Development of an HPLC-PDA Method for the Simultaneous Estimation of Three Antibiotics in Pharmaceutical Formulations and Bovine Milk and Health Risk Assessment

Mohammad Nasir Uddin ¹, Asif Hasnaine ¹, Md Sajedul Haque ¹

⁰Department of Chemistry, University of Chittagong, Chattogram 4331, Bangladesh.

Journal of Food Protection +

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April 17, 2024

View abstract on PubMed

Summary

A new HPLC-PDA method accurately detects tetracycline (TC), oxytetracycline (OTC), and ciprofloxacin (CIP) in pharmaceuticals and milk. Many milk samples exceeded maximum residue levels (MRLs), indicating potential contamination risks.

Area Of Science

Analytical Chemistry
Pharmacology
Food Safety

Background

Antibiotic residues in food products pose significant public health risks.
Accurate and sensitive detection methods are crucial for monitoring these contaminants.
Tetracycline (TC), oxytetracycline (OTC), and ciprofloxacin (CIP) are widely used antibiotics with potential for residue accumulation.

Purpose Of The Study

To develop and validate a simple, rapid, sensitive, and cost-effective HPLC-PDA method for simultaneous determination of TC, OTC, and CIP.
To apply the validated method for quantifying these antibiotics in pharmaceutical formulations and local market milk samples.
To assess the prevalence of TC, OTC, and CIP residues in milk and compare them against established maximum residue levels (MRLs).

Main Methods

High-Performance Liquid Chromatography with Photodiode Array (HPLC-PDA) detection was employed.
Separations were performed on a Shim-pack GIS C18 column using a gradient elution of oxalic acid, acetonitrile, and methanol.
Method validation was conducted according to International Council for Harmonisation (ICH) guidelines, assessing linearity, accuracy, precision, sensitivity, and robustness.

Main Results

The developed HPLC-PDA method demonstrated excellent linearity (r > 0.999) and reproducibility (RSD < 3.22%) for TC, OTC, and CIP.
Simultaneous elution of the three drugs was achieved within 7.39 minutes.
Analysis of milk samples revealed that TC, OTC, and CIP were detected above MRLs in over 50% of the samples analyzed.

Conclusions

The validated HPLC-PDA method is suitable for the routine analysis of TC, OTC, and CIP in pharmaceutical products and milk.
High prevalence of antibiotic residues exceeding MRLs in local milk samples highlights a potential food safety concern in Bangladesh.
Further investigation into the sources of contamination and their toxicological impact is warranted.

Keywords:

Ciprofloxacin HPLC–PDA Oxytetracycline Pharmaceutical and bovine milk Tetracycline