VASP

Voluntary Aflatoxin Sampling Plan

For European Union and Other Markets*

Purpose: To provide an alternative aflatoxin-sampling plan for the almond industry with
an equivalent sensitivity to that currently being used in the European Union (EU). EU
Regulation (EC) No. 401/2006 was used as the guideline for lot size and sample
frequency. The voluntary sampling plan is designed to reduce lots rejected in the EU
market. Sampling for other markets is encouraged, but optional.

Background: It is difficult to get an accurate estimate of the true aflatoxin concentration

in shipments due to the uncertainty (variability) associated with the entire process from
sampling to analytical analysis. Aflatoxin contaminated kernels do not follow a normal
distribution curve and tend to be positively skewed making its distribution within a lot
non-homogenous. As a result of this distribution and the variability, the sampling plan
will falsely accept or reject a portion of the lots as demonstrated by the operating
characteristic curves (OC curves) developed for the almond industry.

• What is an OC curve? The buyers’ risk (bad lots accepted) and sellers’ risk (good
lots rejected) can be predicted based on OC curves. The curves were developed by
determining the variability and distributional characteristics among almond samples
contaminated with aflatoxin. By specifying sample size, number of samples, and
accept/reject levels, the two risks mentioned above can be determined for each
shipment. In general, larger and more frequent samples are more effective in reducing
the buyers’ risk than smaller samples. The OC curves demonstrate how sampling
plans can alter sample size and accept limits, but still be statistically equivalent to the
EU’s protocol. The VASP sampling plan which has been developed is statistically
equivalent to the EU protocol, but uses a smaller sample (15 kg instead of 30 kg) and
a more stringent accept/reject level.

Figure #1 shows how curve # 3 (VASP) closely parallels curve # 5 (EU plan) The
distribution study and subsequent development of the sampling plan was accomplished
by Dr. Tom Whitaker, USDA/Agricultural Research Service located in Raleigh, North
Carolina. Dr. Whitaker is well known throughout the world for developing statistical
sampling plans for aflatoxin analyses for a variety of crops.

*Other markets are encouraged, but optional

 100
Shelled Almonds
90 VCM 100 g Subsample
HPLC 1 aliquot
80
#1 - 1x5 kg <= 2 ng/g
#2 - 2x5 kg <= 2 ng/g
Probability of Accept (%)

70
#3 - 3x5 kg <= 2 ng/g
60 #4 - 4x5 kg <= 2 ng/g
#5 - 3x10 kg <= 4 ng/g
50 Tolerance

40

30
#3 #2 #1
20
#4 #5
10

0
0 5 10 15 20 25 30 35 40
Lot Aflatoxin Concentration (ng/g)

Dr. Whitaker has further defined the variability associated with the entire aflatoxin
analysis from start to finish into 3 major components. The percentage of the total
variability occurs as follows: sampling is 96.2%, sample preparation is 3.6% and
analytical analysis variability is 0.2%. Since the majority of the variability is in the
sampling, attention was focused on the development of a realistic sampling plan that
could be readily implemented by the industry.

USDA has conducted a tremendous amount of research on aflatoxin, and along with
industry data, the correlation of insect damage to aflatoxin is highly documented. In 2001
Dr. Tom Schatzki, of the USDA Western Regional Research Center, published a paper
titled “Dependence of Aflatoxin in Almonds on the Type and Amount of Insect Damage”
(see Attachment 1). A thorough literature search in 2004 conducted by Dr. Bob Sacher,
in a project funded by the Almond Board of California (ABC), reached the same
conclusion (see Attachment 2).

The VASP sampling plan is based on this research and industry data which clearly shows
USDA graded items are the lowest risk for aflatoxin. As initially developed, VASP
focused primarily on consignments which exceeded USDA minimum grades (e.g. ≥ 2%
serious damage). However, in January 2007, the EU proposed “special measures” be
applied to California almonds imported into the EU. This was the result of a Food
and Veterinary Office (FVO) mission which concluded that the aflatoxin control
system for California almonds was inadequate. The EU measures require 100%
surveillance, meaning that every consignment imported from the U.S. will be tested
for aflatoxin. California almonds that have been tested under the VASP protocol
prior to shipment, and accompanied by a uniform VASP certificate, will be subject
to 5% surveillance. Therefore, VASP will require testing of all lots destined for the
EU.

Revised 01/09/07 2
Sampling Plan: This document only addresses the sampling plan for shipments destined
for the EU; however, handlers are encouraged to apply the plan to shipments for other
markets.

