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Article

Toxin Profiling of Amanita citrina and A. sinocitrina: First Report of Buiotenine Detection

by
Yi-Zhe Zhang
1,†,
Yi Yao
2,†,
Kai-Ping Zhang
3,†,
Jia-Qi Liang
4,
Jia-Ju Zhong
1,
Zhong-Feng Li
1,
Hai-Jiao Li
1,* and
Fei Xu
2,5,*
1
State Key Laboratory of Trauma and Chemical Poisoning, National Institute of Occupational Health and Poison Control, Chinese Centre for Disease Control and Prevention, No. 29, Nanwei Road, Xicheng District, Beijing 100050, China
2
School of Public Health, Ningxia Key Laboratory of Environmental Factors and Chronic Diseases Control, Ningxia Medical University, No. 1160, Shengli South Road, Xingqing District, Yinchuan 750004, China
3
Zichuan District National Forest Farm, Zibo 255100, China
4
Center for Disease Control and Prevention of Yiyang, Yiyang 413000, China
5
Physical and Chemical Department, Ningxia Hui Autonomous Region Center for Disease Control and Prevention, Yinchuan 750004, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Toxins 2025, 17(5), 247; https://doi.org/10.3390/toxins17050247
Submission received: 13 April 2025 / Revised: 6 May 2025 / Accepted: 8 May 2025 / Published: 16 May 2025
(This article belongs to the Special Issue Advances in Poisonous Mushrooms and Their Toxins)
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Abstract

:
Amanita species are widely distributed worldwide. Many of these species are poisonous and can cause health problems, resulting in morbidity and mortality. The toxins responsible for poisoning are amatoxins, aminohexadienoic acid, ibotenic acid, muscimol and muscarines, which damage the liver, kidney, central nervous system and parasympathetic nervous system. In recent years, several toxins have been discovered from different poisonous mushrooms. In this study, multiwalled carbon nanotube purification and ultrahigh-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) was used for the sensitive detection and targeted quantitative screening of 12 mushroom toxins (muscarine, two isoxazole derivatives, three tryptamine alkaloids, three amatoxins and three phallotoxins) from Amanita citrina, A. citrina var. grisea and A. sinocitrina. This study found that buiotenine, one of the tryptamine alkaloids, was detected in A. citrina and A. sinocitrina with an average content of 2.90 and 1.19–6.70 g/kg (n = 3) in the dried mushrooms, respectively. None of the 12 common toxins were discovered in A. citrina var. grisea. These results provide reference data for future research on the role of toxins in the evolution of Amanita mushrooms. Future studies should explore the biosynthetic pathways and ecological roles of these toxins in Amanita species.
Keywords:
Amanita; poisonous mushroom; species identification; toxin detection; tryptamine alkaloids
Key Contribution: Based on morphological and molecular biology methods, Amanita citrina, A. citrina var. grisea and A. sinocitrina were successfully identified from China. The mushroom toxins contained in A. citrina, A. citrina var. grisea and A. sinocitrina were analyzed using ultrahigh-performance liquid chromatography–triple quadrupole linear ion trap mass spectrometry (UPLC–MS/MS). Buiotenine was detected as one of the tryptamines in A. citrina and A. sinocitrina.

