Trichothecenes Appendices (JECFA 47, 2001)

Trichothecenes Appendix 1.

Methods for screening for B-trichothecenes and particularly for deoxynivalenol^a

Reference^b	Matrix	Amount of sample (g)	Extraction	Clean-up	Technique
Malone et al. (1998)	Wheat, barley, maize, bran, oats	25	CH₃CN/H₂O	Mycosep	Fluorometric determination after derivatization
Trucksess et al. (1987)	Corn, wheat, barley	50	CH₃CN:H₂O: MeOH (70:25:5) 200ml, shake 60 min	Carbon column C18 column	TLC fluorodensitometric
Trucksess et al. (1984)	Wheat, maize	50	CH₃CN/H₂O	Charcoal/ alumina/celite	TLC
Fernandez et al. (1994)	Wheat	50	CH₃CN/H₂O	Charcoal/ alumina/ celite	TLC
Eppley et al. (1986)	Wheat	50	CH₃CN/H₂O	Charcoal/ alumina/celite	TLC
Eppley et al. (1984)	Wheat	50	CH₃CN/H₂O	Charcoal/ alumina	TLC
Romer (1986)	Foods, feeds		CH₃CN/H₂O	Charcoal/ alumina	TLC
Romer (1999) Accutox^TM			Distilled water		ELISA
Xu et al. (1988)	Wheat, maize		CH₃CN/H₂O	None	ELISA (acetylated)
Ridascreen™ DON (2000)	Cereals, feeds	2	CH₃CN/H₂O		ELISA
Ridascreen™ DON (2000)	Cereals, feeds	2	CH₃CN/H₂O		(acetylated)
Sinha et al. (1995)	Wheat, maize	1	MeOH/ H₂O		ELISA
Abouzied (2000)	Cereals	NA	H₂O	None	ELISA
Veratox™ (1998)	Wheat	10	H₂O	None	CD-ELISA
	Corn
	Barley
	Oat
Schneider et al. (2000)	Wheat	10	CH₃CN/H₂O	None	ELISA (IgY)
Mills et al. (1990)	Wheat	NA	60% MeOH	None	ELISA
Usleber et al. (1992)	Wheat	2	MeOH/ H₂O	Ethyl acetate	ELISA

Reference^b	Matrix	Toxins assayed	LOD/LOQ (ng/g)	Spiking levels (ng/g)	Recovery (%)	CV (%)
Malone et al. (1998)	Wheat, barley, maize, bran, oats	DON	100	500–50 000	NA	NA
Trucksess et al. (1987)	Corn, wheat, barley	DON	NA	200	81–88	2–20
		NIV		100	78–89
		FX		50	61–108
Trucksess et al. (1984)	Wheat, maize	DON	20/40	100–1000	85–93	3–17
			(wheat)	(wheat)
			20/100	100–1000	77–80	6–12
			(maize)	(maize)
Fernandez et al. (1994)	Wheat	DON	40/NA	200	83	NA
				400	82
				800	72
Eppley et al. (1986)	Wheat	DON	50/NA	50	90	30–87
				100	96
				300	95
				1000	78
Eppley et al. (1984)	Wheat	DON	NA/200	200–5000	80	NA
Romer (1986)	Foods, feeds	DON	300/NA	900–1100	99	8
		NIV	500/NA	750–1100	92	15
		FX	300/NA	900–1100	99	9
Romer (1999) Accutox^TM		DON		500
Xu et al. (1988)	Wheat, maize	DON	NA	10–1000	102	12
Xu et al. (1988)	Wheat, maize	DON			100	15
Ridascreen™ DON (2000)	Cereals, feeds	DON	1.25/NA	NA	80–90
		3-AcDON
		15-AcDON
Sinha et al. (1995)	Wheat, maize	DON	250/NA	NA	NA	NA
Sinha et al. (1995)	Wheat, maize	15-AcDON
Abouzied (2000)	Cereals	DON	20/NA	NA	NA	NA
Veratox™ (1998)	Wheat	DON	300/500	NA	NA	NA
	Corn
	Barley
	Oat
Schneider et al. (2000)	Wheat	DON	160/NA	525	104-120	6
Mills et al. (1990)	Wheat	DON	100/NA	NA	NA	NA
Usleber et al. (1992)	Wheat	DON	4.5 ng/ml/200	200–1000	71–79	3.6–4.9

^{a Fordefinitions of abbreviations, see end of Trichothecenes Appendix 6.
^{b The references are listed in the monograph on deoxynivalenol.}Trichothecenes Appendix 2.
Quantitative methods for B-trichothecenes and particularly for deoxynivalenol^a}

