Aflatoxin m1

Glutaraldehyde (GAD, 25% v/v, aqueous solution), acetylcholinesterase (C 3389–500UN), aflatoxin B1, ochratoxin A, aflatoxin M1, acetylthiocholine chloride (ATCh), glycerol, bovine serum albumin, and sodium alginate (from brown algae, viscosity ≥ 2000 cP, 2% (25 °C) (lit.) were purchased from Sigma Aldrich.(Merck, Darmstadt, Germany), aflatoxin B1 was solubilized in methanol (1 mM), followed by dissolution in water.

Aflatoxin M1 (AFM1), AFB1, chloramphenicol (CAP), CAP succinate sodium salt (CAP-SH), florfenicol (FF), hemocyanin keyhole limpet (KLH), carboxymethoxylanmine hemihydrochloride (CMO), 1-ethyl-3-[3-dimethylaminopropyl]-carbodimide (EDC), N-hydroxysuccinimide (NHS), N,N-dicyclohexylohexylcarbodiimide (DCC), sodium bicarbonate, ovalbumin (OVA), bovine serum albumin (BSA), hydrogen tetrachloroaurate, and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, MO, USA). Horseradish peroxidase (HRP) was obtained from Roche (Mannheim, Germany). K-Blue TMB substrate (3,3′, 5,5′-tetramethylbenzidine) was brought from Neogen Corp (Lexington, KY, USA). Sodium carbonate, Tween 20, and ammonium sulfate were obtained from Merck (Darmstadt, Germany). Freund’s complete/incomplete adjuvants, goat anti-rabbit IgG, and microtiter plates were obtained from Thermo (East Grinstead, UK). All strip components were obtained from MDI Technologies (Ambala, India). The Troy Double Axes Programmable Controller (Taichung, Taiwan) was used to draw the test and the control line on the immunostrip. An ELISA reader (Vmax) was from Molecular Devices Co. (Menlo Park, CA, USA). A strip scan reader was purchased from Taiwan Advance Bio-Pharmaceutical Inc. (Taipei, Taiwan).

The following reference standards were purchased: aflatoxin B₁ (AFB₁, > 99%), aflatoxin B₂ (AFB₂, > 99%), aflatoxin G₁ (AFG₁, > 99%), aflatoxin G₂ (AFG₂, > 99%), aflatoxin M₁ (AFM₁, > 98%), and sterigmatocystin (ST, > 99%) all solved in acetonitrile from Sigma-Aldrich Chemie GmbH (Taufkirchen, Germany), aflatoxin M₂ (AFM₂) in acetonitrile (> 98%) and aflatoxicol (AFL, > 99%) from Cfm Oskar Tropitzsch GmbH (Marktredwitz, Germany), and O-methylsterigmatocystin (OMST, > 95%) from Cayman Chemical (Ann Arbor, MI, USA). Of these standards, two standard mixtures were generated: standard mixture 1 (containing 640 nmol/L of AFB₁, AFB₂, AFG₁, and AFG₂ in acetonitrile) and standard mixture 2 (containing 640 nmol/L of AFM₁, AFM₂, ST, and AFL as well as 608 nmol/L of OMST in acetonitrile). All further chemicals and solvents were of analytical grade. Deionized water was obtained from an in-house ultrapure water system (LaboStar, Erlangen, Germany).