Comparison of High-Performance Liquid Chromatography and Capillary Electrophoresis in the Screening of Haemoglobinopathies

Authors

  • Shumaila Asghar Department of Hematology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Asad Mahmood Abbasi Department of Hematology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Jawad Zafar Department of Hematology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Rafia Mahmood Department of Hematology, Combined Military Hospital, Hyderabad/National University of Medical Sciences (NUMS) Pakistan
  • Saima Zahir Department of Hematology, Armed Forces Institute of Pathology/National University of Medical Sciences (NUMS) Rawalpindi Pakistan
  • Maria Khan Department of Hematology, Armed Forces Institute of Transfusion/National University of Medical Sciences (NUMS) Rawalpindi Pakistan

DOI:

https://doi.org/10.51253/pafmj.v74i2.7452

Keywords:

Beta-thalassaemia, Capillary electrophoresis, Haemoglobinopathies, High-Performance liquid chromatography

Abstract

Objective: To compare the results of High-Performance Liquid Chromatography and Capillary Electrophoresis in the
screening of haemoglobinopathies.

Study Design: Cross-sectional study.

Place and Duration of Study: Department of Haematology, Armed Forces Institute of Pathology, Rawalpindi Pakistan, from May 2020 to Nov 2020.

Methodology: Ninety(90) newly diagnosed patients with haemoglobinopathies were included. Patients with a history of recent blood transfusion (<4 weeks) were excluded. Venous blood samples were taken in EDTA tubes, separated into two aliquots and evaluated in parallel on High-Performance Liquid Chromatography and Capillary Electrophoresis analyzers.

Results: Nine different haemoglobinopathies were encountered in the study sample. The most common condition was beta thalassaemia trait found in 50(55.6%) subjects, while Haemoglobin (Hb) (D) homozygous was the least common, detected in only 1(1.1%) subjects. A significant difference was observed between values in cases of Hb(D) Iran trait; Mean Hb(A2) percentage detected by HPLC was 41.96±1.46% compared to 1.5±0.79% detected by CE (p-value<0.001). CE detected a mean of 41.4±1.01% of Hb(D)Iran in the patients of the Hb(D) Iran trait, while High-Performance Liquid Chromatography did not detect this Hb variant (p-value<0.001). A significant difference was also found in cases of Hb(E) trait in percentages of Hb(A2) and Hb(E) variants. High-Performance Liquid Chromatography detected a mean of 31.43±2.84% of Hb(A2) compared to 4.5±0.5% detected by CE (p-value=0.003).
Conclusion: Capillary electrophoresis is capable of identifying haemoglobin variants Hb(D) Iran and Hb(E) in cases of Hb(D) Iran trait and Hb(E) trait/Hb(E) homozygous, which HPLC does not pick up.

Downloads

Download data is not yet available.

References

Nisha TR, Bindu CS. HPLC based evaluation of

Haemoglobinopathies in a tertiary care setting in North Kerala. J

Med Sci Clin Res 2018; 6(1): 32027-32032.

https://doi.org/10.18535/jmscr/v6i1.102.

Shabbir S, Nadeem M, Sattar A, Ara I, Ansari S, Farzana T, et al.

Type and frequency of haemoglobinopathies, diagnosed in the

area of Karachi, in Pakistan, Cogent Med 2016; 3(1): 1-6.

https://doi.org/10.1080/2331205X.2016.1188875.

Piel FB. The present and future global burden of the inherited

disorders of hemoglobin. Hematol Oncol Clin North Am 2016;

(2): 327-341. https://doi.org/10.1016/j.hoc.2015.11.004.

Ghafoor M, Sabar MF, Sabir F. Prevention programmes and

prenatal diagnosis for beta thalassemia in Pakistan: A narrative

review. J Pak Med Assoc 2021; 71(1): 326-331.

https://doi.org/10.47391/jpma.665.

Karadag ME, Akbulut ED, Avci E, Oguz EF, Kader S, Abuşoğlu

G. et al. Evaluation of four different HPLC devices for

hemoglobinopathy screening. Turk J Biochem 2021; 46(1): 39–44.

Canatan D, Delibas S. Report on ten years’ experience of

premarital hemoglobinopathy screening at a center in Antalya,

southern Turkey. Hemoglobin 2016; 40(4): 273-276.

https://doi.org/10.3109/03630269.2016.1170030.

Chakravarthy SN, Ramanathan RS, Smitha S, Valappil V,

Durairaj M. Hide and seek: Efficacy of HPLC and capillary zone

electrophoresis as screening tools for hemoglobin disorders. Int J

Path Lab 2016; 2(1): 6.

