Aflatoxin Control Program for Peanuts 1

J.W. DICKENS, ARS, USDA, North Carolina State University, Raleigh, North Carolina 27607
ABSTRACT A N D S U M M A R Y Under provisions of a USDA Marketing Agreement, an aflatoxin control program for peanuts produced in the United States is administered by the Peanut Administrative C o m m i t t e e c o m p o s e d of peanut growers and shellers. Regulations o f this comm i t t e e c o n t a i n provisions a b o u t the quality of peanuts a c q u i r e d from farmers, storage of unshelled peanuts, a f l a t o x i n testing, quality and disposition of processed lots, and indemnification of handlers for losses caused by lots which test over 25 parts-perbillion aflatoxin. Effects o f the control program on aflatoxin c o n c e n t r a t i o n s in peanut p r o d u c t s are discussed. INTRODUCTION An alfatoxin c o n t r o l program for peanuts produced in the United States is administered b y the Peanut Administrative C o m m i t t e e (PAC) under provisions of a USDA Marketing Agreement for Peanuts (1). The 18-member PAC consists of three grower representatives and three sheller representatives f r o m each o f the three p e a n u t - p r o d u c t i o n areas (Virginia-Carolina, Southeastern, and Southwestern). PAC generally solicits r e c o m m e n d a t i o n s from representatives of p e a n u t - p r o d u c t manufacturers. The committee is financially s u p p o r t e d b y assessment of shellers based upon the volume o f peanuts t h e y purchase. PAC regulates peanut shellers who have signed the Peanut Marketing Agreement, and practically all peanuts p r o d u c e d in the United States are m a r k e t e d through these shellers. They purchase peanuts from the grower (farmers' stock peanuts), store and process these peanuts into marketable lots of in-shell or shelled raw peanuts, and sell them to manufacturers of peanut products, either directly or through brokers. Through PAC, shellers have accepted a major role in the aflatoxin c o n t r o l program o f the peanut industry. They a t t e m p t to channel aflatoxin-contaminated peanuts from the farm to n o n f o o d uses; to prevent ariatoxin c o n t a m i n a t i o n during storage, handling, processing, and shipment; to remove aflatoxin-contaminated kernels during processing; and to deliver aflatoxin-free peanuts to the manufacturer. In this p a p e r provisions of PAC for the 1975 peanut crop and their relation to the aflatoxin control program are discussed. PURCHASING FARMERS' STOCK The aflatoxin-producing Aspergillus flavus group o f fungi exist t h r o u g h o u t the peanut growing areas and m a y produce aflatoxin in peanuts any time that conditions are favorable for fungal growth. During periods of drought, insects and mites in the soil may favor infection by A. flavus and subsequent aflatoxin p r o d u c t i o n before peanuts are dug (2). E x t e n d e d periods of hot, rainy weather while peanuts are in the windrow, i m p r o p e r drying after harvest, and inadequate p r o t e c t i o n from rain during t e m p o r a r y storage and t r a n s p o r t a t i o n are conducive to A. flavus growth. PAC regulations require that all farmers' stock peanuts purchased b y shellers be graded by the Federal-State Inspection Service. The 1800-g grade samples taken for this 1Paper number 4978 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, NC 27607. J. AM. OIL CHEMISTS' SOC., March 1977 (VOL. 54) purpose consist o f p e a n u t pods, peanut kernels which have been shelled during harvesting and handling o p e r a t i o n s prior to sampling ( c o m m o n l y called loose shelled kernels), and foreign material. Kernels shelled from a 500-g sample of the pods are e x a m i n e d for external m o l d or d i s c o l o r a t i o n (damage), and then the cotyledons are split a p a r t for internal e x a m i n a t i o n . These kernels and all loose shelled kernels ( L S K ) from the sample are e x a m i n e d visually for A. flavus infection (3). All suspect kernels ace e x a m i n e d u n d e r 20X or 4 0 X magnification with a s t e r e o m i c r o s c o p e t o determine w h e t h e r or n o t fungal growth is present and whether the color and certain morphological traits are characteristic o f the A. flavus group of fungi. Lots f o u n d t o c o n t a i n kernels suspected to have visible A. flavus g r o w t h are placed in segregation-3 storage. L o t s in which none of these kernels are found b u t with m o r e t h a n 2% damaged kernels or more than 1% concealed damage caused b y rancidity, mold, or decay are placed in segregation-2 storage. Segregation-1 storage receives all o t h e r farmers' s t o c k peanuts. The segregation-3 peanuts are crushed for oil, which is aflatoxin free after refining, and the meal is used for nonfood purposes (4). Segregation-2 peanuts are crushed for oil and the meal is used in animal feed if chemical assay for aflatoxin does n o t indicate otherwise. (PAC m a y allow use of segregation-2 peanuts for edible purposes if there is a shortage o f segregation-1 peanuts, b u t this allowance has never been made.) Peanuts from segregation-1 storage are shelled and used for f o o d purposes provided t h e y are f o u n d acceptable by chemical assay for aflatoxih. These restrictions do n o t a p p l y to peanuts used for seed. STORAGE OF FARMERS' STOCK PEANUTS

