Determination of lean meat powder in pork by UFLC-triple quadrupole mass spectrometry

Importance of the Topic

Analysis of β-agonist residues in livestock is critical for food safety and regulatory compliance. Illegal use of lean meat promoters such as clenbuterol, ractopamine and salbutamol can lead to harmful side effects in humans. Rapid, sensitive and reliable analytical methods are essential to enforce maximum residue limits and protect public health.

Study Objectives and Overview

This study aims to develop and validate an ultra-fast liquid chromatography tandem mass spectrometry method for the simultaneous quantitation of clenbuterol, ractopamine and salbutamol in pork. The approach leverages Shimadzu LC-30A UFLC coupled to LCMS-8030 triple quadrupole mass spectrometer to achieve low limits of quantitation, high precision and broad linear dynamic range suitable for import-export screening.

Methodology and Instrumentation

Sample preparation follows a modified SNT 1924-2007 protocol in which 5 g of pork homogenate is extracted and reduced to 1 mL final volume. Chromatographic separation is performed on a Shim-pack XR-ODSIII column (2.0 mm×50 mm, 1.6 µm) at 40 °C using a binary gradient of 0.1% formic acid in water and acetonitrile at 0.4 mL/min. Mass spectrometric detection uses positive electrospray ionization with MRM transitions optimized for each analyte and corresponding stable isotope internal standards. Calibration is established over 0.05–100 ng/mL with correlation coefficients above 0.999.

Instrumentation Used

• Shimadzu LC-30A Ultra Fast Liquid Chromatograph with DGU-20A5 degasser, SIL-30AC autosampler, CTO-30A column oven
• Shimadzu LCMS-8030 Triple Quadrupole Mass Spectrometer operated in ESI positive MRM mode
• LabSolutions ver. 5.41 chromatography workstation

Key Results and Discussion

• Linearity: All analytes exhibited linear responses from 0.05 to 100 ng/mL with correlation coefficients > 0.999.
• Precision: Repeatability tests (n=6) yielded retention time RSD ≤ 0.18% and peak area RSD ≤ 5.47% across concentration levels.
• Sensitivity: Limits of quantitation for ractopamine and salbutamol met 0.5 µg/kg requirement; clenbuterol LOQ achieved 0.05 µg/kg satisfying Japanese MRL standards.

Benefits and Practical Applications

The proposed method delivers rapid analysis (< 2 min per injection) with excellent sensitivity and precision. It supports high-throughput screening of veterinary drug residues in imported and exported meat and ensures compliance with international regulations. Stable isotope internal standards enhance quantitative accuracy and matrix robustness.

Future Trends and Opportunities

• Expansion to multi-residue panels covering additional β-agonists and other veterinary drugs.
• Integration with automated sample preparation to increase throughput and reduce labor.
• Adoption of high-resolution mass spectrometry for compound confirmation and non-target screening.
• Development of portable LC-MS platforms for field testing at points of entry.

Conclusion

A validated UFLC-MS/MS method using Shimadzu LC-30A and LCMS-8030 enables rapid, precise and sensitive determination of clenbuterol, ractopamine and salbutamol in pork. The method meets or exceeds regulatory LOQ requirements and is suited for routine monitoring of lean meat powder residues in food trade.

Reference

• SNT 1924-2007 Determination of Clenbuterol Ractopamine Salbutamol and Terbutaline residues in Animal Derived Food for Import and Export HPLC-MS/MS Method
• Yao Jingting Shanghai Analysis Center Shimadzu Application Data SSL-CA09-277