Determination of Amatoxins in Lepiota brunneoincarnata and Lepiota venenata by High-Performance Liquid Chromatography Coupled with Mass Spectrometry

Figures & data

Figure 1. Basidiomata of Lepiota species. (A) Lepiota brunneoincarnata (MHHNU 31026); (B) L. brunneoincarnata (MHHNU 31030); (C) L. brunneoincarnata (MHHNU 31032); (D) L. venenata (MHHNU 31031; photos A, B, and D by Zuohong Chen; photo C by Xiaolan He).

Table 1. List of samples analyzed in this study.

Scientific name	Species no.	Geographic origin	Poisoning description	Collection time	ITS GenBank accession no.
Lepiota brunneoincarnata	MHHNU 31026	Guangrao County, Shandong Province N37°01′11.73″, E118°26′44.75″, alt. 592 m	A man poisoned, developed acute hepatic failure	2017.8.16	MK095187
L. brunneoincarnata	MHHNU 31030	Dalian City, Liaoning Province N38°54′38.75″, E121°37′47.70″, alt. 60 m		2017.9.6	MK095188
L. brunneoincarnata	MHHNU 31032	Chendou City, Sichuan Province N30°34′22.68″, E104°03′9.10″, alt. 500 m	A woman poisoned, developed acute hepatic failure	2017.9.6	MK095190
L. venenata	MHHNU 31031	Jingzhou City, Hubei Province N30°20′14.40″, E112°14′5.86″, alt.300 m	Two man poisoned, developed acute hepatic failure	2017.9.10	MK095189

Figure 2. Phylogenetic tree generated from maximum likelihood analysis based on ITS sequences. Maximum likelihood bootstraps over 50% and Bayesian posterior probabilities over 0.90 are reported on the branches. ITS sequences of four Lepoita species in this study are indicated in boldface, and the other ITS sequences of related taxa were retrieved from the GenBank.

Figure 3. HPLC profiles of cyclopeptide toxins extracted from L. brunneoincarnata and L. venenata.

Table 2. Exact masses of compounds in this study.

Peak number	Compounds	True mass (Da)	Observed mass (Da)	Peak identification methods
1	β-Amanitin	919.3382	920.3390 [M + H^+], 942.3330 [M + Na^+]	Based on standard compounds and references
2	α-Amanitin	918.3542	919.3561 [M + H^+], 941.3286 [M + Na^+]	Based on standard compounds and references
3	Amanin	903.3432	904.3535 [M + H^+], 926.3332 [M + Na^+]	Based on references
4	Amaninamide	902.3592	903.3681 [M + H^+], 925.3494 [M + Na^+]	Based on references
5	Unknown	–	573.9142 [M + H^+], 596.4311 [M + Na^+]	Based on references
6	Amanin II	903.3432	904.3535 [M + H^+], 926.3332 [M + Na^+]	Based on references

Table 3. Main amatoxin contents in two Lepiota species (mg g⁻¹dry weight).

Scientific name and species no.	α-AMA	β-AMA	Total
L. brunneoincarnata MHHNU 31026	0.85 (±0.05)	0.72 (±0.02)	1.57
L. brunneoincarnata MHHNU 31030	0.72 (±0.05)	0.57 (±0.02)	1.29
L. brunneoincarnata MHHNU 31032	1.73 (±0.01)	0.94 (±0.01)	2.67
L. venenata MHHNU 31031	1.97 (±0.01)	–	1.97