13_Application_News_No_SCA_210_050 - 第4页

Table 4: Clinical validation results for quantitation of 42 compounds . Percentage recovery from automated sample preparation in plasma . Inter and Intra-assay average accurac y and precision were within acceptable toler…

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Compounds
Precursor ion
Product ion
Retention
time
(min)
Number
total
of MRM
transitions
m/z
Q1
pre
-bias
(V)
Reference
m/z
Collision
energy
(V)
Q3
pre-bias
(V)
m/z
Collision
energy
(V)
Q3
pre
-
bias
(V)
2
-CI
308.00
-
11.0
290.85
-
14.0
-
13.0
275.85
-
24.0
-
29.0
5.03
15
2C
-B
260.05
-
10.0
242.90
-
12.0
-
11.0
227.85
-
22.0
-
23.0
4.58
15
3,4
-Methylenedioxypyrovalerone
276.15
-
10.0
126.10
-
14.0
-
12.0
175.00
-
12.0
-
17.0
5.18
15
4
-MTA
182.10
-
12.0
117.10
-
21.0
-
11.0
165.05
-
12.0
-
10.0
4.45
15
6
-acetylmorphine
328.35
-
12.0
164.95
-
39.0
-
16.0
211.00
-
27.0
-
21.0
3.61
15
6
-acetylmorphine-D3
331.35
-
24.0
165.15
-
43.0
-
17.0
211.10
-
27.0
-
21.0
3.61
2
Amphetamine
136.10
-
10.0
91.00
-
22.0
-
17.0
119.05
-
15.0
-
20.0
3.42
8
Amphetamine
-D5
141.10
-
15.0
93.10
-
18.0
-
17.0
124.15
-
14.0
-
13.0
3.42
2
Anhydroecgonine methyl ester
182.10
-
12.0
118.00
-
23.0
-
11.0
91.05
-
29.0
-
20.0
3.17
15
BDB
194.10
-
13.0
135.00
-
20.0
-
13.0
177.05
-
12.0
-
17.0
4.06
8
Benzoylecgonine
290.15
-
11.0
168.05
-
10.0
-
16.0
77.00
-
29.0
-
13.0
4.57
15
Benzoylecgonine
-D3
293.15
-
14.0
171.20
-
20.0
-
17.0
77.05
-
56.0
-
13.0
4.57
2
Buprenorphine
468.30
-
16.0
54.95
-
52.0
-
20.0
396.00
-
41.0
-
26.0
7.31
15
Buprenorphine
-D4
472.30
-
13.0
59.10
-
50.0
-
22.0
88.10
-
50.0
-
16.0
7.31
2
Cocaethylene
318.15
-
20.0
196.00
-
10.0
-
20.0
76.95
-
32.0
-
30.0
5.49
15
Cocaethylene
-D3
321.15
-
12.0
199.25
-
21.0
-
22.0
85.20
-
32.0
-
16.0
5.49
2
Cocaine
304.15
-
11.0
182.00
-
10.0
-
20.0
76.95
-
30.0
-
29.0
4.94
15
Cocaine
-D3
307.15
-
22.0
185.15
-
19.0
-
20.0
85.25
-
31.0
-
15.0
4.94
2
Codeine
300.15
-
11.0
215.00
-
25.0
-
22.0
151.95
-
62.0
-
28.0
3.56
15
Codeine
-D3
303.15
-
14.0
215.25
-
26.0
-
20.0
181.20
-
37.0
-
17.0
3.56
2
Dextromethorphan
272.20
-
10.0
171.00
-
20.0
-
17.0
215.05
-
12.0
-
14.0
6.43
15
Dihydrocodeine
302.20
-
11.0
198.95
-
33.0
-
19.0
127.95
-
64.0
-
23.0
3.56
15
Dihydrocodeine
-D3
305.20
-
15.0
199.15
-
35.0
-
21.0
128.30
-
55.0
-
25.0
3.56
2
Ecgonine methylester
200.15
-
12.0
182.05
-
18.0
-
18.0
82.05
-
26.0
-
13.0
0.97
15
Ecgonine methylester
-D3
203.15
-
14.0
185.25
-
18.0
-
13.0
85.20
-
26.0
-
30.0
0.97
2
EDDP
278.20
-
10.0
234.00
-
17.0
-
20.0
249.05
-
13.0
-
16.0
6.95
15
EDDP
-D3
281.20
-
19.0
234.30
-
31.0
-
16.0
249.35
-
25.0
-
17.0
6.95
2
Ephedrine
-D3
169.15
-
17.0
151.25
-
14.0
-
16.0
91.20
-
33.0
-
17.0
3.28
2
Ethylmorphine
314.20
-
12.0
152.00
-
65.0
-
14.0
165.00
-
42.0
-
16.0
4.04
15
Hydrocodone
300.15
-
11.0
198.95
-
