Abstract
In this study, we developed and validated two reliable high‐performance liquid chromatography (HPLC) methods for the qualitative detection of six oral β‐lactams, which are commonly used in pediatric patients with acute respiratory infections (ARIs). Two distinct reverse‐phase chromatographic separations of six β‐lactams were obtained. Four β‐lactams (cefadroxil, cephalexin, cefaclor and cefixime) in urine were separated using a gradient program with a mobile phase consisting of K2HPO4 buffer (20 mm, pH 2.8) and acetonitrile on a LichroCART 250 × 4.6 mm, Purospher STAR C18 end‐capped (5 μm) column. Two remained β‐lactams (amoxicillin and cefuroxime) were analyzed using a gradient elution with the mobile phase containing K2HPO4 buffer (20 mm, pH 3.0) and acetonitrile on a LichroCart® Purospher Star C8 end‐capped column (5 μm, 125 × 4.6 mm). Good linearity within the range of 0.3–30 μg/ml for cefadroxil, cephalexin, cefaclor and cefixime, and 0.2–20 μg/ml for amoxicillin and cefuroxime, was attained. The precisions were <14%. The accuracies ranged from 85.87 to 102.8%. The two validated methods were then applied to determine these six antibiotics in 553 urine samples of pediatric patients with ARIs. As a result, 32.2% were positive with one or more of six tested β‐lactams. Cefixime was the most commonly detected agent, accounting for 9.8% of enrolled patients.
Highlights
The two distinct validated methods were applied on the samples collected at hospital admission from pediatric patients with acute respiratory infections (ARIs) who attended to the outpatient clinic of Children's Hospital 1, Ho Chi Minh City (CH1) to investigate the early use of these antibiotics in the community setting
Antibiotic extraction recoveries were measured at three different levels and determined by comparing the peak areas of the analytes spiked in urine after the extraction process with the peak area of an unprocessed standard solution prepared with identical concentrations
Two high‐performance liquid chromatography (HPLC)‐diode array detector (DAD) methods were developed and validated to qualitatively determine six commonly used oral β‐lactams (CFD, CPL, CFO, CFI, AMO and CFU) in 553 urine samples obtained from pediatric patients with ARI
Summary
The two distinct validated methods were applied on the samples collected at hospital admission from pediatric patients with ARIs who attended to the outpatient clinic of Children's Hospital 1, Ho Chi Minh City (CH1) to investigate the early use of these antibiotics in the community setting. Urine samples for CC determination were prepared by diluting (1:10, v/v) the respective working solutions with diluted blank urine (1:5, v/v with pure water) to give eight CC points at 0, 0.3, 0.6, 2.0, 6.0, 12.0, 20.0 and 30.0 μg/ml for CFD, CPL, CFO and CFI, and at 0,. Twenty microliters of each eluate was injected into an equilibrated HPLC system
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