Use of Combined Uncertainty of Pesticide Residue Results For Testing Compliance With Mrls

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Article
Use of Combined Uncertainty of Pesticide Residue
Results for Testing Compliance with MRLs
Zsuzsa Farkas, Andrew Slate, Thomas B. Whitaker, Gabriella Kötelesné Suszter, and Árpád Ambrus
J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/jf505512h • Publication Date (Web): 06 Feb 2015
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Page 1 of 43 Journal of Agricultural and Food Chemistry
1 Use of Combined Uncertainty of Pesticide Residue Results for Testing Compliance
2 with MRLs.
4 Zsuzsa Farkas1, Andrew Slate2, Thomas B. Whitaker2, Gabriella Suszter3, Árpád Ambrus1a
6 1. National Food Chain Safety Office, Budapest Tábornok u 2, 1143 Hungary
7 2. North Carolina State University 124 Weaver Labs, Raleigh, NC 27695-7625, USA
8 3. Wessling Hungary Ltd, Budapest Fóti út 56, 1047 Hungary
9 1a Retired Scientific Adviser, ,
10
11
1
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry Page 2 of 43
12 ABSTRACT
13
14 The uncertainty of pesticide residue levels in crops due to sampling, estimated for 106 individual crops
15 and 24 crop groups from residue data obtained from supervised trials was adjusted with a factor of 1.3 to
16 accommodate the larger variability of residues under normal field conditions. Further adjustment may be
17 necessary in case of mixed lots. The combined uncertainty of residue data including the contribution of
18 sampling is used for calculation of an action limit, which should not be exceeded when compliance with
19 maximum residue limits is certified as part of pre-marketing self control programs. On the contrary, for
20 testing compliance of marketed commodities the residues measured in composite samples should be ≥
21 the decision limit calculated only from the combined uncertainty of the laboratory phase of the residue
22 determination. The options of minimizing the combined uncertainty of measured residues are discussed.
23 The principles described are also applicable to other chemical contaminants.
24
25
26 KEY WORDS: sampling, uncertainty of measurement results, compliance with legal limits, pesticide
27 residues, self control of production, certification of compliance
28
29
2
30 INTRODUCTION
31
32 Distribution of pesticide residues.
33 The distribution of pesticide residues in/on treated crops is affected by many factors including, , for
34 instance, the type and spatial form of plants, cultivation and application method, and weather conditions
35 during application. The field to field variation of the magnitude of pesticide residues and their within
36 field concentration ranges can be expected to vary up to 10000 and 100 fold, respectively.1 Horváth and
37 co-workers have recently reviewed the characteristics of distribution of pesticide residues in crop units
38 or primary samples.2 Their statistical analyses of over 19000 residue data representing combinations of
39 20 crops and 46 pesticides confirmed previous assumptions3,4 that the residue sets obtained from the
40 analyses of crop units can be best described with lognormal distribution having a relative standard
41 deviation, CV, around 0.8. Even if 100-300 natural crop units are collected from one field, the CV of the
42 measured residues would provide only one estimate of the variability of residues. If the random
43 sampling operations are repeated many times the average of CV values approaches the true CV of the
44 parent populations. Consequently, the average of CV values obtained from the analyses of crop units
45 taken from different fields would provide the best estimate for the typical CV of residue values in the
46 given crop units.5 Due to the large variation of pesticide residues in crop units, the average residues in
47 composite samples containing 5-10 or larger number of primary samples6 will also substantially vary.
48 Uncertainty of measurement results.
49 The uncertainty of sampling, as one of the components of the combined uncertainty of the results of
50 pesticide residue analysis, has been estimated based on three independent databases: residues in 21000
51 primary samples5,7, residues in composite samples taken from commercially treated crops8, and residues
52 derived from 25876 samples taken from 12087 independent supervised residue trials.9
53 The contribution of sub-sampling, sample processing and homogenization to the variability of analytical
54 results has been studied by several authors.10-14 It was shown that the efficient comminuting of the
55 samples, especially those with hard peel and soft pulp, is a difficult task, and the result largely depends
3
56 on the crop material, equipment used, as well as the temperature and the length of processing. On the
57 other hand, it is worth to note that, if the residues are stable under the sample processing conditions, the
58 efficiency of sample processing does not depend on the nature of the chemical substances. In addition to
59 homogenization of the sample material, the sample size reduction15 may also contribute to the combined
60 uncertainty with a sub-sampling (CVSS) of 8-12%.
61 Maximum residue limits for chemical contaminants.
62 For assuring the safety of marketed food, regulatory agencies define maximum residue limits (MRL) for
63 pesticides16-19 and maximum limits (ML) for chemical contaminants.17,20,21 In addition, the Codex
64 Alimentarius Commission (CAC) establishes MRLs or MLs to facilitate international trade.22-24 The
65 MRLs and MLs are defined as the maximum legally permissible average concentration of chemical
66 contaminants in a composite sample with specified minimum mass and size in terms of the number of
67 primary samples (single sample increments). The sample sizes defined by the national authorities are
68 generally the same or very similar to those given in the relevant Codex Standards, such as for instance
69 for pesticide residues.5
70 The objective of this paper is to show that available data on the uncertainty (variability) of pesticide
71 residue measurement results may be used in the process of certification of compliance with MRLs. The
72 principles described are also applicable for other chemical contaminants.
73
74 METHODS
75
76 Estimation of combined uncertainty of the measured residues.
77 The combined relative standard uncertainty (expressed as relative standard deviation or coefficient of
78 variation) of the residues measured in composite samples (CVR) can be expressed as25
79 = +

+

+ Equ. 1
4
80 It incorporates the uncertainties of sampling (CVS), sub-sampling (CVSS), sample processing (CVSP)
81 (chopping, mincing and homogenization of analytical sample) and analysis (CVA). The methods for
82 estimation of sampling uncertainty have been described in detail in previous publications,7,8,26 therefore
83 they are not repeated here. Where the contribution of uncertainty of sub-sampling and sample
84 processing is included in the estimated uncertainty of the laboratory phase of measurement (CVL), the
85 two phases of the process can be distinguished as:
86 CV = CV + CV Equ. 2
87 CV = CV

