Automated Direct Sample Analysis (DSA/TOF) for the Rapid Screening and Confirmation of Illicit Street Drugs

A P P L I C AT I O N N O T E
Mass Spectrometry
Authors
Sean Daugherty, Hayley Crowe
PerkinElmer, Inc.
Seer Green, Beaconsfield,
HP9 2FX United Kingdom
Automated Direct
Sample Analysis
(DSA/TOF) for the
Rapid Testing of
Drug Compounds
Introduction
Testing methods for drugs currently require
long chromatographic methods and sample
preparation or immunoassay techniques
(EMIT) with confirmation by GC/MS or
LC/MS/MS. These methods can be timeconsuming and laborious. The majority of
urine testing methods cover only a limited
number of metabolites, since reference
standards are poorly available, making it difficult to keep up with the changing illicit
drug scene. Due to the variety of compounds available, it is important to monitor
the entire spectral range, which is easily performed using a time-of-flight-mass
spectrometer (TOF MS). The ability to analyze samples rapidly is desirable, as drug
seizures may result in thousands of samples. A method was developed using the
AxION® 2 TOF MS integrated with the AxION Direct Sample Analysis™ (DSA™) system
(Figure 1) for testing various classes of street seized, illicit designer drugs, and growth
hormones in solid, liquids as well as metabolites in urine.
This method enables fast screening and confirmation with
minimal or no sample preparation using the AxION DSA
system. We present a real life case study for the analysis of
369 unknown drugs from seized pills, vials, powders and
urine samples (Figure 2). The study shows the suitability of
Figure 1: AxION DSA System
Experimental conditions:
Traditional sample analysis using
SPE/LC/MSMS v. DSA/TOF
Figure 2: Seized street samples and DSA solids and liquids rack (foreground).
this method and instrumentation and also the ease of use and quick
implementation possible of this solution into a laboratory due to
the simple user interface, holistic design and visual reporting (Figure
3). At the end of the first day of installation of the instrument,
sample results were being reported. Rich spectral information for
testing drug compounds takes only 20 seconds, without the need
for classical mass spec training. The presence of the target analytes
was confirmed using AxION Solo™ software by exact monoisotopic
masses of precursor ion and fragment ions. Identification was
performed by reverse automatic database search with set criteria
for mass accuracy, isotopic pattern and abundance. A database
containing all schedule 1-5 drugs as well as other substances to
contain more than 1,000 compounds of illicit and commonly abused
drugs and pharmaceuticals was used. Additional databases added in
minutes. Samples were confirmed using an orthogonal confirmation
methodology to validate the accuracy of this method against current
industry standards.
In addition to the previous advantages described, we can now
perform rapid screening of samples with automated sample
analysis for many different sample types including solid, liquid and
gas. Samples are introduced via disposable mesh and glass capillaries.
The system is fully enclosed, providing decreased noise for excellent
Traditional sample analysis steps - 25 mins:
Wet SPE cartridge à Condition cartridge à Pipette
sample à Dispense sample à Wash salts à Elute
cartridge à Collect eluent à Transfer eluent à
Run analysis à Confirm results
DSA/TOF sample analysis - 25 seconds:
Pipette sample à Load instrument à Acquire spectra
à Confirm results
Sample Prep: Minimal to no sample prep
5 µl for liquid samples was used, no sample prep
(figure 4a - 4b)
Pills and other solids were crushed and analyzed either
directly as solids or after dissolving with MeOH
Urine was diluted 20:1 with 50:50 methanol:water
and directly pipetted onto the mesh
LC conditions: None required using DSA
Pump: PerkinElmer Flexar™ FX-10 or 15 pumps
recommended when running LC-TOF; not required
for DSA
MS conditions:
Mass Spectrometer: PerkinElmer AxION 2 TOF MS
Ionization Source: AxION DSA
Ionization Mode: Positive
Spectral Acquisition Rate: 5 spectra/sec
Capillary Exit Voltage: 100 V
Pulse and TrapPulse Mode: 50-650 m/z (D7:38, D8:52)
Detector Voltage: 5000 and 8000 V
Mass Calibration:
A 2nd order external mass calibration was performed
in positive ionization mode from an infusion of
5 calibration masses.
