Application Note: In-Process Control of Ergoline Psychedelics During Chemical Synthesis by HPTLC

APPLICATION NOTE
In-process control of ergoline psychedelics during

chemical synthesis by HPTLC A-99.1
Keywords
LSD, LSP, LSM, LSZ, mass detection, in-process control, quality control,
HPTLC-MS, scanning densitometry
Introduction
TLC is a simple and convenient tool for monitoring classical organic
syntheses. Standardized HPTLC may additionally provide reliable analytical
endpoints and significantly improved separation. Structure confirmation can
be achieved with HPTLC-MS hyphenation. In addition, HPTLC can be used
as a technique complementary to existing HPLC methods for ensuring a
product of high purity.
Scope
This method is suitable for identification of synthesis products. The CAMAG
TLC-MS Interface 2 is used to directly elute target zones from the HPTLC
plate into the Waters ACQUITY QDa® for mass detection. A second
confirmation can be achieved by recording the UV spectra with the TLC
scanner. A fast way for in-process and quality control during synthesis of
ergoline psychedelics is shown below.
Synthesis of ergoline psychedelics

In a first step lysergic acid (LysS) is produced from ergotamine as starting
material. Then the formation of different amides takes place, with LSD, LSZ,
LSP, and LSM as finished products.
LysS is the starting material for the synthesis of LSD, LSZ, LSP, and
LSM; different side chains are highlighted in red
NOTE: The presented results are to be regarded as examples only!

Please contact CAMAG for more application notes and products!
www.camag.com 1 of 4 02/2016
APPLICATION NOTE
Required or recommended devices

Automatic TLC Sampler 4 or Linomat 5, Automatic Developing Chamber ADC
2, Immersion Device 3, TLC Visualizer, visionCATS software, TLC Scanner,
TLC-MS Interface 2, Waters ACQUITY QDa Detector (Performance),
Empower® or MassLynx® software, optional: UV Cabinet
Sample
Samples from different synthesis steps are applied directly onto HPTLC
plates without further preparation. Depending on the process step the applied
volume can vary as well as the solvent (in most cases dichloromethane
and/or methanol). The purified synthesis products are dissolved in methanol
(1 mg/mL). Samples and purified synthesis products were provided by
Lipomed AG (Arlesheim, Switzerland).
Standards
Methanolic solution of LSD (0.1 mg/mL). Standard was provided by Lipomed
AG (Arlesheim, Switzerland).
Chromatography
Stationary phase: HPTLC Si 60 F254, 20 x 10 cm (Merck).
Sample application: Bandwise application with ATS 4 or Linomat 5, 15 tracks, band length 8 mm,
track distance 11.4 mm, distance from left edge 20 mm, distance from lower
edge 8 mm, application volume between 0.5 und 15 μL for sample solutions,
1 μL for dissolved purified synthesis products and 8 μL for the standard
solution.
Developing solvent: Dichloromethane – methanol – trimethylamine 9:1:0.002 (v/v/v)

In the case of LSZ: dichloromethane – isopropanol – triethylamine 9:1:0.002
(v/v/v)
Development: In the ADC 2 with chamber saturation (with filter paper) 20 min and after
conditioning at 33% relative humidity for 10 min using a saturated solution of
magnesium chloride.
Developing distance: 70 mm (from the lower edge)
Plate drying: Drying 5 min in the ADC 2

Documentation: With the TLC Visualizer under UV 254 and UV 366 nm
Densitometry: Densitometric analyses are performed at 254 nm (absorbance) and

366/>400 nm (fluorescence), slit dimension 5.0 x 0.2 mm, scanning speed
20 mm/s, spectra recording 200 to 500 nm
MS confirmation: The zones to be eluted are marked with a soft pencil under UV 254 nm and/or
UV 366 using the UV cabinet or TLC Visualizer.
Target zones are directly eluted using the TLC-MS Interface 2 with oval
elution head into the ACQUITY QDa Detector at a flow rate of 0.5 mL/min with
methanol (with 0.1% formic acid) – water 90:10 (v/v). For a full scan spectrum
it is recommended to first elute a blank, which can be subtracted from the
spectra of the target zones. For confirmation of substances between 50 and
500 ng per zone are required.

APPLICATION NOTE
+
MS parameter: The ACQUITY QDa Detector is operated in ESI mode using default
parameters. The ESI capillary is set to 0.8 kV, cone voltage to 15 V, and
desolvation temperature at 600 °C. A full scan mass spectrum between m/z
50 and 650 is acquired at a sampling rate of 10.0 points/sec (continuum).
Data processing and evaluation of mass spectra are performed with
Empower. For routine use in quality control Single Ion Recording (SIR) can be
performed.
Results
System Suitability Test (SST):
LSD shows a quenching zone (under UV 254 nm) and a blue fluorescent zone (under UV 366 nm) at
RF ~ 0.43
a) In-process control during synthesis of LSP:
Fig. 1 Chromatograms under UV 254 nm und 366 nm that show the different synthesis steps, including the
purification, 1: LSD as reference, 2: Hydrolysis of lysergic acid from ergotamin, 3: purified lysergic acid (starting
material for the chemical synthesis), 4: crude synthesis product (LSP and side products), 5: Column purification,
6: second column purification, 7: purified LSP; UV spectrum and densitogram of LSP 366/>400 nm (fluorescence)
b) Confirmation of the purified synthesis products:
Fig. 2 HPTLC-MS mass spectra of LSD, LSP, LSM, and LSD, displayed range m/z 250 to 380 and HPTLC Image
Comparison of the purified synthesis products under UV 366 nm (left; tracks 1: LSD as reference; 2: purified LSP,
3: purified LSM, 4: purified LSZ(two product zones due to the formation of the isomers [1]))

APPLICATION NOTE
Acknowledgment
This work was done at Lipomed AG (Arlesheim, Switzerland). We are grateful to Dr. Matthias Grill for
the excellent collaboration.
Literature
[1] David E. Nichols et al., J Med Chem 45 (2002) 4344
[2] Alexander Shulgin et al., TIHKAL: The Continuation, 1997
[3] Matthias Grill and Melanie Broszat, CAMAG Bibliogr Service CBS 115 (2015) 11-12


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APPLICATION NOTE

In-process control of ergoline psychedelics during

Synthesis of ergoline psychedelics

NOTE: The presented results are to be regarded as examples only!

Required or recommended devices

Developing solvent: Dichloromethane – methanol – trimethylamine 9:1:0.002 (v/v/v)

Developing distance: 70 mm (from the lower edge)

Plate drying: Drying 5 min in the ADC 2

Densitometry: Densitometric analyses are performed at 254 nm (absorbance) and

NOTE: The presented results are to be regarded as examples only!

a) In-process control during synthesis of LSP:

b) Confirmation of the purified synthesis products:

NOTE: The presented results are to be regarded as examples only!

NOTE: The presented results are to be regarded as examples only!

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