Reference^b	Matrix	Amount of sample (g)	Extraction	Clean-up	Technique assayed
Veldman et al. (1992)	Feed	20	CH₃CN/H₂O	Celite/cahrcoal/ aluminium oxide	HPLC–UV
Rajakylä et al. (1987)	Wheat, rye	20	CH₃CN	Bond Elut	HPLC–MS
Zabe et al. (2000)	Food	NA	NA	Immunoaffinity	HPLC–NA
Josephs et al. (1998)	Wheat	1	SFE	SFE	GC–ECD
Pineiro et al. (1996)	Liquid culture	50	MeOH	SPE–C18	HPLC–UV
Sano et al. (1987)	Maize, wheat, barley	20	CH₃CN/H₂O	Florisil and CN-	HPLC–FLD
Sano et al. (1987)	Maize, wheat, barley	20	CH₃CN/H₂O	Sep Pak	(methyl acetate)
Schuhmacher et al. (1996)	Wheat	15	CH₃CN/H₂O	Mycosep	HPLC–DAD
Berger et al. (1999)	Wheat	10	CH₃CN/H₂O	Mycosep 227, 216	LC–MS
Cahill et al. (1999)	Wheat	50	H₂O/PEG	DONtest	HPLC–UV
Razzazi-Fazeli et al. (1999)	Wheat	25	CH₃CN/H₂O	Mycosep	HPLC–
Razzazi-Fazeli et al. (1999)	Wheat	25	CH₃CN/H₂O	Mycosep	APCI–MS
Reutter (1999)	Cereals, feed	25	CH₃CN/H₂O	Charcoal/ alumina/ celite/Iimmuno-affinity column	HPLC–UV
Mulders et al. (1986)	Cereals	25	CHCl₃/EtOH	Silica gel	GC–ECD (HFB)
Mulders et al. (1986)	Cereals	25	CHCl₃/EtOH	Silica gel	GC–MS (HFB)
Scott & Kanhere (1986)	Cereals	50	CH₃OH/H₂O	Silica gel	GC–ECD (TMS)
Tacke & Casper (1996)	Wheat, barley, malt	5	CH₃CN/H₂O	C₁₈/alumina	GC–ECD (TMS)
Weingärtner et al. (1997)	Wheat	25	CH₃CN/H₂O	MycoSep‘	GC–ECD (TMS/TBT)
Schollenberger et al. (1998)	Cereals, food, feed	10	CH₃CN/H₂O	Florisil/Bond Elut CBA	GC–MS (TFA)
Croteau et al. (1994)	Maize	50	CH₃CN/H₂O, shake 2.5 h	Charcoal/ alumina	GC–ECD (DMAP, HFBA)
Adler et al. (1992)	Wheat	20	MeOH/ H₂O	Silica gel/C₁₈	GC–ECD (TMS)
Blaas et al. (1984)	Cereals	20	MeOH/ H₂O, hexane	Florisil	GC–MS (TMS)
Seidel et al. (1993)	Wheat, maize	10	MeOH/ H₂O	Florisil/C₁₈	GC–ECD (HFB)
van Egmond (2000)	Wheat	25	CH₃CN/H₂O	Mycosep	GC–FID
Young & Games (1993)	Liquid culture	NA	NA	NA	SFC–UV
Young & Games (1993)	Liquid culture	NA	NA	NA	SFC–MS
Groves et al. (1999)	Maize	5	CH₃CN/H₂O	Aluminium oxide/C18	GC–MS (TMS)
Petterson (1992)	Barley, oats, wheat	20	CH₃CN/H₂O	Charcoal/ aluminia	GC–ECD (TMS)
Stenwig et al. (1992)	Barley, oats	50	CH₃CN/H₂O	Charcoal/ aluminia/ celite	GC–ECD (TMSI)
Lauren & Agnew (1991)	Maize	20	CH₃CN/ MeOH/H₂O	Alumina/ cation resins/celite	HPLC–UV
Lauren & Agnew (1991)	Maize	20	CH₃CN/ MeOH/H₂O	Alumina/ cation resins/celite	GC–ECD (TFAA)
Scudamore et al. (1998)	Maize	30	CH₃CN/4% KCl	Charcoal/ aluminia	GC–MS (TFAA)
Mirocha et al. (1998)	Wheat, barley	25	CH₃CN/H₂O	C18/alumina	GC–MS (TMSI, TCMS)
Liu et al. (1997)	Wheat, barley,oats		CH₃CN/H₂O	Charcoal/ aluminia/ celite	GC–ECD (TMSI)
Park et al. (1991)	Cereals	50	CH₃CN/H₂O	Florisil	GC–MS (TMI/TMA/ TMCS)
Tanaka et al. (1986)	Wheat, barley	20	CH₃CN/H₂O	Florisil/C18	GC–ECD
Tanaka et al. (1986)	Wheat, barley	20	CH₃CN/H₂O	Florisil/C18	(TMS)
Swanson et al. (1986)	Milk	40 mL	Chem tube/ ethyl acetate	C18	GC–ECD (TMS, HFB)
Dahlem et al. (1986)	Urine	9 mL	Clin-Elut/ethyl acetate		GC–ECD (TMS)
Dahlem et al. (1986)	Faeces	12 g	MeOH/H₂O/ Clin-Elut	C18
Kiessling et al. (1984)	Rumen fluid	5 mL	MeOH/0.1% NaCl	C18	GC–FID
Scott et al. (1993)	Beer	50 mL	Chem-Elut/ ethyl acetate/ MeOH	C18	GC–MS
					(HFB)

Tanaka et al. (1990)	Cereals	50	CH₃CN/H₂O	Florisil	GC–ECD
Tanaka et al. (1990)	Cereals	50	CH₃CN/H₂O	Florisil	(TMS)
Scott & Trucksess (1997)	Wheat, maize	NA	NA	Charcoal/C18	GC–MS
Scott & Trucksess (1997)	Wheat, maize	NA	NA	Charcoal/C18	(TMS, HFB)
Scott et al. (1989)	Wheat	50	MeOH/ H₂O	C18/alumina/ carbon	GC–ECD
Scott et al. (1989)	Wheat	50	MeOH/ H₂O	C18/alumina/ carbon	(HFB, TMS)
Luo et al. (1990)	Wheat, maize	40	CH₃CN/H₂O	Florisil	GC–ECD
Luo et al. (1990)	Wheat, maize	40	CH₃CN/H₂O	Florisil	(TMS)
Möller & Gustavsson (1992)	Cereals	50	CH₃CN/ EtOAc/H₂O	Florisil-Sep Pak	GC–ECD
					(Tri-syl/
					TBT)
Onji et al. (1998)	Cereals	15	CH₃CN/H₂O	Bond Elut florisil	GC–MS
Onji et al. (1998)	Cereals	15	CH₃CN/H₂O	Bond Elut florisil	(on-column)
Krska et al. (1997)	Wheat	25	CH₃CN/H₂O	Mycosep	GC–ECD
Krska et al. (1997)	Wheat	25	CH₃CN/H₂O	Mycosep	(Tri-syl/TBT)
Krska (1998)	Wheat	25	CH₃CN/H₂O	Mycosep	GC–ECD
Krska (1998)	Wheat	25	CH₃CN/H₂O	Mycosep	(Tri-syl/TBT)
Josephs (1999)	Wheat	25	CH₃CN/H₂O	Mycosep	GC–ECD
					(Tri-syl/
					TBT)
Kotal et al. (1999)	Cereals	5	CH₃CN/H₂O CH₃CN/ MeOH	GPC (BioBeads S-X3)	GC–ECD
Kotal et al. (1999)	Cereals	5	CH₃CN/H₂O CH₃CN/ MeOH	GPC (BioBeads S-X3)	(TFAA)

