Analysis of Five Mushroom Toxins in Blood by UPLC-HRMS

doi:10.12116/j.issn.1004-5619.2020.301001

Abstract

Abstract: Objective

To develop a method for the simultaneous and rapid detection of five mushroom toxins （α-amanitin， phallacidin， muscimol， muscarine and psilocin） in blood by ultra-high performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）.

Methods

The blood samples were precipitated with acetonitrile-water solution（V_acetonitril∶V_water=3∶1） and PAX powder， then separated on ACQUITY Premier C₁₈ column， eluted gradient. Five kinds of mushroom toxins were monitored by FullMS-ddMS²/positive ion scanning mode， and qualitative and quantitative analysis was conducted according to the accurate mass numbers of primary and secondary fragment ions.

Results

All the five mushroom toxins had good linearity in their linear range， with a determination coefficient （R²）≥0.99. The detection limit was 0.2-20 ng/mL. The ration limit was 0.5-50 ng/mL. The recoveries of low， medium and high additive levels were 89.6%-101.4%， the relative standard deviation was 1.7%-6.7%， the accuracy was 90.4%-101.3%， the intra-day precision was 0.6%-9.0%， the daytime precision was 1.7%-6.3%， and the matrix effect was 42.2%-129.8%.

Conclusion

The method is simple， rapid， high recovery rate， and could be used for rapid and accurate qualitative screening and quantitative analysis of various mushroom toxins in biological samples at the same time， so as to provide basis for the identification of mushroom poisoning events.

Key words: forensic medicine, toxicological chemistry, mushroom toxins, ultra-high performance liquid chromatography-high resolution mass spectrometry, blood, mice

CLC Number:

DF795.1

Wen-qiao LIU, Yan SHI, Ping XIANG, Feng YU, Bing XIE, Mei DONG, Jing HA, Chun-ling MA, Di WEN. Analysis of Five Mushroom Toxins in Blood by UPLC-HRMS[J]. Journal of Forensic Medicine, 2021, 37(5): 646-652.

Figures/Tables 7

References 22

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蘑菇毒素	化学式	保留时间/min	准分子离子	一级定量离子	二级定性子离子
α-鹅膏毒肽	C₃₉H₅₄N₁₀O₁₄S	7.88	[M+H]⁺	919.361 44	901.348 75、259.128 17
羧基二羟鬼笔毒肽	C₃₇H₅₀N₈O₁₃S	9.58	[M+H]⁺	847.329 08	647.259 28、185.092 09
蝇蕈醇	C₄H₆N₂O₂	0.98	[M+H]⁺	115.050 20	98.023 68
毒蝇碱	C₉H₂₀NO₂	1.16	[M]⁺	174.148 86	115.075 16
赛洛新	C₁₂H₁₆N_2O	3.51	[M+H]⁺	205.133 54	58.065 39、160.075 58

蘑菇毒素	化学式	保留时间/min	准分子离子	一级定量离子	二级定性子离子
α-鹅膏毒肽	C₃₉H₅₄N₁₀O₁₄S	7.88	[M+H]⁺	919.361 44	901.348 75、259.128 17
羧基二羟鬼笔毒肽	C₃₇H₅₀N₈O₁₃S	9.58	[M+H]⁺	847.329 08	647.259 28、185.092 09
蝇蕈醇	C₄H₆N₂O₂	0.98	[M+H]⁺	115.050 20	98.023 68
毒蝇碱	C₉H₂₀NO₂	1.16	[M]⁺	174.148 86	115.075 16
赛洛新	C₁₂H₁₆N_2O	3.51	[M+H]⁺	205.133 54	58.065 39、160.075 58

蘑菇毒素	回收率(n=6)			基质效应(n=6)
蘑菇毒素	无	C18	PAX	无	C18	PAX
α-鹅膏毒肽	101.5	102.2¹⁾	81.2¹⁾	106.7	135.1¹⁾	107.4¹⁾
羧基二羟鬼笔毒肽	91.2	109.0¹⁾	86.5¹⁾	107.5	121.2¹⁾	96.3¹⁾
蝇蕈醇	96.5	56.6¹⁾	97.7¹⁾	45.0	23.9¹⁾	41.2¹⁾
毒蝇碱	99.6	111.2¹⁾	88.6¹⁾	111.0	135.9¹⁾	108.2¹⁾
赛洛新	90.5	82.3¹⁾	105.3¹⁾	199.2	83.9¹⁾	104.7¹⁾

蘑菇毒素	回收率(n=6)			基质效应(n=6)
蘑菇毒素	无	C18	PAX	无	C18	PAX
α-鹅膏毒肽	101.5	102.2¹⁾	81.2¹⁾	106.7	135.1¹⁾	107.4¹⁾
羧基二羟鬼笔毒肽	91.2	109.0¹⁾	86.5¹⁾	107.5	121.2¹⁾	96.3¹⁾
蝇蕈醇	96.5	56.6¹⁾	97.7¹⁾	45.0	23.9¹⁾	41.2¹⁾
毒蝇碱	99.6	111.2¹⁾	88.6¹⁾	111.0	135.9¹⁾	108.2¹⁾
赛洛新	90.5	82.3¹⁾	105.3¹⁾	199.2	83.9¹⁾	104.7¹⁾

蘑菇毒素	线性范围/ (ng·mL^-1)	线性方程	决定系数 (R²)	检出限/ (ng·mL^-1)	定量限/ (ng·mL^-1)
α-鹅膏毒肽	10~1 000	y=-794 398+58 316x	0.999 2	10.0	20.0
羧基二羟鬼笔毒肽	10~1 000	y=-638 286+161 201x	0.999 9	10.0	20.0
蝇蕈醇	20~1 000	y=-464 987+112 762x	0.999 0	20.0	50.0
毒蝇碱	0.2~10	y=973 511+6 294 330x	0.996 0	0.2	0.5
赛洛新	0.2~10	y=310 344+4 016 220x	0.999 9	0.2	0.5