PDF herunterladen
Arzneimittelforschung 2008; 58(2): 91-96
DOI: 10.1055/s-0031-1296474
Antibiotics · Antimycotics · Antiviral Drugs · Chemotherapeutics · Cytostatics
Editio Cantor Verlag Aulendorf (Germany)

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

Talita Mota Gonçalves
1   NUDFAC–Núcleo de Desenvolvimento Farmacêutico e Cosmético – NUDFAC-UFPE, Centro de Biodisponibilidade / Bioequivalência, Departamento de Ciências Farmacêuticas, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
,
Danilo César Galindo Bedor
1   NUDFAC–Núcleo de Desenvolvimento Farmacêutico e Cosmético – NUDFAC-UFPE, Centro de Biodisponibilidade / Bioequivalência, Departamento de Ciências Farmacêuticas, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
,
Luís Renato Pires de Abreu
1   NUDFAC–Núcleo de Desenvolvimento Farmacêutico e Cosmético – NUDFAC-UFPE, Centro de Biodisponibilidade / Bioequivalência, Departamento de Ciências Farmacêuticas, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
,
Carlos Eduardo Miranda de Sousa
1   NUDFAC–Núcleo de Desenvolvimento Farmacêutico e Cosmético – NUDFAC-UFPE, Centro de Biodisponibilidade / Bioequivalência, Departamento de Ciências Farmacêuticas, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
,
Clarice M Bueno Rolim
2   CEBIFAR–Centro de Estudos de Bioequivalência e Farmacocinética, Departamento de Farmácia Industrial, Universidade Federal de Santa Maria, Rio Grande do Sul, Brazil
,
Davi Pereira de Santana
1   NUDFAC–Núcleo de Desenvolvimento Farmacêutico e Cosmético – NUDFAC-UFPE, Centro de Biodisponibilidade / Bioequivalência, Departamento de Ciências Farmacêuticas, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Publikationsdatum:
15. Dezember 2011 (online)

    Lizenzen und Reprints

Abstract

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 µg/ml for a plasma sample of 250 µl 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.

