Beyond Healthy Populations: Hospital-Derived Reference Material and Enhanced Dynamic Range Expand Clinical Research Metabolomics Coverage

Clinical research metabolomics requires reference materials and robust workflows for quality control across low-volume samples. NIST SRM 1950 represents healthy adults within a narrow age range and may not reflect the broader metabolites and medications encountered in clinical research. Dried blood spots (DBS) can challenge instruments because of limited volume and low analyte abundance.

We evaluated Oslo-RM, a pooled plasma reference material from nearly 4,000 samples (ages ranging from 0 to 94 years), against NIST SRM 1950 and benchmarked DBS using conventional full-scan and Thermo Scientific Orbitrap Excedion mass spectrometer (MS) Full Scan enhanced dynamic range (eDR) acquisition. Plasma and DBS were analyzed by LC–HRMS with data-dependent MS/MS and matched against mzCloud and an in-house library.

For plasma, mzCloud matches increased with Full Scan eDR from 225 to 276 for NIST SRM 1950 and from 363 to 469 for Oslo-RM. The in-house library identified 93 metabolites in NIST SRM 1950 and 133 in Oslo-RM. In DBS, the Orbitrap Excedion MS detected about 5-fold more compounds than conventional Full Scan MS (6,340 vs 1,199), increased mzCloud-annotated compounds by 85% (222 vs 120), and revealed 13-fold more differentially abundant features than the prior pilot experiment.

Across matrices, eDR improved signal-to-noise (S/N) for low-abundance analytes. Oslo-RM showed more medication signals: 8 by full scan and 18 by eDR, versus gabapentin alone in NIST SRM 1950. Metabolites including bile acids, 3-methoxytyrosine, and 5-hydroxyindoleacetic acid also showed higher peak areas in Oslo-RM. In DBS, up to 5-fold S/N gains were observed for cystine, creatinine, and indole-3-acrylic acid.

These findings show that pooled reference materials and microsampling matrices, paired with Full Scan eDR and intelligent acquisition of the Orbitrap Excedion MS, can broaden metabolite and xenobiotic coverage and support deeper profiling and quality control in clinical research metabolomics.

Learning Objectives:

• Describe how a broadly representative pooled plasma reference material can complement NIST SRM 1950 for clinical research metabolomics quality control.
• Compare metabolite and medication detection efficiencies between conventional workflows and enhanced dynamic range (eDR) acquisition.
• Explain how enhanced dynamic range acquisition can improve detection of low-abundance analytes in complex plasma samples and low-volume microsampling devices like dried blood spots.