+ CV

+ CV Equ. 3
88 It is to be noted that the CVL should always be determined either from retained test portions of the
89 laboratory samples which contain incurred residues,10 or after treatment of the surface of the individual
90 crop units with the test compound.14 Furthermore, the results of collaborative studies or proficiency tests
91 can only provide information on the uncertainty of the analytical procedure (CVA) from the point of
92 extraction and not for the combined uncertainty of the analytical results (CVL), as the appropriate
93 homogeneity of those samples had been carefully checked before distribution to the participants.
94 Principles of control of compliance of food commodities with the legal limits.
95 As the consequence of the definition of MRLs, there are two distinctly different situations in the control
96 of compliance of food commodities with the legal limits. Different sampling plans are required for
97 testing compliance with MRLs of pesticide residues in commodities before and after marketing..
98 Control of commodities on the market (enforcement or monitoring program).
99 As the permissible maximum concentrations of residues and chemical contaminants apply to the average
100 concentration of the substances in the bulk/laboratory sample, the combined uncertainty of the
101 laboratory phase of the determination of the residue concentration (CVL) shall be taken into account in
102 deciding on the compliance or non-compliance of the sampled product. If the samples taken satisfy the
103 minimum requirements for the number of primary samples and the mass of the laboratory/bulk sample,6
104 the average residue determined in the sample provides the basis for deciding on the acceptability of the
5
105 sampled lot. In order to make regulatory action in accordance with the relevant ISO and Codex
106 guidelines,27,28 among other regulatory agencies, the European Commission (EC) regulations clearly
107 indicate that a lot is considered non-compliant if the measured analyte concentration corrected for
108 recovery, where it should specifically be considered, minus two times the expanded uncertainty of the
109 results are above the legal limit. For pesticide residues the default combined relative uncertainty
110 reflecting the CVL value (equ. 3) is defined as 25%.29 This means that a lot could be rejected if the
111 residue measured in the composite sample taken from the lot would be >2×MRL. We call this
112 concentration the Decision Limit (DL). Figure 1 illustrates the situation where the MRL is 0.2 mg/kg
113 and the apple lot (mean of 0.1 mg/kg and CVR of 0.35) would be tested on the market applying the
114 decision limit of 0.4 mg/kg (0.4-2×0.25×0.4=0.2 mg/kg) as defined by the EC. Under this condition a
115 testing laboratory would reject a lot containing residue ≤0.2 mg/kg in 2.3% of the cases, because the
116 dispersion of analytical measurement results can be considered normal.30
117 Premarketing self-control.
118 When the product is tested before placing it on the market, it should be certified that at least a specified
119 proportion of the product in terms of the minimum size and mass of bulk/laboratory sample complies
120 with the legal limit. In this case the combined uncertainty (CVR) including sampling uncertainty shall be
121 taken into account, and the measured value should not be directly compared to the MRL as it may allow
122 placing a product on the market with a substantial proportion containing the contaminant above the
123 permitted limit. Let’s assume that the MRL is 0.1 mg/kg, and the average concentration of a pesticide
124 residue is also 0.1 mg/kg in the sampled lot. If the measured residue in the sample taken from the given
125 lot is < 0.1 mg/kg the sampled lot would be declared to be compliant. However the decision would only
126 be correct in about 56% and would be wrong in 44% of the cases as indicated by the cumulative
127 frequency distribution curve in Figure 2. Therefore a lower concentration must be selected as Action
128 Limit (AL) which should not be exceeded by the residue measured in the sample. The AL can be
129 calculated by taking into account the combined relative uncertainty of the measured value (CVR) (equ. 1)
130 and the targeted level of compliance.
6
131 The relationship of AL and MRL can be described with the following equations:
132 AL + k × CV × AL = MRL Equ. 4

133 AL = Equ. 5

134 The value of k depends on the agreed acceptable violation rate (Pv) or the compliant proportion of the
135 lot, Pc, relative to the minimum sample size specified in the corresponding regulation (1-Pv=Pc). For
136 normal distribution, which may approximately be assumed with samples of size ≥25 for fruits and
137 vegetables, the k is equal to the corresponding standard normal variate, Z. A normal distribution may
138 also be assumed for liquid materials and processed products, such as for instance tomato puree. It is
139 pointed out that in other cases the relative frequency distribution of pesticide residues can be best
140 described with a lognormal distribution.1
141 Calculation of AL for given acceptance probability at MRL.
142 If the distribution of xi can be described by the lognormal function, then the transformed variable yi =
143 Ln(xi) is normally distributed. If the distribution of xi has a mean (m) and standard deviation (s), then the
144 distribution of the transformed variable yi has a mean (M) and standard deviation (S). M and S can be
145 calculated from m and s using the equations:
#$
146 M = Ln " ( Equ. 6
%&$ #$ '
-$
147 ) = *+ ,.$ + 10 Equ. 7
148 Since yi is normally distributed, the probability Pi (0.0 to 1.0) for any value yi occurring from a
149 distribution with mean M and standard deviation S can be calculated from normal distribution tables
150 using the standardized equation:
%23 45'
151 1= Equ. 8

