Supel™ Tox SPE Cartridges: Fast and Simple Cleanup for Mycotoxin Analysis

Fast and Simple Cleanup for Mycotoxin AnalysisSupel™ Tox SPE Cartridgessigma-aldrich.com/supeltoxSupel™ Tox SPE CartridgesSupel™ Tox SPE Cartridgeswhich saves time by eliminating wash steps prior to elution of aflatoxin and zearalenone, deoxynivalenol, and tricothecenes (type A and B), respectively. Cartridges removing interferences associated with analysis of fumonisins (B1 and B2) and ochratoxin A are also available as a part of our Supel Tox product offering. Refer to the table below for further details. Features and Benefits• Remove interferences associated with mycotoxin analysis• Better reproducibility than the current industry standard, immunoaffinity columns• Time associated with sample preparation is up to ten times less than that of immunoaffinity columns• Basic and quick methodology requiring little additional method development• Improved shelf life over immunoaffinity columns due to thermally stable format. No refrigeration required for shipping and storage.The need for a quick, simplistic sample cleanup approach prior to chromatographic mycotoxin analysis has brought about SPE cartridges that significantly decrease sample prep time, increase reproducibility, and are more user friendly as compared to the industry standard immunoaffinity columns. In addition, the Supel Tox SPE approach requires less equipment and fewer consumables, providing an additional cost savings.Unlike the multiple step “bind and elute” strategy implemented when using immunoaffinity columns, the Supel Tox AflaZea, DON and Tricho SPE cartridges employ an “interference removal” strategy SPE Cartridge Analyte(s) Matrix Purification StrategySupel Tox AflaZea Aflatoxin and Zearalenone Grains, feeds, TMR samples, peanuts, peanut products and aqueous solutionsInterference removal Supel Tox DON Deoxynivalenol (DON) Wheat, flour and corn Interference removal Supel Tox Tricho Trichothecenes (Type A and B) Grains and complex matrices Interference removal Supel Tox TrichoBind Trichothecenes (Type A and B) Grains, feeds and other complex matrices Multifunctional bind and eluteSupel Tox FumoniBind Fumonisins (B1 and B2) Grains and cereals Multifunctional bind and eluteSupel Tox OchraBind Ochratoxin A Grain and feed samples Multifunctional bind and eluteSpecialty Products for Mycotoxin AnalysisSample Purification Strategy for the Various Supel Tox SPE Cartridges3Order: 800-325-3010 (U.S.) 314-359-3441 (Global) In the “bind and elute” strategy, the analyte of interest becomes bound to the SPE sorbent. The unwanted matrix is removed, and the analyte is ultimately eluted. In “interference removal,” the unwanted matrix components remain bound to the sorbent, while the purified Comparison of “Bind and Elute” Principle to “Interference Removal” Principle“Bind and Elute” PrincipleOriginal sample (analytes and internal standard in a matrix)Analyte remains on cartridgeUnwanted matrix discarded as waste1. Condition SPE cartridge2. Apply sample to SPE cartridge3. Apply wash solvent4. Apply elution solvent5. Evaporateelution solvent6. ReconstituteDilute analyte fractionPurified and concentratedanalytes and internal standard“Interference Removal ” PrincipleOriginal sample (analytes and internal standard in a matrix)1. Condition SPE cartridge2. Apply sample to SPE cartridge3. Apply elution solventUnwanted matrix remains on cartridge. Cartridge discarded. 