Frömmel C. Newborn Screening for Sickle Cell Disease and

Other Hemoglobinopathies: A Short Review on Classical

Laboratory Methods-Isoelectric Focusing, HPLC, and Capillary

Electrophoresis. Int J Neonatal Screen 2018; 4(4): 39.

https://doi.org/10.3390/ijns4040039.

Greene DN, Pyle AL, Chang JS, Hoke C, Lorey T. Comparison of

SebiaCapillarys Flex capillary electrophoresis with the Bio Rad

Variant II high pressure liquid chromatography in the

evaluation of haemoglobinopathies. Clin Chim Acta 2012;

(15-16): 1232-1238. https://doi.org/10.1016/j.cca.2012.03.027.

Sangkitporn S, Sangkitporn SK, Tanjatham S, Suwannakan B,

Rithapirom S, Yodtup Y. et al. Multicenter validation of fully

automated capillary electrophoresis method for diagnosis of

thalassemias and haemoglobinopathies in Thailand. Southeast

Asian J Trop Med Public Health 2011; 42(4): 1224-1232.

Cotton F, Changying L, Fontaine B. Evaluation of a capillary

electrophoresis method for routine determination of

hemoglobins A2 and F. Clin Chem 1999; 45(2): 237-243.

Bakshi NA, Gulbranson R, Garstka D. Serum free light chain

(FLC) measurement can aid capillary zone electrophoresis in

detecting subtle FLC-producing M proteins. Am J Clin Pathol

; 124(2): 214-218.

https://doi.org/10.1309/xe3u-dark-w1b9-emwm.

Bossuyt X, Lissoir B, Marien G. Automated serum protein

electrophoresis by Capillarys. Clin Chem Lab Med 2003; 41(5):

-710. https://doi.org/10.1515/cclm.2003.107.

Keren DF, Hedstrom D, Gulbransan R, Ou CN, Bak R.

Comparison of sebiacapillarys capillary electrophoresis with the

primus high-pressure liquid chromatography in the evaluation

of haemoglobinopathies. Am J Clin Pathol 2008; 130(5): 824-31.

https://doi.org/10.1309/ajcpqy80hzwhhgzf.

Mohanty PK, Meher S, Dehury S, Bhattacharya S, Das K, Patel S,

et al. Compound heterozygote of Hb D Iran [HBB: c. 67G> C, β

(B4) Glu> Gln] with β0-thalassemia [cds 41/42 (-CTTT)] from

Eastern India Hematol Transfus Cell Ther 2018; 40(1): 82-85.

https://doi.org/10.1016%2Fj.bjhh.2017.09.001.

Chopra P, Bhardwaj S, Negi P, Arora A. Comparison of Two

High-Pressure Liquid Chromatography Instruments Bio-Rad

Variant-II and Tosoh HLC-723G11 in the Evaluation of

Haemoglobinopathies. Indian J Hematol Blood Transfus 2020;

(4): 725-732.

https://doi.org/10.1007/s12288-020-01298-5.

Olivieri NF. The beta-thalassemias. N Engl J Med 1999; 341(2):

-109. https://doi.org/10.1056/nejm199907083410207.

Chhotray GP, Dash BP, Ranjit M. Spectrum of haemoglobinopathies in Orissa, India. Hemoglobin 2004; 28(2): 117-122.

https://doi.org/10.1081/hem-120034244.

Daniel YA, Turner C, Haynes RM. Quantification of hemoglobin

A2 by tandem mass spectrometry. Clin Chem 2007; 53(8): 1448-

https://doi.org/10.1373/clinchem.2007.088682.

Paleari R, Ceriotti F, Harteveld CL, Strollo M, Bakker-Verweij G,

Ter Huurne J, et al. Calibration by commutable control materials

is able to reduce inter-method differences of current highperformance methods for HbA2. Clin Chim Acta 2018; 477: 60-

https://doi.org/10.1016/j.cca.2017.12.001

Downloads

Published

29-04-2024

Issue

Section

Original Articles

How to Cite

1.
Asghar S, Mahmood Abbasi A, Zafar J, Mahmood R, Zahir S, Khan M. Comparison of High-Performance Liquid Chromatography and Capillary Electrophoresis in the Screening of Haemoglobinopathies. Pak Armed Forces Med J [Internet]. 2024 Apr. 29 [cited 2024 Nov. 28];74(2):357-61. Available from: https://pafmj.org/PAFMJ/article/view/7452