Determination of 16 mycotoxins in drug and food homologous products by ultra performance liquid chromatography-tandem mass spectrometry combined with accelerated solvent extraction and QuEChERS

doi:10.3724/SP.J.1123.2019.10034

Abstract

Abstract:

A method was developed for the simultaneous determination of 16 mycotoxins in drug and food homologous products by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with accelerated solvent extraction (ASE) and QuEChERS. The target mycotoxins in drug and food homologous products were extracted by ASE. After concentration, the extracts were purified by QuEChERS. Then, the target compounds were analyzed by UPLC-MS/MS in both positive and negative electrospray ionization and MRM modes. Aflatoxin B1 and fumonisin B1 were quantified by the internal standard method, and the remaining mycotoxins were quantified by the matrix-matched external standard method. The proposed method showed a good linear relationship, with correlation coefficients greater than 0.99. The limits of detection (LODs) and limits of quantification (LOQs) of the 16 mycotoxins ranged from 0.008 μg/kg to 0.3 μg/kg and from 0.03 μg/kg to 1.0 μg/kg, respectively. The blank samples were spiked at three levels, and the recoveries ranged from 70.8% to 118%, with the RSDs being 2.5% to 10.2%. The developed method was successfully applied to mycotoxin analysis in 30 scutellaria, puerarin and sea buckthorn samples bought from local markets. Different levels of mycotoxins were detected in some of the products. The proposed method is simple, rapid and sensitive, and it can be applied to the simultaneous determination of multi-mycotoxins in drug and food homologous products.

Key words: ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), accelerated solvent extraction (ASE), QuEChERS, mycotoxins, drug and food homologous products

FANG Zhen, QU Li, GU Shuqing, CHEN Rouhan, LI You, DENG Xiaojun, GUO Dehua, FENG Feng. Determination of 16 mycotoxins in drug and food homologous products by ultra performance liquid chromatography-tandem mass spectrometry combined with accelerated solvent extraction and QuEChERS[J]. Chinese Journal of Chromatography, 2020, 38(7): 782-790.

Figures/Tables 5

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Mycotoxin	Abbreviation	ESI mode	Retention time/min	Precursor ion (m/z)	Product ion (m/z)	CE/eV	DP/V
CE: collision energy; DP: declustering potential; * quantitative ion.
Aflatoxin B1	AFB1	positive	4.83	313.0	285.0^*	32.99	127
					269.0	40.97	129
Aflatoxin B2	AFB2	positive	4.67	315.0	287.0^*	33.94	129
					259.0	41.99	103
Aflatoxin G1	AFG1	positive	4.65	329.0	243.0^*	32.59	105
					311.0	30.29	149
Aflatoxin G2	AFG2	positive	4.48	331.0	245.0^*	36.99	147
					285.0	38.02	124
Ochratoxin A	OTA	positive	5.80	404.0	239.0^*	19.76	112
					358.0	29.30	118
Ochratoxin B	OTB	positive	5.31	370.2	205.2^*	20.85	49
					324.4	28.87	41
Sterigmatocystin	SMC	positive	6.04	325.0	309.9^*	33.93	123
					280.9	52.91	127
Fumonisins B1	FB1	positive	4.05	722.5	334.3^*	53.07	141
					352.3	56.03	148
Fumonisins B2	FB2	positive	4.41	706.5	336.3^*	51.84	178
					318.2	56.00	130
T-2 toxin	T-2	positive	5.60	484.1	305.0^*	25.82	50
					184.9	19.09	54
HT-2 toxin	HT-2	positive	4.91	442.1	215.0^*	18.87	83
					263.1	14.95	47
Deoxynivalenol	DON	positive	3.25	296.9	249.1^*	18.83	67
					231.1	18.99	67
3-Acetyl-deoxynivalenol	3-AcDON	positive	4.09	339.0	231.0^*	18.90	123
					203.0	20.95	133
15-Acetyl-deoxynivalenol	15-AcDON	positive	4.01	339.1	321.3^*	11.69	127
					137.2	21.06	131
Verruculogen	VER	positive	6.13	512.3	310.0^*	33.92	75
					351.8	19.00	80
Zearalenone	ZEN	negative	5.85	317.0	175.0^*	-35.07	-119
					131.0	-35.31	-125
U-[¹³C₁₇]-Aflatoxin B1	U-[¹³C₁₇]-AFB1	positive	4.81	330.0	255.0	49.40	198
U-[¹³C₃₄]-Fumonisins B1	U-[¹³C₃₄]-FB1	positive	4.04	756.1	356.0	54.09	140

