Determination of Hormones in Fish (Carassius Carassius) by SampliQ- OPT Solid Phase Extraction with High Performance Liquid Chromatography

Significance of the Topic

The consumption of fish contaminated with hormones may pose health risks. Monitoring residues of steroids such as glucocorticoids, estrogens, and androgens in food products is essential to ensure compliance with regulatory limits and protect public health.

Objectives and Study Overview

This application note describes the development and validation of a method integrating SampliQ OPT solid-phase extraction and HPLC-DAD for the extraction and quantitation of 16 hormones in crucian carp (Carassius carassius) meat. It aims to achieve high recoveries, low limits of detection, and good reproducibility.

Used Instrumentation

• Agilent SampliQ OPT SPE cartridges (60 mg, 3 mL)
• Agilent 1200 Series HPLC with diode array detector
• Agilent ZORBAX Eclipse Plus C18 Column (250 × 4.6 mm, 5 μm)
• 0.45 μm PTFE syringe filters

Methodology

The sample preparation involves homogenization of fish meat, methanol extraction, ultrasonic treatment, centrifugation, and SPE cleanup. Cartridges are conditioned with methanol and water, loaded with extract, washed with 30% methanol, and eluted with methanol. The eluate is evaporated, reconstituted, filtered, and injected into HPLC. Chromatographic separation is achieved with a water–acetonitrile gradient at 1.0 mL/min and detection at 230 nm.

Main Results and Discussion

Linearity was established over 1–100 mg/kg for all 16 hormones with correlation coefficients above 0.999. Limits of detection ranged from 0.2 to 1.0 mg/kg. Recovery studies at spiking levels of 2, 5, and 10 mg/kg yielded recoveries between 76.2% and 106.1% with RSDs below 9%. Minimal matrix interferences were observed, demonstrating the method’s specificity and robustness.

Benefits and Practical Applications

• Efficient simultaneous extraction of polar and non-polar hormones using a single SPE cartridge.
• High recoveries and low variability support reliable routine analysis.
• SPE–HPLC workflow is straightforward and suitable for food safety laboratories monitoring hormone residues in fish.

Future Trends and Opportunities

Advancements may include coupling SPE with mass spectrometry for enhanced sensitivity and selectivity, miniaturization of SPE formats for high-throughput screening, and automation to increase sample throughput. Expanding the method to other food matrices and emerging hormones will further support comprehensive residue monitoring.

Conclusion

The optimized SampliQ OPT SPE–HPLC method provides a rapid, reproducible, and sensitive approach for determining multiple hormone residues in crucian carp. Its strong performance in terms of recovery, detection limits, and minimal matrix effects makes it an effective tool for ensuring food safety compliance.