Analysis of Ornithine in Bunashimeji(Beech mushroom)

Importance of the Topic

Ornithine is a non-protein amino acid involved in the urea cycle and has significance in food quality, nutritional assessment, and regulatory compliance. Accurate quantification in edible mushrooms supports safety and labeling standards, aligning with JAS requirements.

Objectives and Study Overview

The study aimed to develop and validate a robust hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC-LC-MS/MS) method for determining ornithine in Bunashimeji (beech mushroom). Key goals included achieving high sensitivity, selectivity, and reproducibility for routine food analysis.

Methodology and Instrumentation

A Shim-pack GIST Amide HILIC column (100 mm×2.1 mm I.D., 3.0 µm) was employed under a gradient elution of 0.3 % formic acid in water (A) and 0.3 % formic acid in acetonitrile (B). The gradient ran from 90 % B to 50 % B over 8 min, held, then returned to initial conditions. Flow rate was 0.30 mL/min and column temperature 40 °C.

The LC system (Nexera™) was coupled to an LCMS-8060NX mass spectrometer via electrospray ionization in positive mode. MRM transition for ornithine was m/z 133 → 70 with collision energy 22 V. Detailed gas flows and temperature settings ensured optimal ionization and signal stability.

Key Results and Discussion

The method delivered a sharp, well-resolved peak for ornithine, as shown by the MS chromatogram of the standard solution. Signal intensities exhibited excellent linearity across the tested range. Matrix effects were minimal, demonstrating reliable quantification in mushroom extracts without significant interference.

Precision and accuracy tests met acceptance criteria, indicating the method’s suitability for routine food analysis. The rapid 13-minute run time supports high sample throughput.

Benefits and Practical Applications

• High specificity and sensitivity for ornithine detection.
• Short analysis time enhances laboratory productivity.
• Minimal sample cleanup due to HILIC retention of polar analytes.
• Compliance with food safety and labeling standards.

Future Trends and Application Opportunities

Emerging directions include integrating isotopic internal standards for absolute quantification, expanding to multi-amino acid profiling, and coupling with high-throughput automation in food QA/QC. Adoption of microflow HILIC and improved MS detectors may further reduce solvent usage and increase sensitivity.

Conclusion

This HILIC-LC-MS/MS approach using Shim-pack GIST Amide column and LCMS-8060NX provides a fast, reliable, and sensitive solution for ornithine analysis in Bunashimeji mushrooms. The validated method meets regulatory standards and offers practical benefits for food quality control laboratories.

Instrument Used

Shim-pack GIST Amide column (100 mm×2.1 mm I.D., 3.0 µm) with Nexera™ HPLC and LCMS-8060NX mass spectrometer in positive ESI mode.

Reference

Shimadzu Application News No. 01-00175 (JP, ENG)