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Integrated chromatograms, stability, and specificity of the liquid chromatography-mass spectrometry (LC-MS/MS) ceramide quantification assay. During the 5-minute run, Mobile Phase A comprised 10 mM ammonium formate and 0.1% formic acid in LC-MS grade water. Mobile Phase B contained 10 mM ammonium formate and 0.1% formic acid in LC-MS grade water/acetonitrile/isopropanol (5%: 47.5%: 47.5%) (A) . Ceramide metabolites and internal standards were separated adequately based on molecular weights and polarities ( B - C ). The stability of ceramide metabolites over three freeze/thaw cycles showed percentage changes represented by blue, orange, and grey bars after 24, 48, and 72 h, respectively. Quality control levels confirmed percentage changes within ± 7% for all metabolites CerC16:0 ( D ), CerC18:0 ( E ), CerC22:0 ( F ), <t>CerC24:0</t> ( G ), and CerC24:1 ( H ). Overlaid chromatograms of blank samples ( I ) confirmed method specificity, showing no interfering peaks at the respective retention times for ceramide metabolites. IS, Internal Standard; %change, stability
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The differential abundance of plasma ceramide species. In plasma, the distribution of ceramides is relatively fluctuating, with <t>C24:0</t> being the most abundant and C18:0 the least concentration ( A ). Likewise, C24:1/C22:0 and C18:0/C24:0 are the highest and lowest ratios of plasma ceramide, respectively ( B ). Independent two-tailed t-test has shown a significant difference between males and females in all measured ceramide species, except for the concentration of C24:0 (ns; not significant P > 0.05) ( C ), and the ratio of C18:0/22:0 ( P > 0.05) ( D ). Data are presented as Mean ± SEM
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Integrated chromatograms, stability, and specificity of the liquid chromatography-mass spectrometry (LC-MS/MS) ceramide quantification assay. During the 5-minute run, Mobile Phase A comprised 10 mM ammonium formate and 0.1% formic acid in LC-MS grade water. Mobile Phase B contained 10 mM ammonium formate and 0.1% formic acid in LC-MS grade water/acetonitrile/isopropanol (5%: 47.5%: 47.5%) (A) . Ceramide metabolites and internal standards were separated adequately based on molecular weights and polarities ( B - C ). The stability of ceramide metabolites over three freeze/thaw cycles showed percentage changes represented by blue, orange, and grey bars after 24, 48, and 72 h, respectively. Quality control levels confirmed percentage changes within ± 7% for all metabolites CerC16:0 ( D ), CerC18:0 ( E ), CerC22:0 ( F ), CerC24:0 ( G ), and CerC24:1 ( H ). Overlaid chromatograms of blank samples ( I ) confirmed method specificity, showing no interfering peaks at the respective retention times for ceramide metabolites. IS, Internal Standard; %change, stability

Journal: Lipids in Health and Disease

Article Title: Circulating ceramide levels and ratios in Emirati youth under 18 years: associations with cardiometabolic risk factors

doi: 10.1186/s12944-024-02080-6

Figure Lengend Snippet: Integrated chromatograms, stability, and specificity of the liquid chromatography-mass spectrometry (LC-MS/MS) ceramide quantification assay. During the 5-minute run, Mobile Phase A comprised 10 mM ammonium formate and 0.1% formic acid in LC-MS grade water. Mobile Phase B contained 10 mM ammonium formate and 0.1% formic acid in LC-MS grade water/acetonitrile/isopropanol (5%: 47.5%: 47.5%) (A) . Ceramide metabolites and internal standards were separated adequately based on molecular weights and polarities ( B - C ). The stability of ceramide metabolites over three freeze/thaw cycles showed percentage changes represented by blue, orange, and grey bars after 24, 48, and 72 h, respectively. Quality control levels confirmed percentage changes within ± 7% for all metabolites CerC16:0 ( D ), CerC18:0 ( E ), CerC22:0 ( F ), CerC24:0 ( G ), and CerC24:1 ( H ). Overlaid chromatograms of blank samples ( I ) confirmed method specificity, showing no interfering peaks at the respective retention times for ceramide metabolites. IS, Internal Standard; %change, stability

Article Snippet: The powder forms of Cer internal standards CerC16:0-d7 [C16:0-ceramide-d7 (d18:1-d7/16:0)], CerC18:0-d7 [C18:0-ceramide-d7 (d18:1-d7/18:0)], CerC24:0-d7 [C24:0-ceramide-d7 (d18:1-d7/24:0)], CerC24:1-d7 [C24:1-ceramide-d7 (d18:1-d7/24:1(15Z))] were also purchased from Avanti Polar Lipids [Alabaster, AL].

Techniques: Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Control

The Multiple Reaction Monitoring (MRM) parameters of ceramide metabolites and corresponding internal standards

Journal: Lipids in Health and Disease

Article Title: Circulating ceramide levels and ratios in Emirati youth under 18 years: associations with cardiometabolic risk factors

doi: 10.1186/s12944-024-02080-6

Figure Lengend Snippet: The Multiple Reaction Monitoring (MRM) parameters of ceramide metabolites and corresponding internal standards

Article Snippet: The powder forms of Cer internal standards CerC16:0-d7 [C16:0-ceramide-d7 (d18:1-d7/16:0)], CerC18:0-d7 [C18:0-ceramide-d7 (d18:1-d7/18:0)], CerC24:0-d7 [C24:0-ceramide-d7 (d18:1-d7/24:0)], CerC24:1-d7 [C24:1-ceramide-d7 (d18:1-d7/24:1(15Z))] were also purchased from Avanti Polar Lipids [Alabaster, AL].

