Institutional members access full text with Ovid®

Share this article on:

High Performance Liquid Chromatography–Mass Spectrometry Assay for Polymyxin B1 and B2 in Human Plasma

Thomas, Tiffany A. PhD*; Broun, Emily C. BS; Abildskov, Kirsten M. MS*; Kubin, Christine J. PharmD; Horan, Jennifer MD, PharmD§; Yin, Michael T. MD, MS; Cremers, Serge PharmD, PhD*,¶

doi: 10.1097/FTD.0b013e31825c827a
Original Article

Background: Polymyxin B is an old antibiotic with increasing clinical relevance in the treatment of multidrug-resistant Gram-negative bacterial infections. However, current dosing regimens are largely empiric as clinical pharmacological characterization of the drug has been hindered by the lack of assays to measure polymyxin B in human plasma.

Methods: A high-performance liquid chromatography–mass spectrometry assay was developed to quantify polymyxin B1 and B2 in human plasma using pure calibrators. After purification with a solid-phase extraction column, polymyxin B1 and B2 were separated on a C18 column by gradient chromatography with an overall runtime of 12 minutes. Polymyxin B1 and B2 were ionized by positive electrospray ionization, and the resulting ions specific to polymyxin B1 and B2 were monitored (selected ion recording).

Results: The dominant ions produced were (M + 2H)2+ at m/z 602.6 and 595.5 for polymyxin B1 and polymyxin B2, respectively. The assay was linear between concentrations of 100 and 2500 ng/mL, with interday precision of 5.9% and 3.4% at 100 ng/mL and 5.3% and 4.0% at 2000 ng/mL for polymyxin B1 and polymyxin B2, respectively. Accuracy was 80.2% and 82.2% at 100 ng/mL and 99.9% and 109.6% at 2000 ng/mL for polymyxin B1 and polymyxin B2, respectively. No interference from other drugs commonly administered with polymyxin B was detected. The performance of the assay is affected by gross hemolysis and hyperlipemia. The method was successfully applied to patient samples. Interestingly, in a single patient the ratio of B1 and B2 did not change over a period of 12 hours after administration of the drug.

Conclusions: A simple method for the simultaneous measurement of polymyxin B1 and polymyxin B2 in human plasma is described, which has the potential to optimize clinical use of this valuable antibiotic by permitting pharmacokinetic studies and therapeutic drug monitoring.

*Irving Inst for Clin and Transl Research

Division of Infectious Diseases, Department of Medicine, Columbia University Medical Center, New York, New York

Clinical Pharmacy, Columbia-Presbyterian Hospital, New York, New York

§Department of Medicine, Duke University, Durham, North Carolina

Departments of Medicine and Pathology, Columbia University Medical Center, New York, New York.

Correspondence: Serge Cremers, PharmD, PhD, Irving Institute for Clinical and Translational Research, Columbia University Medical Center, 630 West 168 Street PH10-105, New York, NY 10032 (e-mail:

Supported by NIH CTSA Grant UL1 RR024156, Doris Duke Charitable Foundation.

The authors declare no conflict of interest.

Received January 15, 2012

Accepted April 26, 2012

© 2012 Lippincott Williams & Wilkins, Inc.