4. Evaporateelution solventPurified analyte eluted5. ReconstitutePurified and concentratedanalytes and internal standardanalyte is eluted. This strategy reduces the number of purification steps, allowing for time savings and increase in sample throughput. The Supel™ Tox AflaZea, DON and Tricho SPE cartridges employ the “interference removal” strategy. sigma-aldrich.com/supeltoxSupel™ Tox SPE CartridgesComparison of Supel™ Tox SPE Cartridges to Immunoaffinity Columns for Matrix Removal Prior to Mycotoxin Analysis• Ten times faster – ten times sample throughput• Vast improvement in reproducibilityThe following is an experiment comparing the use of the Supel Tox AflaZea SPE cartridge to a leading immunoaffinity column for the analysis of aflatoxins B1, B2, G1 and G2 in peanut paste. The results illustrate Supel Tox AflaZea SPE cartridges are superior to immunoaffinity columns in terms of process simplicity, time required for sample preparation, and control of variation. Cleanup of Aflatoxins in Peanut Paste: Immunoaffinity Versus Supel™ Tox (n = 3)G2 G1 B2 B1% RecoveryA?atoxinSupel™ Tox A?aZeaImmunoa?nity120100806040200Immunoaffinity Supel Tox AflaZea SPE Cartridge Sample Prep Time (post-extraction to pre-analysis)• 60 minutes• 8 samples/day (if processing 1 at a time)• 6 minutes• 80 samples/day (if processing 1 at a time)Ease of Use • Large volumes of liquid • Controlled drop rates• Numerous complicated steps• Additional buffer salts required• Must be refrigerated, brought to room temp before use• Small volumes of liquid• Vacuum filtration used• Steps few and not complicated• No additional reagents required • Column does not require special storage conditions Procedure (post-extraction to analysis)Stage 1 (15 minutes) 1. Configure manifold for waste collection 2. Add 1 mL sample to 17 mL of phosphate buffered saline, vortex 3. Uncap/mount/drain cartridges by gravity 4. Apply reservoirs, load sample onto cartridges Stage 2 (15 minutes) 5. Rinse interferences 6. Reconfigure manifold for sample collection 7. Elute/collect sampleStage 3 (30 minutes) 8. Evaporate sample to dryness 9. Reconstitute sample and vortex 10. Transfer 0.2 mL sample to vial 11. Dilute sample and vortexAnalysisPurify and Transfer (6 minutes) 1. Configure manifold for sample collection 2. Mount cartridges 3. Load 2 mL sample 4. Elute and collect under vacuum 5. Transfer 0.2 mL sample to vial 6. Dilute sample and vortexAnalysisComparison of Sample Preparation Methods Using Supel Tox AflaZea SPE vs Immunoaffinity for Aflatoxins in Peanut Paste5Order: 800-325-3010 (U.S.) 314-359-3441 (Global) Analysis of Aflatoxin B1, B2, G1 and G2 in Corn Meal, Wheat Meal and Raw Peanut Paste MatricesThe Supel Tox AflaZea SPE cartridge was used to demonstrate the cleanup of corn meal, wheat meal and raw peanut paste matrices prior to the analysis of Aflatoxin B1, B2, G1 and G2. In the procedure, corn