Internal calibration was performed using m/z
121.05087 and 622.02896 as lock mass ions.
Figure 3: AxION DSA and AxION Solo: a holistic design that enables true one-click
automated sample analysis.
Figure 4a: AxION DSA liquid rack,
5 ul applied.
2
Figure 4b: AxION DSA solid’s rack.
DSA Parameters:
DSA Heater Temperature: 250 oC
Auxiliary Gas: 80 psi
Drying Gas Flow Rate: 3 L/min
Drying Gas Temperature: 25 oC
Corona Needle Voltage: 2200 V
signal to noise, and is holistically designed with simple control
and dedicated target and unknown analysis software. Sources
are easily interchangeable to enable switching from DSA/
TOF to LC/TOF in minutes to allow for a flexible laboratory
workflow. Testing is performed in a single sample analysis of
targeted substances using accurate mass and isotope ratio
supported by the AxION Solo software as well as confirmation
of targeted unknowns, with the AxION EC ID software.
AxION Solo uses accurate mass, isotope ratio and collision
induced dissociation (CID) accurate mass fragments enabling
confirmation by:
•
•
•
•
Monoisotopic mass of the precursor ion
Isotope ratios of the precursor ion
Accurate mass of multiple fragments
Isotope ratios of fragments
Results
369 unknown samples were tested using the AxION DSA/
TOF and AxION Solo. Due to the simple nature of the control
software (Figure 5), the samples can be easily analyzed
without complicated parameter setup and targets screened
automatically. 53 unique substances were detected in this
study, (Table 1). Each sample took less than 25 seconds. This
is a total of 154 minutes or approximately 2.5 hours run and
post data analysis time. No sample prep was required for many
of the samples, but some crushing and MeOH extractions/
dilutions were performed totaling less than 4 hours for analysis
of all samples. All samples were analyzed, confirmed and
reported in a single morning using the AxION Solo Drugs of
Abuse library and post-acquisition data processing software
(Figure 6). As shown in Figure 7, the AxION Solo software easily
screens multiple samples at one time. It provides annotated
mass spectra for all substances detected in the sample that was
run, including mass accuracy in ppm against theoretical. Isotopic
ratio matching for further confirmation and a stop-light visual
summary of substances found/not found against the library,
provide a quick review and confirmation of the drugs detected.
Figure 5: Easy sample submission for the AxION DSA Controller.
Figure 6: AxION Solo Database, extensive list of illicit drugs that are
readily added to in just a few clicks from targeted unknowns determined
using the AxION 2 TOF.
Figure 7: AxION Solo stop light reporting and at-a-glance rich spectral information.
3
