AI-64310-CRFT

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Application Note 570
Precision
Precision was assessed with spiked human urine at
concentrations of 40 and 200 ng/mL. Inter- and intra-
assay CV values at low and high quality-control
concentrations varied between 5.0% and 12.9% (Table 4).
Table 4. Precision data
Figure 6. SRM chromatograms (quantifier: solid line; and qualifier:
dotted line) of the six opioids at LLOQ in spiked human urine
Precision
(%)
Intra
(n=5)
Inter
(n=15)
Intra
(n=5)
Inter
(n=15)
Concentration
(ng/mL)
40 40 200 200
Morphine 12.0 10.8 9.7 7.4
Codeine 6.8 6.4 9.3 8.0
Hydromorphone 7.0 7.7 5.9 5.0
Hydrocodone 8.3 8.2 12.9 10.0
Oxymorphone 14.1 11.4 7.9 6.4
Oxycodone 5.1 6.3 6.7 5.8
Carryover
No carryover was observed.
Solvent Usage
The method used only 5%–10% of the solvent amount
used at a normal flow rate setting (0.5 mL/min). This
dramatically lower solvent use will significantly lower
both initial solvent cost and the cost of disposing of
solvent waste.
Conclusion
We have used a novel approach for sensitive quantitation
of six opioids in urine for forensic toxicology purposes.
This approach used super-dilution to minimize frequently
observed ion suppression in urine samples and used a
microflow LC setup (Ultimate 3000 RSLCnano LC system
and TSQ Vantage mass spectrometer) to compensate for
sensitivity losses from super-dilution. This robust method
was linear between 20 and 5000 ng/mL for the six opioids
and highly accurate and precise. The method used only
5%–10% of the solvent amount used at a normal LC flow
rates, significantly lowering both solvent purchase and
waste disposal costs.
For forensic toxicology use only.