324 / 658
Methods
This method describes the analysis for the determination
of fentanyl and its metabolite, norfentanyl, from a urine
sample. Human urine was used as the test matrix. An
LOQ of 0.5 ng/mL was seen in human urine, with an
LOD below 0.1 ng/mL. Instrumentation used is identified
in Table 1.
Table 1. Instrumentation used in this method
LS-MS/MS:
Aria TLX-4 with Thermo Scientific TSQ Quantum
Access triple quadrupole mass spectrometer
Extraction column:
Thermo Scientific TurboFlow XL C18 P 0.5x50 mm
Analytical column:
Thermo Scientific Hypersyl GOLD aQ 3x50, 5 µm
Experimental Conditions:
A working solution containing fentanyl and norfentanyl at
1000 ng/mL was made. Subsequent dilutions yielded a
curve from 200 ng/mL to 0.5 ng/mL. An internal standard
solution containing both fentanyl-D5 and norfentanyl-D5
was added to all standards. Samples were vortexed and
then centrifuged at 10,000 RCF for 5 minutes and
analyzed immediately.
Results:
The data in Figure 1 shows linear regression for 0.5
ng/mL to 200 ng/mL, with 1/x weighing. Figure 2
demonstrates the limit of quantitation with excellent
signal to noise ratio.
Conclusion:
The Aria TLX-4 system powered by TurboFlow
technology provides a fast, efficient, and automated on-
line separation technology for the extraction and analysis
of fentanyl and its metabolite, norfentanyl. The ability to
run 5.5 minute methods on four channels further
decreases analysis time and increases the efficiency of
the TSQ Quantum Access
™
mass spectrometer. The Aria
TLX-4 coupled with the TSQ Quantum Access can run
one sample every 86 seconds with a 92.9% sample
completion rate with 7.1% re-injection
2
. The method run
time was 5.5 minutes. This system provides a reliable high
throughput method of fentanyl and norfentanyl for
clinical research labs.
References
1. Sauvage et al.
2006
. Therapeutic Drug Monitoring 28(1), pp. 123-130.
2. Crouch, Dennis.
The Analysis of Fentanyl and Norfentanyl using
TurboFlow Column Analyte Isolation and Multiplex-HPLC/MS/MS.
Oral
presentation, AAFS, Washington DC February 17-20
2008
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