Mycotoxin	Abbreviation	ESI mode	Retention time/min	Precursor ion (m/z)	Product ion (m/z)	CE/eV	DP/V
CE: collision energy; DP: declustering potential; * quantitative ion.
Aflatoxin B1	AFB1	positive	4.83	313.0	285.0^*	32.99	127
					269.0	40.97	129
Aflatoxin B2	AFB2	positive	4.67	315.0	287.0^*	33.94	129
					259.0	41.99	103
Aflatoxin G1	AFG1	positive	4.65	329.0	243.0^*	32.59	105
					311.0	30.29	149
Aflatoxin G2	AFG2	positive	4.48	331.0	245.0^*	36.99	147
					285.0	38.02	124
Ochratoxin A	OTA	positive	5.80	404.0	239.0^*	19.76	112
					358.0	29.30	118
Ochratoxin B	OTB	positive	5.31	370.2	205.2^*	20.85	49
					324.4	28.87	41
Sterigmatocystin	SMC	positive	6.04	325.0	309.9^*	33.93	123
					280.9	52.91	127
Fumonisins B1	FB1	positive	4.05	722.5	334.3^*	53.07	141
					352.3	56.03	148
Fumonisins B2	FB2	positive	4.41	706.5	336.3^*	51.84	178
					318.2	56.00	130
T-2 toxin	T-2	positive	5.60	484.1	305.0^*	25.82	50
					184.9	19.09	54
HT-2 toxin	HT-2	positive	4.91	442.1	215.0^*	18.87	83
					263.1	14.95	47
Deoxynivalenol	DON	positive	3.25	296.9	249.1^*	18.83	67
					231.1	18.99	67
3-Acetyl-deoxynivalenol	3-AcDON	positive	4.09	339.0	231.0^*	18.90	123
					203.0	20.95	133
15-Acetyl-deoxynivalenol	15-AcDON	positive	4.01	339.1	321.3^*	11.69	127
					137.2	21.06	131
Verruculogen	VER	positive	6.13	512.3	310.0^*	33.92	75
					351.8	19.00	80
Zearalenone	ZEN	negative	5.85	317.0	175.0^*	-35.07	-119
					131.0	-35.31	-125
U-[¹³C₁₇]-Aflatoxin B1	U-[¹³C₁₇]-AFB1	positive	4.81	330.0	255.0	49.40	198
U-[¹³C₃₄]-Fumonisins B1	U-[¹³C₃₄]-FB1	positive	4.04	756.1	356.0	54.09	140

Mycotoxin	Linear range/(μg/L)	Linear equation	r²	LOD/(μg/kg)	LOQ/(μg/kg)
Y: peak area; Y′: peak area ratio of the analyte to isotope internal standard; X: mass concentration, μg/L.
AFB1	0.05-50	Y′=7.12X+4.85×10^-2	0.9989	0.008	0.03
AFB2	0.05-50	Y=2.87×10⁵X-1.26×10³	0.9960	0.008	0.03
AFG1	0.05-50	Y=3.54×10⁵X+149	0.9907	0.008	0.03
AFG2	0.05-50	Y=1.29×10⁵X-1.73×10³	0.9904	0.008	0.03
OTA	0.05-500	Y=8.02×10⁴X-430	0.9990	0.008	0.03
OTB	0.05-500	Y=3.92×10⁵X+2.83×10³	0.9995	0.008	0.03
SMC	0.5-100	Y=4.17×10⁵X-3.32×10³	0.9965	0.08	0.3
FB1	0.5-1000	Y′=1.41×10^-1X+3.48×10^-2	0.9987	0.08	0.3
FB2	0.5-1000	Y=1.78×10⁴X+1.04×10³	0.9923	0.08	0.3
T-2	0.5-50	Y=3.48×10⁴X+9.59	0.9991	0.1	0.3
HT-2	1.0-100	Y=2.07×10³X+1.97×10³	0.9928	0.3	1.0
DON	0.5-50	Y=2.29×10⁴X+5.02×10³	0.9958	0.1	0.3
3-AcDON	1.0-100	Y=1.53×10⁴X+9.57×10²	0.9967	0.3	1.0
15-AcDON	1.0-100	Y=3.6×10³X+1.44×10⁴	0.9905	0.3	1.0
VER	1.0-100	Y=8.27×10³X+1.92×10³	0.9903	0.3	1.0
ZEN	0.5-100	Y=4.54×10³X+81.7	0.9912	0.1	0.3

