Ultra-fast quantitation of voriconazole in human plasma by coated blade spray mass spectrometry
Introduction
Voriconazole is an antifungal drug from the family of the triazoles. As a derivative of fluconazole, voriconazole possesses a fluoropyrimidine group rather than a triazole moiety in addition to an alpha methylation on the tertiary carbon [1]. In contrast to the first generation of triazole antifungals, voriconazole has a broader spectrum of action [2], [3] and has been demonstrated as effective against fungal infections caused by Aspergillus [4] and Fusarium species [3]. The mechanism of action consists of the inhibition of cytochrome P450 enzyme lanosterol 14-alpha-demethylase, which prevents the conversion of lanosterol to ergosterol, an essential component of cell membranes [5]. Although the effectiveness of this antifungal drug has been widely proven, toxicity risks and a narrow therapeutic concentration window have been reported. Indeed, plasma concentrations above 6 μg/mL have been associated with clinical events such as visual impairment or photopsia, abnormal hepatic function, and higher bilirubin levels [6], [7]. On the other hand, in cases where voriconazole plasma concentrations were below 2 μg/mL, a great proportion of patients suffered a significant progression in their infections [8]. In addition, high pharmacokinetic variability of this drug due to erratic bioavailability and variation in drug metabolism has also been documented [9]. Consequently, the development of an accurate, reliable, fast, and cost-effective analysis of voriconazole in plasma is highly desirable.
Most of the methods for voriconazole determination to date involve the use of liquid chromatography (LC) with either ultra-violet (UV) [1] or mass spectrometry (MS) [10], [11], [12], [13] detection. In addition, methods employing gas chromatography–MS (GC–MS) [14] and capillary electrophoresis (CE) [15] have also been reported. Within the LC–MS applications, the fastest LC separation was attained in 3 min [16] and the sample volume consumption was in the range between 20 μL [10] and 200 μL [16] of plasma. However, when the sample preparation step is considered in the total analysis time, none of the published methodologies can be completed in less than 15 min considering that even in the simplest approaches protein precipitation and centrifugation steps are required in order to have an extract suitable for liquid chromatography [12]. Recent developments in bioanalytical applications using solid phase microextraction (SPME) have demonstrated that this technology is an attractive alternative to conventional methods due to its simplicity, sensitivity and speed of analysis [17]. In SPME, a device coated with a polymeric material is used to extract/enrich analytes from a given sample prior to instrumental analysis. By using novel ultra-thin biocompatible coatings, faster and efficient extractions of small molecules from biofluids can be easily attained [18], [19], [20]. Despite the well-known advantages of using LC, emerging technologies such ambient mass spectrometry [21], [22], [23], [24], [25] and the direct coupling of SPME to very sensitive and selective mass spectrometers have shown outstanding results when aiming to decrease the total analysis time [26]. Among the recently developed SPME-MS technologies, coated blade spray (CBS) [17], SPME-direct analysis in real time (SPME-DART) [27], Bio-SPME-nano-ESI [26], Bio-SPME-OPP [28] and SPME-DBDI [29] excel by the limits of detection achieved. Succinctly, CBS was developed as an ideal compromise between sample preparation and direct coupling to mass spectrometry and it behaves as an SPME device for extraction and as a solid-substrate ESI source when performing ionization. When comparing CBS to other SPME-MS technologies, the former offers additional advantages such as simplicity of the set-up (see Fig. 1), negligible MS carry over (i.e. no-cross talking among experiments), minimal solvent use (≤20 μL), and no-need of expensive parts for the construction of the ionization source [17]. In addition, CBS is the device with the largest coated surface area available for extraction when compare to the other SPME-devices used for direct coupling to MS; therefore, low or even sub parts per billion levels can be easily achieved when rapid extractions (time ≤ 2 min) are performed from small sample volumes (≤300 μL).
In this work, we propose the ultra-fast determination of voriconazole in human plasma by two different CBS-based approaches. In the first strategy, voriconazole is extracted on the CBS device by fully immersion of the coated blade in 300 μL of human plasma during 1 min under vortex agitation. In the second approach, analytes are enriched on the CBS by placing a 10 μL droplet of plasma for 2 min on the coated area. After the completion of the extraction process in both cases, a quick rinsing step is performed in order to remove matrix components potentially adhered to the surface. Once complete, analytes were directly desorbed/ionized from the CBS device and analyzed by mass spectrometry without the need of additional sample preparation or chromatography. Both methodologies were fully validated in terms of linearity, LOD, LOQ, repeatability, reproducibility, relative matrix effects and accuracy employing 5 different lots of plasma and 4 different patients.
