Table 4.
Analytical methods for detection and quantification of psychedelic indole alkaloids in mushrooms.
| Indole Type | Sample (Alkaloids Detected) |
Analytical Technique |
Method | Ref. |
|---|---|---|---|---|
| Tryptamines |
P. cubensis and Copelandia spp. (Psilocin, psilocybin) |
HPLC-DAD | Symmetry RP C18 column (150 × 2.1 mm, 5 µm). MP 10 mM ammonium formate buffer (pH 3.5) and ACN (95:5, v/v), at a flowrate of 0.2 mL/min for 20 min. Detection at 220 nm. | [61] |
|
P. cubensis (Psilocin, baeocystin, psilocybin, aeruginascin) |
HPLC-ESI-MS | Zorbax Eclipse Plus RP C18 column (100 × 2.1 mm, 1.8 μm). MP 10 mmolL−1 ammonium formate with 0.1% (v/v) formic acid and 10 mmolL−1 ammonium formate with 0.1% (v/v) formic acid in methanol. Flow rate 0.25 mL min−1 for 7 min. For MS detection, a triple quadrupole 6460 spectrometer was used with positive ESI ionization in the dynamic multiple reaction monitoring (dMRM) acquisition mode. |
[56] | |
|
P. mexicana (psilocin, baeocystin, psilocybin) |
HPLC-ESI-HRMS | RP C18 column (250 × 4.6 mm, 3 μm) with MP 0.1% C18 column TFA in water and ACN in gradient mode. Flow rate of 0.4 mLmin−1. Detection with HRMS using an exact Orbitrap spectrometer and electrospray ionization in positive mode. | [50] | |
|
Psilocybe spp. and Panaeolus cyanescens (psilocybin) |
HPLC-FL | C18 column (150mm × 4.6 mm, 3 μm). MP 50mM ammonium acetate (AcONH4)–CH3CN (73:27). Isocratic elution and rate flow 1.0 mLmin−1. FL detector at 39 nm (excitation at 321 nm). | [72] | |
| β-carbolines |
P. mexicana (Harmane, harmine, harmol) |
HPLC-ESI-MS | RP C18 column (250 × 2.1 mm, 10 mm ID). MP 0.1% TFA in water and ACN. Flow 2 mLmin−1 with linear gradient with an increase from 10 to 100% ACN for 20 min. |
[50] |
|
Mycena metata (Metatacarbolines) |
HR-MALDI-MS | MALDI-MS imaging of the caps saturated solution in 80% MeOH and 1% TFA, ImagePrep device (Bruker Daltonics). | [71] | |
| Cortinarius brunneus (Brunneins) | HPLC-ESI-MS | RP C18 column (ODS 150 × 2.0 mm i.di, 5 μm). MP water and ACN with FA 0.2% (10:90). Using an isocratic mode for 15 min with flow 0.5 mLmin−1. | [65] | |
| Ergot |
C. purpurea (Lysergic, isolysergic, and paspalic acids) |
CE-UV | P/ACE 2200 CE system with capillary (37 cm × 50 μm ID, 360 μm). The voltage applied was 25 kV, time for the separation was 12 min, and UV detection at 214 nm. | [67] |
|
C. purpurea (Ergometrine, ergotamine, ergosine, ergocryptine) |
LC-MS/MS | RP C18 column (150 × 2.1 mm, 3.5 μm). MP water/0.2 M ammonium bicarbonate and methanol/0.2 M ammonium bicarbonate pH 10 at a flow rate of 0.15 mL/min in gradient mode. Detection with MS using triple quadrupole mass spectrometer with positive electrospray ionization. |
[73] |