A STUDY OF HIGHPERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) IN PATIENTS PRESENTING WITH ANAEMIA AT A TERTIARY CARE HOSPITAL

Abstract

– The most prevalent type of genetically inherited red blood cell disorder worldwide are hemoglobinopathies. Screening and accurate identification of hemoglobin (Hb) variants have become increasingly important in antenatal diagnosis and prevention of Hb disorders. The objective is to identify haemoglobin abnormalities by high performance liquid chromatography (HPLC) for screening the anaemic patients who are visiting the hospital in OPD and admitted in IPD. This is cross sectional retrospective study for a period of 6 months and performed in a tertiary care hospital on patients presenting with anemia in different Departments. In total, high performance liquid chromatography HPLC of 400 patients with anemia was studied. Out of 400 patients, 190 were males and 210 were females. Age group varied from infants to middle-aged adults. The following criteria were used to identify hemoglobinopathies on HPLC patterns as follow: HbS:- 10%-45%: Sickle cell trait, 45%-50%: HbS>HbA-Sickle cell disease, HbS50%: Sickle cell disease, HbA2:- >3.5%: Beta thalassemia trait, HbA2 normal or increase and HbF increase upto 90% : Beta thalassemia major, HbF:- 5% to 30%: hereditary persistence of fetalhemoglobin, HbD Variant: Retention time of unknown peak between 3.90-4.30 minutes. Age and sex was expressed in actual number and percentages. Countinuous variables were presented as mean ± 2SD. Chi square statistics were used to compare categorical variables. P<0.005 was statistically significant. Microsoft excel and graph pad calculator was used for data analysis. 55% cases (total = 220) having hemoglobinopathies. Among those with hemoglobinopathies 120 (55%) were females and 100 (45%) were males. Most of the patients being less than 30 years. HPLC results include 19.75% cases of sickle cell disease, 1.5% of sickle-beta thalassemia, 27.25% of sickle cell trait, 4.5% of beta thalassemia trait, 0.5% of beta thalassemia major, 0.5% of HbD variants, 0.5% of hereditary persistence of fetal hemoglobin and 0.5% of unknown peaks, they have been listed as others. HPLC is very simple, accurate and superior technique in timely detection of various haemoglobin disorders, which helps in early management of patients.