Captiva EMR-Lipid柱净化-超高效液相色谱-串联质谱法快速同时测定谷物及其制品中11种麦角生物碱

doi:10.3724/SP.J.1123.2024.02022

摘要/Abstract

摘要： 麦角生物碱(EAs)是由麦角菌属(Claviceps)产生的一类真菌毒素,当人类摄入被EAs污染的谷物及其制品时,可能会遭受慢性或急性中毒,从而威胁身体健康。鉴于此,本文基于超高效液相色谱-串联质谱(UPLC-MS/MS)建立了谷物及其制品中11种EAs的灵敏、快速测定方法。采用20 mL乙腈-200 mg/L碳酸铵溶液(80∶20, v/v)对谷物及其制品中的11种EAs进行提取,在10000 r/min下离心10 min,用通过式Captiva EMR-Lipid净化柱对上清液进行净化。采用ACQUITY UPLC HSS T3色谱柱(100 mm×3 mm, 1.8 μm)分离,以1 mmol/L乙酸铵溶液和乙腈为流动相进行梯度洗脱。在电喷雾电离(ESI)正离子模式下,采用多反应监测(MRM)模式采集数据,用基质匹配标准曲线进行外标法定量。方法学验证表明,11种EAs在各自的线性范围内具有良好的线性关系,相关系数(r²)为0.9933~0.9999,检出限为0.006~0.2 μg/kg,定量限为0.02~0.6 μg/kg。以小麦粉、薏苡仁、小麦粉制品和玉米粉为基质样品,在低、中、高3个加标水平下,11种EAs的回收率为80.1%~118%,相对标准偏差(RSD)为0.2%~13.3%。将所建方法用于市售谷物及其制品(240份小麦粉、80份玉米粉、30份大米、30份薏苡仁及146份小麦粉制品)中EAs的含量测定。结果表明,每种样品均有EAs检出,11种EAs的检出率为0.57%~20.3%。该方法具有简单、快速、准确等优势,适用于谷物及其制品中多种EAs的快速同时测定。

关键词: 超高效液相色谱-串联质谱, 谷物及其制品, 麦角生物碱, 真菌毒素

Abstract:

Ergot alkaloids (EAs) are mycotoxins produced by Claviceps and are present in cereals and their products; their residues pose significant threats to human health through food consumption, resulting in ergotism and sickness. Herein, a sensitive and rapid method for the determination of 11 EAs in cereals and their products using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) were developed. Eleven EAs were extracted with 20 mL acetonitrile-200 mg/L ammonium acetate solution (80∶20, v/v) for 15 min using the vortex shock method followed by 15 min of ultrasonication. The mixture was subsequently centrifuged at 10000 r/min for 10 min, and the supernatant was purified by a Captiva EMR-Lipid column. Target analytes were separated on an ACQUITY UPLC HSS T3 chromatography column (100 mm×3 mm, 1.8 μm) at a column temperature of 40 ℃ and a flow rate of 0.4 mL/min using an injection volume of 5 μL. Gradient elution was performed using 1 mmol/L ammonium acetate solution and acetonitrile as mobile phases. Data were collected in electrospray positive-ion (ESI⁺) and multi-reaction monitoring (MRM) modes, and quantified using matrix-matched standard curves. The 11 EAs exhibited good linearities in their linear ranges, with correlation coefficients (r²) of 0.9933-0.9999, with limits of detection (LODs) and limits of quantification (LOQs) of 0.002-0.2 and 0.006-0.6 μg/kg, respectively. Recoveries and relative standard deviations (RSDs) of the 11 EAs in matrix samples of wheat flour, coix seed, wheat flour products, and corn flour at low, medium, and high spiked levels were 80.1%-118% and 0.2%-13.3%, respectively. The established method was used to determine EAs in 240 wheat flour, 80 corn flour, 30 rice, and 30 coix seed samples, as well as 146 wheat flour products, with the detection rates of the 11 EAs of 0.57%-20.3%. A maximum total content of EAs of 56.7 μg/kg was recorded for a single sample. The sample pretreatment process used in this method is simple and fast, and the detection method is highly sensitive, with accurate and reliable results obtained. This method is suitable for simultaneously determining various EAs in cereals and their products. The results of this study provide valuable information for future EA risk-assessment studies.

Key words: ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), cereals and their products, ergot alkaloids (EAs), mycotoxins

中图分类号:

O658

刘柏林, 张丹, 赵紫微, 谢继安, 詹子悦, 张琪, 李卫东. Captiva EMR-Lipid柱净化-超高效液相色谱-串联质谱法快速同时测定谷物及其制品中11种麦角生物碱[J]. 色谱, 2025, 43(4): 326-334.