meal, wheat meal and raw peanut paste extracts were spiked with Aflatoxin Standard Mix (Cat. No. 34036) and run through the Supel Tox AflaZea SPE cartridges. The eluate was collected and diluted for HPLC analysis. Results illustrate Supel Tox AflaZea SPE cartridges provide exceptional cleanup of corn meal, wheat meal and raw peanut paste matrices with high analyte recoveries and low %RSD for aflatoxin analysis. G2 G1 B2 B1% RecoveryA?atoxin1009080706050403020100CornWheatPeanut PasteAnalyte Recovery and %RSD of Aflatoxins from Corn, Wheat and Peanut Paste (n = 3)MinA?atoxin B1, B2, G1, G2 StandardSpiked Corn Matrix after Supel Tox CleanupSpiked Wheat Matrix after Supel Tox CleanupSpiked Peanut Paste Matrix after Supel Tox Cleanup1. A?atoxin G22. A?atoxin G13. A?atoxin B24. A?atoxin B1123412340 2 4 6 8 10 12 1411234234 column: Ascentis® Express C18, 10 cm x 2.1 mm I.D., 2.7 µm (53823-U) mobile phase: (A) water; (B) acetonitrile; (C) methanol; (700:120:120,A:B:C) with 0.780 g potassium bromide and 230 µL of nitric acid flow rate: 0.400 mL/min column temp.: 35 °C detector: Fluorescence detector, Excitation: 360 nm, Emission: 440 nm injection: 40 µLAflatoxins in Corn Meal, Wheat Meal and Raw Peanut Paste After Supel Tox CleanupAflatoxin B1, B2, G1, G2 StandardSpiked Corn Matrix after Supel Tox CleanupSpiked Wheat Matrix after Supel Tox CleanupSpiked Peanut Paste Matrix after Supel Tox Cleanup1. Aflatoxin G22. Aflatoxin G13. Aflatoxin B24. Aflatoxin B10 2 4 6 8 10 12 14Minsigma-aldrich.com/supeltoxSupel™ Tox SPE CartridgesAnalysis of Corn Meal and Wheat Meal Matrices for Deoxynivalenol Deoxynivalenol (DON), also referred to as vomitoxin, is a concern primarily in the livestock industry, as it is known to cause a reduction in food intake in farm animals. DON is a type B trichothecene which may occur in a variety of grains.Cleanup of corn meal and wheat meal matrices for the analysis of DON was illustrated with the use of Supel™ Tox DON SPE cartridges. The grain extracts were spiked with deoxynivalenol standard (Cat. No. 34124) and applied to the cartridges. The eluate was collected, evaporated, and reconstituted for HPLC analysis. Blank unspiked matrix samples were also run though cartridges and analyzed. For the recovery calculations, the peak area of DON observed in the blanks was subtracted from that of the spiked samples. The corresponding bar chart and chromatograms illustrate that in addition to effectively removing the unwanted corn and wheat meal matrices, Supel™ Tox DON SPE cartridges also provide high analyte recoveries and reproducibility for the analysis of DON.0%20%40%60%80%100%Corn Wheat% RecoveryAnalyte Recovery and %RSD of Deoxynivalenol (DON) from Corn and Wheat (n = 3) column: Ascentis® Express C18, 10 cm x 2.1 mm I.D., 2.7 µm (53823-U) mobile phase: (A) water; (B) acetonitrile; (C) methanol; (92:4:4, A:B:C) flow rate: 0.400 mL/min column temp.: 35 °C detector: UV 220 nm; 360 nm injection: 40 µLDeoxynivalenol in Corn and Wheat Meal After Supel Tox CleanupCorn MealWheat MealMin0 1.0 2.0 3.0 4.0 5.0 6.0Spiked Corn Matrix after Supel Tox CleanupBlank Corn Matrix after Supel Tox CleanupDeoxynivalenol in