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Representative chromatograms of internal standard and disaccharide peaks from enzymatically digested CSF sample of an Ids KO mouse in a single run. ( A ) internal standard (I.S.) ∆4UA-2S-GlcOEt-6S, ( B ) D0a4, ( C ) D0A0, and ( D ) D0S0. Each species shows two epimers designated as E1 and E2. Shaded orange peaks are the E1s that were used to represent the species for quantification. Blue traces are the authentic standards in a separate run for D0a4, D0A0 and D0S0, respectively. LC-MS conditions refer to experiment section.

Journal: International Journal of Molecular Sciences

Article Title: High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II

doi: 10.3390/ijms21155449

Figure Lengend Snippet: Representative chromatograms of internal standard and disaccharide peaks from enzymatically digested CSF sample of an Ids KO mouse in a single run. ( A ) internal standard (I.S.) ∆4UA-2S-GlcOEt-6S, ( B ) D0a4, ( C ) D0A0, and ( D ) D0S0. Each species shows two epimers designated as E1 and E2. Shaded orange peaks are the E1s that were used to represent the species for quantification. Blue traces are the authentic standards in a separate run for D0a4, D0A0 and D0S0, respectively. LC-MS conditions refer to experiment section.

Article Snippet: D0A0 ∆UA-GlcNAc(IV-A) (HD006); D0S0, ∆UA–GlcNS (IV-S) (HD005); D0a4 ∆UA–GalNAc,4S (∆UA–4S) (CD002); 4UA-2S-GlcNCOEt-6S (HD009) ultrapure (95%+) were purchased from Iduron ( https://iduron.co.uk/ ) on custom order.

Techniques: Liquid Chromatography with Mass Spectroscopy

Calibration curves for D0A0, D0S0 and D0a4 based on the ratio of peak area to I.S. area using undigested human CSF as matrix. ( A – C ) Dynamic range 5–10,000 ng/mL, ( D – F ) inset for dynamic range 5–100 ng/mL. I.S. concentration 20 ng/sample. GAG concentration levels: 5, 10, 20, 100, 1000, 5000, 9000 to 10,000 ng/mL. QCH = 7500 ng/mL, QCM = 300 ng/mL, QCL = 15 ng/mL. Total QCs n = 6. Three (3) µL of standard solutions were used to mimic the mouse CSF sample preparation procedure.

Journal: International Journal of Molecular Sciences

Article Title: High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II

doi: 10.3390/ijms21155449

Figure Lengend Snippet: Calibration curves for D0A0, D0S0 and D0a4 based on the ratio of peak area to I.S. area using undigested human CSF as matrix. ( A – C ) Dynamic range 5–10,000 ng/mL, ( D – F ) inset for dynamic range 5–100 ng/mL. I.S. concentration 20 ng/sample. GAG concentration levels: 5, 10, 20, 100, 1000, 5000, 9000 to 10,000 ng/mL. QCH = 7500 ng/mL, QCM = 300 ng/mL, QCL = 15 ng/mL. Total QCs n = 6. Three (3) µL of standard solutions were used to mimic the mouse CSF sample preparation procedure.

Article Snippet: D0A0 ∆UA-GlcNAc(IV-A) (HD006); D0S0, ∆UA–GlcNS (IV-S) (HD005); D0a4 ∆UA–GalNAc,4S (∆UA–4S) (CD002); 4UA-2S-GlcNCOEt-6S (HD009) ultrapure (95%+) were purchased from Iduron ( https://iduron.co.uk/ ) on custom order.

Techniques: Concentration Assay, Sample Prep

Fit-for-purpose qualification results for Heparan and Dermatan Sulfate Disaccharides in CSF for calibration curves in <xref ref-type= Figure 3 ." width="100%" height="100%">

Journal: International Journal of Molecular Sciences

Article Title: High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II

doi: 10.3390/ijms21155449

Figure Lengend Snippet: Fit-for-purpose qualification results for Heparan and Dermatan Sulfate Disaccharides in CSF for calibration curves in Figure 3 .

Article Snippet: D0A0 ∆UA-GlcNAc(IV-A) (HD006); D0S0, ∆UA–GlcNS (IV-S) (HD005); D0a4 ∆UA–GalNAc,4S (∆UA–4S) (CD002); 4UA-2S-GlcNCOEt-6S (HD009) ultrapure (95%+) were purchased from Iduron ( https://iduron.co.uk/ ) on custom order.

Techniques: Inter Assay, Intra Assay, Injection

A summary of the CSF GAG measurement in MPS patients and animal models.

Journal: International Journal of Molecular Sciences

Article Title: High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II

doi: 10.3390/ijms21155449

Figure Lengend Snippet: A summary of the CSF GAG measurement in MPS patients and animal models.

Article Snippet: D0A0 ∆UA-GlcNAc(IV-A) (HD006); D0S0, ∆UA–GlcNS (IV-S) (HD005); D0a4 ∆UA–GalNAc,4S (∆UA–4S) (CD002); 4UA-2S-GlcNCOEt-6S (HD009) ultrapure (95%+) were purchased from Iduron ( https://iduron.co.uk/ ) on custom order.

Techniques: Concentration Assay, Colorimetric Assay, Inhibition, Enzyme-linked Immunosorbent Assay, Activity Assay, Labeling

HS and DS concentration in FACS-isolated neurons and microglia, CSF, brain, and liver in mice, and CSF of healthy human subjects and MPS II patients (Mean ± SEM).

Journal: International Journal of Molecular Sciences

Article Title: High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II

doi: 10.3390/ijms21155449

Figure Lengend Snippet: HS and DS concentration in FACS-isolated neurons and microglia, CSF, brain, and liver in mice, and CSF of healthy human subjects and MPS II patients (Mean ± SEM).

Article Snippet: D0A0 ∆UA-GlcNAc(IV-A) (HD006); D0S0, ∆UA–GlcNS (IV-S) (HD005); D0a4 ∆UA–GalNAc,4S (∆UA–4S) (CD002); 4UA-2S-GlcNCOEt-6S (HD009) ultrapure (95%+) were purchased from Iduron ( https://iduron.co.uk/ ) on custom order.

Techniques: Concentration Assay