title: A simple and rapid RP-HPLC method for the simultaneous determination of piperacillin and tazobactam in human plasma.
abstract: A rapid and sensitive reverse phase HPLC (RP-HPLC) method for the simultaneous quantitation of piperacillin and tazobactam in human plasma has been developed and validated. The method utilizes a novel, simple and rapid solid phase extraction step, which results in an improved extraction yield of analytes from human plasma, as well as significantly reduced interference from serum components at low UV wavelength detection compared to previously published liquid-liquid extraction methods. Chromatographic separation was carried out on a Hypersil ODS C18, 3μm column using an acetonitrile-trifluoroacetic acid-water gradient elution with dual wavelength quantitation at 254nm for piperacillin and 218nm for tazobactam. Linear relationships between peak area and drug concentration were obtained in the range of 1.0-200μg/mL for piperacillin and 0.78-50μg/mL for tazobactam, with r2=0.9997 and 0.9994 respectively. The assay proved to be sensitive (with a lower limit of quantitation of 1μg/mL for piperacillin and 0.78μg/mL for tazobactam), specific (no interference from plasma components at either 218nm or 254nm), and reproducible (both intra- and inter- day coefficients of variation were ≤6%). With a total process/assay time of less than 30min, the method provides a simple, precise and reproducible assay for monitoring piperacillin and tazobactam plasma levels that can be readily adapted for routine clinical use.
authors.affiliation: Department of Pharmacy Practice, Loma Linda University School of Pharmacy, United States.
authors.forename: John J
authors.initials: JJ
authors.lastname: Veillette
authors.affiliation: Department of Pharmacy Practice, Loma Linda University School of Pharmacy, United States.
authors.forename: S Alexander
authors.initials: SA
authors.lastname: Winans
authors.affiliation: Department of Pharmacy Practice, Loma Linda University School of Pharmacy, United States; Department of Pharmacy, Loma Linda University Medical Center, United States; Loma Linda University School of Medicine, United States.
authors.forename: Steven C
authors.initials: SC
authors.lastname: Forland
authors.affiliation: Department of Pharmaceutical Sciences, Loma Linda University School of Pharmacy, United States. Electronic address: vmaskiewicz@llu.edu.
authors.forename: Victoria K
authors.initials: VK
authors.lastname: Maskiewicz
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006801:Humans; D059625:Liquid-Liquid Extraction / Q000379:methods; D010397:Penicillanic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood; D010878:Piperacillin / Q000032:analysis / Q000097:blood*; D052616:Solid Phase Extraction / Q000379:methods; D000078142:Tazobactam; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D010397:Penicillanic Acid; D000078142:Tazobactam; D010878:Piperacillin
keywords: Hplc; Piperacillin; Solid phase extraction; Tazobactam; Validation