Agilent ASMS 2020 Posters Book

Significance of Topic

Modern analytical challenges in life sciences, environmental testing, food safety, and biopharmaceutical quality require sensitive, robust, and high-throughput mass-spectrometry workflows. Trace quantitation of metals in semiconductor process matrices, multiclass pesticide residues in diverse foods, low-volume plasma metabolomics, peptide allergens in chocolate, food authenticity screening, drug screening in biofluids, sialic-acid profiling in biotherapeutics, vitamin analysis in vaping oils, and native MS of protein complexes each demand tailored sample preparation, chromatography, and MS detection strategies to ensure reliable results.

Objectives and Study Overview

• Develop and validate advanced ICP-MS/MS methods with improved lens and reaction-gas technology (M-lens, NH₃) for ppt-level trace-metal analysis in high-silicon matrices.
• Implement triple-quadrupole LC/MS workflows for targeted quantitation of seven allergenic peptides in dark chocolate, achieving regulatory sensitivity and precision.
• Establish high-resolution GC/Q-TOF classification models for yerba-mate authenticity, combining untargeted feature finding with advanced chemometric software.
• Adapt a low-volume automated plasma sample-prep protocol on a 96-well robotic platform for metabolomics, demonstrating high recovery and reproducibility.
• Integrate LC/MS assays for simultaneous analysis of vitamin E and vitamin E acetate in vaping oils alongside cannabinoids.
• Demonstrate high-flow LC/MS/MS native-mode analysis of proteins and protein assemblies using robust ion sources on Q-TOF instruments.
• Streamline supported-liquid-extraction (SLE) cleanup and triple-quadrupole MS detection for SAMHSA-compliant oral-fluid drug panels.
• Quantify amino acids in spent food-processing media using zwitterionic HILIC columns on LC/TQ platforms.
• Develop high-throughput sialic-acid release and DMB-labeling protocols in 96-well format for UHPLC-FLD/HR-MS quantitation of Neu5Ac and Neu5Gc.
• Deploy enhanced triple-quadrupole LC/MS for 244-pesticide multiresidue screening in heavy food matrices (black tea, oranges).
• Optimize LC/MS/MS parameters and workflow for chickpea powder pesticide analysis, minimizing ion-suppression and maximizing recovery.
• Build DIA LC/Q-TOF drug-screening workflows in whole blood with automated SPE cleanup and novel software tools for rapid, rule-based identification.

Methodology and Instrumentation

• Sample Preparation: Tailored extraction/Cleanup—QuEChERS & EMR-Lipid SPE, Captiva EMR plates, SLE on Agilent Bravo, protein denaturation/digestion on robotic platforms, DMB labeling on thermocyclers.
• Chromatography: UHPLC with superficially porous columns (Poroshell, InfinityLab Poroshell EC-C18, AdvanceBio HILIC-Z), rapid GC for volatiles, fast SEC for native proteins.
• Detection: Agilent 8900/8800 ICP-MS/MS with M-lens; 6470B/6495 LC/TQ with Jet Stream ESI; 6545XT/7250 GC/Q-TOF; 6546/6560 LC/Q-TOF for DIA and native MS; FLD for DMB-sialic acids.
• Software: MassHunter Acquisition & Quantitative Analysis 10.x, MassHunter Qualitative Analysis, BioConfirm, Mass Profiler Professional, Classifier, UniDec for deconvolution, OpenLab CDS, novel LC Screener tool.