Section snippets
Instrumentation
All the experiments were performed using a TSQ Quantiva (Thermo Scientific, San Jose, CA, USA). Data was processed using Trace Finder version 3.0 (Thermo Scientific, San Jose, CA, USA). A home-made coated blade spray interface was built at University of Waterloo, and a thorough description of the operation of this system can be found elsewhere [17].
Chemicals and materials
Methanol (MeOH), acetonitrile (ACN), isopropanol (IPA) and water were all LC–MS grade and purchased from Fisher Scientific. Formic acid was
Results and discussion
Aiming to develop an ultra-fast analysis, while monitoring two MS/MS transitions (i.e. compound of interest and its internal standard), the electrospray event was set to 3 s using a dwell time of 100 ms for each compound. Thus, a minimum of 15 scans per compound is obtained without significantly increasing the total analysis time. Ion chronograms related to the selective reaction monitoring (SRM) of voriconazole at different concentration levels, including the plasma blank, are presented in Fig. 2
Conclusions
In this work, two methodologies for the ultra-fast quantitation of voriconazole in human plasma were developed. Both approaches have all the advantages of SPME such as analyte enrichment and sample clean-up, as well as the speed and simplicity of direct coupling to mass spectrometry offered by CBS. Although the current set-up permits the quantitation of voriconazole down to 0.1 μg/mL in 120 s, in the near future fully automated systems will reduce the total analysis time to only 25 s per sample.
Acknowledgements
The authors thank Thermo Fisher Scientific and the Natural Sciences and Engineering Research Council (NSERC) of Canada for the financial support through the Industrial Research Chair program. We are also very thankful with Thermo Fisher Scientific for lending to our laboratory the triple quadrupole mass spectrometer used in this work (TSQ Quantiva) as part of the Industrial Research Chair program. We acknowledge Waters Corporation for kindly providing the HLB particles used in this study.
References (34)
- et al.
Measurement of voriconazole in serum and plasma
Clin. Biochem.
(2007) - et al.
Fast and simple LC–MS/MS method for quantifying plasma voriconazole
Clin. Chim. Acta
(2012) - et al.
Simultaneous quantitation of azole antifungals, antibiotics, imatinib, and raltegravir in human plasma by two-dimensional high-performance liquid chromatography–tandem mass spectrometry
J. Chromatogr. B: Anal. Technol. Biomed. Life Sci.
(2013) - et al.
Development and validation of a liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay to quantify serum voriconazole
J. Chromatogr. B: Anal. Technol. Biomed. Life Sci.
(2015) - et al.
Determination of voriconazole in human serum and plasma by micellar electrokinetic chromatography
J. Pharm. Biomed. Anal.
(2010) - et al.
A critical review of the state of the art of solid-phase microextraction of complex matrices I. Environmental analysis
TrAC – Trends Anal. Chem.
(2015) - et al.
High throughput paper spray mass spectrometry analysis
Clin. Chim. Acta
(2013) Standard line slopes as a measure of a relative matrix effect in quantitative HPLC–MS bioanalysis
J. Chromatogr. B: Anal. Technol. Biomed. Life Sci.
(2006)- et al.
High throughput quantification of prohibited substances in plasma using thin film solid phase microextraction
J. Chromatogr. A
(2014) - et al.
Development and validation of a HPLC method for the determination of voriconazole in pharmaceutical formulation using an experimental design
Talanta
(2007)
Breakthrough fungal infections in stem cell transplant recipients receiving voriconazole
Clin. Infect. Dis.
Voriconazole treatment for less-common, emerging, or refractory fungal infections
Clin. Infect. Dis.
Efficacy and safety of voriconazole in the treatment of acute invasive aspergillosis
Clin. Infect. Dis.
Voriconazole: a new triazole antifungal
Ann. Pharmacother.
The safety of voriconazole
Clin. Infect. Dis.
A randomized, double-blind, double-dummy, multicenter trial of voriconazole and fluconazole in the treatment of esophageal candidiasis in immunocompromised patients
Clin. Infect. Dis.
Voriconazole therapeutic drug monitoring
Antimicrob. Agents Chemother.
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These authors contributed equally to this work.