Graphical Abstract

1. Introduction

Amanita Pers. species are economically and ecologically important, with >700 accepted species worldwide [1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40]. Based on recent comprehensive phylogenetic studies, Amanita is divided into three subgenera (Amanita, Amanitina and Lepidella) and eleven sections [16,24,39,41]. Interestingly, the species edibility is more or less similar in the different sections [18,19,24,42].
Many Amanita species are famous edible species. For example, Amanita caesarea (Scop.) Pers. from Amanita sect. Caesareae is a common delicious wild mushroom in Europe [20,43,44]. Most species of Amanita sect. Caesareae and some species of sect. Roanokenses and sect. Amanita in China are edible and are widely collected and consumed, such as A. caojizong Zhu L. Yang; Yang-Yang Cui & Qing Cai; A. caesareoides Lj. N. Vassiljeva; A. hemibapha (Berk. & Broome) Sacc.; A. rubromarginata Har. Takah.; A. kitamagotake N. Endo & A. Yamada; A. ochracea (Zhu L. Yang) Yang-Yang Cui, Qing Cai & Zhu L. Yang; A. princeps Corner & Bas; A. pseudoprinceps Yang-Yang Cui, Qing Cai & Zhu L. Yang; A. rubroflava Yang-Yang Cui, Qing Cai & Zhu L. Yang; A. subhemibapha Zhu L. Yang, Yang-Yang Cui & Qing Cai and A. sinensis Zhu L. Yang [18,19,24,45].
In contrast, many Amanita species are poisonous or even deadly, resulting in acute liver and renal failure and psychoneurological disorders [18,19,24,42,46,47,48,49,50,51,52,53,54,55,56,57,58]. The most notorious lethal Amanita are from Amanita sect. Phalloideae, with >70 species producing cyclic peptide toxins, responsible for >70% of all mushroom poisoning fatalities [42,49,50,51,52,53,54,55,56,57,58,59]. Cyclic peptide toxins are classified into three major groups: amatoxins, hallotoxins and virotoxins, with 39 toxins reported [59,60,61,62,63,64,65,66,67,68,69]. The most famous lethal Amanita species in Europe and North America include A. phalloides (Vaill. ex Fr.) Link, A. verna Bull. ex Lam., A. virosa Bertill., A. ocreata Peck, A. bisporigera G.F. Atk., A. suballiacea (Murrill) Murrill and A. tenuifolia (Murrill) Murrill [60,64,70,71,72]. In China, 12 deadly Amanita have been reported, and the most common species in mushroom poisoning outbreaks include A. exitialis Zhu L. Yang & T. H. Li, A. fuliginea Hongo, A. rimosa P. Zhang & Zhu L. Yang, A. subjunquillea S. Imai, A. pallidorosea P. Zhang & Zhu L. Yang, A. subpallidorosea Hai J. Li, A. fuligineoides P. Zhang & Zhu L. Yang and A. subfuliginea Q. Cai, Zhu L. Yang & Y.-Y. Cui [42,52,73,74,75,76].
Thirteen species of mushroom, including A. abrupta Peck, A. boudieri Barla, A. echinocephala (Vittad.) Quél., A. gracilior Bas & Honrubia, A. gymnopus Corner & Bas, A. kotohiraensis Nagas. & Mitani, A. nauseosa (Wakef.) D.A. Reid, A. neoovoidea Hongo, A. oberwinklerana Zhu L. Yang & Yoshim. Doi, A. ovoidea (Bull.) Link, A. proxima Dumée, A. pseudoporphyria Hongo and A. smithiana Bas, produce aminohexadienoic acid (mainly 2-amino-4,5-hexadienoic acid) and cause acute renal failure [54,55,56,57,58,77,78,79,80,81,82,83,84,85]. In China, A. oberwinklerana, A. pseudoporphyria, A. neoovoidea and A. gymnopus are the most common species responsible for mushroom poisoning incidents [42,52,54,55,56,57,58,83,86].
Most species in Amanita sect. Amanita contain neuropsychiatric toxins, including isoxazole derivatives and muscarine [18,19,38,42,52,87]. More than 100 species of sect. Amanita have been reported worldwide ([http://www.amanitaceae.org] on 1 February 2025), and 28 taxa have been recognized in China [18,19,24,38,40]. In Europe and North America, A. muscaria (L.) Lam. and A. pantherina (DC.) Krombh. are the two most common species responsible for poisoning incidents. A. aprica J. Lindgr. & Tulloss, A. gemmata (Fr.) Bertill., A. regalis (Fr.) Michael and A. ibotengutake T. Oda et al. are also poisonous [87,88,89,90,91,92]. In China, >10 species, including A. subglobosa Zhu L. Yang; A. sychnopyramis f. subannulata Hongo; A. concentrica T. Oda, C. Tanaka & Tsuda; A. melleiceps Hongo; A. parvipantherina Zhu L. Yang, M. Weiss & Oberw.; A. pseudosychnopyramis Y.Y. Cui, Q. Cai & Zhu L. Yang; A. rufoferruginea Hongo; A. siamensis Sanmee, Zhu L. Yang, P. Lumyong & Lumyong; A. ibotengutake; A. melleialba Zhu L. Yang, Q. Cai & Y.Y. Cui; A. orientigemmata Zhu L. Yang & Yoshim. Doi; A. pseudopantherina Zhu L. Yang ex Y.Y. Cui, Q. Cai & Zhu L. Yang and A. griseopantherina Y.Y. Cui, Q. Cai & Zhu L. Yang, were discovered in mushroom poisoning incidents [42,52,54,55,56,57,58].
Some species of Amanita sect. Validae also exhibit neurotoxicity. A. citrina Pers. does not contain isoxazole derivatives and muscarine but produces buiotenine at a concentration of 1.6–7.5 mg/g dry weight [59,93,94,95]. Amanita porphyria Alb. & Schwein.: Fr. has been reported to contain 5-hydroxytryptophan [94].
Amanita citrina var. grisea (Hongo) Hongo was first described in Japan and was later reported in China [24,96,97,98]. In 2001, A. sinocitrina, a new species morphologically similar to A. citrina, was described in China [99]. Whether these two taxa also have similar toxins is a fascinating question. While A. citrina has been reported to contain buiotenine, the chemical profiles of var. grisea and A. sinocitrina remain unknown. To determine this, we conducted a study using ultrahigh-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) for toxin profiling and ITS sequencing for phylogenetic analysis, comparing the chemical and genetic diversity among the three species. Our findings showed that buiotenine, one of the tryptamine alkaloids, was detected in A. citrina and A. sinocitrina, and none of the 12 common toxins were discovered in A. citrina var. grisea. The detailed description of species identification and toxin analysis is shown in this study below.

