title: Simultaneous Determination of Eight β-Lactam Antibiotics, Amoxicillin, Cefazolin, Cefepime, Cefotaxime, Ceftazidime, Cloxacillin, Oxacillin, and Piperacillin, in Human Plasma by Using Ultra-High-Performance Liquid Chromatography with Ultraviolet Detection.
abstract: A simple and rapid ultra-high-performance liquid chromatography (UHPLC) method using UV detection was developed for the simultaneous determination of eight β-lactam antibiotics in human plasma, including four penicillins, amoxicillin (AMX), cloxacillin (CLX), oxacillin (OXA), and piperacillin (PIP), and four cephalosporins, cefazolin (CFZ), cefepime (FEP), cefotaxime (CTX), and ceftazidime (CAZ). One hundred-microliter samples were spiked with thiopental as an internal standard, and proteins were precipitated by acetonitrile containing 0.1% formic acid. Separation was achieved on a pentafluorophenyl (PFP) column with a mobile phase composed of phosphoric acid (10 mM) and acetonitrile in gradient elution mode at a flow rate of 500 μl/min. Detection was performed at 230 nm for AMX, CLX, OXA, and PIP and 260 nm for CFZ, FEP, CTX, and CAZ. The total analysis time did not exceed 13 min. The method was found to be linear at concentrations ranging from 2 to 100 mg/liter for each compound, and all validation parameters fulfilled international requirements. Between- and within-run accuracy errors ranged from -5.2% to 11.4%, and precision was lower than 14.2%. This simple method requires small-volume samples and can easily be implemented in most clinical laboratories to promote the therapeutic drug monitoring of β-lactam antibiotics. The simultaneous determination of several antibiotics considerably reduces the time to results for clinicians, which may improve treatment efficiency, especially in critically ill patients.
authors.affiliation: Laboratoire de Pharmacologie-Toxicologie, Assistance Publique des Hôpitaux de Paris, Hôpitaux Universitaires Henri Mondor, Créteil, France tiphaine.legrand@aphp.fr.
authors.forename: Tiphaine
authors.initials: T
authors.lastname: Legrand
authors.affiliation: Service d'Anesthésie et des Réanimations Chirurgicales, Assistance Publique des Hôpitaux de Paris, Hôpitaux Universitaires Henri Mondor, Créteil, France IMIV, CEA, INSERM, CNRS, Université Paris-Sud, Université Paris Saclay, CEA-SHFJ, Orsay, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Vodovar
authors.affiliation: IMIV, CEA, INSERM, CNRS, Université Paris-Sud, Université Paris Saclay, CEA-SHFJ, Orsay, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Tournier
authors.affiliation: Laboratoire de Pharmacologie-Toxicologie, Assistance Publique des Hôpitaux de Paris, Hôpitaux Universitaires Henri Mondor, Créteil, France.
authors.forename: Nihel
authors.initials: N
authors.lastname: Khoudour
authors.affiliation: Laboratoire de Pharmacologie-Toxicologie, Assistance Publique des Hôpitaux de Paris, Hôpitaux Universitaires Henri Mondor, Créteil, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Hulin
mesh_terms: D000658:Amoxicillin / Q000097:blood; D000900:Anti-Bacterial Agents / Q000097:blood*; D002437:Cefazolin / Q000097:blood; D000077723:Cefepime; D002439:Cefotaxime / Q000097:blood; D002442:Ceftazidime / Q000097:blood; D002511:Cephalosporins / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003023:Cloxacillin / Q000097:blood; D016903:Drug Monitoring / Q000379:methods; D006801:Humans; D010068:Oxacillin / Q000097:blood; D010878:Piperacillin / Q000097:blood; D014466:Ultraviolet Rays; D047090:beta-Lactams / Q000097:blood*
chemical_list: D000900:Anti-Bacterial Agents; D002511:Cephalosporins; D047090:beta-Lactams; D000658:Amoxicillin; D000077723:Cefepime; D002442:Ceftazidime; D002437:Cefazolin; D002439:Cefotaxime; D003023:Cloxacillin; D010068:Oxacillin; D010878:Piperacillin