Determining sulfur-containing amino acids by capillary electrophoresis

A fast novel method for total homocyst(e)ine human plasma

Gennaro Vecchione, Maurizio Margaglione, Elvira Grandone, Donatella Colaizzo, Giuseppe Cappucci, Isabella Fermo, Armando D'Angelo, Giovanni Di Minno

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

A high-performance capillary electrophoresis (HPCE) method based on laser-induced fluorescence detection is presented here. It enables the determination of sulfur-containing amino acids within 15 min. Fluorescence of sulfur-containing amino acids in plasma is linear over a range of 50-150 μmol/L for L-methionine, 5-100 μmol/L for L-homocysteine, and 50-200 μmol/L for L-cysteine. For homocysteine, we were able to detect 1 fmol injected, equivalent to a plasma concentration of 10 nmol/L. A similar sensitivity is present for cysteine, an even lower one being found for methionine. The intra- and interassay relative standard deviations are <1%. High-performance liquid chromatography (HPLC) methods are commonly employed for quantifying blood concentrations of sulfur-containing amino acids. A comparative analysis of HPCE and HPLC quantitation of homocysteine has been carried out in 61 blood samples. Plasma concentrations measured by HPCE were in good agreement with those obtained employing an HPLC-based method, a satisfactory correlation being observed between the concentrations obtained by the two methods (r = 0.9972). Thus, the HPCE-based procedure presented here for the measurement of sulfur-containing amino acids in plasma is a simple, fast, accurate, and very sensitive method, suitable for routine determinations in clinical studies.

Original languageEnglish
Pages (from-to)569-574
Number of pages6
JournalElectrophoresis
Volume20
Issue number3
Publication statusPublished - 1999

Fingerprint

Plasma (human)
Sulfur Amino Acids
Capillary electrophoresis
Capillary Electrophoresis
Sulfur
High performance liquid chromatography
Homocysteine
Plasmas
Amino Acids
Methionine
Cysteine
Blood
Fluorescence
High Pressure Liquid Chromatography
Lasers

Keywords

  • Capillary electrophoresis
  • High- performance liquid chromatography
  • Sulfur-containing amino acids

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

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title = "Determining sulfur-containing amino acids by capillary electrophoresis: A fast novel method for total homocyst(e)ine human plasma",
abstract = "A high-performance capillary electrophoresis (HPCE) method based on laser-induced fluorescence detection is presented here. It enables the determination of sulfur-containing amino acids within 15 min. Fluorescence of sulfur-containing amino acids in plasma is linear over a range of 50-150 μmol/L for L-methionine, 5-100 μmol/L for L-homocysteine, and 50-200 μmol/L for L-cysteine. For homocysteine, we were able to detect 1 fmol injected, equivalent to a plasma concentration of 10 nmol/L. A similar sensitivity is present for cysteine, an even lower one being found for methionine. The intra- and interassay relative standard deviations are <1{\%}. High-performance liquid chromatography (HPLC) methods are commonly employed for quantifying blood concentrations of sulfur-containing amino acids. A comparative analysis of HPCE and HPLC quantitation of homocysteine has been carried out in 61 blood samples. Plasma concentrations measured by HPCE were in good agreement with those obtained employing an HPLC-based method, a satisfactory correlation being observed between the concentrations obtained by the two methods (r = 0.9972). Thus, the HPCE-based procedure presented here for the measurement of sulfur-containing amino acids in plasma is a simple, fast, accurate, and very sensitive method, suitable for routine determinations in clinical studies.",
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author = "Gennaro Vecchione and Maurizio Margaglione and Elvira Grandone and Donatella Colaizzo and Giuseppe Cappucci and Isabella Fermo and Armando D'Angelo and {Di Minno}, Giovanni",
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T1 - Determining sulfur-containing amino acids by capillary electrophoresis

T2 - A fast novel method for total homocyst(e)ine human plasma

AU - Vecchione, Gennaro

AU - Margaglione, Maurizio

AU - Grandone, Elvira

AU - Colaizzo, Donatella

AU - Cappucci, Giuseppe

AU - Fermo, Isabella

AU - D'Angelo, Armando

AU - Di Minno, Giovanni

PY - 1999

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N2 - A high-performance capillary electrophoresis (HPCE) method based on laser-induced fluorescence detection is presented here. It enables the determination of sulfur-containing amino acids within 15 min. Fluorescence of sulfur-containing amino acids in plasma is linear over a range of 50-150 μmol/L for L-methionine, 5-100 μmol/L for L-homocysteine, and 50-200 μmol/L for L-cysteine. For homocysteine, we were able to detect 1 fmol injected, equivalent to a plasma concentration of 10 nmol/L. A similar sensitivity is present for cysteine, an even lower one being found for methionine. The intra- and interassay relative standard deviations are <1%. High-performance liquid chromatography (HPLC) methods are commonly employed for quantifying blood concentrations of sulfur-containing amino acids. A comparative analysis of HPCE and HPLC quantitation of homocysteine has been carried out in 61 blood samples. Plasma concentrations measured by HPCE were in good agreement with those obtained employing an HPLC-based method, a satisfactory correlation being observed between the concentrations obtained by the two methods (r = 0.9972). Thus, the HPCE-based procedure presented here for the measurement of sulfur-containing amino acids in plasma is a simple, fast, accurate, and very sensitive method, suitable for routine determinations in clinical studies.

AB - A high-performance capillary electrophoresis (HPCE) method based on laser-induced fluorescence detection is presented here. It enables the determination of sulfur-containing amino acids within 15 min. Fluorescence of sulfur-containing amino acids in plasma is linear over a range of 50-150 μmol/L for L-methionine, 5-100 μmol/L for L-homocysteine, and 50-200 μmol/L for L-cysteine. For homocysteine, we were able to detect 1 fmol injected, equivalent to a plasma concentration of 10 nmol/L. A similar sensitivity is present for cysteine, an even lower one being found for methionine. The intra- and interassay relative standard deviations are <1%. High-performance liquid chromatography (HPLC) methods are commonly employed for quantifying blood concentrations of sulfur-containing amino acids. A comparative analysis of HPCE and HPLC quantitation of homocysteine has been carried out in 61 blood samples. Plasma concentrations measured by HPCE were in good agreement with those obtained employing an HPLC-based method, a satisfactory correlation being observed between the concentrations obtained by the two methods (r = 0.9972). Thus, the HPCE-based procedure presented here for the measurement of sulfur-containing amino acids in plasma is a simple, fast, accurate, and very sensitive method, suitable for routine determinations in clinical studies.

KW - Capillary electrophoresis

KW - High- performance liquid chromatography

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