2. Results

2.1. Mushroom Identification

Based on morphological and phylogenetic analyses, all 18 specimens were successfully identified, including 1 specimen of Amanita citrina, 1 specimen of A. citrina var. grisea and 16 specimens of A. sinocitrina (Table 1; Figure 1 and Figure 2).

2.2. Methodology Examination and Toxin Detection Results

UPLC–MS/MS targeted screening results showed that buiotenine was detected in A. citrina and A. sinocitrina. None of the other 11 toxins, including muscarine, psilocybin, psilocin, ibotenic acid, muscimol, α-amanitin, β-amanitin, γ-amanitin, phallacidin, phallisacin and phalloidin, were detected (Figure 3). In addition, none of the toxins were detected in A. citrina var. grisea. In quantitative analysis, the limits of detection (S/N = 3) and limits of quantification (S/N = 10) of buiotenine in the matrix blank sample were 0.1 and 0.2 mg/kg, respectively. Buiotenine showed good linearity (R2 > 0.9960), the three levels of recoveries (10, 20 and 100 mg/kg) of buiotenine in spiked L. edodes samples were 92.3–96.5% and the relative standard deviations were 3.7–5.6%. This method has certain advantages in the extraction and purification of mushroom toxins. Statistical comparisons were made using one-way analysis of variance. UPLC–MS/MS analyses showed that the average contents of buiotenine were 2.90 ± 0.27 g/kg in A. citrina and ranged from 1.19 ± 0.02 to 6.70 ± 1.36 g/kg in A. sinocitrina (n = 3), respectively, in the Guizhou, Yunnan, Hubei and Hunan regions in China (Table 1).

3. Discussion

Buiotenine, a tryptamine alkaloid, was first described in the 1920s [100,101,102,103] and discovered in different living organisms, such as in the skin secretions of many toads (such as of the Bufo species) [104], plants of the Leguminosae family and fungi [59,103,104,105,106,107]. Buiotenine is present as a part of a complex skin secretion in almost 200 species of the genus Bufo worldwide; this secretion acts as a defense mechanism against predators and has antibacterial and antiviral properties [108]. Since the 1960s, there have been reports of smoking dried toad skin secretions, and since the 1980s, reports of “toad licking” have been increasing [109].
Structurally, buiotenine is very similar to psilocin (the active form of psilocybin), but differs in having a hydroxyl group on the C5 of the indole ring instead of C4 [59,103,107]. The possible hallucinogenic effects noted in the literature are still controversial; due to buiotenine not crossing the blood–brain barrier, it may lack hallucinogenic effects [107,110]. However, buiotenine is a promising candidate for treating rabies [107,111].
More than 100 species in Amanita sect. Validae have been discovered worldwide, and approximately 25 of these species were reported in China [18,19,24,39,40,100]. Previous studies showed that Amanita citrina produces buiotenine [59,93,94,95]. The present study confirmed that one closely related species, A. sinocitrina, also produces buiotenine. Meanwhile, no tryptamine alkaloids were detected in the variety A. citrina var. grisea. Amanita porphyria, a similar species from sect. Validae which is closely related to A. citrina [39,40,98] (Figure 2), does not contain buiotenine but contains 5-hydroxytryptophan [94]. Among these >100 species, how many species are poisonous needs to be studied in the future.

