Stability of Cocaine, Opiates, and Metabolites in Dried Saliva Spots
Abstract
:1. Introduction
2. Results and Discussion
2.1. Optimization of the Stability Protocol
2.2. Assay without Preservative
2.3. Long-Term Stability
3. Materials and Methods
3.1. Reagents and Standards
3.2. Biological Specimens
3.3. Gas Chromatographic and Mass Spectrometric Conditions
3.4. Sample Preparation
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Run Order | Temperature | Light | Preservative | Time |
---|---|---|---|---|
1 | Refrigerator | Presence | Low | 1 Day |
2 | Refrigerator | Presence | Low | 7 Days |
3 | Refrigerator | Presence | High | 7 Days |
4 | Refrigerator | Absence | Low | 1 Day |
5 | Room | Presence | High | 7 Days |
6 | Room | Absence | Low | 7 Days |
7 | Room | Absence | High | 7 Days |
8 | Room | Absence | Low | 1 Day |
9 | Refrigerator | Absence | High | 7 Days |
10 | Room | Absence | High | 1 Day |
11 | Room | Presence | Low | 7 Days |
12 | Refrigerator | Absence | Low | 7 Days |
13 | Room | Presence | High | 1 Day |
14 | Refrigerator | Presence | High | 1 Day |
15 | Room | Presence | Low | 1 Day |
16 | Refrigerator | Absence | High | 1 Day |
Condition | EME | COC | COET | BEG | NCOC | COD | MOR | 6-MAM |
---|---|---|---|---|---|---|---|---|
1 | 0.16 ± 0.008 | 0.17 ± 0.002 | 0.14 ± 0.019 | 0.36 ± 0.050 | 0.03 ± 0.008 | 0.02 ± 0.004 | 0.21 ± 0.015 | 0.03 ± 0.006 |
2 | 0.16 ± 0.011 | 0.18 ± 0.021 | 0.24 ± 0.010 | 0.30 ± 0.005 | 0.06 ± 0.007 | 0.03 ± 0.021 | 0.39 ± 0.099 | 0.03 ± 0.008 |
3 | 0.15 ± 0.034 | 0.18 ± 0.002 | 0.28 ± 0.034 | 0.36 ± 0.04 | 0.07 ± 0.005 | 0.02 ± 0.0006 | 0.21 ± 0.009 | 0.03 ± 0.005 |
4 | 0.18 ± 0.031 | 0.17 ± 0.058 | 0.29 ± 0.016 | 0.28 ± 0.071 | 0.08 ± 0.002 | 0.02 ± 0.009 | 0.46 ± 0.097 | 0.03 ± 0.0002 |
Day | EME | COC | COET | BEG | NCOC | COD | MOR | 6-MAM | |
---|---|---|---|---|---|---|---|---|---|
Without preservative | 1 | 0.16 | 0.23 | 0.29 | 0.31 | 0.09 | 0.02 | 0.35 | 0.03 |
7 | 0.22 | 0.14 | 0.24 | 0.24 | 0.05 | 0.02 | 0.47 | 0.03 | |
With preservative | 1 | 0.54 | 0.35 | 0.34 | 1.23 | 0.13 | 0.04 | 0.88 | 0.04 |
7 | 0.32 | 0.28 | 0.27 | 1.14 | 0.10 | 0.01 | 0.77 | 0.03 |
Analyte | Day | Concentration (ng/mL) * | CV (%) | Variation from Day 1 (%) |
---|---|---|---|---|
EME | 1 | 105.38 ± 0.028 | 5.10 | |