Reference^b	Matrix	Amount of sample (g)	Toxins ng/g	LOD/LOQ levels	Spiking (%) (ng/g)	Recovery	CV (%)
Veldman et al. (1992)	Feed	20	DON	100–500/NA	500–600	60–86	NA
Rajakylä et al. (1987)	Wheat, rye	20	DON	250–1000/NA	NA	75	10
Zabe et al. (2000)	Food	NA	DON	NA	0–5	80	NA
Josephs et al. (1998)	Wheat	1	DON	1600/5350	2350–16 750	53	9.6
Pineiro et al. (1996)	Liquid culture	50	DON	NA	NA	96	NA
Sano et al. (1987)	Maize, wheat, barley	20	DON	20/NA	1000	> 70	NA
			NIV
			FX
Schuhmacher et al. (1996)	Wheat	15	DON	500/2000	1000–50 000	79	9.6
Berger et al. (1999)	Wheat	10	DON	6/50	NA	86	3.6
			FX	3/40		91	3.4
			3-AcDON	3/25		98	2.6
Cahill et al. (1999)	Wheat	50	DON	100/NA	100–10 000	100–81	NA
Razzazi-Fazeli et al. (1999)	Wheat	25	DON	NA/40	50–500	86	3.5–7.7
Razzazi-Fazeli et al. (1999)	Wheat	25	NIV	NA/50		70
Reutter (1999)	Cereals, feed	25	DON	NA/50	NA	NA	NA
Reutter (1999)	Cereals, feed	25		NA/100
Mulders et al. (1986)	Cereals	25	DON	20/NA	100	99	11
Scott & Kanhere (1986)	Cereals	50	DON	20/NA	200	121	NA
Scott & Kanhere (1986)	Cereals	50			1000	93 (barley)
Tacke & Casper (1996)	Wheat, barley, malt	5	DON	50/NA	500–20 000	94–100
Weingärtner et al. (1997)	Wheat	25	DON	35/123	100–2000	102	9.5–18
			3-AcDON	41/143		96
			15-AcDON	35/123		95
			FX	35/124		97
			NIV	33/117		66
Schollenberger et al. (1998)	Cereals, food, feed	10	DON	7/23	50–500	83–98	NA
			FX	6/21		82–120
			15-AcDON	7/22		73–108
			3-AcDON	5/16		88–110
			NIV	12/38		71–107
Croteau et al. (1994)	Maize	50	DON	NA/50	100	73	NA
					250	108
					1000	81
Adler et al. (1992)	Wheat	20	DON	NA	NA	NA	NA
Blaas et al. (1984)	Cereals	20	DON	10/NA	NA	NA	NA
Blaas et al. (1984)	Cereals	20	NIV
Seidel et al. (1993)	Wheat, maize	10	DON	3.5/NA	NA	75–80	5
van Egmond (2000)	Wheat	25	DON	25/75	300	96	4.8
			NIV			96	3.8
			FX			98	2.9
			3-AcDON			102	0.7
Young & Games (1993)	Liquid culture	NA	DON	150/NA	NA	NA	NA
			3-AcDON	100/NA
			ZEN	250/NA
Groves et al. (1999)	Maize	5	DON	500/NA	1000	80–100	NA
			NIV
			FX
			3-AcDON
			15-AcDON
Petterson (1992)	Barley, oats, wheat	20	DON	10/NA	NA	NA	NA
Petterson (1992)	Barley, oats, wheat	20	NIV
Stenwig et al. (1992)	Barley, oats	50	DON	10/NA	NA	90	NA
Lauren & Agnew (1991)	Maize	20	DON	30/NA	NA	55–70	NA
			NIV	20/NA (GC)		73–103

Scudamore et al. (1998)	Maize	30	DON	10/NA	100	72–80	10–16
			NIV	10/NA
			FX	10/NA
Mirocha et al. (1998)	Wheat, barley	25	DON	NA	1 000	97	NA
					5 000	88
					10 000	87
Liu et al. (1997)	Wheat, barley,oats		DON	30/NA	NA	NA	11
				3-AcDON	30/NA
				NIV	50/NA
Park et al. (1991)	Cereals	50	DON	15/NA	1000	83	NA
Park et al. (1991)	Cereals	50	NIV			89
Tanaka et al. (1986)	Wheat, barley	20	DON	2/NA	NA	NA	NA
Tanaka et al. (1986)	Wheat, barley	20	NIV
Swanson et al. (1986)	Milk	40 mL	DON	1 ng/mL/NA	2.5–10 ng/mL	82	10
Dahlem et al. (1986)	Urine	9 mL	DON	50/NA	200–5000 ng/mL	86	8.5
Dahlem et al. (1986)	Faeces	12 g				87	10
Kiessling et al. (1984)	Rumen fluid	5 mL	DON	2/NA	NA	43	NA
Scott et al. (1993)	Beer	50 mL	DON	1.5 ng/mL	5–20 ng/mL	103	NA
			NIV	0.3 ng/mL		90
			ZEN	2 ng/mL/NA		97
Tanaka et al. (1990)	Cereals	50	DON	2/NA	300	88	1.9–11
			NIV	2/NA	300	88
			ZEN	1/NA	500	91
Scott & Trucksess (1997)	Wheat, maize	NA	DON	7–100/NA	500–1000	81–96	NA
Scott & Trucksess (1997)	Wheat, maize	NA	NIV		500–1000	48–73
Scott et al. (1989)	Wheat	50	DON	20–50/NA	200–1000	81–108	NA
Scott et al. (1989)	Wheat	50	NIV	200/NA	200–1000	71–102
Luo et al. (1990)	Wheat, maize	40	DON	5/NA	200	97–103	NA
Luo et al. (1990)	Wheat, maize	40	NIV	5/NA	200
Möller & Gustavsson (1992)	Cereals	50	DON	NA	25–250	86–111	NA
			NIV			72–110
			FX			84–95
			3-AcDON			91–106
Onji et al. (1998)	Cereals	15	DON	100–500/NA	60	75	22
			FX			49	29
			3-AcDON			82	28
Krska et al. (1997)	Wheat	25	DON	NA	100–2000	96	9.5
Krska (1998)	Wheat	25	DON	40/NA	100–2000	96	9.5
Josephs (1999)	Wheat	25	DON	28/57	257–2060	103	4.3–7.8
			NIV	41/82	204–1438	76
			FX	28/56	280–2240	106
			3-AcDON	23/47	255–2040	107
			15-AcDON	23/46	280-2240	107
Kotal et al. (1999)	Cereals	5	DON	NA/50	2000	88	NA
			NIV	NA/40		87
			FX	NA/100		78

