An Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry Method for the Quantification of Vancomycin Requiring Only 2 µL of Rabbit Serum

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 of 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 C 18 (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.

Разработка И Валидация Количественного Вэжх-Мс/Мс Метода Определения Пиперациллина В Плазме Крови

“…Therapeutic drug monitoring with dosing adapted to the altered pharmacokinetics of the individual patient will avoid underdose or overdose. (Barco et al 2015;Schmitt et al 2017;Verhoven et al 2018;Capiau et al 2019).…”

Toxicity Profile and Pharmacokinetic Study of Antibiotic Mixtures, Gentamicin and Vancomycin, in Rat Plasma by Ecofriendly Liquid Chromatography Coupled Tandem Mass Spectrometry

“…HPLC coupled with tandem mass spectrometric (LC–MS/MS) detection technique is the method of choice for the identification and quantification of GTM and VCM in biological fluids because aminoglycoside and glycopeptide lack notable chromophores and fluorophores . Several LC–MS/MS methods have been reported for the determination of GTM − and VCM − in biological fluids. These methods were developed for quantification of the target drugs in positive ion mode either alone or with other antibiotics.…”

A Novel Method for the Determination of Vancomycin in Serum by High-Performance Liquid Chromatography-Tandem Mass Spectrometry and Its Application in Patients with Diabetic Foot Infections

“…So far, reports of determination of vancomycin using Q-trap MS are seldom, Schmitt et al [ 23 ] once established a Q-trap MS method for determination of vancomycin in rabbit serum, but they lacked studies of matrix effects and stability, which caused the incomplete of the method. In this research, a reliable method based on Q-trap-MS was developed to determine vancomycin; the methodological parameters were verified comprehensively.…”