3 | 94.73 ± 0.084 | 17.1 | −10.1 | |
7 | 60.52 ± 0.013 | 4.3 | −42.6 | |
14 | 18.45 ± 0.009 | 9.5 | −82.5 | |
21 | 6.67 ± 0.008 | 20.0 | −93.7 | |
28 | 2.22 ± 0.001 | 10.5 | −97.9 | |
44 | 1.20 ± 0.0007 | 10.7 | −98.9 | |
112 | 0.91 ± 0.0006 | 12.3 | −99.1 | |
121 | 0.29 ± 0.0001 | 9.6 | −99.7 | |
136 | 0.58 ± 0.0002 | 8.1 | −99.4 | |
COC | 1 | 86.28 ± 0.056 | 15.8 | |
3 | 88.25 ± 0.054 | 15.1 | 2.3 | |
7 | 72.27 ± 0.035 | 11.9 | −16.2 | |
14 | 38.90 ± 0.032 | 20.5 | −54.9 | |
21 | 36.01 ± 0.024 | 16.6 | −58.3 | |
28 | 28.48 ± 0.013 | 11.6 | −67.0 | |
44 | 10.93 ± 0.005 | 10.4 | −87.3 | |
112 | 8.72 ± 0.002 | 4.8 | −89.9 | |
121 | 11.00 ± 0.007 | 15.1 | −87.3 | |
136 | 7.67 ± 0.004 | 14.0 | −91.2 | |
COET | 1 | 86.05 ± 0.053 | 16.3 | |
3 | 75.45 ± 0.035 | 12.4 | −12.3 | |
7 | 71.67 ± 0.042 | 15.3 | −16.7 | |
14 | 27.82 ± 0.022 | 20.5 | −67.7 | |
21 | 32.42 ± 0.022 | 17.9 | −62.3 | |
28 | 30.96 ± 0.011 | 9.70 | −64.0 | |
44 | 14.21 ± 0.010 | 17.8 | −83.5 | |
112 | 7.76 ± 0.004 | 13.7 | −91.0 | |
121 | 7.95 ± 0.006 | 19.0 | −90.8 | |
136 | 2.15 ± 0.0008 | 10.2 | −97.5 | |
BEG | 1 | 95.83 ± 0.230 | 18.9 | |
3 | 92.24 ± 0.070 | 6.0 | −3.7 | |
7 | 88.55 ± 0.183 | 16.3 | −7.6 | |
14 | 81.23 ± 0.124 | 12.1 | −15.2 | |
21 | 82.41 ± 0.135 | 13.0 | −14.0 | |
28 | 85.95± 0.174 | 15.9 | −10.3 | |
44 | 77.30 ± 0.034 | 3.5 | −19.3 | |
112 | 78.40 ± 0.072 | 7.2 | −18.2 | |
121 | 79.06 ± 0.055 | 5.4 | −17.5 | |
136 | 82.85 ± 0.145 | 13.8 | −13.5 | |
NCOC | 1 | 117.04 ± 0.018 | 14.8 | |
3 | 98.46 ± 0.019 | 18.1 | −15.9 | |
7 | 98.44 ± 0.013 | 12.4 | −15.9 | |
14 | 73.27 ± 0.014 | 18.6 | −37.4 | |
21 | 58.82 ± 0.010 | 16.8 | −49.7 | |
28 | 49.66 ± 0.004 | 7.9 | −57.6 | |
44 | 29.92 ± 0.003 | 8.8 | −74.4 | |
112 | 13.15 ± 0.002 | 16.2 | −88.8 | |
121 | 18.33 ± 0.003 | 15.2 | −84.3 | |
136 | 6.16 ± 0.0005 | 8.4 | −94.7 | |
COD | 1 | 83.53 ± 0.006 | 15.8 | |
3 | 22.54 ± 0.001 | 13.1 | −73.9 | |
7 | 30.79 ± 0.001 | 11.0 | −64.4 | |
14 | 26.52± 0.002 | 13.8 | −69.3 | |
21 | 27.94 ± 0.002 | 18.1 | −67.7 | |
28 | 27.85 ± 0.002 | 18.4 | −67.8 | |
44 | 29.01 ± 0.002 | 12.3 | −66.5 | |
112 | 24.14 ± 0.0002 | 2.0 | −72.1 | |
121 | 21.67 ± 0.0004 | 4.3 | −75.0 | |
136 | 18.86 ± 0.001 | 15.1 | −78.2 | |
MOR | 1 | 89.11 ± 0.106 | 12.2 | |
3 | 82.63 ± 0.010 | 12.4 | −7.3 | |
7 | 78.45 ± 0.092 | 12.1 | −12.0 | |