^{a Fordefinitions of abbreviations, see end of Trichothecenes Appendix 6.
^{b For references, see the monograph on deoxynivalenol.}Trichothecenes Appendix 3.
Methods for screening for A-trichothecenes and particularly for T-2 and HT-2 toxins^a}

Reference^b	Matrix	Amount of sample (g)	Extraction	Clean-up	Technique
Romer (1986)	Foods, feeds		CH3CN/H2O	Charcoal/ alumina	TLC
Kawamura et al. (1990)	Cereals	20	CH3CN/H2O	Florisil	icELISA
Märtlbauer et al. (1991)	Milk	NA	MeOH	None	ELISA
Barna-Vetro et al. (1997)	Cereals		89% CH3CN	None	ELISA
Yang & Luo (2000)	Wheat	20	CHCl3/EtOH	Charcoal/ alumina	ELISA

Reference^b	Matrix	Amount of sample (g)	Toxins assayed	LOD/LOQ (ng/g)	Spiking levels (ng/g)	Recovery (%)	CV (%)
Romer (1986)	Foods, feeds		T-2	500(TLC)/NA	850-1200	98	12
Romer (1986)	Foods, feeds		HT-2				13
Kawamura et al. (1990)	Cereals	20	T-2	0.2/NA	NA	NA	NA
Märtlbauer et al. (1991)	Milk	NA	T-2	0.5/NA	NA	68-98	5-20
Barna-Vetro et al. (1997)	Cereals		T-2	50/NA	NA	85	3-15
Yang & Luo (2000)	Wheat	20	T-2	NA	NA	NA	NA

From Hack (1994), Gendloff (1987), and Hunter (1985), with sensitivities of 5 pg/ml (0.25 pg/assay), 10 ng/ml (0.5 ng/assay), and 50 ng/assay, respectively. Park & Chu (1996) reported the efficiency of ELISA and RIA for the analysis of T-2 toxin and of 3-AcDON in naturally contaminated maize samples. A homogeneous competition inhibition immunoassay for T-2 toxin was reported by Ligler et al. (1987).

^{a Fordefinitions of abbreviations, see end of Trichothecenes Appendix 6.
^{b For references, see monograph on T-2 and HT-2 toxins}}

Trichothecenes Appendix 4.

Quantitative methods for A-trichothecenes and particularly for T-2 and HT-2 toxins^a

Reference^b	Matrix	Amount of sample (g)	Extraction	Clean-up	Technique
Berger et al. (1999)	Wheat	10	CH₃CN/H₂O	Mycosep 227, 216	LC–MS
Rajakylä et al. (1987)	Wheat, rye	20	CH₃CN	Bond Elut	HPLC–MS
Möller & Gustavsson (1992)	Cereals	50	CH₃CN/ EtOAc/H₂O	Florisil-Sep Pak	GC–ECD (Tri-syl/ TBT)
Rosen & Rosen (1983)	Maize	10	MeOH	Silica gel/C18	GC–MS
Schollenberger et al. (1998)	Cereals, food, feed	10	CH₃CN/H₂O	Florisil/Bond Elut CBA	GC–MS (TFA)
Onji et al. (1998)	Cereals		CH₃CN/H₂O	Bond Elut florisil	GC–MS (on-column)
Croteau et al. (1994)	Maize	50	CH₃CN/H₂O, shake 2.5 h	Charcoal/ alumina	GC–ECD (DMAP, HFBA)
Plattner et al. (1989)	Cultures		Ethyl acetate	charcoal	GC–MS (Tri-syl/TBT)
Scott & Kanhere (1986)	Standards	NA	NA	NA	GC–ECD (TMS, HFB)
Groves et al. (1999)	Maize	5	CH₃CN/H₂O	Aluminium oxide/C18	GC–MS (TMS)
Petterson (1992)	Barley, oats, wheat	20	CH₃CN/H₂O	Charcoal/ alumina	GC–ECD (TMS)
Scudamore et al. (1998)	Maize	30	CH₃CN/4 KCL	Charcoal/ alumina	GC–MS (TFAA)
Hietaniemi & Kumpulainen (1991)	Cereals, feed	20	MeOH/ H₂O	Silica gel/ Florisil/C18	GC–MSD (TMSI)
Park et al. (1991)	Cereals	50	CH₃CN/H₂O	Florisil	GC–MS (TMI/TMA/ TMCS)
Kiessling et al. (1984)	Rumen fluid	5 mL	MeOH/0.1% NaCl	C18	GC–FID (?)
Scott (1997)	Wheat, maize	NA	NA	Charcoal/C18	GC–MS (TMS, HFB)
Scott et al. (1989)	Wheat	50	MeOH/ H₂O	C18/alumina/ carbon	GC–ECD (HFB, TMS)
Luo et al. (1990)	Wheat, maize	40	CH₃CN/H₂O	Florisil	GC–ECD (TMS)
Josephs (1999)	Wheat	25	CH₃CN/H₂O	Mycosep	GC–ECD (HFBI)
Kotal et al. (1999)	Cereals	5	CH₃CN/H₂O	GPC (Bio Beads S-X3)	GC–ECD (TFAA)
Kotal et al. (1999)	Cereals	5	CH₃CN/MeOH		HT-2