152 67 = 8 + 1 × ) Equ. 9
153 In the sampling domain, m takes on the value of the legal limit MRL and xi takes on the value of the AL.
154 Typically xi ≤ m (AL≤MRL), therefore Z will vary from zero to a negative value usually between 0.0
155 and -3.5 based upon the specified acceptance probability (acceptable violation rate), Pv. Once m and s
7
156 are transformed into M and S and the acceptance probability (that will define Z) is specified, yi can be
157 determined from Equation 9. Then xi can be calculated by transforming yi back to xi.
158 97 = exp %67 ' Equ. 10
159 where xi is the AL in the linear scale.
160
161 RESULTS AND DISCUSSION
162
163 Comparison of sampling uncertainties estimated from supervised trials and commercial field
164 samples.
165 Based on the limited data available, the average sampling uncertainty for primary samples, CVSprim of
166 0.78, 0.81, 0.74 and 0.45, were estimated for residues in crop units of a small, medium and large size
167 crops and Brassica leafy vegetables, respectively.7 Additional residue data measured in carrot and
168 parsley enabled the estimation of CVSprim values of 0.59 and 0.60, respectively.8 Duplicate samples taken
169 from 12087 supervised trials carried out in 106 different crops treated with 66 different pesticides
170 provided the best available database for estimation of uncertainty of sampling.31 The supervised trials
171 are carried out usually on small scale with strictly controlled conditions for the application of pesticides.
172 They aim to provide information on the maximum residues which can occur on treated crops under the
173 critical use conditions [shortest interval between last application and harvest (pre-harvest interval),
174 highest permitted dose and application frequency] specified in the registration or use permit
175 documents.32 Consequently, these trials may not reflect the within field residue distribution which occurs
176 where the pesticides are applied according to the general agricultural practice. Table 1 summarizes the
177 estimated sampling uncertainties obtained from the databases consisting of residues in/on primary
178 samples, which were taken from fields treated according to normal farming practice and from supervised
179 trials. The CVS values were calculated for the minimum size of composite samples (12) specified by the
180 Codex Standard for sampling plant products.5 In table 2 the sampling uncertainties calculated from
181 residues in samples taken from commercial fields and from supervised trials are compared. The relative
8
182 sampling uncertainty of residues in field composite samples (CVcs) was calculated from 4 replicate
183 samples and for supervised trials mostly from duplicate samples with range statistics.8 The same
184 tendency was observed in both cases. As the number of datasets from which the CV values were
185 calculated greatly varies and the sample sizes in supervised trials had only sometimes been reported,
186 precise comparison of the variability of residues in supervised trial and field samples could not be done.
187 Therefore the averages of CV ratios (1.15 and 1.19) are considered to be the best indicator of the
188 difference in the variability of residues. Exceptions are papaya, strawberry, cabbage, carrot and parsley
189 for which the ratios of relative sampling uncertainty of composite samples taken from supervised trials
190 and relative sampling uncertainty of residues in field composite samples (CVcs/CVStcs) are within the
191 range of 0.44 and 0.70. The low variability in field trials cannot be explained based on the available
192 information. As it is quite unlikely that supervised trials lead to higher variability of residues than field
193 trials, these values were disregarded.
194 Taking into account the uncertainties of the estimations a rounded correction factor of 1.2 is used to
195 account for the larger variability of residues in field samples.
196 Effect of number of residue values below the LOQ on the estimated sampling uncertainty.
197 The range of residues in primary samples taken from a single field is typically hundred fold. Depending
198 on the average residue and the limit of quantification (LOQ) of the analytical method various
199 proportions of the primary samples may not contain detectable residues. The proportion of residues
200 below LOQ is not known. The effect of the LOQ values in relation to the average of residue data
201 population was studied with experimental supervised trial data1 and synthetic lognormal distributions, as
202 it was found that the within field distribution of residues in crop units (primary samples) can be best
203 described with lognormal distribution.2 Of the supervised trial residue datasets, compiled by the FAO
204 WHO Joint Meeting on Pesticide Residues for estimation of MRLs, those were selected which contained
205 relatively large number of data points. The descriptive statistical parameters of the selected datasets are
206 summarized in Table 3.The selected datasets contained no residue below the LOQ value, except dataset
207 8 where the 27 values below the varying LOQs were replaced with the maximum likelihood estimation
9
208 (MLE) procedure assuming lognormal distribution provided by Villanueva.33 The residue values were
209 also normalized by dividing them with the corresponding average residues of their dataset in order to
210 assist comparison of the spread of residues in the datasets with different mean residues.
211 For studying the effect of LOQ [mg/kg] values on the CV of the data population, the selected data sets
212 were arranged in increasing order. The lowest number of residue values corresponding to the selected
213 percentage of the original data population (10%-90%) were replaced with various LOQ values ranging
214 from 2× reported LOQ to 0.7 average residues in the dataset. These datasets are called adjusted datasets.
215 For instance, in case of dataset 1 the lowest 11 values (0.03, 0.06, 0.07, 0.09, 0.09, 0.1, 0.1, 0.11,
216 0.12,0.12, 0.12) were replaced with 0.04, 0.2, ...1.16) and then the descriptive statistical parameters were
217 recalculated for each adjusted data set. The process was repeated with 20% of the residue values in the
218 original data set, and so on.
219
220 The percentage of residue values below the LOQ and the value of LOQ affect the average and the
221 relative standard deviation of the original residue population (CVo). The relationships of the
222 corresponding CV and average residue of the adjusted datasets are complex, and they are best illustrated
223 graphically. Figure 3 shows the ratios of the CV-s of adjusted dataset (CVAdj) and the original dataset
224 (CVo) as the function of the ratio of LOQ and the average residue (=>' of adjusted dataset (LOQ/=> '.
225 Figure 4 illustrates the CVAdj/CVo relationship as the function of the LOQ/=> residues and the percentage
226 of residues at or below the LOQ obtained from the dataset 8 (Table 3. MLE fitted prochloraz residues).
227 As similar LOQ/=> residues could be obtained from various combinations of LOQ values (Figure 4) and
228 their percentage proportion in the adjusted datasets, the CVAdj values corresponding to LOQ/=> ranges
229 from ≤0.03 to ≤0.7 were grouped and the averages of LOQ/=> and the CVAdj/CVo values being in the
230 given ranges were calculated. The results obtained from the supervised trials and synthetic datasets are
231 shown in Table 4. The results indicate certain variation among datasets which is attributed to the
232 distribution of residues within the individual datasets and sampling uncertainty. As the typical sampling
233 uncertainties were obtained from datasets with detectable residues, they do not represent the situations
10
234 where the various proportions of the treated crops may contain residues below the LOQ. The probability
235 of finding primary samples with non-detectable residues increases as the LOQ/=> increases. However,
236 no general tendency in the relationship between the LOQ/=> and CVAdj/CVo could be established. The
237 grand averages of CVAdj/CVo up to LOQ/=> <0.45 and <0.7 were 1.08 and 1.12, respectively (Table 4).
238 Therefore, it is recommended applying an adjusting factor of 1.1 to account for the potential non-detects
239 in the primary units of the sampled crop.
240 Effect of mixing commodities with different average residues.
241 Mixing commodities containing contamination above the legal limits with lots containing no or low
242 level of contamination is not permitted by law.34 However, it often occurs or might occur in practice.
243 Figure 5 illustrates the residue distribution in mixed and not mixed products. It can be clearly seen that
244 mixing substantially increases the spread of residues.
245 In order to examine the effect of mixing of commodities with substantially different average residues,
246 model experiments were carried out with dataset 9 (Table 3) in its original form (average residue 50.41
247 mg/kg, CV=0.29) and its normalized version having an average of 1 and the same CV as the original
248 dataset. Random samples of 100, 200, 300, 400 and 500 were drawn from the normalized residue data
249 population (representing the low contaminated lot) and 900, 800, 700, 600 and 500 samples were drawn
250 randomly from the original population (representing the contaminated lots). The residue values obtained
251 from the two datasets were mixed and the descriptive statistical parameters of the mixed datasets are
252 summarized in Table 5. The same procedure was repeated with dataset 8 and the normalized dataset 9, in
253 order to model the effect of the different average and CV values of the lots to be mixed. The results are
254 shown in Tables 6 and 7.
255 The mixing reduced the average residue in the mixed lot, but at the same time substantially increased its
256 CV value. The change of the ratios of the average residues of the 'contaminated' and 'non-contaminated'
257 datasets from 50.4 to 278 did not change the tendency of increase of CV values in the mixed dataset.
258 The large difference of the CV values (1.12 for non-contaminated dataset and 0.29 of contaminated
259 dataset, Table 6) did not affect either the dependence on mixing ratio or the increase of CVM/CVo values
11
260 shown in Tables 6 and 7, where CVM is the relative standard deviation of residues in mixed dataset. The
261 percentage of the non-contaminated samples (x %) in the mixed dataset and the ratio of the CVM of the
262 mixed lot and the CVo of the contaminated lot have linear relationship and the linear regression lines
263 practically overlap for the two combinations:
264 Original and normalized dataset 9:
265 CVM/CVo =0.0518x+0.9874; R2= 0.9973 Equ. 11
266 Original dataset 8 and original dataset 9:
267 CVM/CVo =0.05413x+0.9734; R2= 0.9967 Equ. 12
268 The average regression equation is:
269 CVM/CVo =0.05527x+0.9803; R2= 0.995 Equ. 13
270 In practice, the effect of mixing contaminated lot with non-contaminated can only be taken into account
271 in compliance assessment if the producer reveals the mixing ratio. In this case, the appropriate adjusting
272 factor can be calculated with equation 13, and it should be used to multiply the recommended practical
273 sampling uncertainties given in Tables 8 and 9. It should be emphasized that the presented principle is
274 valid and applicable only if the effective mixing of the two lots is possible and it has been done. In
275 practice only liquid products and small grains can be properly mixed, other commodities not.
276 Furthermore it is reemphasized that mixing of contaminated lot with non-contaminated ones are illegal
277 in many countries.
278 Recommended sampling uncertainties for control of pesticide residues in food.
279 Due to the large variability of residues in raw agricultural commodities (e.g. fresh fruits and vegetables)
280 the sampling uncertainty cannot be determined reliably from a few measurements. Therefore, in order to
281 assure that the calculated AL values are correct at least in 95% of the cases, it is recommended that the
282 upper 95% confidence limits of the estimated average sampling uncertainties calculated from residue
283 data obtained from 12087 independent supervised residue trials31 is used (Tables 8 and 9). Taking into
284 account the larger variability expected under field conditions than in supervised trials and the potential
285 effect of non-detectable residues on the relative standard deviation of residues, a combined multiplying
12
286 factor of 1.3 is incorporated in the CVSprim values recommended for use under practical conditions. The
287 CVSprim values are given to enable conveniently determining the expectable sampling uncertainty for
288 various sample sizes (n) with equation 14, when sampling plans are designed for pre-market self control.
@ABC
289 CV? = Equ. 14
√?
290 Farkas and co-workers demonstrated that the relative range of the 95% confidence limits around the
291 sampling uncertainty, estimated from duplicate samples taken from different lots, rapidly decreases up to
292 8 lots and the range gradually decreases with increasing number of lots. The gain above 20 lots is
293 marginal.8 This finding is in line with the ISO Standard,35 which recommends minimum 20 lots to be
294 tested for determination of sampling uncertainty of bulk materials with as many replicate samples as
295 feasible. Therefore, where the number of trials provided the basis for the calculation of sampling
296 uncertainty is less than 20 but equal to or larger than 8, the estimated CVS values should be used with
297 caution. In case of doubt, it is recommended to use the typical sampling uncertainties estimated for the
298 corresponding crop group. Likewise, CVS values estimated for crop groups (Table 9) should be used
299 where the database consists of <8 trials to concord with the conclusions of Lyn and co-workers.36
300 Optimization of sampling and testing plan.
301 Consideration of the uncertainty of the laboratory phase (CVL) for determination of pesticide residues.
302 Though the combined relative uncertainty of sub-sampling and sample homogenization can be at the
303 same magnitude as that of the uncertainty of sampling11,15 laboratories generally pay very little attention
304 to control the variability of these two steps, and the repeatability and reproducibility of the
305 measurements are usually reported based on the recoveries obtained with spiked samples.37-39 The
306 determination of CVL is especially important when only 1-2 g portions are analyzed from the laboratory
307 sample which may amount to 1 to >15 kg for small and medium size and large fruits (e.g. cabbage,
308 watermelon), respectively. As the variability resulted from the test portion and particle size reduction
309 depends on the laboratory equipment used and the sample materials analyzed, no typical values can be
310 estimated. Therefore the CVL should be determined by each laboratory regularly as part of the routine
25
311 operation by reanalysis of retained test portions taken from samples containing incurred residues at
13
312 well detectable concentration. Alternative option is the surface treatment method suggested by
313 Maestroni and co-workers11,12 and applied successfully in other studies.13,14 It is especially important
314 when the test portion size is reduced below 15 g. It should be noted that simple recovery studies with
315 spiked test portions before extraction do not provide any information on the efficiency of sample
316 processing and its consequences on the CVL. Furthermore, neither proficiency test nor collaborative
317 study results provide information on CVL.
318 Minimizing combined uncertainty of the measured residues.
319 Applying equations 1 or 2, the combined uncertainty of the measured result can be simply calculated
320 with an Excel spreadsheet taking into account the best estimates for the sampling uncertainty (Tables 8,
321 9) and the reproducibility CV values of the individual steps of the determination of the residues, which
322 had been obtained from the method validation. The analysts can test the effect of various combinations
323 in advance and select those which fit the purpose of the analyses. As a first step, the largest contributors
324 to the total variance of the results should be reduced if possible. On the other hand, practically no
325 improvement can be achieved by improving the procedure which contributes to the total variance with
326 less than 30% of the largest contributor. Table 10 provides some examples to illustrate the application of
327 the principle.
328 If the task of the laboratory is to test the compliance of the marketed products with the MRL, then only
329 the value of the CVL should be taken into consideration. It should be noted that the default CVL of 25%
330 specified by the European Commission30 (EC) can be comfortably achieved with a CVA of < 20%
331 determined with recovery studies, provided that the CVSp of ≤10% can be maintained. It is pointed out
332 that if a one gram test portion is extracted instead of 15 g applying the same sample processing
333 (homogenization) procedure which is increasingly getting into practice, the CVL can be as large as 150%
334 and the EC requirement cannot be met.29 It should also be noted, that the relevant minimum mass and
335 sample size requirements must be met for market control. With smaller samples justified legal actions
336 cannot be made.
14
337 For pre-marketing control, the sampling uncertainty should always be taken into account. The sampling
338 uncertainty can be reduced with increasing the sample size, provided that the sample processing
339 equipment is suitable for efficiently handling and comminuting large quantity of materials. Alternately, a
340 sample size reduction step to obtain a sample mass which then can be further processed shall be included
341 in the determination process, and its uncertainty shall be determined and taken into account for the
342 calculation of CVL. The latter option is not shown in Table 10.
343 Calculation of action limits and operation characteristic curves.
344 Using the combined uncertainty calculated from the recommended practical sampling uncertainties,
345 CVSprim listed in Tables 8 and 9 and the actual CVL of the laboratory phase of the residue analysis, the
346 AL and the operation characteristic (OC) curves can be calculated based on the underlying theory
347 described with equations 8-10. To facilitate pre-marketing control of pesticide residues an MS Excel
348 template was developed following the basic principles elaborated by Whitaker and co-workers.42,43 The
349 template calculates the AL which should not be exceeded by the measured residues in order to achieve
350 the desired compliance level. If replicate samples are taken, the residues measured in each sample
351 should be below the calculated AL.
352 Figure 6 illustrates the calculated OC curves assuming CVSprim=0.985 , CVL=0.16 (15 g test portion) and
353 CVL 1.5 (1 g test portion; 1 and 3 replicate extraction) and 2% acceptable violation rate. The
354 corresponding AL concentrations of <0.065 mg/kg, <0.011 mg/kg and <0.025 mg/kg clearly show the
355 disadvantage of indiscriminate reduction of test portion size (the residue concentration measured in 1 g
356 test portion should not be larger than 0.011 and 0.025 mg/kg when the MRL is 0.2 mg/kg) for the sake
357 of reduction of the amount of chemicals required. Figure 7 illustrates the relationship of relative and
358 cumulative frequencies of residues in samples taken from a lot and the calculated OC curve. It can be
359 seen that in the given case 98% of the samples contains residues below the MRL and the residues in 2%
360 of the samples would exceed it.
361 It is emphasized that, if a crop has been treated with a pesticide, 100% compliance cannot be certified.
362 At present, there is no information available on an officially acceptable violation rates from any country.
15
363 In order to enable designing a sampling plan, which specifies the number and size of samples to be taken
364 and the corresponding AL, acceptable violation rate should be declared by the responsible national
365 agencies. The BASELINE project carried out within the EU 7th research framework program
366 recommended 98% compliance with the MRLs with 95% probability,9 as performance objective for
367 chemical contaminants, if they do not impose any health risk to consumers. This criterion might be
368 considered as a starting point to establish official acceptable violation rates.
369
370 AUTHOR INFORMATION
371 Corresponding Author:
372 E-mail: farkas.zsuzsa13@gmail.com
373 Notes
374 The authors declare no competing financial interest.
375
376 ABBREVIATIONS USED
377 CV; Coefficient of variation (relative standard deviation);
378 CVA: Relative uncertainty of the analysis phase of the determination of residues including, for instance,
379 extraction, cleanup... and quantitative determination;
380 CVAdj: Relative standard deviation of residues in datasets in which the original residue data were
381 replaced with various LOQ values;
382 CVL; Relative uncertainty of the laboratory phase of the determination of residues including sub-
383 sampling, sample processing and the analysis phase of the determination
384 CVR; Combined relative uncertainty of the measured residues (in general);
385 CVo; Relative standard deviation of residues in the original dataset;
386 CVS; Relative uncertainty of sampling in general;
387 CVSS: Relative uncertainty of the sub-sampling of a large crop or a large bulk (aggregate) sample;
16
388 CVSp: Relative uncertainty of sample processing including chopping, grinding, mincing, mixing the
389 laboratory sample or part of it;
390 CVSprim: Relative sampling uncertainty of residues in primary (incremental samples);
391 CVcs: Relative sampling uncertainty in case of composite samples;
392 CVS1corr; Relative uncertainty of sampling in case of composite samples meeting the minimum sample
393 size requirements of the Codex sampling procedure;5
394 CVStcs: Relative sampling uncertainty of composite samples taken from supervised trials;
395 CVM: Relative standard deviation of residues in mixed dataset;
396 LCL; Lower confidence limit (2.5th percentile);
397 UCL; Upper confidence limit (97.5th percentile);
398 LOQ: limit of quantification of an analyte in a sample matrix;
399 MLE: maximum likely hood estimation procedure;
400 MRL, (ML): maximum concentration of pesticide residues (contaminants) which is legally permitted
401 in/on food and feed;
402 n: Sample size (number of primary samples in a composite sample);
403 => : Average of residues in a dataset;
404 MTp: Mass of portion of sampled material removed for extraction;
405 DL: decision limit, the concentration of pesticide residue in a sample above which the sampled lot can
406 be considered non-compliant;
407 AL: Action Limit, also called accept/reject limit: the maximum residue concentration in a random
408 sample satisfying the minimum requirements of laboratory sample, which can be present if the sampled
409 lot is expected to comply with the MRL at specified probability level;
410 Pv: violation rate; the percent of the sampled lot which does not comply with the MRL, it is also called
411 acceptance probability;
412 Pc: proportion of the sampled crop complying with MRL.
413
17
414 ACKNOWLEDGMENT
415 The authors greatly appreciate the invitation of S. Lehotay, M. Krolskí, J. Sandahl, J. Jenkins and C.
416 Harris, organizers of the Poster Session on Residues in Food and Feed Monitoring Results and Dietary
417 Risk Assessment of the 13th IUPAC International Congress of Pesticide Chemistry, to publish the
418 detailed results of our work in this special issue.
419
420
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557 42. Whitaker, T. B.; Slate, A. B.; Giesbrecht, F. G. Designing Sampling Plans to Detect Foreign
558 Material in Bulk Lots of Shelled Peanuts. Peanut Science 2008, 35, 159-164.
23
559 43. Food and Agriculture Organization of the United Nations. Mycotoxin Sampling Tool, User
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561 30, 2014)
562
563
24
564 FIGURE CAPTIONS
565
566 Figure 1. Distribution of residues in composite samples taken as part of the premarketing self-control
567 (PMc), and distribution of analytical measurement results assuming a CVL of 0.25 and measured
568 residue of 0.4 mg/kg (decision limit DL).
569 Note: The cumulative frequency of analytical measurements (Cum. Freq. Mc) indicates the
570 probability of rejection of the lot based on the random error of the analysis.
571 Figure 2. Distribution of residues in apple composite samples with a true mean of 0.1 mg/kg and
572 CVR of 0.35. MRL = 0.1 mg/kg
573 Figure 3. Relationship of the ratio of the CV values of adjusted (CVAdj) and original (CVO) datasets
574 and the LOQ/average residue in the adjusted datasets of 1-8 and 10.
575 Figure 4. Relationship of CVA/CVo as a function of LOQ/Rave(A) and percent of residues at LOQ in
576 case of dataset 8 (prochloraz residue in cereal grains).
577 Figure 5. Relative and cumulative frequency distributions of phosalone residues in plum fruit
578 samples taken from mixed (upper chart) and not mixed (lower chart) lots.
579 Figure 6. Operation characteristic curves indicating 2% violation rate at the MRL if the residue
580 concentration in the sampled lot was ≤ ALs of 0.065, 0.011 and 0.025. The CVL values and number
581 of test portions in brackets for curves 1, 2 and 3 were 0.16 (1), 1.5 (1) and 1.5 (3), respectively.
582 Figure 7. The relationship of relative and cumulative frequencies of residues in samples taken from a
583 lot and the calculated Operation characteristic curve assuming 98% compliance of the sampled lot.
25
TABLES
Table 1. Comparison of estimated sampling uncertainties from resides measured in primary
samples taken from commercial fields and composite samples derived from supervised trials
Primary field samples Supervised field trials

