Jeffrey Patrick (Presenter)
Prosolia, Inc
Authorship: Justin Wiseman, Joe Kennedy, Jeffrey S. Patrick
Prosolia, INc. INdianapolis, IN USA
| Short Abstract The growth of infections resistant to traditional antibiotics (i.e. MRSA) has created the need to monitor therapeutic concentrations of the drugs and assure their effectiveness. Two of the most common drugs used are Vancomycin and Gentamicin. The active therapeutic windows for these require prompt monitoring of both peak levels due to toxicity and troughs which are ca. 10 mcg/mL. These compounds are currently monitored using immunoassays. Here, a method is developed and characterized using paperspray interfaced to mass spectrometry to monitor both drugs in plasma and whole blood. Limits of detection below 5 mcg/mL of blood or plasma are achieved. This study demonstrates: 1) the first analysis of aminoglycoside antibiotics using paperspray; 2) proof of principle for multiplexing and monitoring more than one of these drugs; 3) proof of principle for analysis of new drugs without assays. |
Long Abstract
The growth of infections resistant to traditional antibiotics (i.e. MRSA) has created the need for testing to monitor therapeutic concentrations of the drugs and assure their effectiveness. Two of the most common drugs used to treat these are Vancomycin and Gentamicin. The active therapeutic windows for these require prompt monitoring of both peak levels due to toxicity and troughs which are ca. 10 mcg/mL. These compounds are currently monitored using immunoassays. Here a method is characterized using paperspray interfaced to mass spectrometry to monitor both drugs in serum, plasma and whole blood. Paperspray (Velox 360) is interfaced to either a triple quadrupole mass spectrometer or to a QExactive Focus mass spectrometer. Detection of the antibiotics is achieved by high resolution exact mass, parallel reaction monitor (targeted high resolution MS/MS), or multiple reaction monitoring analysis. Samples of human blood, serum or plasma are spiked with known concentrations of Vancomycin or Gentamicin. These samples are then used to optimize analytical conditions and sample preparation. Dynamic range, precision and sensitivity are compared for the various analytical approaches. The eluting solvent is optimized and the effect of additives and organic content on the figures of merit are discussed with the addition fluorinated acids providing specific benefit. Limits of detection for the vancomycin and gentamicin of below 5 micrograms/mL in blood or serum are attained. The compatibility of this methodology with other similar compounds is also explored to examine the use of the methodology as a platform. This study demonstrates: 1) the first analysis of aminoglycoside and related antibiotics using paperspray; 2) it serves as proof of principle for multiplexing and monitoring more than one of these drugs; 3) demonstrates the potential for the analysis of new or alternative therapeutics which do NOT yet have immunoassays.
References & Acknowledgements:
| Description | Y/N | Source |
| Grants | no | |
| Salary | yes | Prosolia |
| Board Member | no | |
| Stock | no | |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | yes |