4. Conclusions

This study demonstrated that A. citrina and A. sinocitrina contain buiotenine. To the best of our knowledge, this is the first study to report on buiotenine discovered from A. sinocitrina. Additional research is urgently required to determine the prevalence of buiotenine across various species. Future work should explore the biosynthetic pathways and potential pharmacological activities of buiotenine in other Amanita species.

5. Materials and Methods

5.1. Chemicals and Reagents

A series of certified reference standards, including muscarine, psilocybin, psilocin, buiotenine, ibotenic acid, muscimol, α-amanitin, β-amanitin, γ-amanitin, phalloidin, phallacidin and phallisacin, were acquired from Alta Scientific Co., Ltd. (Tianjin, China) in methanol solution and weresubsequently preserved at −20 °C in a freezer. The QuEChERS-PP purification column, incorporating multiwalled carbon nanotubes as the stationary phase, was supplied by Huaren Health Co., Ltd. (Zhengzhou, China). HPLC-grade solvents and reagents—acetonitrile, methanol, ammonium acetate and formic acid—were sourced from Merck Ltd. (Darmstadt, Germany). For all experimental procedures, ultrapure water (resistivity = 18.2 MΩ/cm, TOC < 3 ppb) was generated using a Milli-Q purification system (Millipore, Billerica, MA, USA).

5.2. Materials

Specimens were collected from Guizhou, Yunnan, Hunan and Hubei Provinces, China (Table 1). All fresh mushrooms were dried at 45 °C, and were then stored in the herbarium in the National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control (NIOHP, China CDC). Toxin analysis was performed using 10 mg aliquots of the lyophilized fungal fruiting bodies.

5.3. Species Identification

Morphological identification was mainly performed as per the methods established in previous studies [18,19,24]. Internal transcribed spacer (ITS) was selected for phylogenetic analysis, and maximum parsimony (MP) analyses were applied to the ITS dataset [24,39,100].

5.4. Sample Preparation for Toxin Detection

A dried mushroom specimen (10 mg) was homogenized with 2 mL of a methanol–water mixture (70:30, v/v) in a 15 mL centrifuge tube using vortex agitation. The resulting suspension was subjected to ultrasonic extraction for 60 min, followed by centrifugation (15,000 rpm, 4 °C, 5 min). The clarified supernatant was then passed through a QuECHERS-PP column to eliminate matrix interferences. Subsequently, 10 μL of the purified extract was diluted with a methanol–water solution (5:95, v/v) to a total volume of 1 mL. Prior to UPLC–MS/MS analysis, the prepared solution was centrifuged at 21,000 rpm for 2 min to ensure clarity [101].
For the qualitative screening of mushroom toxins, 20 µL of mushroom extract was pipetted into a vial and diluted to 1 mL with an initial mobile phase (10% B: acetonitrile). The targeted screening of 12 mushroom toxins was performed under an optimal UPLC–MS/MS detection. The toxins detected in the samples were quantitatively analyzed.
In the quantitative analysis, Lentinula edodes were used as the matrix blank mushroom and spiked samples. The matrix blank mushroom and spiked samples were extracted using the same sample preparation method described above, followed by UPLC–MS/MS analysis. Serial concentrations of positive toxins were added to a matrix blank mushroom sample solution to establish the calibration curves for UPLC–MS/MS analysis.