Crop
Noa CVSprim CVcs Noa CVStcs CVcs/CVStcs
Apple 19 0.82 0.26 636 0.22 1.17
Pear 4 0.88 0.28 316 0.22 1.25
Orange 15 0.90 0.28 328 0.23 1.21
Peach 7 1.08 0.34 409 0.34 1.01
Plum 8 0.91 0.29 344 0.35 0.82
Potato 8 1.06 0.33 104 0.19 1.78
Tomato 4 1.45 0.46 624 0.36 1.29
Medium size crops 1.01 0.32 0.27 1.19
Mango 14 0.73 0.33 494 0.30 1.10
Papaya 4 0.60 0.27 13 0.46 0.58
Squash 4 0.87 0.39 77 0.42 0.92
Grape 14 0.75 0.33 494 0.30 1.13
Cucumber 14 0.80 0.36 111 0.33 1.07
Banana 2 1.03 0.46 47 0.28 1.66
Large crops 0.76 0.34 0.30 1.13
Strawberry 9 0.66 0.21 215 0.33 0.62
Back current 2 0.80 0.25 171 0.18 1.40
Cherry 7 0.94 0.30 189 0.23 1.28
Small fruits 0.80 0.25 0.33 0.76
Lettuce 10 0.75 0.34 769 0.31 1.07
Cabbage 7 0.41 0.18 398 0.35 0.52
26
Carrot 10 0.60 0.19 39 0.43 0.44
Parsley 9 0.65 0.20 2 0.29 0.70
Average 1.15
a
: number of residue datasets used for the calculation of sampling uncertainty;
Note: The sample size of the samples has been recalculated to match the criteria of the minimum requirements of
Codex Sampling procedure.5
27
Table 2. Comparison of estimated sampling uncertainties from resides measured in composite
samples taken from commercial fields and supervised trials
Samples taken from commercial Samples taken from supervised

