Table 2.
Detailed description of the setup of some liquid chromatography (LC)–mass spectrometry (MS) methods for mycotoxin determination
| Reference | Extraction | Cleanup | Analytes | Matrix | LC–MS instrument | LC conditions | MS conditions |
|---|---|---|---|---|---|---|---|
| [39] | CH3CN–H2O (84:16, v/v) | MycoSep 226 AflaZON+, MycoSep 227 (both Romer Labs) | NIV, DON, FUS-X, 3ADON, 15ADON, DAS, HT2, T2, ZEN | Maize | QTRAP MS/MS instrument (Sciex) coupled to 1100 series LC system (Agilent Technologies) | Aquasil RP-18 column (100 mm × 4.6 mm, 3 μm) + C18 guard column; 25 °C, flow rate 1000 μL/min, injection volume 25 μL; eluent A H2O–CH3OH (80:20, v/v), eluent B H2O–CH3OH (10:90, v/v), both containing 5 mM NH4CH3COO-; gradient 0.5 min 0% eluent B, linear gradient to 100% eluent B to 4.5 min, 100% eluent B to 7 min, 7.1 min 0% eluent B, reequilibration 3 min, total run 10 min | APCI± MRM, monitoring of 2 transitions (1 quantifier and 1 qualifier), dwell time 100 ms, polarity switching (2 periods) |
| [40] | CH3CN–H2O (84:16, v/v) | MycoSep 226 AflaZON+ | AFs, NIV, DON, 3ADON, 15ADON, FUS-X, HT2, T2, ZEN, OTA, STER, CIT, verruculogen | Various foods and feed | Quattro Ultima QqQ instrument (Micromass) coupled to Acquity UHPLC system (Waters) | UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); 35 °C, flow rate 300 μL/min, injection volume 5 μL; eluent A ESI+ 10 mM NH4CH3COO-, ESI− 0.1% 0.1% (v/v) aqueous NH3, eluent B CH3OH; gradient initially 20% eluent B, linear increase to 5.5 to 85% eluent B, 100% eluent B within 0.3 min, reequilibration for 2 min at 20% eluent B, total run 10 min | ESI+, ESI−, MRM, 2 chromatographic runs, monitoring of 2 transitions (1 quantifier and 1 qualifier) |
| [41] | 2-step extraction: (1) PBS; (2) 70% CH3OH | AOZFDT2 (VICAM) | AFs, OTA, FBs, DON, ZEA, T2, HT2 | Maize | QTRAP MS/MS (Sciex) instrument coupled to 1100 micro LC system (Agilent Technologies) | Gemini C18 column (150 mm × 2 mm, 5 μm) + Gemini C18 guard column (4 mm × 2 mm, 5 μm); 40 °C, flow rate 200 μL/min, injection volume 20 μL; eluent A H2O, eluent B CH3OH, both containing 0.5% CH3COOH and 1 mM NH4CH3COO-; gradient 3 min at 20% eluent B, jump to 40% eluent B, linear increase to 63% eluent B within 35 min, 63% eluent B for 11 min, reequilibration at 20% eluent B for 10 min, total run 59 min | ESI+, ESI−, dMRM, monitoring of 2 transitions (1 quantifier and 1 qualifier), time window of 1 MRM 0.8 min, cycle time 0.55 s |
| [42] | CH3CN–H2O–CH3COOH (79.5:20:0.5, v/v/v). Evaporation and redissolution in PBS before IAC | Myco6in1+ (VICAM) | AFs, OTA, FBs, DON, ZEN, T2, HT2 | Barley, maize breakfast cereals, peanuts | QTRAP 4500 instrument (Sciex) coupled to a Prominence UFLC XR chromatography system (Shimadzu) | Acquity UPLC HSS T3 end-capped C18 column (100 mm × 2.1 mm, 1.7 μm), 40 °C, flow rate 400 μL/min, injection volume 10 uL; eluents A H2O, eluent B CH3OH, both containing 5 mM NH4CH3COO-; gradient 5% eluent B increased to 50% eluent B in 1 min, linear increase to 100% eluent B within 6 min, 100% eluent B to 8 min, at 8.1 min initial conditions 5% eluent B, reequilibration at 5% eluent B for 2 min, total run 10 min | ESI± MRM, 2 periods, monitoring of 2 transitions (1 quantifier and 1 qualifier), dwell time 50 ms, polarity switching (2 periods) |