31.0
-
20.0
127.90
-
59.0
-
22.0
3.82
15
Hydromorphone
286.15
-
10.0
185.00
-
30.0
-
19.0
157.00
-
42.0
-
15.0
3.24
15
MBDB
208.15
-
20.0
134.95
-
6.0
-
20.0
50.95
-
60.0
-
19.0
4.28
9
m
-CPP
197.10
-
12.0
118.10
-
34.0
-
11.0
154.00
-
20.0
-
15.0
4.38
15
MDA
180.10
-
12.0
105.05
-
21.0
-
22.0
163.05
-
13.0
-
16.0
3.65
15
MDA
-D5
185.10
-
13.0
110.15
-
22.0
-
11.0
168.15
-
13.0
-
18.0
3.65
2
MDEA
208.15
-
11.0
163.00
-
13.0
-
15.0
105.00
-
25.0
-
10.0
4.11
11
MDEA
-D5
213.15
-
23.0
163.15
-
14.0
-
30.0
105.20
-
28.0
-
18.0
4.11
2
MDMA
194.10
-
13.0
163.05
-
15.0
-
28.0
105.05
-
25.0
-
18.0
3.84
12
MDMA
-D5
199.10
-
21.0
165.15
-
15.0
-
18.0
107.15
-
25.0
-
11.0
3.84
2
Mephedrone
178.10
-
13.0
145.05
-
20.0
-
14.0
160.05
-
15.0
-
10.0
3.99
15
Methadone
310.20
-
18.0
310.20
-
8.0
-
21.0
76.95
-
30.0
-
13.0
7.60
14
Methadone
-D9
319.20
-
20.0
268.25
-
20.0
-
20.0
105.05
-
25.0
-
20.0
7.60
2
Methamphetamine
150.15
-
10.0
91.00
-
22.0
-
20.0
119.05
-
16.0
-
21.0
3.63
8
Methcathinone
164.10
-
30.0
131.05
-
21.0
-
23.0
146.05
-
16.0
-
30.0
3.43
13
Methiopropamine
156.10
-
11.0
97.00
-
23.0
-
10.0
58.00
-
12.0
-
23.0
3.39
15
Methylphenidate
234.15
-
20.0
84.00
-
8.0
-
20.0
91.0
-
46.0
-
17.0
4.71
7
Morphine
286.15
-
10.0
152.00
-
60.0
-
15.0
201.00
-
27.0
-
20.0
3.11
15
Morphine
-D3
289.15
-
14.0
152.10
-
59.0
-
26.0
201.15
-
26.0
-
21.0
3.11
2
Naloxone
328.15
-
12.0
310.00
-
21.0
-
21.0
212.00
-
39.0
-
22.0
3.60
14
Naloxone
-D5
333.15
-
12.0
315.20
-
20.0
-
22.0
258.10
-
29.0
-
27.0
3.60
2
Naltrexone
342.15
-
12.0
324.05
-
22.0
-
15.0
270.05
-
28.0
-
28.0
3.75
14
Naltrexone
-D3
345.15
-
16.0
327.15
-
22.0
-
23.0
270.15
-
28.0
-
29.0
3.75
2
Norbuprenorphine
414.25
-
28.0
54.90
-
63.0
-
24.0
83.05
-
50.0
-
14.0
5.50
15
Norephedrine
152.10
-
10.0
134.05
-
15.0
-
13.0
115.05
-
25.0
-
11.0
3.02
11
Norfenfluramine
204.10
-
14.0
159.00
-
20.0
-
15.0
109.05
-
40.0
-
18.0
4.16
15
Noroxycodone
302.15
-
11.0
199.00
-
37.0
-
20.0
196.95
-
26.0
-
20.0
3.56
15
Noroxycodone
-D3
305.15
-
22.0
287.15
-
17.0
-
20.0
190.10
-
25.0
-
20.0
3.56
2
Norpseudoephedrine
152.10
-
10.0
134.05
-
15.0
-
13.0
115.05
-
25.0
-
11.0
3.12
11
Oxycodone
316.15
-
12.0
298.00
-
20.0
-
20.0
240.95
-
29.0
-
24.0
3.73
14
Oxycodone
-D3
319.15
-
23.0
301.10
-
19.0
-
21
259.10
-
26.0
-
27.0
3.73
2
Pholcodine
399.25
-
14.0
114.05
-
36.0
-
11.0
381.05
-
25.0
-
18.0
3.20
5
Ritalinic acid
220.15
-
14.0
84.10
-
22.0
-
14.0
56.05
-
44.0
-
22.0
4.14
10
Table 3: MRM transitions, retention times and the total number of MRM transitions measured using MRM Spectrum mode during the acquisition for
42 compounds and their 20 internal standard compounds. Quantitative data was measured using quantifier ion and reference ion with ion ratio
percentage tolerance of 20%. Additional MRM transitions were used for Library identification.