Reference^b	Matrix	Amount of sample (g)	Toxins assayed	LOD/LOQ (ng/g)	Spiking levels (ng/g)	Recovery (%)	CV (%)
Berger et al. (1999)	Wheat	10	T-2	3/60		87	4.6
Berger et al. (1999)	Wheat	10	HT-2	1/10		90	1.7
Rajakylä et al. (1987)	Wheat, rye	20	T-2	250–1000/NA	NA	89	6.3–7.7
Rajakylä et al. (1987)	Wheat, rye	20	HT-2			90
Möller & Gustavsson (1992)	Cereals	50	T-2	NA	250–2500	76–93	NA
Möller & Gustavsson (1992)	Cereals	50		HT-2			71–94
Rosen & Rosen (1983)	Maize	10	T-2	20/50	111	80	14
Rosen & Rosen (1983)	Maize	10	HT-2	5/20	46	91	8.7
Schollenberger et al. (1998)	Cereals, food, feed	10	T-2	2/6	50–500	87–110	NA
Schollenberger et al. (1998)	Cereals, food, feed	10	HT-2	5/18		74–109
Onji et al. (1998)	Cereals		T-2	100–500/NA	60	98	22
Croteau et al. (1994)	Maize	50	T-2	NA/100	100	70/121	NA
			HT-2	NA/50	250	NA/95
					1000	83/99
Plattner et al. (1989)	Cultures		T-2	NA	NA	NA	NA
Scott & Kanhere (1986)	Standards	NA	T-2	NA	NA	NA	NA
Scott & Kanhere (1986)	Standards	NA	HT-2
Groves et al. (1999)	Maize	5	T-2	500/NA	1000	80–100	NA
Groves et al. (1999)	Maize	5	HT-2
Petterson (1992)	Barley, oats, wheat	20	T-2	50/NA	NA	NA	NA
Petterson (1992)	Barley, oats, wheat	20	HT-2	10/NA
Scudamore et al. (1998)	Maize	30	T-2	10/NA	100	0–103	20–38
			HT-2	10/NA	100	21–53
			DAS	10/NA	100	64–73
Hietaniemi & Kumpulainen (1991)	Cereals, feed	20	T-2	15/NA	NA	72–130	NA
Hietaniemi & Kumpulainen (1991)	Cereals, feed	20		HT-2
Park et al. (1991)	Cereals	50	T-2	15/NA	1000	83	NA
Kiessling et al. (1984)	Rumen fluid	5 mL	T-2	10/NA	NA	67	NA
Kiessling et al. (1984)	Rumen fluid	5 mL	HT-2	5/NA	NA	NA
Scott (1997)	Wheat, maize	NA	T-2	NA	4000	94	NA
Scott (1997)	Wheat, maize	NA	HT-2		1200–2400	88–93
Scott et al. (1989)	Wheat	50	T-2	400/NA	200–4000	66–91	NA
Scott et al. (1989)	Wheat	50	HT-2	100–200/NA	200–4000	72–110
Luo et al. (1990)	Wheat, maize	40	T-2	100/NA	200	97–103	NA
Luo et al. (1990)	Wheat, maize	40	HT-2	100/NA	200
Josephs (1999)	Wheat	25	T-2	10/37	51–1027	100	2.2
Josephs (1999)	Wheat	25	HT-2	10/38	52–1047	117
Kotal et al. (1999)	Cereals	5	T-2	NA/200	2000	91	NA
Kotal et al. (1999)	Cereals	5	NA/100		105

^{a Fordefinitions of abbreviations, see end of Trichothecenes Appendix 6.
^{b For references, see monograph on T-2 and HT-2 toxins}Trichothecenes Appendix 5.
Analytical methods used in surveys summarized in Appendix A of the monograph on

