Quadrupole-time-of-flight mass spectrometric identification of hemoglobin subunits α, β, γ and δ in unknown peaks of high performance liquid chromatography of hemoglobin in β-thalassemias

Uday Y.H. Abdullah, Hishamshah M. Ibrahim, Noraesah B. Mahmud, Mohamad Z. Salleh, Teh L. Kek, Mohd N.F.B. Noorizhab, Haitham M. Jassim, Iekhsan Othman, Syafiq A. Zainal Abidin, Bin Alwi Zilfalil, Prapin Wilairat, Suthat Fucharoen

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Abstract

This is the first report of quadrupole time-of-flight (Q-TOF) mass spectrometric identification of the hemoglobin (Hb) subunits, α, β, δ and γ peptides, derived from enzymatic-digestion of proteins in the early unknown peaks of the cation exchange chromatography of Hb. The objectives were to identify the unknown high performance liquid chromatography (HPLC) peaks in healthy subjects and in patients with β-thalassemia (β-thal). The results demonstrate the existence of pools of free globin chains in red blood cells (RBCs). The α-, β-, δ- and γ-globin peptides were identified in the unknown HPLC peaks. The quantification and role of the free globin pool in patients with β-thal requires further investigation. Identification of all types of Hb subunits in the retention time (RT) before 1 min. suggests that altered Hbs is the nature of these fast-eluting peaks. Relevancy of thalassemias to the protein-aggregation disorders will require review of the role of free globin in the pathology of the disease.

Original languageEnglish
Pages (from-to)182-187
Number of pages6
JournalHemoglobin
Volume43
Issue number3
DOIs
Publication statusPublished - 12 Jul 2019

Keywords

  • High performance liquid chromatography (HPLC)
  • mass spectrometry (MS)
  • peaks
  • quadrupole time-of-flight (Q-TOF)
  • thalassemia

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