| [43] | 2-step extraction: (1) H2O; (2) CH3OH. Evaporation and redissolution in PBS before IAC | Myco6in1+ (VICAM) | AFs, OTA, FBs, DON, ZEN, T2, HT2, NIV | Maize, durum wheat, corn flakes, maize crackers | QTRAP MS/MS instrument (Sciex) coupled to 1100 micro LC system (Agilent technologies) | Gemini C18 column (150 mm × 2 mm, 5 μm) + Gemini C18 guard column (4 mm × 2 mm, 5 μm); 40 °C, flow rate 200 μL/min, injection volume 20 μL; eluent A H2O, eluent B CH3OH, both containing 0.5% CH3COOH and 1 mM NH4CH3COO-; gradient 3 min at 20% eluent B, jump to 40% eluent B, linear increase to 63% eluent B within 35 min, 63% eluent B for 11 min, reequilibration at 20% eluent B for 10 min, total run 59 min | ESI± MRM, monitoring of 2 transitions (1 quantifier and 1 qualifier), dwell time 100 ms, polarity switching (2 periods) |
| [44] | NaCl + H2O–CH3OH (30:70, v/v). Dilution with PBS before IAC | OCHRAPREP + DZT MS-PREP, AOF MS-PREP + DZT MS-PREP, AFLAOCHRA PREP + DZT MS-PREP (R-Biopharm) | OTA + DON, ZEN, T2, HT2; AFs, FBs, OTA + DON, ZEN, T2, HT2; AFs, OTA + DON, ZEN, T2, HT2 | Wholemeal bread, maize and maize-based products including infant foods, oat-based muesli | Acquity TQD tandem QqQ MS instrument (Waters) | Gemini C18 column (150 mm × 2 mm, 5 μm), 40 °C, flow rate 300 μL/min, injection volume 20 μL; eluent A H2O–CH3OH (95:5, v/v), eluent B H2O–CH3OH (98:3, v/v), both containing 0.5% HCOOH and 1 mM NH4HCOO-; gradient 20% eluent B for 0.1 min, to 10 min linear increase to 90% eluent B, 90% eluent B to 15 min, reequilibration at 20% eluent B, total run 20 min | ESI+ MRM, monitoring of 2 transitions (1 quantifier and 1 qualifier), 6 acquisition periods, dwell times from 0.1 to 0.27 s |
| [45] | QuEChERS | AFs, FBs, NIV, DON, 3ADON, 15ADON, FUS-X, HT2, T2, ZEN, OTA, DAS, NEO | Rice, corn, wheat, rye, oat, barley, infant cereals, soya, corn gluten | QTrap 4000 instrument (Sciex) coupled to 1100 series LC system (Agilent Technologies) | Zorbax Bonus-RP column (150 mm × 2.1 mm, 3.5 μm) + Zorbax RB C8 guard column (12.5 mm x 2.1 mm, 3.5 μm), flow rate 250 μL/min, injection volume 40 μL; eluent A 0.15% (v/v) HCOOH + 10 mM NH4HCOO-, eluent B 0.05% HCOOH (v/v) in CH3OH; gradient: 0% eluent B at 1 min, linear increase to 100% eluent B until 15 min, 100% eluent B for 5 min, reequilibration at 0% eluent B for 5 min, total run 25 min | ESI± MRM, monitoring of 2 transitions (1 quantifier and 1 qualifier), 3 acquisition periods | |
| [46] | QuEChERS | AFs, FBs, DON, HT2, T2, ZEN, OTA | Wheat, maize, rice | Micromass Quattro Micro QqQ coupled to Alliance 2695 system (Waters) | Atlantis RP C18 column (150 mm × 2.1 mm, 5 μm), 30 0°C, flow rate 300 μL/min, injection volume 20 μL; eluent A H2O–CH3OH (90:10, v/v), eluent B H2O–CH3OH (10:90, v/v), both containing 5 mM NH4CH3COO-; gradient 20% eluent B for 0.1 min, until 10 min linear increase to 90% eluent B, 90% eluent B to 15 min, reequilibration at 20% eluent B, total run 20 min | ESI± MRM, monitoring of 2 transitions (1 quantifier and 1 qualifier), 3 acquisition periods | |
| [47] | QuEChERS, dilute and shoot | 38 mycotoxins and 288 pesticides | Apple baby food, wheat flour, paprika, black pepper, sunflower seed | QTRAP 5500 instrument (Sciex) coupled to Acquity UHPLC system (Waters) | Acquity UPLC HSS T3 end-capped C18 column (100 mm × 2.1 mm, 1.8 μm), 40 °C, flow rate 350–700 μL/min, injection volume 3 μL; ESI+ eluent A: H2O, eluent B: CH3OH, both containing 0.2% HCOOH + 5 mM NH4HCOO-; ESI− eluent A H2O, eluent B CH3OH, both containing 0.2% HCOOH + 5 mM NH4HCOO-; gradient: 10% eluent B with flow rate 350 μL/min increased to 50% eluent B in 1 min, linear increase to 100% eluent B within 10 min and simultaneous increase of flow rate to 550 μL/min, flow rate 0.7 μL/min at 100% eluent B, reequilibration for 2.5 min at 10% eluent B at 450 μL/min, total run 15.5 min | ESI+, ESI−, dMRM, time window for 1 MRM 0.8 min, cycle time 0.55 s | |