A. flavus g r o w t h during storage m a y be a m a j o r cont r i b u t o r to aflatoxin c o n t a m i n a t i o n in peanuts (5). Possible causes o f A. flavus growth are moisture c o n d e n s a t i o n on roofs and sidewalls, leaking roofs, i m p r o p e r a p p l i c a t i o n o f insecticide sprays or leaking hoses and a p p l i c a t i o n equipment, conveyance o f water from flooded elevator d u m p pits into warehouses, and storage o f peanuts on concrete floors that are damp or have no vapor barriers. A m a j o r p r o b l e m is c o n d e n s a t i o n of moisture that evaporates from peanuts as t h e y dry from an average of a b o u t 9% moisture to a b o u t 7% moisture during storage. This water v a p o r m u s t be p r o p e r l y r e m o v e d from the storage structure t o prevent wetting some o f the peanuts with condensation. PAC regulations require ventilated storage buildings and m a k e provisions to reduce o t h e r causes of wetting. E x c e p t for seed, all peanuts must contain less than 10% moisture (wet basis) when placed in storage. SHELLING AND GRADE REQUIREMENTS Peanut kernels c o n t a m i n a t e d with a f l a t o x i n before harvesting are o f t e n in damaged pods and c o n s e q u e n t l y are more easily shelled by harvesting and handling o p e r a t i o n s than kernels in sound pods. Small, shriveled kernels also occur more f r e q u e n t l y in these damaged pods. Shelled kernels (LSK) and kernels in damaged p o d s are more susceptible than kernels in sound pods to m o l d damage during storage. Consequently, in a f l a t o x i n - c o n t a m i n a t e d farmers' s t o c k peanuts, LSK generally contain higher concentrations of aflatoxin than do unshelled kernels, and the small kernels within each t y p e p r o b a b l y c o n t a i n higher concentrations than do the larger kernels (5). 225A

TABLE I PAC Aflatoxin Testing Program for Shelled Peanuts Step 1 Comminute first 48-1b sample in subsampling mill Step 6 Comminute second 48-1b sample in subsampling mill Step 2 Extract 1100-g subsample
extract

Step 3 Make duplicate analyses (1A & 1 B)

of

Step 4
Let X = a v e r a g e o f

Step 5 PPB Reject if X i> 75 PPB Go to Step 6 if: 16PPB< X < 7 5 PPB
A c c e p t ff X ~< 1 6

l A and 1B

Step 7 Extract 1100-g subsample

Step 8 Make duplicate analyses of extract (2A & 2B)

Step 9 Let Y = average of

1A, 1B, 2A, and

2B Step 12 Extract 1100-g subsample Step 13 Make duplicate analyses of extract (3A & 3B) Step 14 Let Z = average of IA, IB, 2A, 2B, 3A and 3B

