Chromatographic analysis of dithiocarbamate residues and their metabolites in foods employed in dietary exposure studies—a review

Figures & data

Figure 1. Structures of common DTCs.

Table 1. Established ADI of DTCs and their metabolites.

	ADI (mg/kg bw/day)
	JMPR (established year)	EFSA (established year)
DTCs:
Metam		0.001 (2011)
Ferbam, ziram	0–0.003 (1996)	0.006 (ziram only) (2018)^a
Thiram	0–0.01 (1992)	0.01 (2017)
Mancozeb, maneb, metiram, zineb	0–0.03 (1993)	0.023 (mancozeb only) (2019)^b 0.03 (metiram only) (2018)^a
Propineb	0–0.007 (1993)	0.025 (2016)
Dazomet		0.01 (2010)
DTC metabolites:
ETU	0–0.004 (1993)	0.002 (2019)
PTU	0–0.0003 (1999)	0.002 (2016)
MITC		0.004 (2010)
DMTU		0.001 (2011)^c

Note: ADI was not established for milneb and polycarbamate by JMPR or EFSA.

^aADI for public consultation.

^bRapporteur Member State disagreed with the ADI established in 2019. The ADI of 0.05 mg/kg bw/day was established in 2009.

^cThe toxicological end points derived for metam were concluded to be applicable to DMTU.

Table 2. Summary of method performance of liquid chromatographic methods for analysing DTCs published from 2000 to 2020.