SCA_210_050
Table
4: Clinical
validation
results
for quantitation
of 42 compounds
. Percentage
recovery
from automated
sample
preparation
in plasma
. Inter and Intra-assay
average
accuracy
and
precision
were within
acceptable
tolerance
of 85-115% and better
than 15% relative
standard
deviation
(RSD)
SCA_210_050
3.6 3.7 3.8 3.9 4.0
0.0e0
5.0e5
1.0e6
1.5e6
2.0e6
Codeine
RT : 3.756
Compound
Codeine
Formula C18H21NO3
CAS 76-57-3
[M+H]+ 300.15
Q: 300.1 > 152.1
R: 300.1 > 215.1
3.8 3.9 4.0 4.1 4.2
0.0e0
1.0e6
2.0e6
3.0e6
4.0e6
Hydrocodone
RT : 4.018
Compound
Hydrocodone
Formula C18H21NO3
CAS 125-29-1
[M+H]+ 300.15
Q: 300.1 > 199.1
R: 300.1 > 128.1
115.00
127.95
141.00
151.95
165.00
183.00
199.00
215.00
224.95
m/z
125 150 175 200 225
2.0e6
4.0e6
6.0e6
8.0e6
1.0e7
115.00
127.95
141.00
151.95
165.00
183.00
199.00
215.00
224.95
243.00
0.0e0
2.0e5
4.0e5
6.0e5
8.0e5
Acquired MRM spectrum
RT: [3.692 – 3.815] (+)
Codeine MRM spectrum Library Spectrum RT: [3.701] (+) SI: 98
127.90
141.00
151.95
154.95
171.00
182.95
198.95
213.00
241.00
m/z
125 150 175 200 225
2.0e6
4.0e6
6.0e6
8.0e6
1.0e7
127.90
141.00 151.95
152.95
171.00
185.00
198.95
213.00
241.00
0.0e0
5.0e5
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
Acquired MRM spectrum
RT: [3.963 – 4.069] (+)
Hydrocodone MRM spectrum Library Spectrum RT: [4.001] (+) SI: 94
Figure 2: MRM spectrum mode MS/MS data for codeine and hydrocodone extracted from plasma samples using the CLAM-2000
extraction protocol. The identification of isobaric compounds such as codeine and hydrocodone in plasma samples was
confirmed by matching acquired MRM spectrum data with a reference library generated using certified materials.
Compound Codeine
Calibration range 2.5-500ug/L
Q: 300.1 > 152.1
Regression analysis
y = 0.04496957 + 0.01203203
R² = 0.999347
0 50 100 150 200 250 300 350 400 450 500
0
2
4
6
8
10
12
14
16
18
20
22
Area Ratio
Conc.
0 50 100 150 200 250 300 350 400 450 500
0
2
4
6
8
10
12
14
16
18
20
22
24
26
28
Compound Hydrocodone
Calibration range 1-500ug/L
Q: 300.1 > 199.1
Regression analysis
y = 0.05681472x - 0.01050782
R² = 0.9959177
Area Ratio
Conc.
Figure 3: Calibration curve data for codeine and hydrocodone extracted from plasma samples (calibration range 1-500ug/L).
SCA_210_050