title":"Int J Res Med Sci"},{"key":"R9-20250821","doi-asserted-by":"crossref","first-page":"564","DOI":"10.4103\/ijpm.ijpm_2_22","article-title":"Prevalence and spectrum of haemoglobinopathies in females of reproductive age group-a first tertiary care center experience in Punjab, North India","volume":"66","author":"Narang","year":"2023","journal-title":"Indian J Pathol Microbiol"},{"key":"R10-20250821","first-page":"390","article-title":"HPLC \u2013 How necessary is it for haemoglobinopathy diagnosis in India?","volume":"46","author":"Tyagi","year":"2003","journal-title":"Indian J Pathol Microbiol"},{"key":"R11-20250821","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1007\/s12098-007-0117-8","article-title":"HPLC studies in hemoglobinopathies","volume":"74","author":"Colah","year":"2007","journal-title":"Indian J Pediatr"},{"key":"R12-20250821","doi-asserted-by":"crossref","first-page":"294","DOI":"10.4103\/0971-6866.107979","article-title":"Hemoglobinopathies in South Gujarat population and incidence of anemia in them","volume":"18","author":"Patel","year":"2012","journal-title":"Indian J Hum Genet"},{"key":"R13-20250821","first-page":"1","article-title":"Prevalence of haemoglobinopathies in Gujarat, India: A cross-sectional study","volume":"5","author":"Patel","year":"2008","journal-title":"Internet J Hematol"},{"key":"R14-20250821","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1080\/03630269.2022.2142608","article-title":"Spectrum of \u03b2-thalassemia and other hemoglobinopathies in the Saurashtra region of Gujarat, India: Analysis of a large population screening program","volume":"46","author":"Vachhani","year":"2022","journal-title":"Hemoglobin"},{"key":"R15-20250821","first-page":"69","article-title":"A blood transfusion leading to misdiagnosis of beta-thalassaemia carrier status","volume":"8","author":"Wong","year":"2010","journal-title":"Blood Transfus"},{"key":"R17-20250821","doi-asserted-by":"crossref","first-page":"1736","DOI":"10.1373\/clinchem.2004.034991","article-title":"HPLC retention time as a diagnostic tool for hemoglobin variants and hemoglobinopathies: A study of 60000 samples in a clinical diagnostic laboratory","volume":"50","author":"Joutovsky","year":"2004","journal-title":"Clin Chem"},{"key":"R20-20250821","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.phoj.2017.10.002","article-title":"Burden of thalassaemia in India: The road map for control","volume":"2","author":"Colah","year":"2017","journal-title":"Pediatr Hematol Oncol J"},{"key":"R21-20250821","first-page":"81","article-title":"Significance of HbA2 level for diagnosis of beta thalassemia","volume":"4","author":"Sohail","year":"2018","journal-title":"World J Pharm Med Res"},{"key":"R22-20250821","doi-asserted-by":"crossref","first-page":"513","DOI":"10.4103\/0971-5916.73390","article-title":"Spectrum of haemoglobinopathies diagnosed by cation exchange-HPLC and modulating effects of nutritional deficiency anaemias from north India","volume":"132","author":"Rao","year":"2010","journal-title":"Indian J Med Res"},{"key":"R23-20250821","doi-asserted-by":"crossref","first-page":"5414","DOI":"10.1038\/s41598-022-09250-5","article-title":"Significance of borderline HbA(2) levels in \u03b2 thalassemia carrier screening","volume":"12","author":"Colaco","year":"2022","journal-title":"Sci Rep"},{"key":"R24-20250821","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.bcmd.2014.04.005","article-title":"Delineation of the molecular basis of borderline hemoglobin A2 in Chinese individuals","volume":"53","author":"Lou","year":"2014","journal-title":"Blood Cells Mol Dis"},{"key":"R25-20250821","doi-asserted-by":"crossref","first-page":"2069","DOI":"10.1515\/CCLM.2011.696","article-title":"\u03b2-Thalassemia mutations in subjects with borderline HbA\u2082 values: A pilot study in North India","volume":"49","author":"Rangan","year":"2011","journal-title":"Clin Chem Lab Med"},{"key":"R26-20250821","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1111\/ijlh.12948","article-title":"The phenotypic and molecular diversity of hemoglobinopathies in India: A review of 15 years at a referral center","volume":"41","author":"Nadkarni","year":"2019","journal-title":"Int J Lab Hematol"},{"key":"R27-20250821","article-title":"Occurrence of HbA2 Borderline Phenotypes in Areas with High Prevalence of Thalassemia","author":"Mosca","year":"2007"},{"key":"R28-20250821","doi-asserted-by":"crossref","first-page":"1380","DOI":"10.3324\/haematol.12840","article-title":"Significance of borderline hemoglobin A2 values in an Italian population with a high prevalence of beta-thalassemia","volume":"93","author":"Giambona","year":"2008","journal-title":"Haematologica"},{"key":"R29-20250821","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1111\/ijlh.12246","article-title":"Impact of iron deficiency on hemoglobin A2% in obligate \u03b2-thalassemia heterozygotes","volume":"37","author":"Sharma","year":"2015","journal-title":"Int J Lab Hematol"},{"key":"R30-20250821","doi-asserted-by":"crossref","first-page":"472","DOI":"10.3324\/haematol.2011.052357","article-title":"Iron deficiency does not compromise the diagnosis of high HbA (2) \u03b2 thalassemia trait","volume":"97","author":"Passarello","year":"2012","journal-title":"Haematologica"},{"key":"R31-20250821","doi-asserted-by":"crossref","first-page":"858294","DOI":"10.1155\/2013\/858294","article-title":"Hemoglobin A2 lowered by iron deficiency and \u03b1 -Thalassemia: Should screening recommendation for \u03b2 -thalassemia change?","volume":"2013","author":"Denic","year":"2013","journal-title":"ISRN Hematol"},{"key":"R32-20250821","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1007\/BF03194650","article-title":"Genotyping of alpha-thalassemia in microcytic hypochromic anemia patients from North India","volume":"47","author":"Sankar","year":"2006","journal-title":"J Appl Genet"},{"key":"R35-20250821","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1309\/AJCP28QKSOPHYOBC","article-title":"Quantification of HbA (2) in patients with and without beta-thalassemia and in the presence of HbS, HbC, HbE, and HbD Punjab hemoglobin variants: comparison of two systems","volume":"131","author":"Higgins","year":"2009","journal-title":"Am J Clin Pathol"},{"key":"R36-20250821","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1309\/73WJV2E0RTVQH0BB","article-title":"Spurious hemoglobin barts caused by bilirubin: A common interference mimicking an uncommon hemoglobinopathy","volume":"125","author":"Howanitz","year":"2006","journal-title":"Am J Clin Pathol"}],"container-title":["International Journal of Applied &amp; Basic Medical Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ovid.com\/10.4103\/ijabmr.ijabmr_70_25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T13:22:41Z","timestamp":1787836961000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ovid.com\/10.4103\/ijabmr.ijabmr_70_25"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7]]},"references-count":31,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2025]]}},"URL":"https:\/\/doi.org\/10.4103\/ijabmr.ijabmr_70_25","relation":{},"ISSN":["2229-516X","2248-9606"],"issn-type":[{"value":"2229-516X","type":"print"},{"value":"2248-9606","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7]]},"assertion":[{"value":"2025-02-06","name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-06-30","name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}}]}}