Analysis of Acetyl Progesterones in Pork, Eggs, and Milk Using the Agilent Captiva EMR—Lipid by LC/MS/MS

Significance of the Topic

Accurate detection of synthetic acetyl progesterone residues in pork, eggs, and milk is critical for ensuring food safety, meeting regulatory requirements, and protecting consumer health. These compounds, when illicitly used as growth promoters, pose potential risks and require sensitive analytical methods for reliable surveillance.

Objectives and Study Overview

The study aimed to develop and validate a streamlined workflow combining Agilent QuEChERS extraction with Captiva EMR–Lipid cleanup followed by LC/MS/MS to quantify four acetyl progesterones (flurogestone acetate, megestrol acetate, melengestrol acetate, and chlormadinone acetate) in various animal-derived food matrices.

Methodology and Used Instrumentation

• Sample Preparation: Homogenized samples were extracted with acetonitrile using the Agilent Bond Elut Vet Drug QuEChERS kit, followed by lipid removal on Captiva EMR–Lipid cartridges, nitrogen drying, and reconstitution in ACN/water.
• Liquid Chromatography: Agilent 1290 Infinity II system with Poroshell 120 SB-C18 column (3.0×100 mm, 2.7 µm), 0.4 mL/min flow, 40 °C, gradient elution from 60% to 100% organic mobile phase over 6 min.
• Mass Spectrometry: Agilent 6470 triple quadrupole with Jet Stream ESI in positive mode, gas temperature 250 °C, MRM transitions optimized for each analyte, data acquired using MassHunter software.

Main Results and Discussion

The method demonstrated satisfactory linearity and minimal matrix effects across pork, liver, kidney, egg, and milk. Recoveries ranged from approximately 85% to 115% with RSDs below 10% at spiking levels of 1–10 ng/g. Calibration curves showed strong correlation in all matrices, confirming method robustness and reproducibility.

Benefits and Practical Applications

This approach offers a rapid, sensitive, and reproducible solution for routine monitoring of acetyl progesterones in food. The simplified cleanup and fast UHPLC run enable high throughput analysis, making it suitable for quality control laboratories, regulatory agencies, and research settings.

Future Trends and Opportunities for Application

Future developments may focus on broadening the method to additional steroid hormones, integrating online sample preparation for automation, applying high-resolution mass spectrometry for confirmatory analysis, and exploring its use in complex environmental or biological matrices.

Conclusion

The validated method utilizing QuEChERS extraction and Captiva EMR–Lipid cleanup coupled with LC/MS/MS provides a reliable, efficient workflow for quantifying synthetic acetyl progesterones in meat, eggs, and milk, supporting food safety programs and regulatory compliance.