CVcs/CVStcs
fields trials
Crop Noa CVcs LCL UCL Nob CVStcsc LCL UCL
Apple 137 0.34 0.17 0.77 636 0.22 0.20 0.24 1.52
Gooseberry 104 0.32 0.11 0.73 171 0.18 1.78
Lettuce 42 0.28 0.00 1.21 769 0.31 0.29 0.34 0.90
Carrot 29 0.44 0.20 0.67 36 0.43 0.35 0.55 1.03
Parsley 24 0.21 0.02 0.40 2 0.29 0.15 1.83 0.70
Average 1.19
a
: number of crop - pesticide combinations; (in each case four replicate samples were taken from one field)
b
: number of replicate residue datasets
c
: The relative standard deviations of residues in composite samples were adjusted to the minimum sample size
requirements of the Codex sampling procedure31
28
Table 3. Descriptive statistical parameters of data sets used for modeling the effect of LOQ values
Datasets with original residue values
1 2 3 4 5 6 7 8 9 10 11
Count 106 78 85 36 36 64 111 121 1000 1000 1000
LOQ 0.02 0.02 0.01 0.02 0.01 0.05 0.05 0.01 0.01 0.01 0.01
<LOQ % 0 0 0 0 0 0 0 0 0 0 0
Min 0.03 0.03 0.01 0.02 0.27 0.70 0.09 0.01 20.46 0.10 0.03
Ave 1.66 0.26 0.66 0.10 2.73 15.52 7.56 0.18 50.41 1.01 1.03
P0.95 5.0 0.7 1.4 0.3 7.5 47.0 20.0 0.5 77.7 2.4 3.3
Max 6.90 0.85 4.80 0.45 17.00 86.00 30.00 1.20 112.52 4.86 18.42
CV 0.94 0.75 0.97 1.06 1.25 1.06 0.75 1.12 0.29 0.69 1.39
LOQ/=> 0.012 0.076 0.015 0.207 0.004 0.003 0.007 0.055 0.001 0.010 0.010
Datasets with normalized residue values
1 2 3 4 5 6 7 8 9 10 11
Min 0.02 0.11 0.01 0.21 0.10 0.05 0.15 0.04 0.41 0.09 0.03
Ave 1.00 1.00 1.00 1.00 1.00 1.00 1.04 1.00 1.00 1.00 1.00
P0.95 3.0 2.8 2.1 3.0 2.7 3.0 2.6 2.9 1.5 2.3 3.2
Max 4.15 3.24 7.29 4.66 6.22 5.54 3.97 6.63 2.23 4.83 17.86
CV 0.94 0.75 0.97 1.06 1.25 1.06 0.71 1.12 0.29 0.69 1.39
LOQ/=> 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.000 0.010 0.010
Datasets:
1. Wheat, barley straw pyraclostrobin; 2. Tomato endosulfan; 3. Cucurbits propamocarb; 4. Potato aldicarb; 5.
Soya beans glyphosate; 6. Propamocarb lettuce; 7. Cereal grains prochloraz; 8. MLE fitted prochloraz in wheat
and barley straw; 9. Synthetic lognormal distribution with mean of 50.41 and CV of 0.29; 10. Synthetic lognormal
distribution with mean of 1.01 and CV of 0.69; 11. Synthetic lognormal distribution with mean of 1.03 and CV of
1.39.
29
The selected datasets contained no residue below the LOQ value, except dataset 8 where the 27 values below the
varying LOQs of 0.1 mg/kg (6), 0.05 mg/kg (19) and 0.02 mg/kg (2) were replaced with the maximum likelihood
procedure assuming lognormal distribution provided by Villanueva.34
30
F GHI GJKLGMK CVAdj/CVo values

