108139-1 Study Title Data Requirements Author Study ... - IR-4 Project

Bayer Corporation 108139-1
Agriculture Division
Study Title
A Modified Analytical Method for the Determination of Cyfluthrin Residues in Various Plant
Matrices by Chemical Ionization – GC/MS
Data Requirements
EPA Ref.: OPPTS 860.1340, Residue Analytical Method – Plants
Author
S. M. Moore, C. V. Lam and Chris Nuessle
Study Completion Date
March 27, 2002
Performing Laboratory
Bayer Corporation
Agricultural Division
Environmental Research Section
Bayer Research Park
17745 South Metcalf Avenue
Stilwell, KS 66085
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Bayer Corporation 108139-1
Agriculture Division
Table Of Contents
6
Page No.
Study title ................................................................................................................................... 1
Data requirements...................................................................................................................... 1
Author ........................................................................................................................................ 1
Study completion date................................................................................................................ 1
Performing laboratory................................................................................................................. 1
Statement of data confidentiality ................................................................................................ 2
Certification of good laboratory practice ..................................................................................... 3
Certification of availability of raw data ........................................................................................ 4
Certification of authenticity ......................................................................................................... 5
Inquires ...................................................................................................................................... 5
Table of contents........................................................................................................................ 6
1.0 Summary........................................................................................................... 22
2.0 Introduction ....................................................................................................... 25
3.0 Experimental ..................................................................................................... 25
3.1 Location............................................................................................................ 25
3.2 Materials ........................................................................................................... 25
3.2.1 Apparatus.......................................................................................................... 25
3.2.2 Reagents/supplies............................................................................................. 27
3.3 Reference standards......................................................................................... 28
3.3.1 Cyfluthrin standard solutions ............................................................................. 28
3.3.1.1 Primary standard............................................................................................... 28
3.3.2 Secondary standards in toluene ........................................................................ 28
3.3.3 [ 2 H6]Cyfluthrin standard solution........................................................................ 29
3.3.3.1 Primary deuterated standard............................................................................. 29
3.3.3.2 Deuterated secondary standards in toluene ...................................................... 29
3.3.4 Linearity standard solutions............................................................................... 30
3.3.4.1 Linearity standards............................................................................................ 30
3.3.5 Quantification standard ..................................................................................... 31

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3.4 Analytical method.............................................................................................. 31
3.4.1 Matrix preparation ............................................................................................. 31
3.4.2 Procedure (apples and pears only) ................................................................... 31
3.4.2.1 Extraction.......................................................................................................... 31
3.4.2.2 Florisil clean-up................................................................................................. 32
3.4.2.3 Final extract preparation for analysis................................................................. 32
3.4.3 Procedure (all matrices except nutmeat, apple, and pears)............................... 33
3.4.3.1 Extraction.......................................................................................................... 33
3.4.3.2 Extraction (nutmeat).......................................................................................... 34
3.4.3.3 ACN / hexane partition ...................................................................................... 35
3.4.3.4 Florisil clean-up................................................................................................. 35
3.4.3.5 Final extract preparation for analysis................................................................. 36
3.4.4 Analysis by gas chromatography / mass spectroscopy (GC/MS)....................... 36
3.4.5 Disposal of solutions ......................................................................................... 39
3.5 Method Validation ............................................................................................. 40
3.5.1 Requirements.................................................................................................... 40
3.5.2 Recovery procedure.......................................................................................... 40
4.0 Results and discussion...................................................................................... 40
4.1 Sample extraction (Except nutmeat, apple, and pear), liquid / liquid
partition, and florisil clean-up............................................................................. 41
4.1.1 Sample extraction (apple and pear only), and florisil clean-up........................... 41
4.1.2 Sample extraction (nutmeat only), ACN / hexane partition,
and florisil clean-up ........................................................................................... 41
4.2 Addition of internal standard solution................................................................. 42
4.3 GC/MS of cyfluthrin ........................................................................................... 42
4.4 Recovery of cyfluthrin........................................................................................ 42
4.5 Limits of quantitation and detection (except apple and pears) ........................... 42
4.6 Linearity of detector response ........................................................................... 43
4.7 Time interval of the analytical method ............................................................... 43
4.8 Conclusions ...................................................................................................... 43
5.0 Bibliography ...................................................................................................... 44
Tables
1. Recovery of cyfluthrin from cherry fresh fruit measured as part
of the field residue program............................................................................... 46
2. Recovery of cyfluthrin from peach fresh fruit measured as part
of the field residue program............................................................................... 47
3. Recovery of cyfluthrin from plum fresh fruit measured as part
of the field residue program............................................................................... 48
7

