Editorial
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Methodol. Mar 26, 2016; 6(1): 20-24
Published online Mar 26, 2016. doi: 10.5662/wjm.v6.i1.20
Cation-exchange high-performance liquid chromatography for variant hemoglobins and HbF/A2: What must hematopathologists know about methodology?
Prashant Sharma, Reena Das
Prashant Sharma, Reena Das, Department of Hematology, Level 5, Research Block A, Postgraduate Institute of Medical Education and Research, Chandigarh 160012, India
Author contributions: Sharma P and Das R both made substantial contributions to conception of this article; Sharma P drafted the article; Das R made critical revisions related to important intellectual content; all authors gave final approval of the version of the article submitted.
Conflict-of-interest statement: Both authors of the paper declare that they have no conflicting interests (including commercial, personal, political, intellectual or religious interests) that are related to the work submitted for consideration for publication.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Dr. Prashant Sharma, MD, DNB, DM, Associate Professor, Department of Hematology, Level 5, Research Block A, Postgraduate Institute of Medical Education and Research, Sector 12, Chandigarh 160012, India. sharma.prashant@pgimer.edu.in
Telephone: +91-88-72016123
Received: August 28, 2015
Peer-review started: September 1, 2015
First decision: December 7, 2015
Revised: January 28, 2016
Accepted: February 23, 2016
Article in press: February 24, 2016
Published online: March 26, 2016
Abstract

Cation-exchange high-performance liquid chromatography (CE-HPLC) is a widely used laboratory test to detect variant hemoglobins as well as quantify hemoglobins F and A2 for the diagnosis of thalassemia syndromes. It’s versatility, speed, reproducibility and convenience have made CE-HPLC the method of choice to initially screen for hemoglobin disorders. Despite its popularity, several methodological aspects of the technology remain obscure to pathologists and this may have consequences in specific situations. This paper discusses the basic principles of the technique, the initial quality control steps and the interpretation of various controls and variables that are available on the instrument output. Subsequent sections are devoted to methodological considerations that arise during reporting of cases. For instance, common problems of misidentified peaks, totals crossing 100%, causes of total area being above or below acceptable limits and the importance of pre-integration region peaks are dealt with. Ultimately, CE-HPLC remains an investigation, the reporting of which combines in-depth knowledge of the biological basics with more than a working knowledge of the technological aspects of the technique.

Keywords: Anemia, Diagnosis, Hematological disorders, Hematopathology, Hemoglobin, Hemoglobinopathies, High-performance liquid chromatography, Laboratory instrumentation, Red blood cells, Thalassemia

Core tip: Interpretation of cation-exchange high-performance liquid chromatography requires in-depth knowledge of the biological basics of the disorders of hemoglobin with knowledge of the technological aspects of the technique. Pathologists may be unaware of the nuances of the technique, the rigorous quality control required and the approach to pitfalls that may be encountered. Here we list the most common of these, and based on literature and our experience, attempt to guide novices in this exciting and useful technology.