Table 4. Relationship of average LOQ/E
LOQ/=> Average of CVAdj/CVo values in datasets
Range Average 1-8 &10 11
<0.03 0.02 1.17
0.031-0,06 0.04 1.24
0.060- 0.083 0.07 1.12
<0.1 0.10 1.02
0.11-0.2 0.16 1.07 1.24
0.21-0.3 0.26 1.07
0.21-0.3 0.27 1.10
0.31-0.4 0.36 1.07
0.31-0.4 0.39 0.99
0.41-0.5 0.43 1.30
0.41-0.5 0.46 0.92
0.51-0.6 0.53 0.83
0.51-0.6 0.55 0.87
0.61-0.7 0.64 0.93
Grand average 1.08 1.12
Table 5. Descriptive statistical parameters of the mixed dataset obtained from the commingled
datasets 9 (D9) and its normalized version (ND9).
Count Min Ave P0.95 Max CV LOQ/=> CVM/CVo
D9 1000 20.5 50.4 77.7 112.5 0.29 0.000
ND9 1000 0.41 1 1.54 2.2 0.29 0.01
ND9_1:D9_9 1000 0.49 45.8 77.0 111.4 0.447 0.0002 1.54
ND9_2:D9_8 1000 0.48 40.9 77.2 112.5 0.584 0.0002 2.01
ND9_3:D9_7 1000 0.45 35.8 73.8 111.4 0.727 0.0003 2.51
31
ND9_4:D9_6 1000 0.47 30.6 72.8 111.4 0.874 0.0003 3.01
ND9_5:D9_5 1000 0.41 25.5 71.5 112.5 1.053 0.0004 3.63
Notes:
D9: identification of dataset (see Table 3); ND9: normalized form of D9 dataset
ND9_1:D9_9 indicates a mixture containing 10% of ND9 and 90% of D9
dataset D8 and the original version of dataset D9
Count Min Ave P0.95 Max CV LOQ/Ave CVM/CVo
D9 1000 20.5 50.4 77.7 112.5 0.29 0.0
D8 121 0.0 0.18 0.5 1.2 1.12 5.6
D8_1:D9_9 1000 0.0 45.7 77.0 111.4 0.45 0.0 1.56
D8_2:D9_8 1000 0.0 40.8 77.2 112.5 0.59 0.1 2.04
D8_3:D9_7 1000 0.0 35.6 73.8 111.4 0.74 0.1 2.56
D8_4:D9_6 1000 0.0 30.3 72.8 111.4 0.90 0.2 3.09
D8_5:D9_5 1000 0.0 25.0 71.5 112.5 1.08 0.2 3.74
Note: D8_1:D9_9 indicates a mixture containing 10% of D8 and 90% of D9
32
datasets D8 and its normalized version (ND8)
Count Min Ave P0.95 Max CV LOQ/Ave CVM/CVo
D8 121 0.01 0.18 0.53 1.2 1.12 0.055
ND8 1000 0.04 1.00 2.93 6.63 1.12 0.010
D8_1:ND8_9 1000 0.01 0.94 3.26 6.63 1.20 0.011 1.08
D8_2:ND8_8 1000 0.01 0.85 2.93 6.63 1.31 0.012 1.19
D8_3:ND8_7 1000 0.01 0.77 2.76 6.63 1.30 0.013 1.18
D8_4:ND8_6 1000 0.01 0.65 2.6 6.63 1.41 0.015 1.28
D8_5:ND8_5 1000 0.01 0.65 2.76 6.63 1.54 0.015 1.40
Note: D8_1:ND8_9 indicates a mixture containing 10% of D8 and 90% of ND8
33
Table 8. Summary of sampling uncertainties of pesticide residues in individual crop units or
primary sample increments recommended for practical use
Crop name ka CVSprim Crop name ka CVSprim
Foods
Almond 60 0.85 Melon 74 1.74
Apple 636 0.98 Mint leaves 18 1.15
Artichoke 8 0.49 Mustard green 345 0.72
Avocado 10 3.51 Nectarine 9 1.18
Banana 47 1.46 Onion (bulb) 44 2.32
Barley (grain) 63 1.02 Onion (green) 29 1.96
Basil (dry) 9 1.07 Orange 328 1.06
Basil (fresh) 12 0.47 Paddy rice (husked grain) 62 0.93
Bean 72 1.38 Papaya 13 2.96
Bean (Dry) 137 2.30 Pea (dry) 68 1.43
Beetroot (root) 8 1.83 Pea (edible podded) 18 2.43
Blackberry 15 1.06 Pea (succulent seeds) 36 1.70
Blueberry 74 0.95 Peach 409 1.52
Broccoli 205 1.24 Peanut 52 0.91
Brussels sprout 22 1.95 Pear 316 1.02
Cabbage, head 398 1.59 Pecan 39 1.17
Cantaloupe 64 1.96 Pepper 503 1.62
Carrot 39 2.30 Pineapple 21 2.03
Cauliflower 73 1.53 Plum 344 1.58
Celery 267 0.93 Potato 104 0.92
Cherry 189 1.09 Radish 61 1.28
Coffee (dry bean) 18 3.44 Radish (root) 28 1.58
Cotton (undelinted seed) 21 3.28 Rape greens 39 1.30
Cotton seed 130 1.66 Raspberry 22 1.18
Cranberry 55 0.75 Rice (grain) 64 0.69
Cucumber 111 1.61 Sorghum (grain) 27 0.95
Egg plants 11 1.02 Soya bean 13 1.72
34
Endive 16 0.55 Soybean (dry) 128 1.75
Grape 494 1.32 Spinach 304 1.24
Grapefruit 99 1.53 Squash 77 2.11
Hop (cones, dried) 87 1.15 Strawberry 215 1.55
Hops (Fresh) 17 1.76 Sugar beet root 51 2.10
Japanese apricot 8 0.90 Sugar cane 15 4.27
Leaf Lettuce 769 1.38 Sunflower seed 21 2.04
Lemon 195 1.19 Tea (Fresh) 52 3.76
Lima beans 15 2.38 Tomato 624 1.58
Lima beans (dry) 13 2.35 Turnip 9 1.41
Mandarin 36 2.10 Wheat (grain) 86 1.45
Mango 32 1.11
Animal feeds
Alfalfa (forage) 48 2.54 Pea hay 28 1.28
Alfalfa (hay) 64 1.84 Peanut fodder 13 1.67
Almond (hull) 238 1.14 Peanut hulls 89 0.72
Barley fodder (hay and straw) 93 1.42 Rice (shoot panicle) 16 0.62
Barley straw 85 1.14 Rice (straw) 66 1.86
Bean (Green) Plant 40 2.25 Rye straw 12 2.28
Bean forage 31 0.69 Sorghum (fodder) 72 1.67
Bean hay 14 2.32 Soybean (forage) 184 0.97
Beetroot (top) 8 0.82 Soybean (hay) 199 1.10
Cotton gin trash 58 1.22 Sugar beet (top) 261 1.33
Maize straw 110 1.74 Wheat (forage) 191 1.08
Oat foliage 9 1.07 Wheat (straw) 353 1.08