Corn Meal After Supel Tox Cleanupcolumn: Ascentis® Express C18, 10 cm x 2.1 mm I.D., 2.7 µm (53823-U)mobile phase: (A) water; (B) acetonitrile; (C) methanol; (92:4:4, A:B:C) ?ow rate: 0.400 mL/mincolumn temp.: 35 °Cdetector: UV 220 nm; 360 nminjection: 40 µLDONDONMin0 1.0 2.0 3.0 4.0 5.0 6.0Deoxynivalenol in Wheat Meal After Supel Tox CleanupSpiked Wheat Matrix after Supel Tox CleanupBlank Wheat Matrix after Supel Tox Cleanupcolumn: Ascentis® Express C18, 10 cm x 2.1 mm I.D., 2.7 µm (53823-U)mobile phase: (A) water; (B) acetonitrile; (C) methanol; (92:4:4, A:B:C) ?ow rate: 0.400 mL/mincolumn temp.: 35 °Cdetector: UV 220 nm; 360 nminjection: 40 µLDON7Order: 800-325-3010 (U.S.) 314-359-3441 (Global) Supel Tox SPE CartridgesDescription Qty. Cat. No. Supel Tox AflaZea SPE Cartridge, 6 mL 30 55314-USupel Tox DON SPE Cartridge, 6 mL 30 55316-USupel Tox Tricho SPE Cartridge, 6 mL 30 55308-USupel Tox TrichoBind SPE Cartridge, LRC 25 55307-USupel Tox FumoniBind SPE Cartridge, LRC 25 55315-USupel Tox OchraBind SPE Cartridge, LRC 25 55318-UFeatured and Related ProductsDescription Qty. Cat. No. Ascentis Express HPLC Columns (2.7 µm particles)C18, 10 cm x 2.1 mm I.D. 1 53823-UC18, 15 cm x 2.1 mm I.D. 1 53825-URP-Amide, 10 cm x 2.1 mm I.D. 1 53913-URP-Amide, 15 cm x 2.1 mm I.D. 1 53914-UF5, 10 cm x 2.1 mm I.D. 1 53569-UF5, 15 cm x 2.1 mm I.D. 1 53571-UAscentis Express Guard Columns (2.7 µm particles)Holder 1 53500-UC18 Cartridge, 5 mm x 2.1 mm I.D. 3 53501-URP-Amide Cartridge, 5 mm x 2.1 mm I.D. 3 53514-UF5 Cartridge, 5 mm x 2.1 mm I.D. 3 53594-ULaboratory EquipmentCoors™ Büchner Funnel, capacity 186 mL 1 Z247324Individual Filter Adapters, size 3 1 Z254274Visiprep™ Solid Phase Extraction Manifold, 12-port Model 1 57030-UVisiprep Solid Phase Extraction Manifold, 24-port Model 1 57250-UVisiprep DL Solid Phase Extraction Manifold, 12-port Model 1 57044Visiprep DL Solid Phase Extraction Manifold, 24-port Model 1 57265Disposable Valve Liners, PTFE 100 57059Visi™-1 Single SPE Tube Processor 1 57080-USPE Tube Adapters for 1, 3, 6 mL tubes 12 57020-UMycotoxin Reference Materials (Neats)3-Acetyldeoxynivalenol 5 mg 3292715-Acetyldeoxynivalenol 5 mg 32928Aflatoxin B1 5 mg 32754*Aflatoxin B2 5 mg 32755Aflatoxin G1 5 mg 32756Aflatoxin G2 5 mg 32757*Deoxynivalenol 5 mg 32943Fumonisin B1 5 mg 32936Fusarenon X 5 mg 33438Ochratoxin A 5 mg 32937T-2 Toxin 5 mg 33947*Zearalenone 5 mg 32939Mycotoxin Single Component Standard Solutions3-Acetyldeoxynivalenol, 100 µg/mL in acetonitrile 2 mL 3413215-Acetyldeoxynivalenol, 100 µg/mL in acetonitrile 2 mL 34133Aflatoxin B1, 2 µg/mL in acetonitrile 2 mL 34029Aflatoxin B2, 0.5 µg/mL in acetonitrile 2 mL 34034Aflatoxin G1, 2 µg/mL in acetonitrile 2 mL 34032Aflatoxin G2, 0.5 µg/mL in acetonitrile 2 mL 34033Deoxynivalenol, 100 µg/mL in acetonitrile 2 mL 34124Fumonisin B1, 50 µg/mL in acetonitrile/water 2 mL 34139Fumonisin B2, 50 µg/mL in acetonitrile/water 2 mL 34142Fumonisin B3, 50 µg/mL in acetonitrile/water 1 mL 32606Fusarenon X, 100 µg/mL in acetonitrile 2 mL 34130Supel™ Tox Product OfferingDescription Qty. Cat. No. Nivalenol, 100 µg/mL in acetonitrile 2 mL 34131Ochratoxin A, 10 µg/mL in acetonitrile 2 mL 