Simplified hemoglobin chain detection by capillary electrophoresis

Shihabi, Zak K.; Hinsdale, Mark E.

doi:10.1002/elps.200410043

Cited by 20 publications

(5 citation statements)

References 12 publications

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“…By simplifying sample preparation, Shihabi and Hinsdale [72] have used highionic strength buffers and high pH after red cell lysis to denature the hemoglobin molecule, dissociating the a and b chains, while simultaneously releasing heme. This method achieved results similar to the more complicated sample preparation using urea and acetone-HCl extraction.…”

Section: Hemoglobinsmentioning

confidence: 99%

Capillary electrophoresis and the clinical laboratory

et al. 2006

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Over the past 15 years, CE as an analytical tool has shown great promise in replacing many conventional clinical laboratory methods, such as electrophoresis and HPLC. CE's appeal was that it was fast, used very small amounts of sample and reagents, was extremely versatile, and was able to separate large and small analytes, whether neutral or charged. Because of this versatility, numerous methods have been developed for analytes that are of clinical interest. Other than molecular diagnostic and forensic laboratories CE has not been able to make a major impact in the United States. In contrast, in Europe and Japan an increasing number of clinical laboratories are using CE. Now that automated multicapillary instruments are commercially available along with cost-effective test kits, CE may yet be accepted as an instrument that will be routinely used in the clinical laboratories. This review will focus on areas where CE has the potential to have the greatest impact on the clinical laboratory. These include analyses of proteins found in serum and urine, hemoglobin (A1c and variants), carbohydrate-deficient transferrin, forensic and therapeutic drug screening, and molecular diagnostics.

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Section: Hemoglobinsmentioning

confidence: 99%

Capillary electrophoresis and the clinical laboratory

et al. 2006

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“…The developed application represents an effective tool for the identification of globin variants for diagnostic purposes. The obtained separation time of around 3 min is several times shorter than separation times presented in literature for related CE determinations of globin variants .…”

Section: Resultsmentioning

confidence: 63%

Rapid determination of globin chains in red blood cells by capillary electrophoresis using INSTCoated fused‐silica capillary

Tůma

2014

J of Separation Science

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A laboratory-made INSTCoated fused-silica capillary has been newly used for CE separation of four mixtures of proteins in sodium phosphate BGEs at pH 3.0 and 2.5, respectively. The obtained separation efficiencies range from 145,000 theoretical plates per meter for myoglobin to 1,216,000 m(-1) for lysozyme. A total of 49-89% of the number of theoretical plates was obtained in a commercial polyvinyl alcohol coated capillary compared to the INSTCoated capillary under the same experimental conditions, 0-86% was obtained in a laboratory polyacrylamide-coated capillary, and only 0-6% was obtained in an uncoated fused-silica capillary. The RSD values for the intraday repeatability for an INSTCoated capillary were 0.1-1.0% (migration time) and 0.3-2.4% (peak area); RSD values for the interday repeatability in the same capillary are 0.6-1.4% (migration time) and 2.4-5.5% (peak area); RSD values for interday repeatability between different capillaries equaled 1.7-2.1% (migration time) and 2.8-10.9% (peak area). The INSTCoated capillary has been further used for rapid determination of globin chains isolated from red blood cells. A separation of α and β chains prepared from adult blood has been completed in 3 min with a peak resolution of 1.3, and the separation of α and (G)γ chains prepared from newborn blood took 3 min with a peak resolution of 3.6.

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“…Hemoglobin variants have already been analyzed by CE to solve different clinical issues such as thalassemia [24,25]. Recent reviews pointed out virtues and vices of CE techniques with regard to biomolecule detection [26] or biomarker discovery [27].…”

Section: Giuseppe E De Benedetto Matteo Fanigliulomentioning

confidence: 99%

A new CE‐ESI‐MS method for the detection of stable hemoglobin acetaldehyde adducts, potential biomarkers of alcohol abuse

Benedetto

Fanigliulo

2009

Electrophoresis

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A new CE-ESI-MS method was developed to provide a simple way to study changes to hemoglobin (HbA) induced by acetaldehyde (Ach) in vitro. Instrumental parameters were univariately optimized in order to maximize the sensitivity of the CE-ESI-MS method. The electrophoretic separations were carried out in poly-E323-coated capillaries using 60 mM formic acid raised to pH 3.0 with ammonia and containing 5% 2-propanol while the sheath liquid, 2-propanol/water (30:70) with 0.1% formic acid, was delivered at 1.0 microL/min through a coaxial sheath flow electrospray interface. The HbA was incubated with Ach for intervals up to 24 h at concentration varying in the window 0.2-20 mM. Four stable Ach-hemoglobin adducts in the hemoglobin tryptic digest were observed at the submillimolar Ach concentration and characterized by MS/MS experiments: although the alpha and beta N-amino terminal modifications were expected, the two internal ones arising, respectively, from the condensation of Ach molecules on the histidine residue in position 4 in alpha4 (i.e. the fourth peptide after tryptic digestion of alpha chain starting from amino terminal) and on the asparagine residue in position 2 in beta3, were identified for the first time. During the in vitro experiments higher concentrations of Ach were also used; however, it was not possible to identify any other stable modification of hemoglobin. Interestingly, those stable modifications are the only ones in vivo identified in the hemoglobin of moderate alcohol drinkers.

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Simplified hemoglobin chain detection by capillary electrophoresis

Cited by 20 publications

References 12 publications

Capillary electrophoresis and the clinical laboratory

Capillary electrophoresis and the clinical laboratory

Rapid determination of globin chains in red blood cells by capillary electrophoresis using INSTCoated fused‐silica capillary

A new CE‐ESI‐MS method for the detection of stable hemoglobin acetaldehyde adducts, potential biomarkers of alcohol abuse

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