Table 1: Identified Substance from Testing of Drug Compounds
Opioids
ELEMENTAL
FORMULA
ION
OBSERVED
THEORETICAL
MASS
MEASURED
MASS
ERROR
(PPM)
SYNTHETIC OPIOID
C21H27NO
[M+H]+
310.2165
310.2171
-1.93
SEMI-SYNTHETIC OPIOIDS
C25H33NO4
[M+H]+
412.2477
412.2491
-3.40
METHYLDESORPHINE
OPIOID ANALGESIC
C18H21NO2
[M+H]+
284.1645
284.1653
-2.82
DIHYDROMORPHINE
SYNTHETIC OPIOID
C17H21NO3
[M+H]+
288.1594
288.1606
-4.16
HYDOMORPHINOL
OPIATE ANALOGUE
C17H21NO4
[M+H]+
304.1543
304.1549
-1.97
MORPHINE
OPIATE ANALGESIC
C17H19NO3
[M+H]+
286.1438
286.1441
-1.05
OXYCODONE
SEMI-SYNTHETIC OPIOIDS
C18H21NO4
[M+H]+
316.1543
316.1552
-2.85
CODEINE
OPIATE
C18H21NO3
[M+H]+
300.1594
300.1587
2.33
DIHYDROCODEINE
SYNTHETIC OPIOID
C18H23NO3
[M+H]+
302.1751
302.1756
-1.65
6-ACETYLMORPHINE 6-MAM
HEROIN METABOLITE
C19H21NO4
[M+H]+
328.1543
328.1535
2.44
EDDP
METHADRON METABOLITE
C20H23N
[M+H]+
278.1903
278.1905
-0.72
ELEMENTAL
FORMULA
ION
OBSERVED
THEORETICAL
MASS
MEASURED
MASS
ERROR
(PPM)
DRUG
DESCRIPTION
METHADONE
ETORPHINE/M99
Steroids
DRUG
DESCRIPTION
TESTOSTERONE PROPIONATE
STEROID HORMONE DERIVATIVE
C22H32O3
[M+H]+
346.2458
346.2446
3.47
TESTOSTERONE DECANOATE
STEROID HORMONE DERIVATIVE
C29H46O3
[M+H]+
443.3520
443.3523
-0.68
NANDROLONE DECANOATE
ANABOLIC STEROID
C28H44O3
[M+H]+
429.3363
429.3362
0.23
STANOZOLOL
ANABOLIC STEROID DERIVATIVE
C21H32N2O
[M+H]+
329.2587
329.2577
3.04
BOLDENONE UNDECYLENATE
ANABOLIC STEROID DERIVATIVE
C30H44O3
[M+H]+
453.3363
453.3344
4.19
METHANDROSTENOLONE
ANABOLIC STEROID
C20H28O2
[M+H]+
301.2162
301.2153
2.99
TESTOSTERONE ENANTHATE
STEROID HORMONE DERIVATIVE
C26H40O3
[M+H]+
401.3050
401.3049
0.25
MESTEROLONE
ANABOLIC STEROID DERIVATIVE DHT
C20H32O2
[M+H]+
305.2475
305.2477
-0.66
TRENBOLONE
LIVESTOCK STEROID
C18H22O2
[M+H]+
271.1693
271.1694
-0.37
NANDRALONE PHENYLPROPIONATE
ANABOLIC STEROID
C27H34O3
[M+H]+
407.2581
407.2591
-2.46
TRENBOLONE ENATHATE
LIVESTOCK STEROID
C25H34O3
[M+H]+
383.2581
383.2584
-0.78
ELEMENTAL
FORMULA
ION
OBSERVED
THEORETICAL
MASS
MEASURED
MASS
ERROR
(PPM)
C10H15NO
[M+H]+
166.1226
166.1228
-1.20
Stimulant, Amphetamine and Other Psychoactive Substances
4
DRUG
DESCRIPTION
4-METHOXYAMPHETAMINE
SEROTONERGIC DRUG AMPHETAMINE CLASS
FLUOROAMPHETAMINE
PSYCHOACTIVE DRUG AMPHETAMINE
C9H12FN
[M+H]+
154.1027
154.1025
1.30
TCP
DISSOCIATIVE ANESTHETIC DRUG
C15H23NS
[M+H]+
250.1624
250.1619
2.00
PARAHEXYL
SYNTHETIC HOMOLOGUE THC
C22H32O2
[M+H]+
329.2475
329.2471
1.21
MDMA
EMPATHOGENIC AMPHETAMINE CLASS
C11H15NO2
[M+H]+
194.1176
194.118
-2.06
N-ETHYLAMPHETAMINE
STIMULANT DRUG AMPHETAMINE CLASS
C11H17N
[M+H]+
164.1434
164.1439
-3.05
TETRAHYDROCANNABINOL (THC)
PSYCHOACTIVE CANNABIS
C21H30O2
[M+H]+
315.2319
315.2330
-3.49
CAFFEINE
STIMULANT
C8H10N4O2
[M+H]+
195.0877
195.0878
-0.51
NRG-2
SYNTHETIC CATHINONE
C12H17NO
[M+H]+
