Achieving Maximum Protein and Peptide Recovery, Sensitivity, and Reproducibility using QuanRecovery Vials and Plates

Importance of the topic

High selectivity, sensitivity, and reproducibility are essential for quantitative LC-MS assays of proteins and peptides. Non-specific binding (NSB) on vial and plate surfaces can drastically reduce analyte recovery, compromise assay sensitivity, distort calibration linearity, and increase variability if not mitigated early in method development.

Objectives and study overview

This white paper evaluates QuanRecovery vials and plates with MaxPeak High Performance Surfaces (HPS) to suppress hydrophobic NSB without using carrier proteins. Peptide recoveries were compared across various container materials, sample matrices, storage conditions, and handling practices to identify strategies for maximizing sensitivity and reproducibility.

Methodology and instrumentation

Standard peptides (50 pg/mL–100 ng/mL) were prepared in water/acetonitrile matrices with varied organic content, pH, and additives. Samples were stored in polypropylene vials/plates, glass vials (silanized and TruView), commercial low-bind plates, and QuanRecovery products under different temperatures, durations, and sealing options. Recoveries were determined by UPLC–MS/MS using a CORTECS UPLC C18+ column (2.1×50 mm) on an ACQUITY UPLC I-Class PLUS system with water/ACN gradient (0.1% formic acid) and detection by Xevo TQ-S in SRM mode. Reference solutions included 0.1% rat plasma as a recovery standard.

Instrumentation used

• ACQUITY UPLC I-Class PLUS System
• CORTECS UPLC C18+ Column (2.1×50 mm)
• Xevo TQ-S Tandem Quadrupole Mass Spectrometer

Main results and discussion

• Glass and unmodified polypropylene containers showed poor recovery (<10%) for hydrophobic peptides (glucagon, insulin, melittin).
• QuanRecovery vials/plates and a commercial low-bind plate greatly improved recovery; QuanRecovery performed best for highly hydrophobic analytes.
• Higher organic content in the sample matrix improved recovery; QuanRecovery enabled full recovery at ≥10% ACN versus ≥25%–30% for other containers.
• Varying acid or base additives and pH (1.3–11.5) affected recovery in standard plates but not in QuanRecovery products, which maintained ≥90% recovery.
• QuanRecovery surfaces resisted harsh cleaning (1 M HNO₃ or 1 M NaOH) without loss of performance.
• Standard plates exhibited concentration-dependent losses that distorted calibration; QuanRecovery produced linear calibration curves from 50 pg/mL to 100 ng/mL.
• Recovery stability over time (up to 51 h), temperature (4 °C–25 °C), sample volume (200–500 µL), and various sealing options was superior with QuanRecovery.
• Residual volumes were minimal (5–8 µL) in QuanRecovery vials/plates versus >50 µL in other low-bind plates, optimizing sample utilization.

Benefits and practical applications

• Up to 50× increase in peak area and CV reduction from 41.8% to 1.7% for challenging peptides.
• Enhanced reproducibility and reliable quantitation without carrier proteins.
• Reduced method redevelopment and resource waste.
• Compatible with diverse pre- and post-analytical workflows in proteomics, peptidomics, QA/QC, and biomarker assays.

Future trends and opportunities

As LC-MS assays evolve toward lower detection limits and more complex matrices, engineered vial and plate surfaces will become increasingly important. Future advances may include tailored polymer chemistries for specific analyte classes, integration with automated platforms to eliminate manual contamination, and predictive modeling of NSB using machine learning. Novel sample formats such as microfluidic cartridges and dried matrix spots will benefit from optimized container–analyte interfaces to sustain assay robustness.

Conclusion

QuanRecovery vials and plates with MaxPeak HPS effectively mitigate hydrophobic NSB, delivering consistent high recovery, linear quantitation, minimal residual volume, and resistance to extreme pH and temperature variations. Implementing these products in LC-MS workflows enhances sensitivity, reproducibility, and efficiency in demanding proteomic and peptidomic analyses.

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