Main Results and Discussion

• ICP-MS/MS with NH₃ mass-shift removed Si interferences, achieving sub-ppt detection of 38 elements in 10 and 100 ppm Si matrices with robust stability (RSD <6%).
• Triple-quadrupole LC/TQ quantified seven allergen peptides in dark chocolate at LOQs down to 2.5 mg/kg, meeting VITAL 3.0 and AOAC SMPR criteria with R²>0.99 and recoveries of 75–102%.
• High-res GC/Q-TOF plus chemometric modeling (PLS-DA, SIMCA) classified yerba-mate brands and adulteration levels, identifying flavor markers and PAH contaminants.
• Automated low-volume (25 µL) plasma extraction on Bravo achieved >80% median recovery across 32 metabolites, RSD ~5%, facilitating ppt-level metabolomics.
• Rapid LC/MSD iQ workflow resolved vitamin E and vitamin E acetate in complex vaping oil cannabinoid matrices, with LOQs ~0.01 µg/mL and spectral matching library confidence >98%.
• Native MS of protein complexes (ADH, β-gal, NISTmAb) on LC/Q-TOF at 0.1 mL/min achieved preserved oligomer states, broad charge envelopes, and minimal denaturation with a single ion-source setting.
• SLE cleanup of oral fluid prior to 1290 LC/TQ analysis of SAMHSA drug panel (16 compounds) delivered S/N>10 at 0.4 ng/mL, R²>0.99 across 0.1–125 ng/mL curves and acceptable interday precision.
• ZIC-HILIC LC/TQ quantified 24 amino acids in spent media extraction with LOQs 5–20 µg/L, R²>0.99 and QC recoveries 75–102% (RSD <11%), enabling fermentation monitoring.
• DMB labeling 96-well workflow separated six sialic acids by RP-UHPLC/FLD in 10 min; calibration R²>0.999, LOD ~0.012 pmol, enabling Neu5Ac/Neu5Gc quantitation at 0.25 pmol on-column.
• 1290 LC/TQ multiresidue screening of 244 pesticides in tea and oranges achieved LLOQs <10 ng/g for >95% of analytes, R²>0.99, recovery 80–120% for ~50% of analytes, and acceptable precision.
• Chickpea powder pesticide method with QuEChERS cleanup plus 5× dilution on 6470 LC/TQ delivered LOQs 10 ng/kg, R²>0.995, and recoveries >80% for most analytes.
• DIA LC/Q-TOF drug screening in whole blood with EMR-Lipid and Bravo automation gave <10% matrix effects for 77% of 153 analytes, >70% recoveries, baseline separation of isobars in 10 min, and fast review by LC Screener tool.

Benefits and Practical Applications

• Enhanced uptime and robustness via vent-free ICP-MS, automated SPE cleanup, and low-fouling chromatographic/divert valves.
• High throughput: 10 min runs for multi-class assays, 6 min native MS, 96-well automated workflows reduce labor and variability.
• Improved data confidence through mass-shift reactions, high-resolution TOF library matching, rule-driven Screener review, and fragment coelution criteria.
• Regulatory compliance: Meets or exceeds sensitivity, precision, and recovery guidelines for semiconductor QA, allergen labeling, pesticide MRLs, drug-of-abuse thresholds, and biologic characterization.
• Flexible method updates: Q-TOF DIA and tMRM libraries allow rapid addition of new analytes without revalidation of chromatography or acquisition methods.

Future Trends and Possibilities

• Integration of automated sample prep with real-time data-review dashboards and AI-driven anomaly detection for lab-wide QC.
• Expansion of native MS workflows to intact viral vectors and high-mass assemblies on faster Q-TOF platforms with ion-mobility filtering.
• Coupling high-res GC/Q-TOF food authenticity screening with isotopic or elemental fingerprinting databases for fraud detection.
• Microfluidic and microscale sample prep for ultra-low volume metabolomics in clinical trials and single-cell studies on automated platforms.
• Advance of real-time LC/TQ quantitation with inline SPE and dynamic MRM updates in regulated environments.

Conclusions

A wide array of state-of-the-art LC/MS and GC/MS/MS technologies combined with robotic sample preparation, reaction-cell enhancements, high-res TOF acquisition, and advanced software tools now enable laboratories to meet stringent requirements across industries—achieving ppt to ppm sensitivity, robust performance in challenging matrices, and truly high-throughput workflows for routine and research applications.

References

• MassHunter Software Reference Manual, Agilent Technologies.
• Marty MT et al. Bayesian Deconvolution for MS and IM Data. Anal Chem. 2015.
• McCurdy E, et al. ICP-MS/MS in Reaction Mode. Spectroscopy. 2019.
• Kowalski D, Laine D. Rapid Cannabinoid LC-DAD Method. AOAC 2018.
• Van Aernum ZL. Rapid Online Buffer Exchange for Native MS. ChemRxiv 2019.
• AOAC SMPR 2016.002 for Food Allergen Analysis.
• Regulation (EC) 396/2005 on Pesticide MRLs. European Commission.
• Blount BC et al. Vitamin E Acetate in EVALI Cases. N Engl J Med. 2020.
• Kaneko Y et al. Anti-Inflammatory IgG Activity from FC Sialylation. Science. 2006.