Premium
Development of a validated HPLC method for the determination of four penicillin antibiotics in pharmaceuticals and human biological fluids
Author(s) -
Samanidou Victoria F.,
Evaggelopoulou Evaggelia N.,
Papadoyannis Ioannis N.
Publication year - 2006
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200600081
Subject(s) - chromatography , chemistry , dicloxacillin , repeatability , cloxacillin , detection limit , penicillin , high performance liquid chromatography , solid phase extraction , quantitative analysis (chemistry) , antibiotics , biochemistry
A quantitative method for the determination of four penicillin antibiotics, amoxicillin (AMO), oxacillin (OXA), cloxacillin (CLO), and dicloxacillin (DICLO), has been developed. Separation was achieved on an Inertsil ODS‐3 (250×4 mm, 5 μm) column after selective extraction of penicillin drugs from biological matrices by means of SPE. Gradient elution with a mobile phase consisting of 0.1% TFA (pH 1) and ACN, and PDA detection with monitoring at 240 nm was applied. Salicylic acid (5 ng/μL) was used as the internal standard. RP‐8 Adsorbex Merck cartridges provided high absolute recoveries (98–101%). The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability, and sensitivity. Repeatability ( n = 8) and between‐day precision ( n = 8) revealed RSD <10%. Recoveries from biological samples ranged from 91 to 103%. The detection limits were estimated as 3.3 ng for AMO, OXA, and CLO, and 6.6 for DICLO in blood plasma. LOD in whole blood and urine was 6.6 ng. Injection volume was 20 μL. The method was applied to commercially available AMO containing pharmaceuticals and spiked biological matrices. The method was also applied to biological samples after AMO oral administration, where the drug was successfully identified and quantified.