deoxynivalenol and Appendices A and B of the monograph on T-2 and HT-2 toxins^a}

Reference^b	Matrix	Amount of sample (g)	Extraction	Clean-up	Method of quantification	Toxins assayed	LOD/ LOQ (ng/g)	Spiking levels (ng/g)	Recovery (%)	RSD_r	RSD_R
Abouzied et al. (1991)	Grain foods	25	CH₃CN:H₂O 9:1		ELISA	DON	1000
Ali et al. (1998)	Maize	20	CH₃CN:H₂O 3:1 100 ml	Florisil column	GC–MS	DON	10/NA
Ali et al. (1998)	Maize	20	CH₃CN:H₂O 3:1 100 ml	Florisil column	TMS derivatives	NIV	10/NA
Eskola et al. Cereals (2000a,b)			Mycosep 227	GC–MS column	DON	NA/10
					TMS derivatives	3-AcDON
						NIV
						FX
						DAS
						HT-2	NA/20
						T-2	NA/20
FAO/WHO (2000)	Cereals	20	H₂O 8 ml	Petroleum ether:ethanol: H₂O	TLC
	Cereal products		Chloroform+ ethanol (8+2) 100 ml
Fernández et al. (1994) (modified from Trucksess et al., 1984)	Wheat	50	CH₃CN:H₂O 84:16 200 ml	Charcoal/celite/ alumina column	HPTLC Chloroform: acetone:2- propanol 8:1:1	DON	40/NA	200	83
								400	82
								800	72
Furlong & Valente Soares (1995)	Wheat, maize, wheat products		Methanol:4% KCl 9:1	Chloroform	GC–FID	DON	200/NA		85
				Charcoal/celite/ alumina column	HFB derivatives	NIV	100/NA		87
						DAS	100/NA		83
						T-2	100/NA		87
						HT-2	500/NA		91
						T-2 triol	300/NA
						T-2 tetrol	400/NA
Golinski et al. (1996)	Wheat	20	Tanaka et al. (1985)	Tanaka et al. (1985)	HPLC C-R4A C-18 column UV (229 nm) methanol water 20%	DON	10/NA
Golinski et al. (1996)	Wheat	20	Tanaka et al. (1985)	Tanaka et al. (1985)	HPLC C-R4A C-18 column UV (229 nm) methanol water 20%	NIV
Groves et al. (1999)	Maize, maize products	5	CH₃CN:H₂O 84:16, 20 ml shake 60 min	Mixture of neutral aluminum oxide and C-18 packing material	GC–MS TMS derivatives	17 toxins	500/NA	1000	80–100
Hietaniemi & Kumpulainen (1991)	Grains, feeds	20	CH₃CN:H₂O 95:5, 100 ml, homogenize 5 min	Hexane isobutanol Silica gel column Florisil or Sep Pak C18 column	GC–MS	DON	5		72–128
					TMS derivatives	DAS	5		(mean, 100)
						3-AcDON	5
						FX	5
						NIV	5
						T-2	15
						HT-2	15
Home Grown Cereals Authority (1999, 2000)	Wheat	20	CH₃CN:H₂O 84:16, 100 mL shake 120 min	Mycosep No. 225 column	GC–MS	DON	20/NA
					TMS derivatives	T-2	20/NA
					TMS derivatives	HT-2	20/NA
Kamimura et al. (1981)	Maize	100	CH₃OH:H₂O 95:5, 200 mL shake 30 min	1-Amberlite XAD-4 column 2-Florisil	TLC Developing solvents: 1-CHCl₃: methanol 93:7 2-Toluene: acetone: methanol 5:3:2	DON	20–50/NA	1000	93
						T-2	100–500/	2000	83
						T-2	NA
						HT-2	100–500/	2000	92
						HT-2	NA
						NIV	20–50/NA	1000	82
						FX	20–50/NA	1000	90
						ZEN	10–50/NA	2000	88
						DAS	100–500/	2000	102
						DAS	NA
						NEO	100–500/	2000	108
						NEO	NA
						Butenolide	100–500/	2000	92
						Butenolide	NA
						Moniliform	200–500/	2000	76
						Moniliform	NA
Kim et al. (1993) modified from Tanaka, 1985)	Barley, maize	40	CH₃CN:H₂O 3:1, 160 ml 30 min	Hexane Florisil column	GC–MS TMS derivatives	DON	5	200	92–105 (barley)
						3-AcDON	5		87–95 (maize)
						15-AcDON	5
						NIV	5
						4-ANIV	5
Langseth (2000a,b)	Cereals	25	CH₃CN:H₂O 84:16, homogenize	Mycosep 225	GC–MS PFPA derivatives	DON	20–30	400	90
						3-AcDON
						NIV		400	72
						FX
						T-2	30	400	90
						HT-2	20	400	91
						Scirpentriol
						MAS
						DAS
Langseth & Claasen (1992)	Cereals	25	CH₃CN:H₂O 84:16, homogenize	Charcoal/ celite/ alumina column	GC–ECD TMS derivatives	DON	10/30	400	80–110
						3-AcDON		400
						NIV		400
						FX		400
Langseth & Elen (1996)	Cereals	25	CH₃CN:H₂O 84:16, homogenize	Charcoal/ celite/ alumina column, hexane, silica cartridge	GC–ECD TMS derivatives
Lauren & Agnew (1991)	Grains	20	CH₃CN:CH₃OH: H₂O, 80:5:15 100 ml, 2 h	Alumina–carbon cation exchange resin, hydrolysis	HPLC C-8 GC–ECD TFAA derivatives	DON	10*
Lew et al. (2000)	Maize	25	CH₃CN:H₂O 84:16, 100 mL shake 80 min	Mycosep No. 227 column	GC–ECD Type B, TMS derivatives Type A, HFBI derivatives	DON	50
						NIV	50
						3-AcDON	50
						15-AcDON	50
Luo et al. (1990)	Maize, wheat	40	CH₃CN:H₂O 3:1, 160 ml shake 30 min	Hexane Florisil column	GC–MS TMS derivatives	DON	5		97–103
						NIV	5		97–104
						T-2	100		97–105
						15-AcDON	100		97–106
						HT-2	100		97–107
Mallman (2000)	Cereals	50	CH₃CN:H₂O 84:16, 100 mL	Mycosep No. 224, automatic	HPLC C-18	DON	100
Marasas et al. (1979)	Maize	50	Hexane, 100 mL methanol:1% KCl , 55:45 200 mL	Hexane chloroform	GC–MS TMS derivatives	DON	100/NA		60
						DAS	100/NA		90
						T-2	100/NA		87
Marx et al. (1995)		20	CH₃CN:H₂O 3:1, 200 mL	Hexane Florisil Sep-pak	HPTLC	DON	50
Möller & Gustavsson (1992)	Cereals	50	Ethyl acetate: acetonitrile 4:1 250 ml + 10 ml water, shake 30 min	Hexane Sep Pak Florisil cartridge	GC–ECD TMS derivatives	DON		250	89
						FX		250	87
						NIV		250	72
						3-AcDON		250	93
						DAS		250	89
						Tricho- thecins		250	115
						NEO		250	97
						HT-2		250	94
						T-2		250	84
Möller & Gustavsson (1992)	Cereals	50	Ethyl acetate: acetonitrile 4:1 250 ml + 10 ml water, shake 30 min	Hexane Sep Pak Florisil cartridge	GC–ECD TMS derivatives	DON		250	86
						FX		250	84
						NIV		250	68
						3-AcDON		250	89
						DAS		250	105
						Tricho- thecenes		250	81
						NEO		250	93
						HT-2		250	77
						T-2		250	79
Neira et al. (1997)	Wheat products	Pacin et al. (1997)	Pacin et al. (1997)	Pacin et al. (1997)	GC–ECD HFB derivatives	DON	6/17
Niessen et al. (1993)	Beer	20 ml			ELISA	DON	50 ng/ml	50	45
								500	77
								1000	83
Pacin et al. (1997) (modified Romer® MY 8402s)	Wheat products (with fat)	25	1-n-hexane, 2-acetonitrile:H₂O 84:16, 100 mL blend 3 min	Mycosep No. 225	TLC	DON	25/48	100	102
								250	87
								500	92.4

