Simple high-performance liquid chromatography determination of ampicillin in human serum using solid-phase extraction...

Area of Science:

• Analytical Chemistry
• Pharmacology

Background:

• Accurate quantification of antibiotics in biological matrices is crucial for effective patient treatment.
• Therapeutic drug monitoring (TDM) ensures optimal drug efficacy and minimizes toxicity.
• Lenampicillin hydrochloride is an oral prodrug of ampicillin.

Purpose of the Study:

• To develop and validate a simple, reproducible method for ampicillin analysis in human serum.
• To establish a reliable assay for therapeutic drug monitoring of ampicillin.

Main Methods:

• Serum samples were extracted using solid-phase extraction (SPE) with styrene divinyl/benzene sorbent cartridges.
• Ampicillin was separated and quantified using reversed-phase C18 high-performance liquid chromatography (HPLC) with UV detection at 220 nm.
• A mobile phase of acetonitrile-10 mM NaH2PO4 (6.5:93.5, v/v) was employed.

Main Results:

• The SPE method achieved a high recovery rate of 98.4+/-5.6% from human serum.
• The quantitation limit was determined to be 0.19 microg/ml using 0.5 ml of serum.
• Linear calibration curves (0.19–9.41 microg/ml) with excellent correlation coefficients (r=0.999) were obtained.

Conclusions:

• The developed HPLC method is simple, reproducible, and accurate for ampicillin analysis in human serum.
• This assay is suitable for therapeutic drug monitoring of ampicillin, particularly after oral administration of lenampicillin hydrochloride.
• The method provides reliable quantification for optimizing antibiotic dosage regimens.