Matrix	DTCs	Extraction	Clean-up	Quantification techniques	Recoveries (%)	Precision (%)	LOQ	References
Fruit, vegetables	MDTC DMDTC EBDTC PBDTC	Extraction with alkaline buffer (sodium hydrogen phosphate/EDTA/ tetrabutylammonium hydrogen sulphate) (pH 11) containing sulphite	–	LC-UV (286 nm) or EC Column: C18 (150 x 4 mm, 5 μm) Mobile phase: methanol/extraction medium (gradient)	72–111	1–12	UV: 0.013–0.018 mg CS2/L EC: 0.009–0.012 mg CS2/L	van Lishaut and Schwack (2000)
Fruit	Neutral DTCs: dazomet thiram Metabolites: ETU PTU	Matrix solid-phase dispersion with Carbograph, elution with dichloromethane/methanol (80:20, v/v)	–	LC-APCI-MS (positive-ion mode) Column: C8 (150 × 4.6 mm, 5 μm) Mobile phase: methanol/water (gradient)	Unsatisfactory for fruits with high acid content (orange and lemon), thiram not recovered	4–21	Dazomet 0.5 mg/kg thiram 2.5 mg/kg ETU 0.25 mg/kg PTU 0.25 mg/kg	Blasco et al. (2004)
Fruit, vegetables	DMDTCs EBDTCs PBDTC	Extraction with alkaline buffer (sodium hydrogen carbonate/dl-penicillamine)	–	LC-ESI-MS (negative-ion mode) Column: ZIQ-pHILIC (150 × 4.6 mm, 5 μm) Mobile phase: acetonitrile/10 mM ammonia (gradient)	90–100	2	DMDs 0.04 mg/kg EBDs 0.05 mg/kg PBDs 0.04 mg/kg	Crnogorac and Schwack (2007)
Fruit, vegetables	EBDTCs	1. Treatment with alkaline EDTA/cysteine 2. Derivatisation with dimethyl sulphate	dSPE (MgSO4, PSA)	LC-ESI-MS^2 (positive-ion mode) Column: C18 (50 × 2.1 mm, 1.9 μm) Mobile phase: methanol/0.1% formic acid (gradient)	82–101	1.8–9.2	0.8 mg/kg	Hayama and Takada (2008)
Fruit, wheat, vegetables	EBDTCs	1. Treatment with alkaline EDTA/cysteine 2. Derivatisation with methyl iodide	C18 SPE cartridge	LC-ESI-MS^2 (positive-ion mode)	79–104	–	0.01 mg/kg	Brewin et al. (2008)
Fruit, vegetables	DMDTCs EBDTCs PBDTC	Extraction with alkaline buffer (sodium hydrogen carbonate/dl-penicillamine)	–	LC-ESI-MS^2 (negative-ion mode) Column: ZIQ-pHILIC (150 × 4.6 mm, 5 μm) Mobile phase: acetonitrile/10 mM ammonia (gradient)	97–101	3	0.003–0.005 mg/kg	Crnogorac et al. (2008)
Fruit, vegetables, cereals	DMDTCs EBDTCs PBDTC ETU PTU	1. Treatment with alkaline EDTA 2. Derivatisation with methyl iodide	–	LC-APCI-MS (positive ion mode) Column: Ncleodur C8 (150 × 4.6 mm, 5 μm) Mobile phase: methanol/water (gradient)	61–117	–	0.03–0.1 mg/L	Kim et al. (2010)
Tomato	Thiram Mancozeb Propineb	1. Treatment with alkaline EDTA/cysteine 2. Derivatisation with iodomethane	–	LC-UV (272 nm) Column: C18 (250 × 4.6 mm, 5 μm) Mobile phase: acetonitrile/water/methanol (25:65:35)	92–102	2–15	Thiram 0.05 mg/kg mancozeb and propineb 0.2 mg/kg	Jafari et al. (2012)
Fruit, vegetables	DMTDCs	Extraction with sulphite solution	–	LC-ESI-MS (negative-ion mode) Column: ZIQ-pHILIC (150 × 4.6 mm, 5 μm) Mobile phase: acetonitrile/10 mM ammonia (gradient)	81–106	1–13	0.002 mg/kg	Ringli and Schwack (2013)
Fruit, vegetables	DMDTCs EBDTCs PBDTC Thiram	Extraction with alkaline buffer (sodium hydrogen carbonate/dl-penicillamine) (pH 12)	–	LC-ESI-MS^2 (negative-ion mode) Column: ZIQ-pHILIC (150 × 4.6 mm, 5 μm) Mobile phase: acetonitrile/10 mM ammonia (gradient)	39–161	4–41	LOD = 0.03-1.19 mg/kg	Schmidt et al. (2013)
Apple, tomato, cucumber, rice	Thiram Mancozeb Propineb Metriam	1. Treatment with alkaline EDTA/cysteine 2. Derivatisation with iodomethane	–	LC-UV (272 nm) Column: C18 (250 × 4.6 mm, 5 μm) Mobile phase: acetonitrile/water (gradient)	71–105	0.6–13	0.011–0.105 mg/kg	Zhou et al. (2013)
Fruit, vegetables	Thiram N,N-diethyl-N′,N′-dimethylthiuram disulphide	Extraction with chloroform Exchange solvent to acetonitrile	–	LC-EC cell Column: C18 (50 × 4.6 mm, 2.6 μm) Mobile phase: acetonitrile/phosphate buffer (pH 5)	96.7–104.2	0.1–3.7	0.074–0.076 mg/L	Charoenkitamorn et al. (2015)
Vegetables	DMDTCs EBDTCs PBDTC Dazomet Metam	1. Treatment with alkaline EDTA 2. Derivatisation with iodomethane	–	LC-UV (272 nm) followed by AAS Column: C18 (250 × 4.6 mm, 5 μm) Mobile phase: acetonitrile/water (gradient)	92–100	0.6–1.6	0.13–0.66 mg/kg	Al-Alam et al. (2017)
Beverage	DMDTCs EBDTCs PBDTC Milneb Metiram Thiram	1. Extraction with NaHCO3/dithiothreitol 2. Derivatisation with dimethyl sulphate	dSPE (PSA)	LC-ESI-MS^2 (negative-ion mode) Column: BEH C18 (120 × 2.1 mm, 1.7 μm) Mobile phase: acetonitrile/0.1% formic acid (gradient)	97–108	3.2–9.6	0.23-6.97 mg/kg	Kakitani et al. (2017)
Banana	Propineb Propylenediamine PTU	Propineb: extraction with water in presence of vitamin C, tin chloride, hydrochloric acid and n-hexane Metabolite: extraction with acetonitrile	dSPE (GCB)	LC-ESI-MS^2 (positive-ion mode) Column: Poroshell 120 SB-AQ (120 × 2.1 mm, 2.7 μm) Mobile phase: acetonitrile/water (55:45)	75–103	1.3–9.7	0.005 mg/kg	Song et al. (2018)
Baby food, infant formula	Propineb PTU	1. Treatment with alkaline EDTA/cysteine 2. Derivatisation with dimethyl sulphate	dSPE (MgSO4, PSA, C18)	LC-ESI-MS^2 (positive-ion mode) Column: C18 (120 × 2.1 mm, 1.8 μm) Mobile phase: methanol/10 mM ammonia (gradient)	85–120	1.1–12	0.003–0.01 mg/kg	Vaclavik et al. (2018)
Fruit, mushrooms, vegetables	EBDTC PBDTC	1. Treatment with alkaline EDTA/cysteine 2. Derivatisation with dimethyl sulphate	–	LC-MS^2 (positive ion mode)	85–107		0.0004–0.0015 mg/kg	Li et al. (2019)

van Lishaut H, Schwack W. 2000. Selective trace determination of dithiocarbamate fungicides in fruits and vegetables by reversed-phase ion-pair liquid chromatography with ultraviolet and electrochemical detection. J AOAC Int. 83(3):720–727.