LIU Bolin, ZHANG Dan, ZHAO Ziwei, XIE Ji’an, ZHAN Ziyue, ZHANG Qi, LI Weidong. Rapid and simultaneous determination of 11 ergot alkaloids in cereals and their products by ultra performance liquid chromatography-tandem mass spectrometry combined with Captiva EMR-Lipid column purification[J]. Chinese Journal of Chromatography, 2025, 43(4): 326-334.

图/表 6

参考文献

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Compound	Retention time/ min	Parent ion (m/z)	Product ions (m/z)	CE/eV	CVs/V
Ergometrine maleate (马来酸麦角新碱)	1.01	326.3	223.2^*, 208.0	38	24, 28
Ergosine (麦角辛)	1.89	548.4	223.2^*, 268.2	34	32, 24
Ergotamine (麦角胺)	2.20	582.4	223.2^*, 208.1	36	30, 44
Ergocornine (麦角考宁)	4.59	562.4	268.2^*, 208.1	40	26, 44
Dihydroergocristine (双氢麦角汀)	4.70	612.4	270.2^*, 168.1	40	30, 60
Ergocryptine (麦角隐亭)	4.79	576.4	223.2^*, 268.2	40	35, 26
Ergocristine (麦角克碱)	4.83	610.4	223.2^*, 208.1	40	36, 48
Ergosinine (麦角辛宁)	4.97	548.4	223.2^*, 277.2	40	32, 26
Ergocorninine (麦角异柯宁碱)	5.17	562.4	277.2^*, 223.2	38	26, 34
Ergocryptinine (麦角异卡里碱)	5.30	576.4	223.2^*, 305.2	40	36, 28
Ergocristinine (麦角异克碱)	5.34	610.4	305.2^*, 325.2	40	28, 28

Compound	Retention time/ min	Parent ion (m/z)	Product ions (m/z)	CE/eV	CVs/V
Ergometrine maleate (马来酸麦角新碱)	1.01	326.3	223.2^*, 208.0	38	24, 28
Ergosine (麦角辛)	1.89	548.4	223.2^*, 268.2	34	32, 24
Ergotamine (麦角胺)	2.20	582.4	223.2^*, 208.1	36	30, 44
Ergocornine (麦角考宁)	4.59	562.4	268.2^*, 208.1	40	26, 44
Dihydroergocristine (双氢麦角汀)	4.70	612.4	270.2^*, 168.1	40	30, 60
Ergocryptine (麦角隐亭)	4.79	576.4	223.2^*, 268.2	40	35, 26
Ergocristine (麦角克碱)	4.83	610.4	223.2^*, 208.1	40	36, 48
Ergosinine (麦角辛宁)	4.97	548.4	223.2^*, 277.2	40	32, 26
Ergocorninine (麦角异柯宁碱)	5.17	562.4	277.2^*, 223.2	38	26, 34
Ergocryptinine (麦角异卡里碱)	5.30	576.4	223.2^*, 305.2	40	36, 28
Ergocristinine (麦角异克碱)	5.34	610.4	305.2^*, 325.2	40	28, 28

Analyte	Linear range/(μg/L)	Linear equation	r²	LOD/(μg/kg)	LOQ/(μg/kg)
Ergometrine maleate	0.01-50	y=2.80×10⁵x+4.18×10²	0.9969	0.06	0.2
Ergosine	0.01-50	y=9.24×10⁴x+2.12×10²	0.9988	0.06	0.2
Ergotamine	0.002-10	y=2.02×10⁵x-7.54×10¹	0.9991	0.013	0.04
Ergocornine	0.002-10	y=2.99×10⁵x+1.64×10²	0.9998	0.013	0.04
Dihydroergocristine	0.002-10	y=3.82×10⁵x+3.76×10²	0.9994	0.013	0.04
Ergocryptine	0.002-10	y=4.06×10⁵x+4.78×10²	0.9994	0.013	0.04
Ergocristine	0.002-10	y=1.63×10⁵x+4.98×10²	0.9977	0.013	0.04
Ergosinine	0.001-5	y=6.89×10⁵x+2.43×10²	0.9998	0.006	0.02
Ergocorninine	0.001-5	y=3.88×10⁵x+4.41×10²	0.9993	0.006	0.02
Ergocryptinine	0.001-5	y=4.78×10⁵x+5.23×10²	0.9995	0.006	0.02
Ergocristinine	0.001-5	y=2.10×10⁵x+6.04×10²	0.9996	0.006	0.02