Key words

Ampicillin, bioequivalence, tandem mass spectrometry - Antibiotics - CAS 69-53-4
 

  • References

  • 1 Budavari S. The Merck Index. An Encyclopedia of Chemicals, Drugs and Biologicals. 12th ed New Jersey: Susan Buavari; 1996
    Google Scholar
  • 2 Belal F, El-Kerdawy MM, El-Ashry SM, El-Wasseef DR. Kinetic spectrophotometric determination of ampicillin and amoxicillin in dosage forms. Farmaco. 2000; 55: 680-686
    CrossrefPubMedGoogle Scholar
  • 3 Straub RF, Voyksner RD. Determination of penicillin G, ampicillin, amoxicillin, cloxacillin and cephapirin by high-performance liquid chromatography-electrospray mass spectrometry. J Chromatogr. 1993; 647: 167-181
    CrossrefPubMedGoogle Scholar
  • 4 Gilman AG, Goodman LS, Rall TW, Murad F. As Bases Farmacológicas da Terapêutica. 10th ed Rio de Janeiro: Mc Graw-Hill; 2003
    Google Scholar
  • 5 Labaune JP, Barre J, Flouvat B, Hermann Ph, Houin G, Trovin JH (1991) Propriétés pharmacocinétiques des medicaments. Paris: Masson; 1991
    Google Scholar
  • 6 Erah PO, Barrett DA, Shaw PN. Reversed-phase high-performance liquid chromatographic assay methods for the analysis of a range of penicillins in in vitro permeation studies. J Chromatogr B. 1998; 705: 63-69
    PubMedGoogle Scholar
  • 7 Luo W, Ang CYW, Thompson HCJr. Rapid method for the determination of ampicillin residues in animal muscle tissues by high-performance liquid chromatography with fluorescence detection. J Chromatogr B. 1997; 694: 401-407
    PubMedGoogle Scholar
  • 8 Lee H, Lee JS, Lee HS. Simultaneous determination of ampicillin and metampicillin in biological fluids using high-performance liquid chromatography with column switching. J Chromatogr B. 1995; 664: 335-340
    PubMedGoogle Scholar
  • 9 Qureshi SZ, Qayoom T, Helalet MI. Simultaneous spectrometric and volumetric determinatiom of amoxicillin, ampicillin and cloxacillin in drung formulations: reaction mechanism in the base catalysed hydrolysis followed by oxidation with iodate in dilute acid solution. J Pharm Biomed. 1999; 21: 473-482
    CrossrefPubMedGoogle Scholar
  • 10 Xu L, Wang H, Xiao Y. Spectrophotometric determination of ampicillin sodium in pharmaceutical products using sodium 1,2-nafphthoquinone-4-sulfonic as the chromogentic reagent. Spectrochim. Acta A. 2004; 60: 3007-3012
    CrossrefPubMedGoogle Scholar
  • 11 Mahgoub H, Aly FA. Spectrophotometric determination of ampicillin sodium and sulbactam sodium in two-component mixtures. J Pharm Biomed. 1998; 17: 1273-1278
    CrossrefPubMedGoogle Scholar
  • 12 Pajchel G, Pawlowski K, Tyski S. Simultaneous determination of amoxicillin / clavulanic acid and ampicillin/salbactam in pharmaceutical formulations for injections. J Pharm Biomed. 2002; 29: 75-81
    CrossrefPubMedGoogle Scholar
  • 13 Carceles CM, Espuny A, Vicente MS, Diaz MS, Escudero E. Single-dose pharmacokinetics of ampicillin/sulbactam (2:1) combination after intravenous administration to sheep and goats. Res Vet. 1996; 61: 143-146
    PubMedGoogle Scholar
  • 14 Sorensen LK, Snor LK, Elkaer T, Hansen H. Simultaneous determination of seven penicillins in muscle, liver and kidney tissues from cattle and pigs by a multiresidue high-performance liquid chromatography method. J Chromatogr B. 1999; 734: 307-318
    PubMedGoogle Scholar
  • 15 Verdon E, Fuselier R, Hurtaud-Pessel D, Couedor P, Cadieu N, Laurentie M. Stability of penicillin antibiotic residues in meat during storage: Ampicillin. J Chromatogr A. 2000; 882: 135-143
    CrossrefPubMedGoogle Scholar
  • 16 Haginaka J, Wakai J, Yasuda H, Uno T, Takahashi V, Katagi T. High-performance liquid chromatography determination of ampicillin and its metabolites in rat plasma, bile and urine by post-column degradation with sodium hypochlorite. J Chromatogr A. 1987; 400: 101-111
    CrossrefPubMedGoogle Scholar
  • 17 Ishida M, Kobayashi K, Awata NN, Sakamoto F. Simple high-performance liquid chromatography determination of ampicillin in human serum using solid-phase extraction disk cartridges. J Chromatogr B. 1998; 727: 245-248
    PubMedGoogle Scholar
  • 18 Akhtar MJ, Khan S, Salan KMA. Determination of ampicillin in human plasma by high-performance liquid chromatography using ultraviolet detection. J Pharm Biomed Anal. 1993; 11: 375-378
    CrossrefPubMedGoogle Scholar
  • 19 Miyazaki K, Ohtani K, Sunada K, Arita T. Determination of ampicillin, amoxicillin, cephalexin, and cepharadine in plasma by high-performance liquid chromatography using fluorometric detection. J. Chromatogr B Biomed.Sci Appl. 1983; 276: 478-482
    PubMedGoogle Scholar
  • 20 Lanbéck-Vallén K, Carlquist J, Nordgren T. Determination of ampicillin in biological fluids by coupled-column liquid chromatography and post-column derivatization. J Chromatogr B Biomed. 1991; 567: 121-128
    PubMedGoogle Scholar
  • 21 Horimoto S, Mayumi T, Aoe K, Nishimura N, Sato T. Analysis of β-lactam antibiotics by high performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry using bromoform. J Pharm Biomed. 2002; 30: 1093-1102
    CrossrefPubMedGoogle Scholar
  • 22 Pozo OJ, Guerrero C, Sancho JV, Ibáñez M, Pitarch E, Hogendoorn E et al. Efficient approach for the reliable quantification and confirmation of antibiotics in water using online solid-phase extraction liquid chromatography/tandem mass spectrometry. J Chromatogr A. 2006; 1103: 83-93
    CrossrefPubMedGoogle Scholar
  • 23 Goto T, Ito Y, Yamada S, Matsumoto H, Oka H. High-throughput analysis of tetracycline and penicillin antibiotics in animal tissues using electrospray tandem mass spectrometry with selected reaction monitoring transition. J Chromatogr A. 2005; 1100: 193-199
    CrossrefPubMedGoogle Scholar
  • 24 Becker MM, Zittlau E, Petz M. Residue analysis of 15 penicillins and cephalosporins in bovine muscle, kidney and milk by liquid chromatography-tandem mass spectrometry. Anal. Chim Acta. 2004; 520: 19-32
    CrossrefPubMedGoogle Scholar
  • 25 Lagerwerf FM, van Dongen WD, Steenvoordem RJJM, Honing M, Jonkman JHG. Exploring the boundaries of bioanalytical quantitative LC–MS–MS. Trends Anal Chem. 2000; 19: 418-427
    CrossrefPubMedGoogle Scholar
  • 26 Schellen A, Ooms B, Van de Lagemaat D, Vreeken R, Van Dongen WD. Generic solid phase extraction-liquid chromatography– tandem mass spectrometry method for fast determination of drugs in biological fluids. J Chromatogr B. 2003; 788: 251-259
    CrossrefPubMedGoogle Scholar