71 research outputs found
Evaluation of Coupling Reversed Phase, Aqueous Normal Phase, and Hydrophilic Interaction Liquid Chromatography with Orbitrap Mass Spectrometry for Metabolomic Studies of Human Urine
In this study, we assessed three liquid chromatographic
platforms:
reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction
(HILIC) for the analysis of polar metabolite standard mixtures and
for their coverage of urinary metabolites. The two zwitterionic HILIC
columns showed high-quality chromatographic performance for metabolite
standards, improved separation for isomers, and the greatest coverage
of polar metabolites in urine. In contrast, on the reversed phase
column, most metabolites eluted very rapidly with little or no separation.
Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic
platform, approximately 970 metabolite signals with repeatable peak
areas (relative standard deviation (RSD) ≤ 25%) could be putatively
identified in human urine, by elemental composition assignment within
a 3 ppm mass error. The ability of the methodology for the verification
of nonmolecular ions, which arise from adduct formation, and the possibility
of distinguishing isomers could also be demonstrated. Careful examination
of the raw data and the use of masses for predicted metabolites produced
an extension of the metabolite list for human urine
Evaluation of Coupling Reversed Phase, Aqueous Normal Phase, and Hydrophilic Interaction Liquid Chromatography with Orbitrap Mass Spectrometry for Metabolomic Studies of Human Urine
In this study, we assessed three liquid chromatographic
platforms:
reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction
(HILIC) for the analysis of polar metabolite standard mixtures and
for their coverage of urinary metabolites. The two zwitterionic HILIC
columns showed high-quality chromatographic performance for metabolite
standards, improved separation for isomers, and the greatest coverage
of polar metabolites in urine. In contrast, on the reversed phase
column, most metabolites eluted very rapidly with little or no separation.
Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic
platform, approximately 970 metabolite signals with repeatable peak
areas (relative standard deviation (RSD) ≤ 25%) could be putatively
identified in human urine, by elemental composition assignment within
a 3 ppm mass error. The ability of the methodology for the verification
of nonmolecular ions, which arise from adduct formation, and the possibility
of distinguishing isomers could also be demonstrated. Careful examination
of the raw data and the use of masses for predicted metabolites produced
an extension of the metabolite list for human urine
Evaluation of Coupling Reversed Phase, Aqueous Normal Phase, and Hydrophilic Interaction Liquid Chromatography with Orbitrap Mass Spectrometry for Metabolomic Studies of Human Urine
In this study, we assessed three liquid chromatographic
platforms:
reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction
(HILIC) for the analysis of polar metabolite standard mixtures and
for their coverage of urinary metabolites. The two zwitterionic HILIC
columns showed high-quality chromatographic performance for metabolite
standards, improved separation for isomers, and the greatest coverage
of polar metabolites in urine. In contrast, on the reversed phase
column, most metabolites eluted very rapidly with little or no separation.
Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic
platform, approximately 970 metabolite signals with repeatable peak
areas (relative standard deviation (RSD) ≤ 25%) could be putatively
identified in human urine, by elemental composition assignment within
a 3 ppm mass error. The ability of the methodology for the verification
of nonmolecular ions, which arise from adduct formation, and the possibility
of distinguishing isomers could also be demonstrated. Careful examination
of the raw data and the use of masses for predicted metabolites produced
an extension of the metabolite list for human urine
Evaluation of Coupling Reversed Phase, Aqueous Normal Phase, and Hydrophilic Interaction Liquid Chromatography with Orbitrap Mass Spectrometry for Metabolomic Studies of Human Urine
In this study, we assessed three liquid chromatographic
platforms:
reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction
(HILIC) for the analysis of polar metabolite standard mixtures and
for their coverage of urinary metabolites. The two zwitterionic HILIC
columns showed high-quality chromatographic performance for metabolite
standards, improved separation for isomers, and the greatest coverage
of polar metabolites in urine. In contrast, on the reversed phase
column, most metabolites eluted very rapidly with little or no separation.
Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic
platform, approximately 970 metabolite signals with repeatable peak
areas (relative standard deviation (RSD) ≤ 25%) could be putatively
identified in human urine, by elemental composition assignment within
a 3 ppm mass error. The ability of the methodology for the verification
of nonmolecular ions, which arise from adduct formation, and the possibility
of distinguishing isomers could also be demonstrated. Careful examination
of the raw data and the use of masses for predicted metabolites produced
an extension of the metabolite list for human urine
Evaluation of Coupling Reversed Phase, Aqueous Normal Phase, and Hydrophilic Interaction Liquid Chromatography with Orbitrap Mass Spectrometry for Metabolomic Studies of Human Urine
In this study, we assessed three liquid chromatographic
platforms:
reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction
(HILIC) for the analysis of polar metabolite standard mixtures and
for their coverage of urinary metabolites. The two zwitterionic HILIC
columns showed high-quality chromatographic performance for metabolite
standards, improved separation for isomers, and the greatest coverage
of polar metabolites in urine. In contrast, on the reversed phase
column, most metabolites eluted very rapidly with little or no separation.
Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic
platform, approximately 970 metabolite signals with repeatable peak
areas (relative standard deviation (RSD) ≤ 25%) could be putatively
identified in human urine, by elemental composition assignment within
a 3 ppm mass error. The ability of the methodology for the verification
of nonmolecular ions, which arise from adduct formation, and the possibility
of distinguishing isomers could also be demonstrated. Careful examination
of the raw data and the use of masses for predicted metabolites produced
an extension of the metabolite list for human urine
Major metabolites in midgut.
<p>(A) Relatively abundant metabolites in midgut. Data are presented as a heat-map of peak areas, from red (>1×10<sup>7</sup>) to blue (<5×10<sup>3</sup>). (B) Relatively abundant lipids in the midgut. Hist = acyl histamine. Red = area >10<sup>7</sup> Yellow area >2×10<sup>4</sup> Blue <10<sup>3</sup>.</p
Extracted ion chromatograms showing GSH and its precursors extracted from Drosophila cuticle.
<p>Extracted ion chromatograms showing GSH and its precursors extracted from Drosophila cuticle.</p
Major metabolites in testes.
<p>(A) Testes relatively abundant polar metabolites. *Gives guanine as an in source fragment ion. **Isomeric with adenosine which but elutes earlier gives guanine as an in source fragment ion. Data are presented as a heat-map of peak areas, from red (>1×10<sup>7</sup>) to blue (<5×10<sup>3</sup>). (B) Relatively abundant lipids in testes. Data are presented as a heat-map of peak areas, from red (>1×10<sup>7</sup>) to blue (<1×10<sup>3</sup>).</p
The top 25 most abundant metabolites by area response in <i>Drosophila</i> whole fly and in ten tissues from <i>Drosophila</i>.
<p>Red = area >10<sup>8</sup> Yellow area >10<sup>7</sup> Blue = area <10<sup>5</sup>.</p
Extracted ion traces for decarboxy S-adenosylmethione and its fragment methylthioadenosine, a metabolite unique to accessory glands.
<p>Extracted ion traces for decarboxy S-adenosylmethione and its fragment methylthioadenosine, a metabolite unique to accessory glands.</p
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