5.5. LC–MS/MS Conditions

UPLC–MS/MS was carried out with a Waters ACQUITY I-Class UPLC system coupled with a Waters Xevo TQ-S MS/MS system (Waters, Milford, MA, USA). The mass spectrometry acquisition parameters of mushroom toxins and the specific parameters of UPLC were chosen according to our previous work [101]. Buiotenine and psilocin were isomers ([M + H]+ = 205.1) with the same mass spectrum parameters. The mass spectrum parameters of the 12 mushroom toxins are in Table 2.
Briefly, the conditions were as follows: the chromatographic separation was carried out using an Atlantis T3 analytical column (2.1 × 100 mm, 3 μm; Waters, USA) and the mobile phase consisted of two components: (A) an aqueous solution of ammonium acetate (10 mmol/L) and (B) acetonitrile, delivered at a constant flow rate of 0.3 mL/min. For mass spectrometric detection, an electrospray ionization (ESI) source was operated in positive-ion mode (ESI+). Data acquisition was conducted via multiple reaction monitoring (MRM). The ionization parameters were optimized as follows: the source temperature was maintained at 550 °C and the spray voltage was set to 5500 V.

Author Contributions

All authors contributed to the study conception and design. Conceptualization, H.-J.L.; Data curation, Y.-Z.Z., Y.Y. and K.-P.Z.; Formal analysis, Y.-Z.Z., Y.Y. and K.-P.Z.; Funding acquisition, H.-J.L. and F.X.; Investigation, Y.-Z.Z., Y.Y., K.-P.Z., J.-Q.L. and Z.-F.L.; Methodology, F.X.; Project administration, J.-Q.L. and J.-J.Z.; Resources, J.-J.Z.; Software, J.-Q.L., J.-J.Z. and Z.-F.L.; Writing—original draft, Y.-Z.Z. and Y.Y.; Writing—review and editing, H.-J.L. and F.X. All authors have read and agreed to the published version of the manuscript.

Funding

This study was supported by the Independent Research Fund of State Key Laboratory of Trauma and Chemical Poisoning (2025SKLCDC-02); the National Natural Science Foundation of China (32270021); the Science and Technology Support Program of Yinchuan, Ningxia (No. 2024SF001); and the Key Research and Development Program of Ningxia (No. 2024BEH04071).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding authors.

Acknowledgments

Special thanks are due to Ya-Juan Zhou (Guizhou CDC) and Jin-Jun Liang (Hunan CDC) for help in field collection.

Conflicts of Interest

The authors declare no conflicts of interest.