a
: number of residue datasets used for the estimation of sampling uncertainty
Table 9. Summary of estimated sampling uncertainties for crop groups recommended for
practical use
35
Crop groups Noa UCL of CVStcs Practical CVSprim
Small fruitsb 768 0.35 1.43
Medium-sized fruitsb 2139 0.28 1.14
Large-sized fruitsb 560 0.32 1.32
Medium-sized vegetablesb 1211 0.38 1.57
Bush berries 171 0.20 0.83
Legume vegetablesc 211 0.38 1.55
Brassica vegetablesc 698 0.35 1.43
Fruiting vegetables, cucurbitsc 337 0.40 1.66
Leafy vegetablesc 1872 0.31 1.27
Root and tuber vegetablesc 256 0.34 1.39
Stalk and stem vegetablesc 276 0.22 0.92
Pulsesc 346 0.45 1.83
Cereal grainsc 340 0.23 0.96
Grasses, for sugar or syrup productionc 15 1.06 4.37
Tree nutsc 101 0.23 0.95
Oilseedsc 247 0.37 1.52
Seeds for beverages and sweets 22 0.77 3.18
Legume forage and fodder 288 0.31 1.29
Straw, hay (of legume feeds) 523 0.31 1.26
Cereal forage, fodder and straw 1176 0.32 1.30
Grass forage 19 0.32 1.32
Grass hay 18 0.23 0.93
Dried herbs 99 0.27 1.11
By-products for animal feedc 391 0.25 1.02