| [48] | Dilute and shoot | No | 39 mycotoxins | Wheat, maize | QTRAP 4000 instrument (Sciex) coupled to 1100 series LC system (Agilent Technologies) | Gemini C18 column (150 mm × 2 mm, 5 μm) + Gemini C18 guard column (4 mm × 2 mm, 5 μm); 40 °C, flow rate 1000 μL/min, injection volume 5 μL; eluent A CH3OH–H2O–CH3COOH (10:89:1, v/v/v), eluent B CH3OH–H2O–CH3COOH (97:2:1, v/v/v), both containing 5 mM NH4CH3COO-; gradient 2 min at 100% eluent A, linear increase to 100% eluent B within 12 min, held at 100% eluent B for 3 min, reequilibration at 100% eluent A for 4 min, total run 19 min | ESI+, ESI−, dMRM, dwell time 100 ms, pause time 5 ms |
| [36] | Dilute and shoot | No | 295 analytes | Apple puree, hazelnut, maize, green pepper | QTRAP 5500 instrument (Sciex) coupled to 1290 series LC system (Agilent Technologies) | Gemini C18 column (150 mm × 2 mm, 5 μm) + Gemini C18 guard column (4 mm × 2 mm, 5 μm); 40 °C, flow rate 1000 μL/min, injection volume 5 μL; eluent A CH3OH–H2O–CH3COOH (10:89:1, v/v/v), eluent B CH3OH–H2O–CH3COOH (97:2:1, v/v/v), both containing 5 mM NH4CH3COO-; gradient: 2 min at 100% eluent A, linear increase to 50% eluent B within 3 min, linear increase zo 100% eluent B within 9 min, hold at 100% eluent B for 4 min, reequilibration at 100% eluent A for 2.5 min, total run 20.5 min | ESI+, ESI−, dMRM, MRM window ±27 s for positive mode, ±42 s for negative mode, scan time 1 s |
| [49] | Raw extract, SIDA | No | AFs, FBs, DON, HT2, T2, OTA, ZEN | Maize, cereal-based products | 6490 triple-quadrupole instrument coupled to 1290 series UHPLC system (both Agilent Technologies) | Zorbax RRHL Eclipse Plus C18 column (100 mm × 2.1 mm, 1.8 μm); 30 °C, flow rate 350 μL/min, injection volume 3 μL; eluent A H2O–HCOOH (99.9:0.1, v/v), eluent B CH3OH–HCOOH (99.9:0.1, v/v) both containing 5 mM NH4HCOO-; gradient: 0.5 min at 30% eluent B, linear increase to 100% eluent B in 7.5 min, hold at 100% eluent B for 1.5 min, at 9.6 min back to 30% eluent B, reequilibration at 30% eluent B for 2 min, total run 11.5 min | ESI±, dMRM, monitoring of 2 transitions (1 quantifier and 1 qualifier) |
| [50] | CH3CN–H2O (84:16, v/v), SIDA | Bond Elut Mycotoxin SPE cartridges (Agilent Technologies) | NIV, DON, FUS-X, DON-3-Glc, 3ADON, 15ADON, HT2, T2, ENNs, BEA, ZEN | Barley, malt, oat, wheat, maize | QTRAP 4000 instrument (Sciex) coupled to LC-20A Prominence system series LC system (Shimadzu) | Hydrosphere RP-C18 column (100 mm × 3 mm, 3 μm) + C18 guard column; 40 °C, flow rate 200 μL/min, injection volume 10 μL; eluent A H2O–HCOOH (99.9:0.1, v/v), eluent B CH3OH–HCOOH (99.9:0.1, v/v); gradient ESI− 2 min at 10% eluent B, linear increase to 99% eluent B in 6 min, hold at 99% eluent B for 7.5 min, for 2 min back to 10% eluent B, reequilibration at 10% eluent B for 9.5 min, total run 25 min; ESI+ 2 min at 10% eluent B, linear increase to 87% eluent B in 6 min, hold at 87% eluent B for 7 min, increase to 100% eluent B in 5 min, hold at 100% eluent B for 3.5 min, for 2 min back to 10% eluent B, reequilibration at 10% eluent B for 9.5 min, total run 34.5 min | ESI−, ESI+, dMRM, 2 single chromatographic runs, monitoring of 2 transitions (1 quantifier and 1 qualifier) |