14 | 73.28 ± 0.024 | 3.4 | −17.8 | |
21 | 59.83 ± 0.047 | 8.1 | −32.9 | |
28 | 49.25 ± 0.035 | 7.4 | −44.7 | |
44 | 39.75 ± 0.044 | 11.4 | −55.4 | |
112 | 21.63 ± 0.032 | 15.2 | −75.7 | |
121 | 28.19 ± 0.025 | 8.9 | −68.4 | |
136 | 31.34 ± 0.052 | 17.0 | −64.8 | |
6-MAM | 1 | 108.96 ± 0.003 | 8.0 | |
3 | 94.91 ± 0.005 | 17.5 | −12.9 | |
7 | 90.84 ± 0.005 | 16.7 | −16.6 | |
14 | 62.66 ± 0.002 | 9.8 | −42.5 | |
21 | 52.75 ± 0.003 | 19.5 | −51.6 | |
28 | 35.48 ± 0.002 | 16.1 | −67.4 | |
44 | 24.01 ± 0.0005 | 6.8 | −78.0 | |
112 | 10.64 ± 0.0001 | 3.3 | −90.2 | |
121 | 13.38 ± 0.0007 | 16.2 | −87.7 | |
136 | 13.19 ± 0.0007 | 16.0 | −87.9 |
Analyte | Retention Time (min) | Transitions (m/z) | Collision Energy (eV) | Dwell Time (µs) |
---|---|---|---|---|
EME | 8.51 | 271.5–83.1 * 181.3–82.0 | 5 10 | 50 |
EME-d3 | 8.50 | 274.5–86.1 | 10 | 50 |
COC | 11.97 | 182.5–82.2 * 182.5–150.1 | 10 5 | 50 |
COC-d3 | 11.96 | 184.1–85.0 | 10 | 50 |
COET | 12.22 | 196.5–82.0 * 196.5–150.2 | 10 5 | 50 |
BEG | 12.25 | 239.6–82.2 * 239.6–122.2 | 15 15 | 50 |
BEG-d3 | 12.21 | 241.9–85.1 | 15 | 50 |
NCOC | 12.33 | 178.1–105.1 * 178.1–135.1 | 15 10 | 50 |
COD | 13.08 | 369.8–229.2 369.8–354.3 * | 20 20 | 50 |
COD-d3 | 13.08 | 374.0–374.0 | 5 | 50 |
MOR | 13.31 | 235.4–146.1 * 235.4–220.2 | 10 5 | 50 |
6-MAM | 13.66 | 397.9–287.4 * 397.9–340.5 | 20 20 | 50 |
6-MAM-d3 | 13.65 | 402.4–402.4 | 5 | 50 |
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Almeida, E.; Soares, S.; Gonçalves, J.; Rosado, T.; Fernández, N.; Rodilla, J.M.; Passarinha, L.A.; Barroso, M.; Gallardo, E. Stability of Cocaine, Opiates, and Metabolites in Dried Saliva Spots. Molecules 2022, 27, 641. https://doi.org/10.3390/molecules27030641
Almeida E, Soares S, Gonçalves J, Rosado T, Fernández N, Rodilla JM, Passarinha LA, Barroso M, Gallardo E. Stability of Cocaine, Opiates, and Metabolites in Dried Saliva Spots. Molecules. 2022; 27(3):641. https://doi.org/10.3390/molecules27030641
Chicago/Turabian StyleAlmeida, Ema, Sofia Soares, Joana Gonçalves, Tiago Rosado, Nicolás Fernández, Jesus M. Rodilla, Luís A. Passarinha, Mário Barroso, and Eugenia Gallardo. 2022. "Stability of Cocaine, Opiates, and Metabolites in Dried Saliva Spots" Molecules 27, no. 3: 641. https://doi.org/10.3390/molecules27030641