Techniques: Targeted Proteomics

The liquid chromatography-mass spectrometry (LC-MS/MS) ceramide quantification assay validation results

Journal: Lipids in Health and Disease

Article Title: Circulating ceramide levels and ratios in Emirati youth under 18 years: associations with cardiometabolic risk factors

doi: 10.1186/s12944-024-02080-6

Figure Lengend Snippet: The liquid chromatography-mass spectrometry (LC-MS/MS) ceramide quantification assay validation results

Article Snippet: The powder forms of Cer internal standards CerC16:0-d7 [C16:0-ceramide-d7 (d18:1-d7/16:0)], CerC18:0-d7 [C18:0-ceramide-d7 (d18:1-d7/18:0)], CerC24:0-d7 [C24:0-ceramide-d7 (d18:1-d7/24:0)], CerC24:1-d7 [C24:1-ceramide-d7 (d18:1-d7/24:1(15Z))] were also purchased from Avanti Polar Lipids [Alabaster, AL].

Techniques: Chromatography, Biomarker Discovery, Concentration Assay

Comparative results of the liquid chromatography-mass spectrometry (LC-MS/MS) ceramide quantification assay in this study and other relevant studies

Journal: Lipids in Health and Disease

Article Title: Circulating ceramide levels and ratios in Emirati youth under 18 years: associations with cardiometabolic risk factors

doi: 10.1186/s12944-024-02080-6

Figure Lengend Snippet: Comparative results of the liquid chromatography-mass spectrometry (LC-MS/MS) ceramide quantification assay in this study and other relevant studies

Article Snippet: The powder forms of Cer internal standards CerC16:0-d7 [C16:0-ceramide-d7 (d18:1-d7/16:0)], CerC18:0-d7 [C18:0-ceramide-d7 (d18:1-d7/18:0)], CerC24:0-d7 [C24:0-ceramide-d7 (d18:1-d7/24:0)], CerC24:1-d7 [C24:1-ceramide-d7 (d18:1-d7/24:1(15Z))] were also purchased from Avanti Polar Lipids [Alabaster, AL].

Techniques: Chromatography, Extraction, Solvent, Injection, Modification

Median and 95% reference ranges of plasma ceramide levels and ratios in individuals aged 6–17 years

Journal: Lipids in Health and Disease

Article Title: Circulating ceramide levels and ratios in Emirati youth under 18 years: associations with cardiometabolic risk factors

doi: 10.1186/s12944-024-02080-6

Figure Lengend Snippet: Median and 95% reference ranges of plasma ceramide levels and ratios in individuals aged 6–17 years

Article Snippet: The powder forms of Cer internal standards CerC16:0-d7 [C16:0-ceramide-d7 (d18:1-d7/16:0)], CerC18:0-d7 [C18:0-ceramide-d7 (d18:1-d7/18:0)], CerC24:0-d7 [C24:0-ceramide-d7 (d18:1-d7/24:0)], CerC24:1-d7 [C24:1-ceramide-d7 (d18:1-d7/24:1(15Z))] were also purchased from Avanti Polar Lipids [Alabaster, AL].

Techniques: Clinical Proteomics

The differential abundance of plasma ceramide species. In plasma, the distribution of ceramides is relatively fluctuating, with C24:0 being the most abundant and C18:0 the least concentration ( A ). Likewise, C24:1/C22:0 and C18:0/C24:0 are the highest and lowest ratios of plasma ceramide, respectively ( B ). Independent two-tailed t-test has shown a significant difference between males and females in all measured ceramide species, except for the concentration of C24:0 (ns; not significant P > 0.05) ( C ), and the ratio of C18:0/22:0 ( P > 0.05) ( D ). Data are presented as Mean ± SEM

Journal: Lipids in Health and Disease

Article Title: Circulating ceramide levels and ratios in Emirati youth under 18 years: associations with cardiometabolic risk factors

doi: 10.1186/s12944-024-02080-6

Figure Lengend Snippet: The differential abundance of plasma ceramide species. In plasma, the distribution of ceramides is relatively fluctuating, with C24:0 being the most abundant and C18:0 the least concentration ( A ). Likewise, C24:1/C22:0 and C18:0/C24:0 are the highest and lowest ratios of plasma ceramide, respectively ( B ). Independent two-tailed t-test has shown a significant difference between males and females in all measured ceramide species, except for the concentration of C24:0 (ns; not significant P > 0.05) ( C ), and the ratio of C18:0/22:0 ( P > 0.05) ( D ). Data are presented as Mean ± SEM

Article Snippet: The powder forms of CerC16:0 [C16:0-ceramide (d18:1/16:0)], CerC18:0 [C18:0-ceramide (d18:1/18:0)], CerC22:0 [C22:0-ceramide (d18:1/22:0)], CerC24:0 [C24:0-ceramide (d18:1/24:0)], CerC24:1 [C24:1-ceramide (d18:1/24:1(15Z))] were purchased from Avanti Polar Lipids [Alabaster, AL].

Techniques: Clinical Proteomics, Concentration Assay, Two Tailed Test