Analyte	Linear range/(μg/L)	Linear equation	r²	LOD/(μg/kg)	LOQ/(μg/kg)
Ergometrine maleate	0.01-50	y=2.80×10⁵x+4.18×10²	0.9969	0.06	0.2
Ergosine	0.01-50	y=9.24×10⁴x+2.12×10²	0.9988	0.06	0.2
Ergotamine	0.002-10	y=2.02×10⁵x-7.54×10¹	0.9991	0.013	0.04
Ergocornine	0.002-10	y=2.99×10⁵x+1.64×10²	0.9998	0.013	0.04
Dihydroergocristine	0.002-10	y=3.82×10⁵x+3.76×10²	0.9994	0.013	0.04
Ergocryptine	0.002-10	y=4.06×10⁵x+4.78×10²	0.9994	0.013	0.04
Ergocristine	0.002-10	y=1.63×10⁵x+4.98×10²	0.9977	0.013	0.04
Ergosinine	0.001-5	y=6.89×10⁵x+2.43×10²	0.9998	0.006	0.02
Ergocorninine	0.001-5	y=3.88×10⁵x+4.41×10²	0.9993	0.006	0.02
Ergocryptinine	0.001-5	y=4.78×10⁵x+5.23×10²	0.9995	0.006	0.02
Ergocristinine	0.001-5	y=2.10×10⁵x+6.04×10²	0.9996	0.006	0.02

Analyte	Spiked level/ (μg/kg)		Recoveries/%								RSDs/%
Analyte	Spiked level/ (μg/kg)		Wheat flour		Coix seed		Wheat flour product		Corn flour		Wheat flour		Coix seed		Wheat flour product		Corn flour
Ergometrine maleate		0.2		91.7		93.9		106		103		2.2		0.7		4.1		7.0
		40		83.5		88.6		90.2		102		1.1		1.3		1.3		1.2
		200		96.3		93.3		91.8		97.5		0.2		1.6		3.2		0.7
Ergosine		0.2		90.7		90.8		81.7		93.0		1.2		0.8		5.8		1.5
		40		98.1		101		95.9		96.8		1.2		1.5		1.4		0.5
		200		94.3		98.8		105		90.7		1.5		1.9		0.9		1.5
Ergotamine		0.04		90.0		95.8		87.3		100		6.2		4.2		4.9		2.0
		8		92.8		99.7		83.7		97.4		0.2		0.4		3.6		0.8
		40		92.3		98.9		102		96.0		1.3		0.3		12.2		0.7
Ergocornine		0.04		89.7		94.7		87.0		92.0		5.5		9.9		1.1		4.7
		8		102		98.5		118		96.7		1.6		1.8		1.6		1.0
		40		101		101		115		92.7		0.7		0.8		0.8		0.5
Dihydroergocristine		0.04		98.7		80.1		102		92.9		1.4		7.0		2.1		3.4
		8		89.1		99.1		103		82.9		0.4		1.0		1.6		1.1
		40		89.4		80.8		97.4		100		0.5		5.4		1.7		1.4
Ergocryptine		0.04		87.3		90.7		86.7		80.3		4.8		5.7		1.3		3.8
		8		95.8		84.8		111		90.5		1.2		2.0		2.2		1.1
		40		91.7		82.9		103		83.1		1.0		1.9		0.4		1.7
Ergocristine		0.04		88.0		82.3		100		96.4		10.8		3.1		2.6		5.6
		8		99.9		93.5		112		95.1		0.7		1.7		3.7		0.9
		40		97.6		88.9		111		90.2		1.4		0.7		1.5		2.9
Ergosinine		0.02		86.7		102		107		85.0		12.5		1.6		2.7		5.1
		4		83.3		95.1		104		89.9		0.9		6.9		1.0		2.9
		20		82.0		87.8		99.4		82.7		2.2		4.8		6.9		1.7
Ergocorninine		0.02		110		103		107		82.9		6.6		2.2		1.9		6.7
		4		115		82.0		111		89.1		1.5		10.4		4.2		2.4
		20		114		83.4		105		94.9		3.2		7.2		13.1		1.6
Ergocryptinine		0.02		82.7		94.7		81.7		85.9		13.3		6.5		4.7		6.5
		4		96.1		80.7		97.5		89.1		1.2		7.6		1.4		2.9
		20		100		87.8		112		92.0		7.9		10.2		3.5		0.9
Ergocristinine		0.02		85.3		85.0		85.0		87.3		8.9		5.9		11.8		9.5
		4		102		84.8		97.3		91.6		1.5		3.4		1.6		3.6
		20		103		80.3		108		91.9		2.7		11.8		6.0		0.7