a
: number of replicate sample sets used for estimation of CVStcs
b
: crops are grouped according to the size of crop units specified by the Codex Sampling procedure6
36
Table 10. Examples for the contribution of the individual steps of the analytical procedure to the
combined uncertainty (CVR) of the results
Percentage contribution to total
CVSprim n Noa CVSp MTp Nob CVA CVL CVR variance of results
Vs VSp VA VL
1.43 10 1 0.1 15 1 0.12 0.16 0.477 89% 4% 6% 11%
1.43 10 1 1.5 1 1 0.12 1.50 1.571 8% 91% 1% 92%
1.43 10 2 1.5 1 1 0.12 1.06 0.877 15% 84% 1% 85%
1.43 10 3 1.5 1 1 0.12 0.87 0.675 21% 77% 1% 79%
1.43 10 3 1.5 1 2 0.12 0.87 0.674 21% 78% 1% 79%
1.43 25 3 1.5 1 1 0.06 0.87 0.576 10% 90% 0% 90%
1.43 25 3 0.3 5 1 0.06 0.18 0.304 71% 26% 3% 29%
0.81 10 1 0.1 15 1 0.12 0.16 0.300 73% 11% 16% 27%
0.81 10 1 0.3 5 1 0.12 0.32 0.412 39% 53% 8% 61%
0.81 10 2 0.3 5 1 0.12 0.23 0.303 52% 36% 12% 48%
0.81 10 2 0.3 5 2 0.12 0.22 0.300 56% 38% 6% 44%

a
: number of test portions withdrawn from the homogenized laboratory sample for extraction
b
: number of replicate analyses performed from the extract of the test portion
37
FIGURE GRAPHICS
10 1
Relative frequency of residues
9 DL 0.9
Probability of rejection
8 0.8
7 MRL 0.7
6 0.6
5 0.5
4 0.4
3 0.3
2 0.2
1 0.1
0 0
0 0.1 0.2 0.3 0.4 0.5 0.6
Residues [mg/kg] in composite samples
Rel frequ. PMc Rel frequ Mc Cum. Freq. Mc
Figure 1.
0.8 100%
90%
Cumulative frequency of residues
Relative frequency of residues
0.7
80%
0.6
70%
0.5 60%
rel frequ.
0.4 50%
Cum frequ
0.3 40%
30%
0.2
20%
0.1 10%
0 0%
0 0.1 0.2 0.3 0.4
Residues [mg/kg]
Figure 2.
38
3.0
Datasets of 1-8 and 10
2.5
2.0
CVAdj /Cvo
1.5
1.0
0.5
0.0
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
LOQ/Average residue
Figure 3.
2
CVAdj/CV0
0
0.06 0.06 0.13 0.12 0.17 0.27
0.31
0.51
0.49
LOQ/Rave(Adj)
Figure 4.
39
16 120%
14
Relative frequency [%] 100%
Cumulative frequency
12
80%
10
8 60%
6
40%
4
20%
2
0 0%
0 0.5 1 1.5 2 2.5 3 3.5 4
Normalised residues
16 120%
14
100%
Relative frequency [%]
Cumulative frequency
12
80%
10
8 60%
6
40%
4
20%
2
0 0%
0 0.5 1 1.5 2 2.5 3 3.5 4
Normalised reidues
Figure 5.
40
100
90
80
Probability of Accepting Lot (%)
1
70 MRL
60
50 2
40
30
20
3
10
0
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4
Lot Pesticide Concentration (mg/kg)
Figure 6.
100 100%
90 90%
80 80%
Relative frequency of residues [%]
70 70%
60 MRL OC % 60%
OC curve [%]
AL
50 Rel freq.% 50%
40 Cumulative 40%
30 30%
20 20%
10 10%
0 0%
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4
Lot pesticide concentration [mg/kg]
Figure 7.
41
42
GRAPHIC FOR TABLE OF CONTENTS
100 100%
90 90%
80 80%
Relative frequency of residues [%]

70 70%
60 Action MRL
OC %
60%
OC curve [%]
50 Limit Rel freq.% 50%
40 Cumulative 40%
30 30%
20 20%
10 10%
0 0%
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4
Lot pesticide concentration [mg/kg]
43

Use of Combined Uncertainty of Pesticide Residue Results For Testing Compliance With Mrls

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Journal of Agricultural and Food Chemistry is published by the American Chemical

1 Use of Combined Uncertainty of Pesticide Residue Results for Testing Compliance

6 1. National Food Chain Safety Office, Budapest Tábornok u 2, 1143 Hungary

8 3. Wessling Hungary Ltd, Budapest Fóti út 56, 1047 Hungary

9 1a Retired Scientific Adviser, ,

23 The principles described are also applicable to other chemical contaminants.

27 residues, self control of production, certification of compliance

32 Distribution of pesticide residues.

48 Uncertainty of measurement results.

60 uncertainty with a sub-sampling (CVSS) of 8-12%.

61 Maximum residue limits for chemical contaminants.

69 for pesticide residues.5

72 principles described are also applicable for other chemical contaminants.

76 Estimation of combined uncertainty of the measured residues.

79 =  + 

85 two phases of the process can be distinguished as:

86 CV = CV + CV Equ. 2

94 Principles of control of compliance of food commodities with the legal limits.

98 Control of commodities on the market (enforcement or monitoring program).

116 dispersion of analytical measurement results can be considered normal.30

117 Premarketing self-control.

130 and the targeted level of compliance.

132 AL + k × CV × AL = MRL Equ. 4

140 described with a lognormal distribution.1

141 Calculation of AL for given acceptance probability at MRL.

145 calculated from m and s using the equations:

150 using the standardized equation:

158 97 = exp %67 ' Equ. 10

159 where xi is the AL in the linear scale.

161 RESULTS AND DISCUSSION

193 trials, these values were disregarded.

195 account for the larger variability of residues in field samples.

218 original data set, and so on.

239 in the primary units of the sampled crop.

240 Effect of mixing commodities with different average residues.

244 mixing substantially increases the spread of residues.

254 shown in Tables 6 and 7.

263 practically overlap for the two combinations:

264 Original and normalized dataset 9:

265 CVM/CVo =0.0518x+0.9874; R2= 0.9973 Equ. 11

266 Original dataset 8 and original dataset 9:

267 CVM/CVo =0.05413x+0.9734; R2= 0.9967 Equ. 12

268 The average regression equation is:

269 CVM/CVo =0.05527x+0.9803; R2= 0.995 Equ. 13

277 in many countries.

278 Recommended sampling uncertainties for control of pesticide residues in food.

300 Optimization of sampling and testing plan.

317 study results provide information on CVL.

318 Minimizing combined uncertainty of the measured residues.

327 the principle.

336 cannot be made.

343 Calculation of action limits and operation characteristic curves.

351 should be below the calculated AL.

360 of the samples would exceed it.

368 considered as a starting point to establish official acceptable violation rates.

370 AUTHOR INFORMATION

371 Corresponding Author:

372 E-mail: farkas.zsuzsa13@gmail.com

374 The authors declare no competing financial interest.

376 ABBREVIATIONS USED

377 CV; Coefficient of variation (relative standard deviation);

379 extraction, cleanup... and quantitative determination;

79 = +

86 CV = CV + CV Equ. 2

132 AL + k × CV × AL = MRL Equ. 4