Page 60 - Clinical-Medical Solution for Clinical Chemistry

A Novel Platform of On-line Sample Pre-treatment and LC/MS/MS
Analysis for Screening and Quantitation of Illicit Drugs in Urine

Performance of MRM-based Quantitative Method

Linearity of the calibration curves with both IS method (14 Process Ef ciency (P.E) was evaluated based on the peak
analytes) and external standard method (4 analytes) were area (external standard) or peak ratios (IS method) of
constructed using the standard samples prepared by pre-spiked samples and neat-spiked sample at all
pre-spiked in urine matrix are shown in Figure 4. The concentrations. The results shown in Table 3 indicate the
method parameters are summarized in Table 3. It can be P.E obtained for the 18 analytes are between 62~122%
seen that good linearity with R greater than 0.995 was except four analytes with higher values, Norbuprenorphine,
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obtained for the eighteen illicit drugs in the range from 20 Morphine, MAM, and Methadone. This could be due to
ng/mL to 200 ng/mL in urine. interference from urine, which causes ion enhancement.

Accuracy of the quantitation method was evaluated with Speci city of the method for detection and con rmation
pre-spiked standard samples at all concentrations. The of the eighteen illicit drugs was evaluated (Figure 5). The
results are shown in Table 3, which indicate that reliable con rmation criteria for each target include quanti er
quantitation accuracy was obtained, except Methadone at MRM peak, its ratios with reference MRM transitions as
20 ng/mL with an accuracy of 130%. well as retention time.

Area Ratio Area Ratio Area Ratio Area Ratio Area Ratio Area Ratio
3 1.5
2.0
1.5 0.50 1.5
2 1.5 1.0
1.0 1.0 1.0
1 0.5 0.25
0.5 0.5 0.5
0 0.0 0.0 0.0 0.00 0.0
0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio
NIME / NITRA-d5 KET / KET-d4 NORKET / NORKET-d4 BU / BU-d4 NORBU / NORBU-d4 MORP / MORP-d3
Area Ratio Area Ratio Area Ratio Area Area Area
2.0 3000000 3000000
1000000
1.5 2 2000000 2000000
1.0 500000
1 1000000 1000000
0.5
0.0 0 0 0 0
0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 25.0 Conc. 0.0 25.0 Conc. 0.0 25.0 Conc.
COD / COD-d3 MAM / MAM-d6 BE / BE-d3 PCP METQ METD
Area Area Ratio Area Ratio Area Ratio Area Ratio Area Ratio
2000000 4
2
2 2
1500000 3 1.5
1000000 1 2 1.0
1 1
500000 1 0.5
0 0 0 0 0 0.0
0.0 25.0 Conc. 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio 0.0 1.0 Conc. Ratio
PROP AMPH / AMPH-d5 MDEA / MDEA-d6 MDA / MDA-d5 MDMA / MDMA-d6 METH / METH-d8

Figure 4: Calibration curves of 14 illicit drugs with isotope-labelled internal standards and 4 illicit drugs
with external standard in human urine on LCMS-8040. Details are shown in Table 3.

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