Analysis of 6 Alkaloids Common to Psychedelic Mushrooms on Force Biphenyl by HPLC-UV

	Peaks	t_R (min)
1.	N,N-desmethyl Psilocybin (Norbaeocystin)	1.314
2.	N-desmethyl Psilocybin (Baeocystin)	1.750
3.	Aeruginascin	2.173
4.	Psilocybin	2.324
5.	4-hydroxy-N-Methyltryptamine (Norpsilocin)	2.784
6.	4-hydroxy-N,N-Dimethyltryptamine (Psilocin)	3.296

Column

Force Biphenyl (cat.# 962931E)

Dimensions:

100 mm x 3.0 mm ID

Particle Size:

3 µm

Pore Size:

100 Å

Guard Column:

Force Biphenyl EXP Guard Cartridge 5 mm, 3.0 mm ID (cat.# 962950253)

Temp.:

50 °C

Standard/Sample

Diluent:

Water

Conc.:

50 ppm

Inj. Vol.:

1 µL

Mobile Phase

Water, 10 mM ammonium formate, 0.1% formic acid

Methanol, 0.1% formic acid

Time (min)	Flow (mL/min)	%A	%B
0.00	0.8	95	5
4.00	0.8	65	35
4.01	0.8	5	95
5.00	0.8	5	95
5.01	0.8	95	5
8.00	0.8	95	5

Detector	UV/Vis @ 222 nm
Flow Cell Size:	500 nL
Instrument	Waters ACQUITY UPLC H-Class
Sample Preparation	Standards were prepared in a 2.0 mL amber short-cap vial (cat.# 21142) with a 250 µL glass insert (cat.# 24518) and capped with a 9 mm short cap (cat.# 24497).
Notes	System and column were passivated using LC Passivation Solution (cat.# 32475). The outlined method conditions were used to perform five injections of passivation solution (5 µL) on column, diverting to waste. Five warm-up injections were completed prior to analysis. Storage Conditions: Flush column using acetonitrile with 0.1% formic acid for 5 min.

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