34037T-2 Toxin, 100 µg/mL in acetonitrile 2 mL 34071*Zearalenone, 50 µg/mL in acetonitrile 1 mL 46916-UZearalenone, 100 µg/mL in acetonitrile 2 mL 34126Mycotoxin Mixture Standard Solutions for Multi-analyte DetectionAflatoxin Mix 4: B1, 2 µg/mL in acetonitrile G1, 2 µg/mL in acetonitrile B2, 0.5 µg/mL in acetonitrile G2, 0.5 µg/mL in acetonitrile2 mL 1 mL34036Aflatoxin Mix 4: B1, 20 µg/mL in acetonitrile G1, 20 µg/mL in acetonitrile B2, 20 µg/mL in acetonitrile G2, 20 µg/mL in acetonitrile 2 mL 33415Japanese Aflatoxin Mix: B1, 25 µg/mL in acetonitrile G1, 25 µg/mL in acetonitrile B2, 25 µg/mL in acetonitrile G2, 25 µg/mL in acetonitrile 5 x 1 mL 40139-UFumonisin Mix: FB1, 50 µg/mL in acetonitrile/water FB2, 50 µg/mL in acetonitrile/water 2 mL 34143B-Trichothecene Mix: DON, 100 µg/mL in acetonitrile NIV, 100 µg/mL in acetonitrile 3-AcDON, 100 µg/mL in acetonitrile 15-AcDON, 100 µg/mL in acetonitrile 2 mL 34134Certified Matrix Reference Materials for Mycotoxin Analysis (CRMs)Compound Feed (Aflatoxin blank) Aflatoxin B1, <1 µg/kg50 g BCR375Compound Feedingstuff (Aflatoxins, very low level) Aflatoxin B1, 2.6 µg/kg Aflatoxin B2, 0.2 µg/kg Aflatoxin G1, 0.4 µg/kg Aflatoxin G2, <0.2 µg/kg 2 x 75 g ERMBE375Compound Feedingstuff (Aflatoxins, high level) Aflatoxin B1, 12.9 µg/kg Aflatoxin B2, 0.68 µg/kg Aflatoxin G1, 0.5.2 µg/kg 2 x 75 g ERMBE376Maize (Zearalenone, very low level) Zearalenone, <5 µg/kg60 g ERMBC716Maize (Zearalenone, low level) Zearalenone, 83 µg/kg60 g ERMBC717Maize Flour (Deoxynivalenol, blank) Deoxynivalenol, <50 µg/kg150 g BCR377Peanut Butter (Aflatoxins, low level) Aflatoxin B1, <0.2 µg/kg Aflatoxin B2, <0.2 µg/kg Aflatoxin G1, <0.2 µg/kg Aflatoxin G2, <0.2 µg/kg 1 ea. BCR401RWheat (Ochratoxin A, blank) Ochratoxin A, <0.6 µg/kg55 g BCR471Mycotoxin Solutions (CRMs)Aflatoxin B1, 3.79 µg/g in acetonitrile 4 mL ERMAC057Aflatoxin B2, 3.80 µg/g in acetonitrile 4 mL ERMAC058Aflatoxin G1, 3.78 µg/g in acetonitrile 4 mL ERMAC059Aflatoxin G2, 3.80 µg/g in acetonitrile 4 mL ERMAC0604-Deoxynivalenol, 25.1 µg/g in acetonitrile 1 ea. IRMM315Nivalenol, 24.0 µg/g in acetonitrile 1 ea. IRMM316Zearalenone, 9.95 µg/g in acetonitrile 4 mL ERMAC699*Not available in the U.S.A.PFK11644 / T4130261023Order/Customer Service (800) 325-3010 • Fax (800) 325-5052 Technical Service (800) 325-5832 • sigma-aldrich.com/techservice Development/Custom Manufacturing Inquiries (800) 244-1173 Safety-related Information sigma-aldrich.com/safetycenterWorld Headquarters 3050 Spruce St. St. Louis, MO 63103 (314) 771-5765 sigma-aldrich.comEnabling Science to Improve the Quality of LifeSigma-Aldrich® Worldwide OfficesArgentinaFree Tel: 0810 888 7446 Tel: (+54) 11 4556 1472 Fax: (+54) 11 4552 1698AustraliaFree Tel: 1800 800 097 Free Fax: 1800 800 096 Tel: (+61) 2 9841 0555 Fax: (+61) 2 9841 0500AustriaTel: (+43) 1 605 81 10 Fax: (+43) 1 605 81 20BelgiumTel: (+32) 3 899 13 01 Fax: (+32) 3 899 13 11BrazilFree Tel: 0800 701 7425 Tel: (+55) 11 3732 3100 Fax: (+55) 11 5522 9895CanadaFree Tel: 1800 565 1400 Free Fax: 1800 265 3858 Tel: (+1) 905 829 9500 Fax: (+1) 905 829 9292ChileTel: (+56) 2 495 7395 Fax: (+56) 2 495 7396 People’s Republic of ChinaFree