LC-MS/MS based quantitation of ciprofloxacin and its application to antimicrobial resistance study in Balb/c mouse plasma, urine, bladder and kidneys

• Published in: Analytical methods 2018-01, Vol.1 (1), p.1237-1246
• Main Authors: Matta, Murali K, Chockalingam, Ashok, Gandhi, Adarsh, Stewart, Sharron, Xu, Lin, Shea, Katherine, Patel, Vikram, Rouse, Rodney
• Format: Article
• Language: English
• Subjects: Acceptance criteria; Acetonitrile; Ammonium; Antiinfectives and antibacterials; Antimicrobial agents; Antimicrobial resistance; Bladder; Calibration; Ciprofloxacin; Drug resistance; Elution; Flow rates; Flow velocity; Formic acid; Kidneys; Liquid chromatography; LITAF protein; Matrices (mathematics); Mice; Quantitation; Spectrometry; Urinary bladder; Urine
• ISSN: 1759-9660; 1759-9679
• DOI: 10.1039/c7ay02923c

The authors proposed a simple, high throughput liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of ciprofloxacin in small sample volumes of plasma, urine, bladder and kidneys of mice. d 8 -Ciprofloxacin was used as an internal standard (IS) which efficiently tracked the matrix effect in different matrices. The method employed 20 μL of plasma/30 μL of urine or tissue homogenate for sample processing using a simple protein precipitation technique. The processed samples were chromatographed on a C 18 column by using a mixture of 5 mM ammonium formate (0.1% formic acid)-acetonitrile as the mobile phase at a flow rate of 0.6 mL min −1 with a gradient elution for 3 min. The calibration curve obtained was linear over the concentration range of 100-5000 ng mL −1 with r 2 ≥ 0.99 in all matrices. Method validation was performed as per FDA guidelines and the results met the acceptance criteria. The multiple reaction-monitoring mode (MRM) was used for the quantification of ion transitions at m / z 332.1/230.8 and 340.1/296.1 for the analyte and the IS, respectively. The validated method was used for preclinical studies of antimicrobial resistance in mice. The authors proposed a simple, high throughput liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of ciprofloxacin in small sample volumes of plasma, urine, bladder and kidneys of mice.