|Year : 2019 | Volume
| Issue : 3 | Page : 136-140
Rapid screening of nine illicit drugs in human blood and urine by direct analysis in real-time mass spectrometry
Ying Zhang1, Wenfang Zhang1, Guobin Xin1, Li Liu1, Xiaokun Duan2, Charles Liu2
1 Forensic Science Service of Beijing Public Security Bureau, Key Laboratory of Forensic Toxicology, Ministry of Public Security, Beijing, China
2 ASPEC Technologies Limited, Beijing, China
|Date of Web Publication||18-Sep-2019|
No. 1, Long-Gang Road, Qing-He, Hai-Dian District, Beijing
Source of Support: None, Conflict of Interest: None
We aimed to establish for the rapid detection of morphine, O6-monoacetylmorphine, heroin, codeine, cocaine, methamphetamine, ketamine, methadone, and dolantin in human blood and urine by direct analysis in real-time coupled with tandem mass spectrometry (DART-MS/MS). These samples were extracted by acetonitrile-methanol (V/V = 4:1), using DART 12 Dip-it automatic sampling system. They were injected at 400°C, and analyzed by positive ion and multiple reaction monitoring mode. The detection limits of morphine, O6-Monoacetylmorphine, heroin, codeine, cocaine, methamphetamine, ketamine, methadone, and dolantin were 100, 50, 50, 100, 20, 20, 10, 1, and 0.01 ng/mL, respectively. The practical cases contained methamphetamine, codeine, cocaine, and O6-monoacetylmorphine were detected accurately and rapidly. The method has the advantages of high sensitivity and good accuracy. The sample processing is simple and can be analyzed in a short time. This method is suitable for the analysis of morphine, O6-monoacetylmorphine, heroin, codeine, cocaine, methamphetamine, ketamine, methadone, and dolantin in some practical cases.
Keywords: Direct analysis in real-time mass spectrometry, forensic toxicological analysis, illicit drugs, rapid screening
|How to cite this article:|
Zhang Y, Zhang W, Xin G, Liu L, Duan X, Liu C. Rapid screening of nine illicit drugs in human blood and urine by direct analysis in real-time mass spectrometry. J Forensic Sci Med 2019;5:136-40
|How to cite this URL:|
Zhang Y, Zhang W, Xin G, Liu L, Duan X, Liu C. Rapid screening of nine illicit drugs in human blood and urine by direct analysis in real-time mass spectrometry. J Forensic Sci Med [serial online] 2019 [cited 2019 Oct 17];5:136-40. Available from: http://www.jfsmonline.com/text.asp?2019/5/3/136/267150
| Introduction|| |
Mass spectrometry (MS) plays a vital role in analyzing illicit drugs and toxicants of forensic applications. With the development of the ionization approach, people have been focused on the ambient ionization methods recently. Rapid and accurate screening for illicit drugs is an important topic in a forensic toxicological analysis. MS is one of the most useful techniques for this purpose due to its high sensitivity, specificity, and accuracy. However, in forensic laboratory, the complex biological samples such as blood, urine, and tissues are generally encountered. Using gas chromatography (GC)-MS or liquid chromatography (LC)-MS to analysis, these samples generally required tedious sample preparations to achieve good identifiable accuracy and instrumental sensitivity. Direct analysis in real-time-MS (DART-MS) is an ambient noncontact ionization MS and has emerged as a practical analysis method for a variety of samples due to no or little sample pretreatment was involved, which is attractive for forensic toxicological analysis., Herein, we introduced a rapid screening approach, utilizing DART ambient ionization source integrated with a tandem quadrupole mass spectrometer, for the detection of nine general illicit drugs in human blood and urine samples with minimum sample preparation., In addition, this DART-MS/MS screening method was evaluated by analyzing the positive samples confirmed by GC-MS from real forensic cases. The results by the new method are in consistent with those by GC-MS.
| Materials and Methods|| |
A DART SVP ionization source (IonSense), coupled with a QTRAP 4000 mass spectrometer (SCIEX) through a VAPUR (IonSense) interface was employed in this study. Measurements were performed in positive ion mode with multiple reaction monitoring (MRM) of transitions with optimized MS conditions. Unless otherwise specified, the DART gas (He) flew at 2.5 L/min; gas beam temperature was at 400°C; and the sliding speed of the 12-Dip-It module was 0.6 mm/s. The spectral acquisition was conducted with a unit resolution. The MRM scanning was achieved at 100 ms dwell time for each of the pairs. The Q-trap 4000 was scanned at a mass range of m/z 50–1000. Data acquisition and analysis were conducted using the analyst sozftware (SCIEX) IonSense (USA) SCIEX (USA).