Almond shipments destined for the EU will have to be tested by taking an aggregate
sample totaling 15 kg. The aggregate sample will be divided into three 5 kg sub-samples;
when analyzed, each sub-sample must be ≤ 2 ppb total aflatoxin for the lot to be
accepted. The VASP plan provides for the designation of a lot which failed the standards
for the EU to be shipped to other markets – provided the lots do not exceed the specific
market’s tolerance. Failed lots may also be reprocessed (e.g. manual or electronic
sorting, blanching) to remove the serious damage and then be retested to determine if
they comply with EU or other market requirements. Figure # 2 provides a flow diagram
of the VASP decision process. This is the minimum requirement, more robust
sampling plans, such as following the EU’s protocol of 30kg sample is acceptable.

Finished Product Sampling: Sampling needs to be consistent within the industry in

order to reduce variability of results. Each outgoing bin or pallet is to be sampled by
taking incremental samples from throughout the lot. The total aggregate weight from the
incremental samples needs to be 15 kg. Figure #3 provides recommended sampling
patterns for bins and cartons. Each bin or pallet needs to be samples at least once,
utilizing the sample locations indicated, ensuring that a 15kg sample is collected. If
probing sealed bins or cartons, it is important to seal the probe hole, and initial and date it
so the customer will know it has been sampled and not tampered with.

• In-line sampling should be conducted at a rate to yield a total sample of 15 kg or 33

pounds which is representative of the entire lot. This may be done manually or by
automatic in-line samplers.

• Sampling Manufactured Items follows the same protocol as above. Research has
shown that reprocessing such as sorting (electronic and manual) and blanching will
reduce aflatoxin contamination..

Lot Size/Sublot: The VASP plan focused on a day’s production run or a container load.
The EU definition of a lot is “an identifiable quantity of a food commodity delivered at
one time and determined by the official to have common characteristic such as origin,
variety, type of packaging, packer, consignor or markings. Based on this requirement, if
each container is sampled according to the VASP plan, it will be consistent with this
regulation.

Sample Preparation: A qualified individual must be used to collect representative

incremental samples of the lot to equal the aggregate weight of 15 kg. The representative
incremental samples collected from the lot must all be mixed together to be sure that each
subsample contains portions of the whole lot. Sub-dividing the total sample into the three
5 kg sub-samples should be done by the laboratory prior to grinding and analysis.

Revised 01/09/07 3
Analytical Analysis: Each of the 5 kg samples is individually ground and presented for
analytical analysis. Grinding should be accomplished by a method which not only
reduces the particle size but also is effective in thoroughly mixing the particles to a
homogenous grind. No specific method or laboratory is specified; however, for a VASP
certificate to be issued for a shipment to Europe, analyses must be done by a USDA-
approved laboratory, following specific analytical parameters and procedures which are
consistent with EU Regulation (EC) No. 401/2006, annex II, section 4.3. Analysis results
must be documented on the VASP certificate.

Records Verification: - Accurate records must be maintained to demonstrate

reprocessing of failed lots.

Voluntary Program: In order to monitor the impact of the VASP program and compare
results with European surveillance, handlers who participate in the VASP program will
be asked to sign a Memorandum of Understanding (MOU) agreeing to follow the testing
and sampling protocols of the program. As part of the agreement handlers agree to allow
the Almond Board staff to periodically review records to ensure that the handler is
performing in accordance with the agreement.

Revised 01/09/07 4
Figure #2
Figure # 1

Eu and Other Markets Aflatoxin Sampling Plan

Finished Product

Other EU or Other No US Std Sheller Run or Yes EU or Other

Follow Market Other Follow Market
Markets Better? Markets
Regulations Regulations
Serious Damage < =2%

EU
EU

Almond Shipments to Europe Divert to

Appropriate
Non -EU
Market

3 X 5 KG
Reprocess**
@ 2 ppb Total

All 3 < =
2 ppb Total No
Aflatoxin*

NOTES:
Yes * Each of the 3 individual samples has to be less
than or equal to 2 ppb total aflatoxin. If one
sample is greater than 2 ppb total aflatoxin the
entire lot fails eventhough the other 2 tests are
less than 2ppb total aflatoxin .
Ship EU

** Reprocessing refers to steps taken to reduce

serious damage levels.Action taken, such as
manual or electronic sorting and blanching would
EU be considered reprocessing.Testing is mandatory
after reprocessing to confirm aflatoxin levels are
less than or equal to 2 ppb total aflatoxin.
Shipments

Revised 1/9/07

Revised 01/09/07 5
Figure #3
Figure # 2

Probe Sampling Each Bin or Pallet

3
2
1

5
4

9
8
7

Repeat position 1 Overall probe position

for the 10th bin Make an X

9 5
8 4
3
10 2 7
1 6
Bin

Note: Each bin or pallet in the container is 9 5

to be sampled in this method. 8 4
3
2 7
1 6
Pallet

Revised 4/28/06

Revised 01/09/07 6