Step 10 Accept if Y ~<22 PPB Reject if Y ~> 38 PPB Go to Step 11 if: 22PPB< Y < 38PPB Step 15 Accept if Z =~25 PPB Reject if Z > 25 PPB

Step 11 Comminute third 48-1b sample in subsampling mill

1.0 0.9 LLI (D Z O8

1:1_ 0.7 LLI (D 0.6 Ii 0 0.5

peanuts and damaged kernels or a total of 3.0% unshelled peanuts, damaged kernels, and kernels with m i n or defects. Moisture co n t en t o f the peanuts must not exceed 10% (wet basis) in the Virginia-Carolina p e a n u t - p r o d u c t i o n area and 9% in the other areas. Foreign material must not exceed 0.1% for some grades and 0.2% for o t h er grades. All kernels which fail to ride screens with designated slot sizes are called "fall-through." Fall-through must not exceed 3% of the lot except 4% is allowed for U.S. No. 2 Virginia grade peanuts and only 2% is allowed for certain grades of Spanish peanuts with splits.

A F L A T O X I N INDEMNI FICATION PROGRAM

The PAC indemnifies shellers for most of the losses related to aflatoxin c o n t a m i n a t i o n in lots of edible peanuts which meet grade requirements. PAC regulations prohibit the sheller from negotiating the sale o f peanuts on the basis of aflatoxin content. The b u y e r (manufacturer) is guaranteed that the peanuts wig test " n e g a t i v e " (not more than 25 parts-per-biUion) by the official PAC aflatoxin-testing program, but the sheller is n o t allowed to make any other specification in regard to aflatoxin. Manufacturers who complete tests within 36 hr after the lot is sampled may accept lots of peanuts on the basis o f their o w n aflatoxin analyses, but only those lots which test positive in PAC approved laboratories may be returned to the sheller because of aflatoxin c o n t a m i n a t i o n . 10 20 30 40 50 60 70

>t-- Q4 J 1313 Q5 <I: 133 0 Q2 It" 13_ 0.1

PAC A F L A T O X I N TESTING PROGRAM

All shelled peanuts sold for h u m a n c o n s u m p t i o n must be positive-lot-identified and sampled by the Federal-State Inspection Service for aflatoxin tests. The aflatoxin testing program is outlined in Table I. One 48-1b sample is cornm i n u t ed in a subsampling mill (6) and a 1100-g subsample is sent to a PAC approved laboratory. (Except for two independent laboratories, PAC approved laboratories are operated by federal or state government.) The entire 1100-g subsample is ex t r act ed in 3000 ml methanol-water (55:45) and 1000 ml hexane or technical-grade petroleum ether. Duplicate 50-ml portions o f the m e t h a n o l - w a t e r extract are analyzed for aflatoxin by Method II o f AOAC (7). The two independent determinations o f aflatoxin concentration by thin layer c h r o m a t o g r a p h y (TLC) are averaged. If the average is 16 parts-per-billion (ppb) or less, an "aflatoxinnegative" certificate is issued and the lot is accepted. If the average is more than 75 ppb, the average co n ce nt r a t i on is shown on the certificate, and the lot is rejected. Otherwise, a second 48-1b sample is analyzed by the same procedure, and the aflatoxin determinations f r o m the first and second 48-1b samples are averaged. If the average of the four determinations is 22 p p b or less, an aflatoxin-negative certificate J. AM. OIL CHEMISTS' SOC., March 1977 (VOL. 54)

LOT CONCENTRATION (PPB)