 PubMed Web of Science ®Google Scholar

Blasco C, Font G, Pico Y. 2004. Determination of dithiocarbamates and metabolites in plants by liquid chromatography–mass spectrometry. J Chromatogr A. 1028(2):267–276. doi:10.1016/j.chroma.2003.12.002

 PubMed Web of Science ®Google Scholar

Crnogorac G, Schwack W. 2007. Determination of dithiocarbamate fungicide residues by liquid chromatography/mass spectrometry and stable isotope dilution assay. Rapid Commun Mass Spectrom. 21(24):4009–4016. doi:10.1002/rcm.3312

 PubMed Web of Science ®Google Scholar

Hayama T, Takada M. 2008. Simple and rapid method for the determination of ethylenebisdithiocarbamate fungicides in fruits and vegetables using liquid chromatography with tandem mass spectrometry. Anal Bioanal Chem. 392(5):969–976. doi:10.1007/s00216-008-2346-8

 PubMed Web of Science ®Google Scholar

Brewin S, Miller C, Khoshab A. 2008. The LC-MS/MS ethylene bisdithiocarbamate (EBDC) analytical method – a comparison with the CS2 analysis technique on wheat samples generated from field trials. 7th European Pesticide Residues Workshop (EPRW), Book of Abstracts, Berlin, Germany, 2008, p. 142.

 Google Scholar

Crnogorac G, Schwack W, Schmauder S. 2008. Trace analysis of dithiocarbamate fungicide residues on fruits and vegetables by hydrophilic interaction liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom. 22(16):2539–2546. doi:10.1002/rcm.3646

 PubMed Web of Science ®Google Scholar

Kim HY, Choi HJ, Eom JY, Seo EC, Choi SH, Cheong SY, Choi SH, Lee HJ, Choi JC. 2010. Determination of dithiocarbamates in agricultural products circulated in Korea Korean. J Food Sci Technol. 42:1–7.

 Google Scholar

Jafari A, Shoeibi S, Amini M, Amirahmadi M, Rastegar H, Ghaffarian A, Ghazi-Khansari M. 2012. Monitoring dithiocarbamate fungicide residues in greenhouse and non-greenhouse tomatoes in Iran by HPLC-UV. Food Addit Contam Part B Surveill. 5(2):87–92. doi:10.1080/19393210.2012.657693

 PubMed Web of Science ®Google Scholar

Ringli D, Schwack W. 2013. Selective determination of thiram residues in fruit and vegetables by hydrophilic interaction LC-MS. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 30(11):1909–1917. doi:10.1080/19440049.2013.833669

 PubMed Web of Science ®Google Scholar

Schmidt B, Christensen HB, Petersen A, Sloth JJ, Poulsen ME. 2013. Method validation and analysis of nine dithiocarbamates in fruits and vegetables by LC-MS/MS. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 30(7):1287–1298. doi:10.1080/19440049.2013.801083

 PubMed Web of Science ®Google Scholar

Zhou L, Xu J, Luan L, Ma J, Gong Y, Qin D, Pan C. 2013. Optimization and validation of a method based on derivatization with methylating agent followed by HPLC–DAD for determining dithiocarbamates residues. Acta Chromatogr. 25(4):613–625. doi:10.1556/AChrom.25.2013.4.2

 Web of Science ®Google Scholar

Charoenkitamorn K, Chailapakul O, Siangproh W. 2015. Development of gold nanoparticles modified screen-printed carbon electrode for the analysis of thiram and their derivative in food using ultra-high performance liquid chromatography. Talanta 132:416–423. doi:10.1016/j.talanta.2014.09.020

 PubMed Web of Science ®Google Scholar

Al-Alam J, Bom L, Chbani A, Fajloun Z, Millet M. 2017. Analysis of dithiocarbamate fungicides in vegetable matrices using HPLC-UV followed by atomic absorption spectrometry. J Chromatogr Sci. 55(4):429–435. doi:10.1093/chromsci/bmw198

 PubMed Web of Science ®Google Scholar

Kakitani A, Yoshioka T, Nagatomi Y, Harayama K. 2017. A rapid and sensitive analysis of dithiocarbamate fungicides using modified QuEChERS method and liquid chromatography–tandem mass spectrometry. J Pesticide Sci. 42(4):145–150. doi:10.1584/jpestics.D17-025

 PubMed Web of Science ®Google Scholar

Song S, Wei J, Chen Z, Lei Y, Zhang Y, Deng C, Tan H, Li X. 2018. Determination of propineb and its metabolites propylenethiourea and propylenediamine in banana and soil using gas chromatography with flame photometric detection and LC–MS/MS analysis. J Environ Sci Health B. 53(3):153–160. doi:10.1080/03601234.2017.1399765

 PubMed Web of Science ®Google Scholar

Vaclavik L, Shippar JJ, Koesukwiwat U, Mastovska K. 2018. Method development and validation for low-level propineb and propylenethiourea analysis in baby food, infant formula and related matrices using liquid chromatography-tandem mass spectrometry. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 35(12):2387–2399. doi:10.1080/19440049.2018.1539529

 PubMed Web of Science ®Google Scholar

Li J, Dong C, Yang Q, An WJ, Zheng ZT, Jiao BN. 2019. Simultaneous Determination of Ethylenebisdithiocarbamate (EBDC) and Propylenebisdithiocarbamate (PBDC) Fungicides in Vegetables, Fruits, and Mushrooms by Ultra-High-Performance Liquid Chromatography Tandem. Food Anal Methods. 12(9):2045–2055. doi:10.1007/s12161-019-01538-z

 Web of Science ®Google Scholar