A Simple Liquid Chromatography–Tandem Mass Spectrometry Method for Determination of Plasma Fentanyl Concentration in Rats and Patients with Cancer Pain

Abstract

A fentanyl patch is widely used for the treatment of cancer pain. Its few adverse effects include constipation and drowsiness. The absorption volume of transdermally applied fentanyl may differ according to its site of application and variability in patch adhesion. Since fentanyl is predominantly metabolized by the drug-metabolizing enzyme cytochrome P450 (CYP) 3A4 in the liver, its concentration may vary in cases of physiologically reduced CYP3A4 activity in the liver (liver disease and aging) or on co-administration of drugs. The clinical significance of measuring plasma concentration of fentanyl is high, but conventional methods require complicated processes such as solid-phase extraction and liquid-liquid extraction before the sample is injected into an HPLC system. In this study, a simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for determining plasma fentanyl concentrations by deproteinization with acetonitrile. A recovery test was conducted using an absolute calibration curve to confirm the method's linearity and inter-and intra-day reproducibility. The required plasma volume for detection was reduced from 1 mL in the conventional method to 20 µL in the present study, and a good calibration curve was obtained in the concentration range from 0.05 to 5 ng/mL. These findings suggest that the method for sample preparation and quantification developed in this study are appropriate for measuring fentanyl concentration in human plasma in clinical settings.

Simultaneous quantification of fentanyl, sufentanil, cefazolin, doxapram and keto‐doxapram in plasma using liquid chromatography–tandem mass spectrometry

“…Regarding cefazolin, multiple assays have been reported with comparable performance (Carlier et al, ; Kiriazopoulos et al, ; Lillico et al, ; Parker et al, ). The reported fentanyl assays required larger sample volumes except for Hisada et al (), who needed only 20 μL and reached a lower LLOQ of 0.05 μg L −1 compared with our 0.1 μg L −1 .…”

“…Concerning doxapram, four of the five reported assays date from the 1990s (Aranda et al, 1988;LeGatt et al, 1986;Nichol et al, 1980;Robson & Prescott, 1977) and are inferior to our assay with respect to the use of a different drug for internal standard, sample preparation which requires an evaporation step, higher LLOQ, larger sample volume required, a longer run time and two assays not being able to mea- assays have been reported with comparable performance (Carlier et al, 2012;Kiriazopoulos et al, 2017;Lillico et al, 2016;Parker et al, 2016). The reported fentanyl assays required larger sample volumes except for Hisada et al (2013), who needed only 20 μL and reached a lower LLOQ of 0.05 μg L −1 compared with our 0.1 μg L −1 .…”

“…In general, for all analytes, our assay performed better than or comparable to prior reported assays, even in comparison with assays measuring only a single analyte, which makes it easier to achieve good performance on run time, required sample volume and matrix effects (Aranda et al, 1988;Carlier et al, 2012;Clavijo et al, 2011;Fernandez Mdel et al, 2013;Hisada et al, 2013;Kiriazopoulos et al, 2017;LeGatt et al, 1986;Lillico et al, 2016;Mahlke et al, 2014;Nichol et al, 1980;Nosseir et al, 2014;Palleschi et al, 2003;Parker et al, 2016;Robson & Prescott, 1977;Saari et al, 2012;Suzuki et al, 2017). Furthermore, most assays use a different drug as an internal standard, whereas we used a deuterated form of fentanyl, which shows better comparable behavior to the analytes that are measured than using a different drug.…”

Quantitative determination of fentanyl in newborn pig plasma and cerebrospinal fluid samples by HPLC‐MS/MS

“…Capillary voltage was set at 0.8 kV. FEN and 13 C 6 FEN were detected by multiple reaction monitoring (MRM) mode with a dwell time of 0.20 s. The following transitions were monitored in ESI +: m / z 337.0 → m / z 188.0 – in accordance with the values reported in the literature – using a cone voltage (CV) of 35 V and a collision energy (CE) of 25 eV for FEN quantification, m / z 337.0 → m / z 105.0 using a CV of 25 V and a CE of 45 eV for FEN confirmation and m / z 343.0 → m / z 188.0 using a CV of 45 V and a CE of 25 eV for 13 C 6 FEN.…”

“…Lower sample volumes were used by Chang et al . and Hisada et al ., using an LLE procedure and a simple protein precipitation method respectively.…”