Mycotoxin	Linear range/(μg/L)	Linear equation	r²	LOD/(μg/kg)	LOQ/(μg/kg)
Y: peak area; Y′: peak area ratio of the analyte to isotope internal standard; X: mass concentration, μg/L.
AFB1	0.05-50	Y′=7.12X+4.85×10^-2	0.9989	0.008	0.03
AFB2	0.05-50	Y=2.87×10⁵X-1.26×10³	0.9960	0.008	0.03
AFG1	0.05-50	Y=3.54×10⁵X+149	0.9907	0.008	0.03
AFG2	0.05-50	Y=1.29×10⁵X-1.73×10³	0.9904	0.008	0.03
OTA	0.05-500	Y=8.02×10⁴X-430	0.9990	0.008	0.03
OTB	0.05-500	Y=3.92×10⁵X+2.83×10³	0.9995	0.008	0.03
SMC	0.5-100	Y=4.17×10⁵X-3.32×10³	0.9965	0.08	0.3
FB1	0.5-1000	Y′=1.41×10^-1X+3.48×10^-2	0.9987	0.08	0.3
FB2	0.5-1000	Y=1.78×10⁴X+1.04×10³	0.9923	0.08	0.3
T-2	0.5-50	Y=3.48×10⁴X+9.59	0.9991	0.1	0.3
HT-2	1.0-100	Y=2.07×10³X+1.97×10³	0.9928	0.3	1.0
DON	0.5-50	Y=2.29×10⁴X+5.02×10³	0.9958	0.1	0.3
3-AcDON	1.0-100	Y=1.53×10⁴X+9.57×10²	0.9967	0.3	1.0
15-AcDON	1.0-100	Y=3.6×10³X+1.44×10⁴	0.9905	0.3	1.0
VER	1.0-100	Y=8.27×10³X+1.92×10³	0.9903	0.3	1.0
ZEN	0.5-100	Y=4.54×10³X+81.7	0.9912	0.1	0.3

Mycotoxin	Spiked level/(μg/kg)	Flos lonicerae		Pueraria lobata		Seabuckthorn
Mycotoxin	Spiked level/(μg/kg)	Recovery/%	RSD/%	Recovery/%	RSD/%	Recovery/%	RSD/%
AFB1	0.1	109	4.2	113	5.0	92.8	6.4
	0.5	92.8	2.5	105	3.8	96.5	4.6
	1.0	97.1	2.8	104	4.2	96.3	3.0
AFB2	0.1	115	6.9	80.9	4.7	108	7.4
	0.5	108	5.5	84.5	5.4	87.9	7.3
	1.0	83.6	5.9	81.4	7.5	79.4	5.5
AFG1	0.1	87.5	4.4	85.6	6.9	83.5	3.4
	0.5	85.3	4.8	85.9	4.4	80.6	4.1
	1.0	79.8	2.8	75.5	3.9	79.8	3.8
AFG2	0.1	75.4	4.9	79.5	7.7	79.5	3.9
	0.5	85.3	5.4	83.8	6.4	80.4	5.0
	1.0	87.9	4.3	82.7	4.1	82.7	5.7
OTA	0.1	82.4	7.2	115	6.9	75.0	5.9
	0.5	79.6	6.9	109	5.2	79.6	6.1
	1.0	83.3	5.4	103	4.1	83.4	3.7
OTB	0.1	73.0	9.7	85.3	8.4	79.3	7.4
	0.5	79.4	5.2	78.6	6.6	76.2	4.9
	1.0	78.9	5.9	82.8	4.7	74.5	6.3
SMC	0.5	76.4	7.9	77.7	8.4	71.6	6.2
	1.0	82.5	6.2	72.9	5.3	74.7	7.1
	5.0	79.6	5.1	79.8	5.9	78.7	4.8
FB1	0.5	93.8	5.8	89.4	7.3	113	4.8
	1.0	97.6	4.7	103	4.0	104	5.1
	5.0	102	3.7	94.8	3.4	107	3.4
FB2	0.5	72.4	5.3	72.1	7.3	75.6	4.3
	1.0	76.9	3.9	73.5	7.9	75.1	3.4
	5.0	74.5	4.4	77.4	5.4	74.8	5.1
T-2	1.0	77.6	6.0	72.9	5.9	72.1	7.3
	5.0	74.5	5.6	77.3	6.1	78.7	4.7
	10	79.3	5.2	72.4	4.7	80.1	5.9
HT-2	1.0	79.5	10.2	74.3	8.5	70.8	7.9
	5.0	75.2	7.7	77.3	5.6	73.2	8.2
	10	74.1	6.1	74.0	5.4	75.1	3.4
DON	0.5	118	6.2	71.9	6.9	112	5.7
	1.0	83.5	6.8	79.7	6.1	84.9	6.1
	5.0	80.9	4.5	75.8	4.7	88.5	4.3
3-AcDON	1.0	76.0	9.3	72.6	8.1	109	6.3
	5.0	73.9	6.0	77.4	4.7	114	5.2
	10	75.2	5.4	75.8	3.4	107	7.5
15-AcDON	1.0	72.8	8.6	71.9	5.6	73.7	5.9
	5.0	75.9	7.9	72.5	6.3	80.1	5.1
	10	74.3	3.9	73.9	4.1	79.4	4.0
VER	1.0	71.8	5.2	114	3.9	117	3.6
	5.0	75.3	4.7	109	5.4	113	5.0
	10	79.3	4.6	112	5.6	108	3.6
ZEN	0.5	111	6.9	74.3	6.7	76.9	5.1
	1.0	108	6.3	78.4	3.4	78.6	4.9
	5.0	94.5	4.4	73.5	5.3	74.7	3.4