An HPLC-Florescence Detection (FLD) method was developed and validated for the determination of sixteen polycyclic aromatic hydrocarbons (PAHs) in fish fillets. The method employs a quick, easy, cheap, effective, rugged and safe (QuEChERS) multiresidue sample preparation procedure adopted from the Association of Analytical Communities (AOAC) Official method 2007.01 for extraction and cleanup. The analytes were separated on an Agilent ZORBAX Eclipse PAH HPLC column (4.6 mm × 50 mm, 1.8 µm) by gradient elution with a binary system of acetonitrile - water and subse- quent fluorescence detection set at appropriate excitation and emission wavelengths. The analyte recoveries ranged from 83.4% to 101% with relative standard deviations ranging from 0.6 to 1.9% at three different fortification levels. The limits of detection and quantification ranged from 0.04 to 0.84 and 0.1 to 2.80 ng/g, respectively.
The document presents an analytical method developed and validated by researchers at Rhodes University for detecting polycyclic aromatic hydrocarbons (PAHs) in fish tissue. The study utilizes High-Performance Liquid Chromatography with Fluorescence Detection (HPLC-FLD). Sample preparation involves the Agilent Bond Elut QuEChERS AOAC Extraction kit and Dispersive SPE kit to extract and clean up the analytes from fish fillets.
The method was validated by analyzing spiked samples at three different fortification levels. Results showed that analyte recoveries ranged from 83.4% to 101%, with relative standard deviations between 0.6% and 1.9%. The limits of detection (LOD) and quantification (LOQ) for the sixteen target PAHs were determined, ranging from 0.04 ng/g to 0.84 ng/g and 0.1 ng/g to 2.80 ng/g, respectively.
| Flow rate | 0.8 mL/min |
|---|---|
| Column temperature | 18 °C |
| Injection volume | 1000 pL (1 mL) |
| Sample mass | 5.0 g |
| Centrifugation speed | 4000 rpm |
| Recovery range | 83.4% to 101% |
Sixteen polycyclic aromatic hydrocarbons (PAHs) including naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, 1,2-benza[a]anthracene, chrysene, benzo[e]pyrene, benzo[e]acenaphthylene, benzo[k]fluoranthene, dibenzo[a,h]anthracene, benzo[g,h,i]perylene and indeno[1,2,3-cd]pyrene.
The limit of detection for Fluoranthene is 0.04 ng/g.
The limit of quantification for Dibenzo[a,h]anthracene is 2.80 ng/g.