FIG. 1. Probability of accepting aflatoxin-contaminated lots of shelled peanuts with the PAC aflatoxin testing program. PAC requires removal of all LSK from farmers' stock peanuts prior to shelling. Whole LSK that ride screens with designated slot sizes ( 1 6 / 6 4 x 3/4-in. for runner type, 1 5 / 6 4 x 3/4-in. for Spanish and Valencia type, 1 5 / 6 4 x 1-in. for Virginia type) m a y be included with other peanuts prepared for h u m an c o n s u m p t i o n . All o th e r LSK must be designated for crushing, b u t t h e y m a y be used for wildlife feed or rodent bait if they do n o t test m o r e than 25 partsper-billion aflatoxin. Shelled peanuts sold for h u m a n c o n s u m p t i o n must meet r e q u i r e m e n t s about kernel size, damaged kernels, moisture c o n t e n t , and foreign material. F o r compliance with these requirements, each lot must be sampled and graded by the Federal-State Inspection Service. The lots must be "positive-lot i d e n t i f i e d " by means of l o t - n u m b e r e d Federal-State Inspection Service tags sewn in the closure o f bags or by means of official seals on bulk containers. Edible peanuts must n o t contain m o r e than a total o f 1.5% unshelled 226A

TABLE II Distribution According to Aflatoxin Concentration of Shelled-Peanut Lots Produced for Edible Purposes 5 % of total lots above indicated PPB in 1973 % of accepted lots above indicated PPB in 1973 % of total lots above indicated PPB in 1974 % o f accepted lots above indicated PPB in 1974 53.00 49.20 32.80 31.50 10 32.00 27.00 15.20 13.50 15 20.00 13.80 7.30 5.50 20 12.40 6.40 3.50 2.50 Aflatoxin concentrations (PPB) 25 30 35 7.70 2.70 1.70 0.69 4.80 1.10 0.85 0.27 3.00 0.38 0.42 0.08 40 1.90 0.12 0.21 0.02 45 1.20 0.04 0.10 0.00 50 0.70 0.01 0.05 0.00 55 0.46 0.00 0.03 0.00

is issued. If the average is 38 ppb or mr, re, the average concentration is shown on the certificate, and the lot is rejected. Otherwise, a third 48-1b sample is analyzed, and when the six determinations average 25 ppb or less, a negative certificate is issued. Otherwise, the average concentration is shown on the certificate and the lot is rejected. The results of all aflatoxin analyses are made available to the buyer of the peanuts. As indicated previously, the manufacturer located in the production area may elect to c o n d u c t his own 36-hr ariatoxin analyses. In this case, the Federal-State Inspection Service will provide the manufacturer's laboratory with two 1100-g subsamples from the first 48-1b sample and hold two subsamples in reserve. A f te r analysis of his subsamples, the manufacturer may accept the lot or request the official PAC test outlined above. Whitaker and Dickens (unpublished data) have estimated the probability of accepting lots with various concentrations of alfatoxin when the official PAC testing program is used. As shown in Figure 1, there is over 99% probability of accepting lots with 10 p p b and very little probability o f accepting lots with 70 ppb. A p p r o x i m a t e l y 65% of the lots with 25 ppb are accepted.

official documents sent to the purchaser o f the peanuts.

E V A L U A T I O N OF THE A F L A T O X I N CONTROL PROGRAM

The average aflatoxin c o n c e n t r a t i o n in all lots o f peanuts accepted by the PAC aflatoxin testing program and the risk of accepting lots with high concentrations depends u p o n the co n d i t i o n of the peanut crop after shelling. Whitaker and Dickens (unpublished data) have estimated the distribution according to aflatoxin concentrations for all lots of edible-grade shelled peanuts p r o d u c e d from b o t h the 1973 and 1974 crops and for those lots accepted from each crop by the PAC testing program. All lots contained an average aflatoxin co n cen t r at i o n of 10 ppb for the 1973 crop and 5 ppb for the 1974 crop. As shown in Table II, ca. 7.70% of the lots produced and 2.70% of the lots accepted f r o m the 1973 crop contained over 25 ppb aflatoxin c o m p a r e d to 1.70% and 0.69%, respectively, for the 1974 crop. Only 0.01% o f the lots accepted from the 1973 crop and n o n e of the lots accepted from the 1974 crop contained m o r e than 50 ppb aflatoxin. The manufacturer has the final responsibility for producing wholesome peanut products for the c o n s u m e r market. The National Peanut Council has published a voluntary code of good practices for purchasing, handling, storage, processing, and testing of shelled peanuts (8). The voluntary code r e c o m m e n d s the removal from the processing stream before a n d / o r after blanching of those peanut kernels most likely to contain aflatoxin. These kernels include moldy, discolored, shriveled, and damaged kernels; kernels that resist blanching or splitting; and those that have a dark color after roasting. Destruction by roasting of from 30 to 50% of any aflatoxin remaining in the peanuts also reduces aflatoxin in the finished p r o d u c t (9). The alfatoxin control program o f the PAC, the procedures outlined in the National Peanut Council code o f g o o d practices, the destruction of aflatoxin during roasting, and thorough aflatoxin testing o f the finished p r o d u c t help provide the consumer with safe p e a n u t products.