| [51] | CH3CN–H2O (84:16, v/v), evaporation, reconstitution in CH3OH and H2O | SPE (Oasis HLB columns) | AFs, OTA, DON, ZEN, T2, HT2 | Wheat flour, barley flour, crisp bread | Accela HPLC system, Exactive HRMS instrument (Thermo Fisher Scientific); 1100 micro-LC system (Agilent Technologies), QTRAP instrument (Applied Biosystems) | Kinetex C18 column (100 mm × 2.1 mm, 2.6 μm); 40 °C, flow rate 200 μL/min, injection volume 20 μL; eluent A H2O, eluent B CH3OH, both containing 0.5% CH3COOH and 1 mM NH4CH3COO-; gradient 10% eluent B start, until 4 min linear increase to 40% eluent B, 60% eluent B in 27 min, keep for 5 min, reequilibration at 10% eluent B for 7 min, total run 20 min | HESI-II (heated-electrospray, ESI+, HCD fragmentation (in-source fragmentation) |
| [52] | QuEChERS (2 g sample, 10 mL 0.1% HCOOH in H2O, 3 min shaking, 10 mL CH3CN, 3 min shaking, 4 g MgSO4, 1 g NaCl, shaking) | No additional cleanup | 3ADON, 15ADON, DON, DON-3-Glc, FUS-X, NIV, HT2, T2, DAS, NEO, AFs, OTA, FBs, STER, ZEN, penitrem A, BEA, Alternaria toxins, ergot alkaloids | barley | Accela HPLC system, Exactive HRMS instrument (Thermo Fisher Scientific) | Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm); 40 °C, flow rate 300 μL/min, injection volume 5 μL; eluent A H2O with 5 mM NH4HCOO- and 0.1% HCOOH, eluent B CH3OH; gradient: start with 5% eluent B, increase to 50% eluent B in 6 min, increase to 95% eluent B within 4 min, keep until 15 min of the run, reequilibration at 5% eluent B for 3 min | HESI-II, ESI+/ESI− |
| [53] | QuEChERS (2 g sample, 10 mL 0.1% HCOOH in H2O, 3 min shaking, 10 mL CH3CN, 3 min shaking, 4 g MgSO4, 1 g NaCl, 0.5 trisodium citrate dihydrate, shaking) | No additional cleanup | 3ADON,15ADON, DON, DON-3-Glc, FUS-X, NIV, HT2, T2, DAS, NEO, AFs, OTA, FBs, STER, ZEN, mycophenolic acid, MON, BEA, Alternaria toxins, ergot alkaloids, culmorins | malting barley | Acquity UHPLC system (Waters), Q-Exactive system (Thermo Fisher Scientific) | Atlantis T3 column (100 mm × 2.1 mm, 3 μm); 30oC, flow rate 300 μL/min, injection volume 2 μL; eluent A CH3CN–H2O–CH3COOH (95:4.9:0.1, v/v/v), eluent B H2O–CH3COOH (99.9:0.1, v/v), both containing 5 mM NH4CH3COO-; gradient 5% eluent A start for 1 min, increase to 15% eluent A in 14 min, increase to 100% eluent A in next 15 min, kept at 100% eluent A for 3 min, reequilibration for 4.4 min | HESI-II (positive, negative) |
15ADON 15-acetyldeoxynivalenol, 3ADON 3-acetyldeoxynivalenol, AFs aflatoxins, APCI atmospheric pressure chemical ionization, BEA beauvericin, CIT citrinin, DON-3-Glc deoxynivalenol 3-glucoside, DAS diacetoxyscirpenol, dMRM dynamic multiple-reaction monitoring, DON deoxynivalenol, ENNs enniatins, ESI electrospray ionization, FBs fumonisins B, FUS-X fusarenon X, HCD high-energy collisional dissociation, HPLC high-performance liquid chromatography, HRMS high-resolution mass specrometry, HT2 HT-2 toxin, IAC immunoaffinity chromatography, MON moniliformin, MRM multiple-reaction monitoring, NEO neosolaniol, NIV nivalenol, OTA typo in ocratoxin A, PBS phosphate-buffered saline, QqQ triple quadrupol, QuEChERS quick, easy, cheap, effective, rugged, and safe, SIDA stable isotope dilution assay, SPE solid-phase extraction, STER sterigmatocystin, T2 T-2 toxin, UFLC ultrafast liquid chromatography, UHPLC ultrahigh-performance liquid chromatography, UPLC ultraperformance liquid chromatography, ZEA zearalanol, ZEN zearalenone