Tel: 800 819 3336 Tel: (+86) 21 6141 5566 Fax: (+86) 21 6141 5567Czech RepublicTel: (+420) 246 003 200 Fax: (+420) 246 003 291DenmarkTel: (+45) 43 56 59 00 Fax: (+45) 43 56 59 05FinlandTel: (+358) 9 350 9250 Fax: (+358) 9 350 92555FranceFree Tel: 0800 211 408 Free Fax: 0800 031 052 Tel: (+33) 474 82 28 88 Fax: (+33) 474 95 68 08GermanyFree Tel: 0800 51 55 000 Free Fax: 0800 64 90 000 Tel: (+49) 89 6513 0 Fax: (+49) 89 6513 1169HungaryTel: (+36) 1 235 9055 Fax: (+36) 1 235 9068IndiaTelephone Bangalore: (+91) 80 6621 9400 New Delhi: (+91) 11 4358 8000 Mumbai: (+91) 22 4087 2364 Pune: (+91) 20 4146 4700 Hyderabad: (+91) 40 3067 7450 Kolkata: (+91) 33 4013 8000Fax Bangalore: (+91) 80 6621 9550 New Delhi: (+91) 11 4358 8001 Mumbai: (+91) 22 2579 7589 Pune: (+91) 20 4146 4777 Hyderabad: (+91) 40 3067 7451 Kolkata: (+91) 33 4013 8016IrelandFree Tel: 1800 200 888 Free Fax: 1800 600 222 Tel: +353 (0) 402 20370 Fax: + 353 (0) 402 20375IsraelFree Tel: 1 800 70 2222 Tel: (+972) 8 948 4222 Fax: (+972) 8 948 4200ItalyFree Tel: 800 827 018 Tel: (+39) 02 3341 7310 Fax: (+39) 02 3801 0737JapanTel: (+81) 3 5796 7300 Fax: (+81) 3 5796 7315KoreaFree Tel: (+82) 80 023 7111 Free Fax: (+82) 80 023 8111 Tel: (+82) 31 329 9000 Fax: (+82) 31 329 9090LuxembourgTel: (+32) 3 899 1301 Fax: (+32) 3 899 1311MalaysiaTel: (+60) 3 5635 3321 Fax: (+60) 3 5635 4116MexicoFree Tel: 01 800 007 5300 Free Fax: 01 800 712 9920 Tel: (+52) 722 276 1600 Fax: (+52) 722 276 1601The NetherlandsTel: (+31) 78 620 5411 Fax: (+31) 78 620 5421New ZealandFree Tel: 0800 936 666 Free Fax: 0800 937 777 Tel: (+61) 2 9841 0555 Fax: (+61) 2 9841 0500NorwayTel: (+47) 23 17 60 00 Fax: (+47) 23 17 60 10PolandTel: (+48) 61 829 01 00 Fax: (+48) 61 829 01 20PortugalFree Tel: 800 202 180 Free Fax: 800 202 178 Tel: (+351) 21 924 2555 Fax: (+351) 21 924 2610RussiaTel: (+7) 495 621 5828 Fax: (+7) 495 621 6037SingaporeTel: (+65) 6779 1200 Fax: (+65) 6779 1822SlovakiaTel: (+421) 255 571 562 Fax: (+421) 255 571 564South AfricaFree Tel: 0800 1100 75 Free Fax: 0800 1100 79 Tel: (+27) 11 979 1188 Fax: (+27) 11 979 1119SpainFree Tel: 900 101 376 Free Fax: 900 102 028 Tel: (+34) 91 661 99 77 Fax: (+34) 91 661 96 42SwedenTel: (+46) 8 742 4200 Fax: (+46) 8 742 4243SwitzerlandFree Tel: 0800 80 00 80 Free Fax: 0800 80 00 81 Tel: (+41) 81 755 2511 Fax: (+41) 81 756 5449ThailandTel: (+66) 2 126 8141 Fax: (+66) 2 126 8080United KingdomFree Tel: 0800 717 181 Free Fax: 0800 378 785 Tel: (+44) 1747 833 000 Fax: (+44) 1747 833 313United StatesToll-Free: 800 325 3010 Toll-Free Fax: 800 325 5052 Tel: (+1) 314 771 5765 Fax: (+1) 314 771 5757VietnamTel: (+84) 8 3516 2810 Fax: (+84) 8 6258 4238Internet sigma-aldrich.com©2013 Sigma-Aldrich Co. LLC. All rights reserved. SAFC, SIGMA-ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries. Ascentis is a registered trademark of Sigma-Aldrich Co. LLC. Solutions within and Supel are trademarks of Sigma-Aldrich Co. LLC. Supelco brand products are sold through Sigma-Aldrich, Inc. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see product information on the Sigma-Aldrich website at www.sigmaaldrich.com and/or on the reverse side of the invoice or packing slip.