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Figure 1. Basidiomata of Amanita species studied in the present paper: (a) A. citrina (150913-24), (b) A. citrina var. grisea (WX20170922-067), (c) A. sinocitrina (150914-18), (d) A. sinocitrina (171012-23). Bars = 1 cm.
Figure 1. Basidiomata of Amanita species studied in the present paper: (a) A. citrina (150913-24), (b) A. citrina var. grisea (WX20170922-067), (c) A. sinocitrina (150914-18), (d) A. sinocitrina (171012-23). Bars = 1 cm.
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Toxins 17 00247 g002
Figure 2. Phylogenetic tree inferred from MP analysis based on internal transcribed spacer (ITS) sequences. Only bootstrap values > 50% are reported. Sequences from present study are shown in bold.
Figure 2. Phylogenetic tree inferred from MP analysis based on internal transcribed spacer (ITS) sequences. Only bootstrap values > 50% are reported. Sequences from present study are shown in bold.
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Toxins 17 00247 g003
Figure 3. MRM chromatograms of buiotenine in A. citrina (a) and A. sinocitrina (b).
Figure 3. MRM chromatograms of buiotenine in A. citrina (a) and A. sinocitrina (b).
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Table 1. The buiotenine contents and species identification information in different mushroom specimens used in the present study.
Table 1. The buiotenine contents and species identification information in different mushroom specimens used in the present study.
SpeciesSpecimen VoucherCollection DateLocationITS NumbersBuiotenine (g/kg)
A. citrina150913-2413-Sep-2015China: GuizhouOR3451882.90 ± 0.27
A. citrina var. griseaWX20170922-06722-Sep-2017China: YunnanOR345187<LOD
A. sinocitrina141025-0525-Oct-2014China: Guizhou-2.70 ± 0.19
150914-0214-Sep-2015China: GuizhouOR3451841.19 ± 0.02
150914-1814-Sep-2015China: GuizhouOR3451773.42 ± 0.66
150914-2714-Sep-2015China: GuizhouMN6470234.10 ± 0.95
171012-1912-Oct-2017China: GuizhouOR3451794.00 ± 0.73
171012-2312-Oct-2017China: GuizhouOR3451862.66 ± 0.52
171012-3812-Oct-2017China: GuizhouOR3451786.70 ± 1.36
DL20170920-01120-Sep-2017China: GuizhouOR3451851.22 ± 0.08
GZHS20200703-0403-Jul-2020China: GuizhouOR3451822.97 ± 0.31
GZHS20200703-0603-Jul-2020China: GuizhouOR3451834.48 ± 0.97
GZRH20200701-0401-Jul-2020China: GuizhouOR3451744.20 ± 0.85
GZRH20200709-0209-Jul-2020China: GuizhouOR3451803.85 ± 0.68
GZSN20200617-00617-Jun-2020China: GuizhouOR3451752.88 ± 0.16
HB20190708-02 08-Jul-2019China: HubeiOR3451813.70 ± 0.69
HNZJJCL20200624-0224-Jun-2020China: HunanOR3451765.26 ± 1.15
HNZJJCL20200924-0624-Sep-2020China: HunanOR3451732.66 ± 0.65
Table 2. The mass spectrum parameters of 12 mushroom toxins.
Table 2. The mass spectrum parameters of 12 mushroom toxins.
AnalytePrecursor Ion
(Q1, m/z)		Product Ion
(Q3, m/z)		DP
(V)		CE
(V)		Retention Time
(min)
α-amanitin919.485.7 a, 259.3 b85, 85107, 601.92
β-amanitin920.685.6 a, 259.5 b103, 103104, 591.53
γ-amanitin903.686.1 a, 243.1 b125, 125123, 603.21
Phalloidin789.7157.2 a, 85.7 b108, 108108, 815.13
Phallacidin847.4157.2 a, 330.3 b101, 10180, 634.53
Phallisacin863.6156.8 a, 85.8 b101, 101117, 882.79
Muscarine174.257.1 a, 97.2 b73, 7032, 261.30
Ibotenic acid158.9113.2 a, 142.1 b50, 5016, 170.87
Muscimol115.198.0 a, 68.1 b43, 4017, 200.89
Psilocybin285.258.1 a, 205.3 b82, 8253, 251.32
Buiotenine205.258.1 a, 160.1 b60, 6933, 193.26
Psilocin205.158.1 a, 160.1 b40, 4532, 233.51
Notions: a quantified ions; b qualitative ions.
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Zhang, Y.-Z.; Yao, Y.; Zhang, K.-P.; Liang, J.-Q.; Zhong, J.-J.; Li, Z.-F.; Li, H.-J.; Xu, F. Toxin Profiling of Amanita citrina and A. sinocitrina: First Report of Buiotenine Detection. Toxins 2025, 17, 247. https://doi.org/10.3390/toxins17050247

AMA Style

Zhang Y-Z, Yao Y, Zhang K-P, Liang J-Q, Zhong J-J, Li Z-F, Li H-J, Xu F. Toxin Profiling of Amanita citrina and A. sinocitrina: First Report of Buiotenine Detection. Toxins. 2025; 17(5):247. https://doi.org/10.3390/toxins17050247

Chicago/Turabian Style

Zhang, Yi-Zhe, Yi Yao, Kai-Ping Zhang, Jia-Qi Liang, Jia-Ju Zhong, Zhong-Feng Li, Hai-Jiao Li, and Fei Xu. 2025. "Toxin Profiling of Amanita citrina and A. sinocitrina: First Report of Buiotenine Detection" Toxins 17, no. 5: 247. https://doi.org/10.3390/toxins17050247

APA Style

Zhang, Y.-Z., Yao, Y., Zhang, K.-P., Liang, J.-Q., Zhong, J.-J., Li, Z.-F., Li, H.-J., & Xu, F. (2025). Toxin Profiling of Amanita citrina and A. sinocitrina: First Report of Buiotenine Detection. Toxins, 17(5), 247. https://doi.org/10.3390/toxins17050247

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