APPEAL TESTING FOR AFLATOXIN

Manufacturers may appeal official PAC aflatoxin tests if positive-lot identification has been maintained for the lot. A 144-1b sample is c o m m i n u t e d and duplicate analyses are made on three 1100-g subsamples. If the average of the six determinations is 25 ppb or less, the lot must be accepted. If the average is more than 25 ppb, the manufacturer may reject the lot. The m a n u f a c t u r e r pays all costs of appeal testing, and there is no limit on the number of appeals made on a lot.

DISPOSITION OF A F L A T O X I N CONTAMINATED LOTS

Lots of shelled peanuts which test positive for aflatoxin may be remilled or blanched in an a t t e m p t to remove the afiatoxin. Remilling may include the following procedures: (a) screening t o r e m o v e small kernels, (b) treatments to remove low-density kernels and foreign material, ( c ) e l e c tronic color sorting to r e m o v e discolored kernels, and (d) hand picking to remove discolored kernels. The blanching process consists of removing the skin or testa from the kernels followed by color sorting and hand-picking to remove the discolored kernels and those kernels that retain their skins. Aft er blanching or remilling, the lots are considered to be new lots and are subjected to the PAC aflatoxin testing program outlined above. There is no limit on the n u m b e r of times a lot may be remilled, and the lot may be blanched following remilling. Eventually, all lots which fail to pass the PAC aflatoxin testing program must be restricted f r o m food or feed except for the oil which is aflatoxin-free after proper refining. Exported peanuts that do not test negative must show aflatoxin concentrations on J. AM. OIL CHEMISTS' SOC., March 1977 (VOL. 54)

ACKNOWLEDGMENT Advice was received from W.I. Yarbrough, Manager, Peanut Administrative Committee. REFERENCES 1. "1975 Marketing Agreement for Peanuts," Peanut Administrative C o m m i t t e e , P.O. Box 18856, Atlanta, GA 30326.
2. Dickens, J.W., and J.B. Satterwbite, Journal o f the A m e r i c a n Peanut Research and Education Association 5:48 ( 1 9 7 3 ) .

3. Dickens, J.W., and J.B. Satterwhite, Oleagineux 25:321 (1971). 4. Parker, W.A., and D. Melnick, JAOCS 42:471 (1966). 5. Dickens, J.W., Journal o f the A m e r i c a n Peanut Research and Education Association 7:54 (1975). 6. Dickens, J.W., and J.B. Satterwhite, Food Technol. 23:90 (1969). 227A

7. "Official Methods of Analysis of the Association of Analytical Chemists," Chapter 26, Twelfth Edition, AOAC, Washington, DC, 1975. 8. "Voluntary Code of Good Practices for Peanut Product Manufacturers," Eleventh Edition, 1976, National Peanut Council,

7900 Westpark Drive, McLean, VA 22101. 9. Waltking, Arthur E., J. Ass. Offie. Anal. Chem. 54:533 (1971). [ R e c e i v e d J u l y 6, 1976]

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