Analysis of Chloramphenicol and Metabolites in Royal Jelly and Honey Using SPE Coupled with LC/MS/MS

Význam tématu

The monitoring of antibiotic residues in bee products such as royal jelly and honey is critical for ensuring food safety, protecting consumer health, and complying with international regulatory standards. Chloramphenicol and its analogues pose a risk due to their potential toxicity and strict residue limits. A robust analytical approach enables reliable detection of trace-level contaminants, supports quality assurance in apiculture, and underpins regulatory enforcement.

Cíle a přehled studie

This study aimed to develop and validate a sensitive, reproducible, and high-throughput method for simultaneous quantitation of chloramphenicol, thiamphenicol, florfenicol, and the metabolite florfenicol amine in royal jelly and honey. The workflow combined enzymatic treatment, solid-phase extraction (SPE) using Agilent Bond Elut Plexa PCX cartridges, and tandem LC/MS/MS detection on an Agilent 6470 system.

Použitá metodika

Sample preparation involved:

• Weighing 2 g royal jelly or 5 g honey (dissolved in water).
• Adding trypsin digestion (0.25 g) and incubation at 40 °C for 2 h in darkness.
• Extracting with acetonitrile containing 2% ammonium hydroxide.
• Centrifugation and nitrogen drying of the organic phase.
• Reconstitution to a fixed volume and SPE cleanup on Plexa PCX cartridges (200 mg or 500 mg).
• Conditioning, loading, washing, elution with methanol/ammonium hydroxide, and final filtration.

Použitá instrumentace

The analytical platform comprised:

• Agilent 1260 Infinity II LC system with Poroshell 120 EC-C18 column (100 × 3.0 mm, 2.7 µm).
• Mobile phase A: water + 0.1% formic acid; B: acetonitrile; gradient from 10% to 100% B in 7 min.
• Agilent 6470 triple quadrupole MS with JetStream ESI source, operated in positive and negative ion modes.
• Agilent MassHunter software for acquisition and quantitation.

Hlavní výsledky a diskuse

Method performance was demonstrated by recoveries ranging from 72% to 117% across spiking levels (1–5 ng/g) and RSDs below 7%. Chloramphenicol and florfenicol showed the highest recoveries, while thiamphenicol and florfenicol amine exhibited slightly lower but acceptable values. The dual‐mode detection enhanced sensitivity and selectivity for all analytes.

Přínosy a praktické využití metody

This validated workflow offers:

• Low detection limits suitable for regulatory thresholds.
• High throughput sample handling via SPE automation.
• Reliable quantitation for quality control in food testing laboratories.

Budoucí trendy a možnosti využití

Future developments may include extension to other animal-derived matrices, integration with high-resolution mass spectrometry for non-target screening, and miniaturization of the SPE step for faster turnaround. Data‐driven automation could further improve reproducibility and lab productivity.

Závěr

The SPE‐LC/MS/MS method using Bond Elut Plexa PCX cartridges and an Agilent 6470 system delivers precise, accurate, and sensitive analysis of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in royal jelly and honey. This approach supports stringent monitoring of antibiotic residues in apicultural products.