Determination of ampicillin in human plasma by solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) and its use in bioequivalence studies.
A simple, fast, sensitive and selective solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) method for the quantitative analysis of ampicillin (CAS 69-53-4) in human plasma was developed using amoxicillin as internal standard, and sample extraction by solid-phase extraction (SPE). Extracts were separated by reversed-phase C18 with aqueous mobile phase (acetonitrile, 80:20, v/v) with 0.1% formic acid. The method was validated and successfully applied in a bioequivalence study of capsules 500 mg of ampicillin. Using a short running time of 2.5 min, the lower limit of quantification (LLOQ) for obtained ampicillin was 0.1 microg/ml for a plasma sample of 250 microl and a recovery of 94.38% +/- 4.05. Bioequivalence between the products was determined by calculating 90% confidence intervals (CI) for the ratio of Cmax, AUC0-t and AUC0-inf values for the test and reference products, which were within the 0.80-1.25 interval proposed by FDA and EMEA. It is concluded that the two formulations are bioequivalent in their rate and extent of absorption, and thus, may be used interchangeably.
Gonçalves
Talita Mota
TM
NUDFAC-Núcleo de Desenvolvimento Farmacêutico e Cosmético, NUDFAC-UFPE, Centro de Biodisponibilidade / Bioequivalência, Departamento de Ciências Farmacêuticas, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil.
Bedor
Danilo César Galindo
DC
de Abreu
Luís Renato Pires
LR
de Sousa
Carlos Eduardo Miranda
CE
Rolim
Clarice M Bueno
CM
de Santana
Davi Pereira
DP
D000293:Adolescent; D000328:Adult; D000658:Amoxicillin / Q000097:blood; D000667:Ampicillin / Q000097:blood* / Q000493:pharmacokinetics*; D000900:Anti-Bacterial Agents / Q000097:blood* / Q000493:pharmacokinetics*; D002138:Calibration; D002853:Chromatography, Liquid; D018592:Cross-Over Studies; D004311:Double-Blind Method; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D011786:Quality Control; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry; D013810:Therapeutic Equivalency
D000900:Anti-Bacterial Agents; D007202:Indicators and Reagents; D000667:Ampicillin; D000658:Amoxicillin