TITLE:
An Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Vancomycin Requiring Only 2 µL of Rabbit Serum
AUTHORS:
Veronika Schmitt, András Szeitz, Tara L. Klassen, Urs O. Häfeli
KEYWORDS:
Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry, Vancomycin, Rabbit, Serum, Pharmacokinetics
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.8 No.9,
September
6,
2017
ABSTRACT: A highly sensitive ultra-high performance liquid
chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for
the quantification of vancomycin (VAN) in low volumes of rabbit serum. For each
analysis, 2 μL rabbit serum was precipitated with methanol that contained the
internal standard teicoplanin (TEI). The supernatant was transferred into a 384
well-plate, diluted with water, covered with a pierceable silicone mat and 5 μL
was analyzed in positive ionization mode. The UHPLC-MS/MS consisted of an
Agilent 1290 Infinity UHPLC system connected to an AB Sciex QTrap® 5500 hybrid linear ion-trap triple quadrupole mass
spectrometer equipped with a Turbo Spray source. Chromatographic separation was
achieved using a Waters Acquity UPLC BEH C18 (1.7 μm, 2.1 mm × 100
mm) column, a VanGuard (1.7 μm, 2.1 × 5 mm) guard column and a mobile phase of
water and methanol both containing 5 mM ammonium acetate with 0.1% formic acid.
VAN was quantified with multiple reaction monitoring using the transitions of m/z 725.5/144.2, and TEI was monitored
at m/z 940.6/316.4. The accuracy,
precision, linearity, range and lower limit of quantification (LLOQ) were
determined. The accuracy was ≤9.93% and the precision was ≤10.6%. The range was
established as 0.1 to 40 μg·mL-1. The LLOQ was 0.1 μg·mL-1 VAN requiring 2 μL of sample with an accuracy of -20.2% and precision of 8.39%.
The method was applied successfully to determine the VAN concentrations in
rabbit serum after the i.v. administration of VAN via implanted ear catheters.