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Introduction
Aflatoxin (AF) is the strongest known naturally occurring chemical structures of B1, B2, G1, G2, M1, and M2
carcinogen. Animal feed and food products are strictly aflatoxins.
inspected for AF contamination. Figure 1 shows the
Figures 2 to 4 show chromatograms of AF found in ppb). As shown in Figure 6, the sample was pretreated by
naturally contaminated peanut meal, corn, and poultry an acetone extraction of AF.
feed. The samples were prepared for HPLC injection
according to the CB Method of the Association of Official Table 1 shows the percent recovery of aflatoxins in milk
Analytical Chemists (AOAC). and dairy products using this method. The percent
recovery was low when solvents other than methanol and
Figure 5 shows the analysis of contaminated cheese the mixed methanol/acetone solvents were used.
containing aflatoxins M1 (0.1 – 0.9 ppb) and M2 (0.01
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Analytical Conditions
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Analytical Conditions
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Analytical Conditions
Figure 4 – Chromatogram of mixed feed for chicken (aflatoxins B1, B2, G1, and G2, 33 ppb each added)
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Analytical Conditions
Column : Silica gel (5.5µm particle size), 4mm i.d. x 100mm
Mobile phase : Toluene-ethylacetate-formic acid-methanol
(90 : 5 : 2.5 : 2.5)
Flow rate : 1.1ml/min
Column temp. : 40°C
Detection : Shimadzu spectrofluorophotometer
(Em=365nm, Ex=425nm)
(a) Standard of aflatoxins : AFB1, B2, G1 and G2,
5ng each; AFM1 and M2 10 ng each.
(b) Aflatoxins from contaminated cheese :
injection amount, 20 µl
Figures 5 – Chromatograms of aflatoxins extracted from contaminated cheese
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Sample (75g)
water
acetone (215ml)
Blended for 30 min
saturated sodium sulfate (10ml)
diet earth (10g)
Stirred
Filtered
Filtrate (200ml)
5% sodium chloride (100ml)
hexane (100ml)
Shaken for 5 min
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Recovery of aflatoxins (%)
AFB1 AFB2 AFG1 AFG2 AFM1 AFM2
Milk 98 90 86 93 93 95
Butter 88 92 99 96 92 94
Cheese 60 80 67 85 93 97
This Application Note was edited with the Food Research Institute, Ministry of Agriculture,
cooperation of Dr. Tetsuya Goto of the National Forestry, and Fisheries, Japan.
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