Collection of the samples
We have selected 20 samples from positive cases that we met in routine work. The specimens in this study were from autopsy. They are heart blood or urine. Samples were stored in refrigerators at −20°C. This study was performed in accordance with the Chinese law and the relevant ethical standards.
The blood and urine samples were processed by adding 2 volumes acetonitrile-methanol (V/V = 4:1) to precipitate the proteins. After being centrifuged, the upper clear solution was directly submitted to DART-MS/MS analysis.
Three replicates of each sample were carried out automatically using 12-Dip-It™ sampler module (IonSense). The closed end of a sampling glass rod dipped into the sample solution and immersed for 1 s. The sampling rod was inserted between the DART ® gun exit and VAPUR for full scanning, product ion scanning, and MRM. [Table 1] shows the optimization mass results of the nine illicit drugs, concluding precursor ion, production, desorption voltage, and collision energy.
| Results and Discussions|| |
A fast screening approach for the detection of nine illicit drugs (morphine, o6-monoacetylmorphine, heroin, codeine, cocaine, methamphetamine, ketamine, methadone, and dolantin) in blood and urine matric were established, using a tandem quadrupole MS integrated with DART ionization source (DART-MS/MS). These samples were extracted by acetonitrile, methanol, ethanol, and solvents mixed in different proportions. Moreover, the optimizing result of the solvent was acetonitrile-methanol (V/V = 4:1). The temperature of the ionization source was at 400°C generally. Nine illicit drugs were studied, and the limited detection was setup. Their detection limits (morphine, O6-monoacetylmorphine, heroin, codeine, cocaine, methamphetamine, ketamine, methadone, and dolantin) were 100, 50, 50, 100, 20, 20, 10, 1, and 0.01 ng/mL, respectively. In addition, this DART-MS/MS screening method was evaluated by analyzing the positive samples confirmed by GC-MS from real forensic cases. The results by the new method are in consistent with those by GC-MS [Table 2].
|Table 2: The LOD and 20 positive cases results of the nine illicit drugs|
Click here to view
The standard substance of 100 ng/ml was used to optimize its MS condition with a tandem quadrupole MS. We chose two or three precursor ions/product ions that were optimized. Two or three ion pairs were selected for each substance, and each ion pair was optimized with DP and CE to obtain the optimization data.
In 20 positive cases that we selected, six detected morphine, three detected O6-monoacetylmorphine, five detected codeine, two detected cocaine, and four detected methamphetamine. The results were consistent with GC/MS [Figure 1], [Figure 2], [Figure 3], [Figure 4].
|Figure 2: Screening morphine, O6-monoacetylmorphine, and codeine in human blood from the positive cases|
Click here to view
|Figure 4: Screening methamphetamine in human urine from the positive cases|
Click here to view
The four below representative MRM spectra of morphine, codeine, O6-monoacetylmorphine, cocaine, methamphetamine in human blood or urine samples are from positive cases.
| Conclusion|| |
Postmortem forensic toxicological analysis of human blood, urine and other body fluid is always challenging. Forensic analysis commonly requires the detection of drugs in human blood, urine, or other body fluids that are difficult to analyze. DART-MS/MS provides sufficient sensitivity for analysis of nine illicit drugs. This method has a simple sample preparation and is much faster than GC-MS or liquid chromatography-mass spectrometry. Its sensitivity is also sufficient for complex matrices such as blood or urine. Preliminary, most published studies on the DART-MS technique have focused on in vitro samples ,,,,, and only a few reports have documented its use for analytes in human blood or urine specimens ,,. In some reports, the sensitivity of DART-Time of Flight-TOF-mass spectrometry was not sufficient. Later-by the development of the mass spectrometry and the DART source, a series of experiments shows that the sensitivity is high . In this study, we evaluated DART-MS/MS for the analysis of nine illicit drugs in blood and urine samples. The target can be easily extracted out from the complicated matrix by preliminary optimization experiments. The limit of detection was set up that most are below 100ng/mL. We also applied DART-MS/MS to the analysis of some confirmed positive case samples and found that the results were consistent with those from GC-MS. Our results demonstrate that DART-MS/MS is a simple, rapid, and cost-effective tool for high-throughput sample analysis in real time. The method requires only several seconds per sample and minimal quantities of samples are consumed. It does not require complicated sample preparation, and cleanup of proteins can be achieved with acetonitrile-methanol (4:1, v/v)which is two volumes of samples. The results demonstrated this rapid screening method is suitable for forensic toxicological screening of suspected illicit drugs in blood and urine. However, many aspects need to be investigated in future studies. Supplementary techniques are required to confirm the results in the absence of the standard substances. And it needs to establish the database for the compounds. During using of the DART source, We found that in the process of using DART source-The implementation of a DART source with fixed flow and pressure, as well as good thermal control enabled improvements in sampling reliability during analysis.
