**Reagents**

*Aflatoxin B1 Occurrence, Detection and Toxicological Effects*

**A.1 Acetylation of cellophane tubings [134]**

Visking dialysis tubings (Sigma- Aldrich).

(iv) A mixture of benzene, acetic anhydride and pyridine in the ratio

(ii) Reagent B—0.5% CuSO4.5H2O in 1% Sodium Potassium tartrate

(iv) Folin-Ciocalteu's phenol reagent (Sigma-Aldrich Chemie GmbH, Fluka Biochemika) diluted with distilled water in the ratio 1:1(V/V). This is

5 ml of reagent C was added to 1 ml of the test sample. This was thoroughly mixed and left at room temperature for 10 min. Thereafter, 0.5 ml of reagent D was added and allowed to remain at room temperature for 30 min. Absorbance was

Serial dilutions of Bovine serum albumin (Sigma) were treated likewise and used to plot standard graph. The unknown protein value in each test sample is

(iii) Reagent C—50 ml of reagent A mixed with 1 ml of reagent B

The cellophane tubings were filled with distilled water and soaked in distilled water for 24 hours. The tubings were then soaked in turn, for 30 min each time in 50% ethanol, absolute ethanol and diethyl ether successively. The tubings were thereafter soaked in the mixture of benzene, acetic anhydride, and pyridine, prepared as described above, for 18 hours. Each tubing was then properly rinsed in distilled water and stored in 10% KCl solution at 4°C until

(v) 10% KCl (10 g of KCl in 100 ml distilled water)

(i) Aqueous ethanol (50% V/V)

**A.2 Protein content determination [131]**

labeled reagent D.

(i) Reagent A—2% Na2CO3 in 0.1 N NaOH

meant to be extrapolated from the standard graph.

(ii) Absolute ethanol

5:4:2 (V/V)

**Procedure**

required.

**Reagents**

**Procedure**

determined at 620 nm.

**A.3 Iodine solution**

(0.3% Iodine in 3% KI)

(iii) Diethyl ether

**A. Appendix**

**Material**

**Reagents**

**64**