192.1383
192.1385
-1.04
PHENMETRAZINE
STIMULANT MORPHOLINE
C11H15NO
[M+H]+
178.1226
178.1228
-1.12
MEPHEDRONE
STIMULANT AMPHETAMINE/CATHINONE
C11H15NO
[M+H]+
178.1226
178.1229
-1.68
MDPV
PSYCHOACTIVE STIMULATE NDRI
C16H21NO3
[M+H]+
276.1594
276.1589
1.81
DIETHYLCATHINONE
STIMULANT DRUG CATHINONE CLASS APPETITE SUPRESSANT
C13H19NO
[M+H]+
206.1539
206.1527
5.82
FLUOROMETHCATHINONE FMC
STIMULANT AMPHETAMINE/CATHINONE
C10H12FNO
[M+H]+
182.0976
182.097
3.29
COCA LEAVES/COCAINE*
STIMULANT SNDRI TROPANE ALKALOID
C17H21NO4
[M+H]+
304.1543
304.1547
-1.32
Other Identified Substances
ELEMENTAL
FORMULA
ION
OBSERVED
THEORETICAL
MASS
MEASURED
MASS
ERROR
(PPM)
TOPICAL ANALGESIC METABOLITE OF COCAINE
C16H19NO4
[M+H]+
290.1387
290.1389
-0.69
BARBITURATE
C11H18N2O3
[M+H]+
227.1390
227.1381
3.96
TADALAFIL/CIALIS
ERECTILE DYSFUNCTION
C22H19N3O4
[M+H]+
390.1448
390.1459
-2.82
CARBAMAZEPIN
ANTICONVULSANT MOOD STABILZER
C15H12N2O
[M+H]+
237.1023
237.1029
-2.53
AMITRIPTYLINE
TRICYCLIC ANTIDEPRESSIVE
C20H23N
[M+H]+
278.1903
278.1901
0.72
SIBUTRAMINE
ORAL ANOREXIANT
C17H26CLN
[M+H]+
280.1827
280.1839
-4.28
SILDENAFIL/VIAGRA
ERECTILE DYSFUNCTION
C22H30N6O4S
[M+H]+
475.2122
475.2129
-1.47
RISPERIDONE
ANTIPSYCHOTICS SCHIZOPHRENIA
C23H27FN4O2
[M+H]+
411.2191
411.2182
2.19
QUININE
ANTI MALARIAL
C20H24N2O2
[M+H]+
325.1911
325.1923
-3.69
ZOLPIDEM
INSOMNIA MED
C19H21N3O
[M+H]+
308.1758
308.1767
-2.92
CLENBUTEROL
BRONCHIAL DILATOR PERFORMANCE ENHANCING
C12H18CI2N2O
[M+H]+
277.0869
277.0867
0.72
DIAZEPAM
BENZODIAZEPINE
C16H13CLN2O
[M+H]+
285.0789
285.0787
0.70
KETAMINE
NMDA RECEPTOR ANTAGONIST
C13H16CINO
[M+H]+
238.0993
238.0989
1.68
LIDOCAINE
ANESTHETIC USED TO CUT COCAINE
C14H22N2O
[M+H]+
235.1805
235.1796
3.83
BENZOCAINE
SURFACE ANESTHETIC
C9H11NO2
[M+H]+
166.0863
166.0862
0.60
DRUG
DESCRIPTION
BENZOYLECGONINE
PENTOBARBITAL
Conclusion
The AxION DSA/TOF system provides a new solution for the testing of illicit drugs in seconds compared to lengthy sample
preparation and chromatographic run times of 20-60 min. Testing of unknown drugs in samples is performed in seconds
using monoisotopic mass of the precursor ion, isotope ratios of the precursor ion, accurate mass of multiple fragment ions and
isotope ratios of fragment ions using AxION Solo software. AxION EC ID software provides a solution that can identify exact mass
of precursor and fragments of unknown compounds in seconds. These compounds can be screened against public or user-defined
databases and added easily and quickly to your targeted library. If chromatography is desired, AxION DSA can switch to LC/TOF in
a few minutes to adapt to any workflow that you require in your lab.
For Research Use Only. Not for Use in Diagnostic Procedures.
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