Application

Note: 432
Identification of Lysergic Acid Diethylamide (LSD)
in Candy by UHPLC/MS
Jason R. Stenzel, Washington State Patrol – Crime Laboratory Division, Cheney, WA, USA
Guifeng Jiang, Thermo Fisher Scientific, San Jose, CA, USA

Goal Experimental Conditions

Key Words Positively identify trace amounts of lysergic acid 1. Standard and Sample Preparation
• Accela™ UHPLC diethylamide (LSD) in sugar candy quickly, with minimal
A 1000 mg/L solution of LSD in methanol was purchased
sample preparation and no chemical derivatization.
• MSQ Plus™ from Alltech (State College, PA, USA) and diluted to
about 5 mg/L with methanol.
• Hypersil Introduction
The sugar cubes and candy hearts were purchased
GOLD™ PFP Lysergic acid diethylamide (LSD) is a controlled substance from a local grocery store. The candy hearts and sugar
in forensic chemistry that is notorious for being difficult to cubes were treated with 3-5 drops of this LSD solution and
• Forensic Analysis
identify. Its myriad evidentiary forms include paper tabs, allowed to stand for 24 hours prior to use. Ten (10) mg
• LSD eye drops, sugar cubes and small sugary candies such as scrapings from the sugar cube or candy heart were added
sweet tarts, valentine hearts or mints. Because it is such a to 2 mL methanol. This mixture was vortexed for 30 sec,
potent hallucinogen, typical street doses require only 40 to allowed to settle for 1 min, and the supernatant was
120 µg of LSD. The small personal-use amounts seized by filtered through a cotton-plugged Pasteur pipette. The
state and local law enforcement often lack sufficient drug filtrate was centrifuged for 90 sec, and the supernatant
to allow both forensic analysis by traditional means and was filtered through a second cotton-plugged Pasteur
archiving of some of the evidence for follow-up testing. pipette and transferred to the autosampler vial.
Most forensic laboratories confirm the presence of LSD
by using gas chromatography with mass spectrometry 2. Chromatographic Conditions
(GC/MS). LSD is extracted from the evidence with an Chromatographic analyses were performed using the
organic solvent, derivatized, and determined by GC/MS. Accela UHPLC system (Thermo Fisher Scientific, San Jose,
GC/MS resolves LSD from other compounds and provides CA). The chromatographic conditions were as follows:
structural information that can be compared to reference
spectra in a searchable library. Column: Hypersil GOLD PFP (perfluorinated phenyl) 1.9 µm,
The disadvantages of GC/MS are its requirements 100 x 2.1 mm
for extensive sample preparation, including chemical Flow Rate: 1 mL/min
derivatization of LSD to a more volatile form, and its Mobile Phase: A: Water with 0.06 % acetic acid
impaired performance with analytes that are polar, B: Acetonitrile (ACN) with 0.06% acetic acid
thermally labile, or nonvolatile. LSD has a high affinity C: Methanol with 0.06% acetic acid
for active sites in liners that can spoil chromatographic Gradient: T (min) A% B% C%
resolution. LSD-doped sugar cubes or candy can foul the 0.00 95.0 0.0 5.0
1.00 95.0 0.0 5.0
GC with sugars, increasing the burden of instrument 1.50 90.0 5.0 5.0
maintenance and hindering throughput. 2.70 70.0 10.0 20.0
An alternative method to positively identify LSD in 3.00 5.0 15.0 80.0
complex food matrices is to use ultra high performance 7.00 5.0 0.0 95.0
liquid chromatography with mass spectrometric detection 7.10 95.0 0.0 5.0
8.00 95.0 0.0 5.0
(UHPLC/MS). UHPLC/MS offers a threefold benefit
Column Temperature: 45 °C
compared to GC/MS; simpler sample preparation, no
Injection: 2 µL partial loop injection, 25 µL loop size
derivatization, and less time wasted baking out or cleaning Syringe Speed: 8 µL/sec
the instrument. This application note demonstrates how a Flush Speed: 100 µL/sec
working forensic laboratory uses UHPLC/MS to analyze Flush Volume: 400 µL
sugar candies for LSD. LSD in doped sugar cubes and Wash Volume: 100 µL
candy hearts is simply extracted, separated within 5 minutes Flush/Wash source: Bottle with methanol
on a Hypersil GOLD PFP 1.9 µm column, and confirmed
by a fast scanning single quadrupole mass spectrometer.
3. Mass Spectrometer Conditions In addition to these
offices, Thermo Fisher
MS analysis was carried out on a MSQ Plus single
Scientific maintains
quadrupole LC/MS detector (Thermo Fisher Scientific,
a network of represen-
San Jose, CA). The MS conditions were as follows:
tative organizations
throughout the world.
Ionization: Electrospray (ESI)
Polarity: Positive
Probe Temperature: 500 °C
Cone Voltage: 90 V
Scan Mode: Full scan with mass range of m/z = 125-425 amu
ESI Voltage: 3.5 kV Africa
Scan Time: 0.2 s +43 1 333 5034 127
Australia
+61 2 8844 9500
Austria
+43 1 333 50340
Results
Belgium
LSD elutes at 4.49 min and is detected by using full scans +32 2 482 30 30
(m/z = 125 - 425) of the single quadrupole mass spectrometer. Canada
+1 800 530 8447
The extracted ion chromatograms from m/z 324 ± 0.5 are
China
displayed in Figure 1A. The MS spectrum of the LSD +86 10 8419 3588
standard shows two molecular ion signals: [M+H]+ at Denmark
m/z 324.1, and [M+ACN+Na]+ at m/z 387.1. The MS +45 70 23 62 60
Figure 1: Extracted ion chromatograms at m/z = 324 ± 0.5 amu obtained by Europe-Other
spectrum of LSD also shows two fragment ion signals at UHPLC/MS on a Hypersil GOLD PFP column: (A) LSD standard; (B) methanol +43 1 333 5034 127
m/z 223.3 and 281.3 (Figure 2A). extract of LSD-doped candy heart; (C) methanol extract of LSD-doped sugar France
The methanol extracts from the candy hearts and sugar cube. See text for details. +33 1 60 92 48 00
cubes, doped with LSD, were analyzed using the same Germany
+49 6103 408 1014
UHPLC/MS method as for the standard LSD (Figure 1B, 1C).
India
Positive confirmation of LSD in the samples is assured +91 22 6742 9434
both by retention time matching and MS spectra matching Italy
+39 02 950 591
of the samples (Figure 2B-C) with the LSD standard.
Japan
+81 45 453 9100
Conclusion Latin America
+1 608 276 5659
UHPLC/MS can positively identify trace amounts of
Middle East
lysergic acid diethylamide (LSD) in sugar candy in 8 min, +43 1 333 5034 127
after a simple 10 min sample prep involving no chemical Netherlands
derivatization. +31 76 579 55 55
South Africa
+27 11 570 1840
Spain
+34 914 845 965
Sweden / Norway /
Finland
Figure 2: MS spectra of LSD +46 8 556 468 00
obtained by UHPLC/MS on a Switzerland
Hypersil GOLD PFP column: +41 61 48784 00
(A) LSD standard; (B) methanol UKg
extract of LSD-doped candy +44 1442 233555
heart; (C) methanol extract of USA
LSD-doped sugar cube. +1 800 532 4752
See text for details. www.thermo.com
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