title: Multiple response optimization of a liquid chromatographic method for determination of fluoroquinolone and nitroimidazole antimicrobials in serum and urine.
abstract: OBJECTIVE
  Development and validation of a sensitive, selective and robust SPE assisted HPLC method for the quantification of fluoroquinolones and nitroimidazoles in human serum and urine using design of experiments methodology.
  
  
  METHODS
  Design of experiments was employed for method optimization (Box-Behnken design) and robustness testing (Plackett-Burman design). Sample preparation involved a simple solid phase extraction, which offered a satisfactory recovery (≥94%). Analytes were separated on a phenyl hexyl column with mobile phase comprising water, acetonitrile and triethyl amine in ratio of 74:26:0.15 v/v, with a flow rate of 1.1mL/min.
  
  
  RESULTS
  Calibration curves were linear over selected range (≥0.995) for all the analytes. The method was sensitive with detection limits of 0.06-0.16μg/mL in serum and urine samples. Inter and intra-day precision data (in terms of %RSD) was found to be less than 7%. Stability studies were carried out to assess freeze thaw, short term and long term stability and all analytes were found to be stable. The method was successfully applied for determination of antimicrobial drugs in spiked serum and urine.
  
  
  CONCLUSIONS
  The obtained results corroborated the potential of the proposed method for determination of all the four antimicrobial drugs in therapeutic drug monitoring, bioequivalence and drug-drug interaction studies.
authors.affiliation: Department of Pharmacy, Faculty of Engineering and Technology, Annamalai University, Annamalai Nagar, Tamil Nadu 608002, India. Electronic address: reddysince1989@gmail.com.
authors.forename: Sai Sandeep
authors.initials: SS
authors.lastname: Mannemala
authors.affiliation: Department of Pharmacy, Faculty of Engineering and Technology, Annamalai University, Annamalai Nagar, Tamil Nadu 608002, India. Electronic address: vallikonnect@gmail.com.
authors.forename: Valliappan
authors.initials: V
authors.lastname: Kannappan
mesh_terms: D000890:Anti-Infective Agents / Q000097:blood* / Q000652:urine*; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D016903:Drug Monitoring / Q000592:standards*; D024841:Fluoroquinolones / Q000032:analysis*; D064368:Healthy Volunteers; D006801:Humans; D009593:Nitroimidazoles / Q000032:analysis*; D012015:Reference Standards; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*
chemical_list: D000890:Anti-Infective Agents; D024841:Fluoroquinolones; D009593:Nitroimidazoles
keywords: Antimicrobial bioanalysis; Design of experiments; Fluoroquinolones; High performance liquid chromatography; Nitroimidazoles