Park et al. (1992)	Barley	Luo et al. (1990)	Luo et al. (1990)	Florisil silica cartridge	GC–ECD TMS derivatives	DON	2		88–90
Park et al. (1992)	Barley	Luo et al. (1990)	Luo et al. (1990)	Florisil silica cartridge	GC–ECD TMS derivatives	NIV
Pettersson (1993)	Barley, oats, wheat	20	CH₃CN:H₂O 84:16	Charcoal/ alumina column	GC–ECD TMS derivatives	DON	10
						NIV
						T-2	20–50
						HT-2	10/30
						DAS
						3-AcDON
Romer Labs Inc. (1994); Lops et al. (1998)	Wheat	25	CH₃CN:H₂O 84:16, 100 ml blend 3 min	Mycosep No. 224	TLC, tolueno: ethyl acetate: formic acetate 5:4:1	DON	500/NA		> 85
Ryu et al. (1996)	Barley, maize	20	CH₃CN:H₂O 3:1, 200 ml shake 30 min	Hexane Florisil column	GC–MS TMS derivatives	DON	5		83.8
						T-2	5		88.2
						HT-2	5		91.3
						NIV	5		91.8
						FX	5		90.9
						3-AcDON	5		87.9
Schollenberger et al. (1998)	Cereals, cereal foods	10	Tanaka et al. (1985)	Hexane Florisil cation exchange cartridge	GC–MS TFAA derivatives	DON	7/23	50–500	83–98
						T-2	2/6	50–500	87–110
						HT-2	5/18	50–500	74–109
						NIV	12/38	50–500	71–107
						FUS	6/21	50–500	82–120
						15-AcDON	7/22	50–500	73–108
						3-AcDON	5/16	50–500	88–110
						DAS	7/22	50–500	82–116
Schwadorf & Müller (1991)	Barley, oats, wheat	20	Tanaka et al. (1985)	Tanaka et al. (1985)	GC–MS TFA derivatives	NIV	15		84.7
						DON	15		89.1
						3Ac-DON	15		81.3
						FX	15
						HT-2	15		79.6
						T-2	15		80.2
						DAS	15		77.8
						15-AcDON	15		83.2
Scott et al. (1981)	Cereals	50	Methanol: H₂O 1+1, 250 mL	Partition: aqueous ammonium sulfate 30%/ ethyl acetate, silica gel column	GC–ECD HFBI derivatives	DON	10	100 (wheat	72, 73,
								flour)	86, 70
								1000 (wheat	85, 59,
								flour)	78, 68
								100 (wheat)	101, 102
								500 (wheat)	77, 88
								1000 (wheat)	78, 86
Scott et al. (1986)	Cereals	50	Methanol: H₂O 7:3, 150 ml	Partition: aqueous ammonium sulfate 30%/ ethyl acetate, silica gel column	GC–ECD TMS derivatives	DON	150/NA
Scott et al. (1986)	Cereals	50	Methanol: H₂O 7:3, 150 ml		GC–ECD TMS derivatives	NIV
Scott et al. (1989)	Cereals	50	Methanol: H₂O 7:3, 150 mL	Partition: aqueous ammonium sulfate 30%/ ethyl acetate, silica gel column	GC–ECD TMS derivatives: DON and NIV HFB derivatives: other trich-othecenes	DON	20/50	800 (wheat)	81
						NIV	200
						T-2	200/400	4000 (wheat)	86
						HT-2	50/200	2000 (wheat)	82
						DAS	200
Scott et al. (1993)	Beer	10	Chem Elut CE 1020, Varian	C-18 SPE (Supel-clean, ) Supelco	GC–MS HFB derivatives	DON	0.1–1.5	5–20 ng/ml	103
Scott et al. (1993)	Beer	10	Chem Elut CE 1020, Varian	C-18 SPE (Supel-clean, ) Supelco	GC–MS HFB derivatives	NIV	0.01–0.3
Solovey et al. (1999) (modified Romer® products MY 8402s)	Maize, wheat. maize and wheat	25	CH₃CN:H₂O 84:16, 100 mL blend 3 min	Mycosep No. 225	TLC	DON	25/48	100	82
Solovey et al. (1999) (modified Romer® products MY 8402s)	Maize, wheat. maize and wheat	25	CH₃CN:H₂O 84:16, 100 mL blend 3 min	Mycosep No. 225				300	86
Spanjer (2000)	Cereals, cereal products	25	PEG:H₂O 5 g/400 ml	immunoaffinity column DON test^TM HPLC (VICAM)	HPLC–UV RP18	DON	16/32	146–1070	92–103	13
Stratton et al. (1993)	Wheat, barley	50	Acetonitrile:H₂O 21:4, 250 ml shake 2 h	n-Hexane LC–NH₂ solid phase extraction column	HPLC C-18 column UV	T-2	150/NA	400 (wheat)	84.7
						DON	50/NA	100 (wheat)	89.8
						HT-2	120/NA	400 (wheat)	84.9
						DAS	70/NA
				Charcoal/celite/ alumina column	DON, 220 nm T-2, HT-2, 200 nm, mobile phase, methanol: H₂O
Sydenham & Thiel (1987)	Maize, oats	50	Methanol: water 1:1, shake 30 min	Extrelut 20 column Silica gel cartridge	GC–ECD HFBI derivatives	T-2	100/NA		88.0	5
Sydenham & Thiel (1987)	Maize, oats	50	Methanol: water 1:1, shake 30 min	Extrelut 20 column Silica gel cartridge	GC–ECD HFBI derivatives	DAS	100/NA		88.2	4.4
Tanaka et al. (1985)	cereal	20	CH₃CN:H₂O 3:1, 200 ml shake 30 min	Hexane Florisil Sep-pak	GC–ECD TMS derivatives	DON	2/NA	300	87
Tanaka et al. (1985)	cereal	20	CH₃CN:H₂O 3:1, 200 ml shake 30 min	Hexane Florisil Sep-pak	GC–ECD TMS derivatives	NIV	2/NA	300	86
Trucksess et al. (1984); Eppley et al. (1986)	Wheat, maize	50	CH₃CN:H₂O 84:16, 200 mL	Charcoal/ celite/ alumina column	TLC chloroform: acetone: 2-propanol 8:1:1	DON	50/NA		77–93	50 ppb, 45	87
										300 ppb, 39	40
										1000 ppb, 64	64
Trucksess et al. (1987)	Wheat	50	Acetonitrile: H₂O: methanol 70:25:5, 200 mL shake 60 min	Lead acetate Charcoal/ celite/ alumina column Octa-decylsilane column	HPTLC 1-chloroform: acetone: 2-propanol 8:1:1 2-chloroform: acetone:2- propanol 7:1.5:1.5	DON	50	50	61.3
								100	85
								200	81.5
						NIV
						FX
Trucksess et al. (1996)	White flour	25	CH₃CN:H₂O 84:16, 100 mL blend 3 min or shake 30 min	Mycosep No. 225 column	HPLC Novapack C₁₈column	DON	20/500	500	95	7.52	11.46
								1000	84	5.72	18.98
								2000	85	3.14	12.72
Tutelyan et al. (1990)	Wheat	25	CH₃CN:H₂O 84:16, 125 mL	Charcoal/ alumina column	HPLC, silica gel column, UV (224 nm) mobile phase, hexane:isopro- panol: H₂O 75:25:1.5	DON	50/NA	500	90
Vega et al. (1994) (Romer method®)	Wheat	25	CH₃CN:H₂O 84:16, 100 mL blend 3 min	Mycosep No. 227	HPTLC tolueno:ethyl acetate: formyl acetate 6:3:1	DON	10
						DAS
						HT-2
						T-2
Veratox® (1998); Trucksess et al. (1995)	Wheat, barley	50	H₂O, 250 ml blend 2 min		ELISA (Veratox method)	DON	300/500	1000	65
								2000	70
								4000	78
								1000	40
								2000	70
								4000	68
Vicam ® (1996); Lops et al. (1998)	Wheat	25	Polyethy-lene-glycol 5 g/400 mL H₂O	Immuno-affinity column, DON test^TM HPLC (VICAM)	HPLC Supelcosil LC-18 DAD 220 nm acetonitrile: H₂O 10:90	DON	50	500	75
								1000	84
								2000	85
Vrabcheva et al. (1996)	Wheat	2	CH₃CN:H₂O 84:16, 10 mL stir 120 min		ELISA (Ridascreen DON method) acetylate toxin before assay	DON	50		70–110
		2	Methanol: H₂O 70:30, 8 mL, stir 60 min CH₃OH: PBS 70:30, stir 30 min			3-AcDON
						15-AcDON
						T-2	>40
						DAS
Weingärtner et al. (1997)	Wheat	25	CH₃CN:H₂O 84:16, 100 mL shake 90 min	Mycosep No. 225 column derivatives	GC–ECD TMS	DON	36/124	DON 100		98	3.95
						NIV	33/144	DON 1000		80
						3-AcDON	41/123	DON 2000		81
						15-AcDON	35/124
						FX	35/117
Yang et al. (1992)	Cereals, cereal products	20	H₂O, 8 ml Chloroform + ethanol (8 + 2) 100 ml	Charcoal/ alumina column	ELISA	T-2

