A sensitive assay of amoxicillin in mouse serum and broncho-alveolar lavage fluid by liquid-liquid extraction and reversed-phase HPLC.
A sensitive and simple high-performance liquid chromatography (HPLC) method was developed and validated for the analysis of amoxicillin in mouse serum and broncho-alveolar lavage (BAL) fluid. One hundred microlitres of sample were needed for the assay. Sample processing was carried out with liquid-liquid extraction. Cefadroxil was used as an internal standard. The chromatographic separation was achieved on a C18 reversed-phase column with a mobile phase consisting of phosphate buffer, 1-octanesulphonic acid sodium salt and acetonitrile. The detection was conducted at 210 nm. The ranges of the standard curves were 0.2-20 and 0.05-5 microg/ml for serum and BAL samples, respectively. The recoveries of amoxicillin from serum and normal saline were 87 and 88%, respectively. The coefficients of variation were 1.78-6.13% for intra-day and 0.82-6.42% for inter-day analyses. The accuracy was within 100+/-6%. This method was successfully applied to analyze amoxicillin in mouse serum and BAL samples from a pharmacokinetic study.
Du
Xiaoli
X
Center for Anti-Infective Research and Development, Hartford Hospital, 80 Seymour Street, Hartford, CT 06102, USA.
Li
Chonghua
C
Sun
Heather K
HK
Nightingale
Charles H
CH
Nicolau
David P
DP
D000658:Amoxicillin / Q000032:analysis* / Q000097:blood* / Q000493:pharmacokinetics; D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics*; D001992:Bronchoalveolar Lavage Fluid; D002138:Calibration; D002434:Cefadroxil / Q000032:analysis; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004345:Drug Industry / Q000379:methods; D051379:Mice; D008813:Mice, Inbred ICR; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D013997:Time Factors; D014466:Ultraviolet Rays
D000900:Anti-Bacterial Agents; D002434:Cefadroxil; D000658:Amoxicillin