Financial support and sponsorship
This research was supported by the Project for Strengthening the Police Force with Science and Technology (project number 2018GABJC29).
Conflicts of interest
There are no conflicts of interest.
| References|| |
Vaiano F, Busardo F P, Palumbo D, et al.
A novel screening method for 64 new psychoactive substances and 5 amphetamines in blood by LC-MS/MS and application to real cases[J]. Journal of Pharmaceutical and Biomedical analysis, 2016, 129:441-449.
Lesiak A D, Shepard J R E. Recent advances in forensic drug analysis by DART-MS[J]. Bioanalysis,2014,6(6):819-842.
Jeri D. Ropero-Miller, Peter R. Scout. Forensic Toxicology Research and Development Evaluation of New and Novel Direct Sample Introduction, Time of Flight Mass Spectrometry(AccuTOF-DART) Instrument for Postmortem Toxicology Screening, Final Report. Award Number: 2006-DN-BX-K014. Document No: 224522.
EshwarJagerdeo and Mohamed Abdel-Rehim. Screening of Cocaine and Its Metabolites in Human Urine Samples by Direct Analysis in Real-Time Source Coupled to Time-of-Flight Mass Spectrometry After Online Preconcentration Utilizing Microextraction by Packed Sorbent. J. Am. Soc. Mass Spectrom.2009, 20, 891-899.
Andrew H. Grange and G. Wayne Sovocool. Detection of illicit drugs on surfaces using direct analysis in real time (DART) time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom., 2011, 25, 1271-1281.
W. C. Samms, Y. J. Jiang, M. D. Dixon et al.
Analysis of Alprazolam by DART-TOF Mass Spectrometry in Counterfeit and Routine Drug Identification Cases, J. Forensic Sci., 2011, 56, 993-998.
S. E. Edison, L. A. Lin and L. Parales. Practical considerations for the rapid screening for pesticides using ambient pressure desorption ionisation with high-resolution mass spectrometry. Food Additives and Contaminants., 2011, 25, 127–139.
S. E. Edison, L. A. Lin and B. M. Gamble et al.
Surface swabbing technique for the rapid screening for pesticides using ambient pressure desorption ionization with high-resolution mass spectrometry, Rapid Commun. Mass Spectrom., Vol. 28, No. 10, October 2011, 1393–1404.
Wang L, Pan CP et al.
Direct analysis in real time mass spectrometry for the rapid identification of four highly hazardous pesticides in agrochemicals. Rapid Commun. Mass Spectrom, 2012,26:1859-1867.
Tiina J. Kauppila, Ville Arvola et al.
Direct analysis of illicit drugs by desorption atmospheric pressure photoionization. Rapid Commun. Mass Spectrom., 2008, 22:979-985.
Eshwar Jagerdeo and Mohamed Abdel-Rehim. Screening of Cocaine and Its Metabolites in Human Urine Samples by Direct Analysis in Real-Time Source Coupled to Time-of-Flight Mass Spectrometry After Online Preconcentration Utilizing Microextraction by Packed Sorbent. J. Am. Soc. Mass Spectrom.2009, 20, 891-899.
Joseph LaPointe, Brian Musselman, Teresa O' Neill et al.
Detection of “Bath Salt” Synthetic Cathinones and Metabolites in Urine via DART-MS and Solid phase Microextraction. J. Am. Soc. Mass Spectrom.2015, 26, 159-165.
Wilcox F. Duvivier, Teris A. van Beek et al.
Rapid analysis of Δ-9-tetrahydrocannabinol in hair using direct analysis in real time ambient ionization orbitrapmass spectrometry. Rapid Commun. Mass Spectrom.2014, 28, 682-690.
Ashton D Lesiak et al.
Recent advances in forensic drug analysis by DART-MS. Bioanalysis.2014, 6(6), 819-842.
[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2]