* Limit of detection

^{a Fordefinitions of abbreviations, see end of Trichothecenes Appendix 6.
^{b For references, see monographs on deoynivalenol and T-2 and HT-2 toxins}}

Trichothecenes Appendix 6.

Sampling methods used in surveys summarized in Appendix A of the monograph on
deoxynivalenol and Appendices A and B of the monograph on T-2 and HT-2 toxins

Reference^a	Grain	No. of samples	Site of collection	Lot size	Method of collection
Apro et al. (1987)	All	Proportional to total production of each area			Portions 50 g Automatic sampling
Eskola (2000a,b)	Cereals		Mills and malt houses	4–1000 t	Industry: Automatic sampling equipment: 100–200 g every 50–1000 kg
Eskola (2000a,b)					Farmers: Grain handling equipment. Several subsamples
Fonseca (1991)	Peanuts, maize, rice, beans	Proportional to size of lot	Bags or in bulk	20 t	Minimun no. of bags or points (bulk) to be sampled is a function (square root) of the lot size, portions of 200 g
Langseth & Elen (1997)	Barley, oats, wheat	Proportional to production and level of Fusarium infection	Grain loads, silos		Grain loads and
Olavarría (1992)	Maize, wheat, rice	Proportional to total production of each area	Harvest	8–12 t truck	Grain handling equipment (metal probe, 130 cm); 12–15 portions 300 g
Quiroga et al. (1995)	Wheat	Proportional to total production of each area	Harvest	max 2000 ha	Portions 50 g Automatic sampling or grain-handling equipment
Resnik & Pacin (2000)	Wheat, maize	Proportional to total production of each area	Harvest	max 2000 ha	Portions 50 g Automatic sampling or grain-handling equipment
Spanjer (2000)	Wheat, rye, buck-wheat, maize, rice, oats, millet, barley, grain products	Imported and domestic	Mills, silos, retail shops		Automatic and hand sampling

Reference^a	Grain	Sample size (kg)
Reference^a	Grain	Sample	Combined first sub- sample	Combined second sub- sample	Laboratory sample	Analysed sample
Apro et al. (1987)	All	Small grains (wheat, sorghum, oat, maize) = 10; peanut, soya bean, sunflower=50; cotton = 60	3 (wheat) 5 (maize)	1 (wheat) 2 (maize)	0.2	0.025
Eskola (2000a,b)	Cereals	3–100			0.5–1
Eskola (2000a,b)		0.5–1
Fonseca (1991)	Peanuts, maize, rice, beans	Homogenized and quartered into 4-kg samples
Langseth & Elen (1997)	Barley, oats, wheat				0.3–0.8	0.025
Olavarría (1992)	Maize, wheat, rice	5–6				0.025
Quiroga et al. (1995)	Wheat	10 Homogenized and quartered	3 14-mesh sieve	1 20-mesh sieve	0.2	0.025
Resnik & Pacin (2000)	Wheat, maize	10 Homogenized and quartered	3 (wheat) 5 (maize) 14-mesh sieve Romer mill			0.05
Spanjer (2000)	Wheat, rye, buck-wheat, maize, rice, oats, millet, barley, grain products	3–5 kg for grains	Slurry prepared with water in ratio 2:3		0.0625 of slurry	0.025

Abbreviations for Trichothecenes Appendixes 1–6: AcDON, acetylated deoxynivalenol; 4-ANIV, 4-acetylnivalenol; APCI, atmospheric pressure chemical ionization; CBA, carboxylic acid; CD, competitive direct; DAD, diode array detection; DAS, diacetoxyscirpenol; DMAP, 4-dimethylaminopyridine; DON, deoxynivalenol; ECD, electron capture detection; ELISA, enzyme-linked immonosorbent assay; FID, flame ionization detection; FLD, fluorimetric detection; FX, fusarenon X; GC, gas chromatography; GPC, gel permeation chromatography; HFB, heptafluorobutanoyl; HFBA, heptafluorobutyric anhydride; HPLC, high-performance liquid chromatography; HPTLC, high-performance thin-layer chromatography; ic, intracerebrally; Ig, immunoglobulin; LC, liquid chromatography; LOD, limit of detection; LOQ, limit of quanitification; MAS, monoacetoxyscirpenol; MS, mass spectroscopy; NEO, neosolaniol; NIV, nivalenol; NA, not available; PEG, polyethylene glycol; RSD_r, relative standard deviation for repeatability; RSD_R, relative standard deviation for reproducibility; SFC, supercritical fluid chromatography; SFE, supercritical fluid extraction; SPE, solid-phase extraction; TBT, tributyl tin; TCMS, trichloromethylsilane; TFA, trifluoroacetic acid; TFAA, trifluoroacetic anhydride; TLC, thin-layer chromatography; TMA, trimethylaluminium; TMCS, trimethylchlorosilane; TMI, trimethylindium; TMS, trimethylsilyl; TMSI, trimethylsilylimidazole; UV, ultra-violet radiation; ZEN, zearalenone

^{a For references, see monographs on deoynivalenol and T-2 and HT-2 toxins}

    See Also:
       Toxicological Abbreviations