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Agriculture Division
4. Recovery of cyfluthrin from potato fresh fruit measured as part
of the field residue program .................................................................................. 49
5. Recovery of cyfluthrin from orange fresh fruit measured as part
of the field residue program .................................................................................. 50
6. Recovery of cyfluthrin from lemon fresh fruit measured as part
of the field residue program .................................................................................. 51
7. Recovery of cyfluthrin from grapefruit fresh fruit measured as part
of the field residue program .................................................................................. 52
8. Recovery of cyfluthrin from tomato fresh fruit measured as part
of the field residue program .................................................................................. 53
9. Recovery of cyfluthrin from pepper fresh fruit measured as part
of the field residue program .................................................................................. 54
10. Recovery of cyfluthrin from squash fresh fruit measured as part
of the field residue program .................................................................................. 55
11. Recovery of cyfluthrin from cucumber fresh fruit measured as part
of the field residue program .................................................................................. 56
12. Recovery of cyfluthrin from cantaloupe fresh fruit measured as part
of the field residue program .................................................................................. 57
13. Recovery of cyfluthrin from carrot roots measured as part
of the field residue program................................................................................... 58
14. Recovery of cyfluthrin from radish roots and tops measured as part
of the field residue program .................................................................................. 59
15. Recovery of cyfluthrin from head lettuce heads with wrapper leaves and heads
without wrapper leaves measured as part of the field residue program ................. 60
16. Recovery of cyfluthrin from leaf lettuce fresh leaves measured as part
of the field residue program .................................................................................. 61
17. Recovery of cyfluthrin from grape fresh fruit measured as part
of the field residue program................................................................................... 62
18. Recovery of cyfluthrin from dried pea dry pea measured as part
of the field residue program................................................................................... 63
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Agriculture Division
19. Recovery of cyfluthrin from dried bean dry seed measured as part
of the field residue program................................................................................... 64
20. Recovery of cyfluthrin from mustard green leaves measured as part
of the field residue program............................................................................... 65
21. Recovery of cyfluthrin from broccoli flowering heads and stems
measured as part of the field residue program .................................................. 66
22. Recovery of cyfluthrin from cabbage heads with wrapper leaves and heads
without wrapper leaves measured as part of the field residue program ............. 67
23. Recovery of cyfluthrin from celery trimmed leaf stalks and untrimmed
leaf stalks measured as part of the field residue program.................................. 68
24. Recovery of cyfluthrin from spinach leaves measured as part
of the field residue program............................................................................... 69
25. Recovery of cyfluthrin from tobacco whole green leaves and flue cured
leaves measured as part of the field residue program ....................................... 70
26. Recovery of cyfluthrin from peanut nutmeat and hay measured as part
of the field residue program............................................................................... 71
27. Recovery of cyfluthrin from apple fresh fruit measured as part
of the field residue program............................................................................... 72
28. Recovery of cyfluthrin from pear fresh fruit measured as part
of the field residue program............................................................................... 73
29. LODs of cyfluthrin from crop matrices ............................................................... 74
30. Cyfluthrin recovery ranges ................................................................................ 75
Figures
1. Chemical name, acronymns, storage conditions, and structures
for cyfluthrin and [ 2 H6]cyfluthrin ......................................................................... 76
2. Flow diagram of the residue analytical method for apple and pear matrices...... 77
3. Flow diagram of the residue analytical method for cyfluthrin in all matrices
except apple, pear, and nutmeat ....................................................................... 78
4. Flow diagram of the residue analytical method for cyfluthrin in nutmeat ............ 79
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Agriculture Division
5. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 80
6. GC/MS chromatogram of a typical cherry fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 81
7. GC/MS chromatogram of a typical cherry fresh fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ........................................ 82
8. GC/MS chromatogram of a typical cherry fresh fruit 0.5 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ........................................ 83
9. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 84
10. GC/MS chromatogram of a typical peach fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 85
11. GC/MS chromatogram of a typical peach fresh fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ........................................ 86
12. GC/MS chromatogram of a typical peach fresh fruit 0.4 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ........................................ 87
13. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 88
14. GC/MS chromatogram of a typical plum fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 89
15. GC/MS chromatogram of a typical plum fresh fruit 0.01 ppm recovery
/ 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................................... 90
16. GC/MS chromatogram of a typical plum fresh fruit 0.1 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ........................................ 91
17. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 92
18. GC/MS chromatogram of a typical potato fresh tuber control /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 93
19. GC/MS chromatogram of a typical potato fresh tuber 0.01 ppm
cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ......................... 94
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20. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 95
21. GC/MS chromatogram of a typical orange fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 96
22. GC/MS chromatogram of a typical orange fresh fruit 0.01 ppm
cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ......................... 97
23. GC/MS chromatogram of a typical orange fresh fruit 0.1 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ........................................ 98
24. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution......................................................... 99
25. GC/MS chromatogram of a typical lemon fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 100
26. GC/MS chromatogram of a typical lemon fresh fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 101
27. GC/MS chromatogram of a typical lemon fresh fruit 0.1 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution...................................... 102
28. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 103
29. GC/MS chromatogram of a typical grapefruit fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 104
30. GC/MS chromatogram of a typical grapefruit fresh fruit
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 105
31. GC/MS chromatogram of a typical grapefruit fresh fruit
0.1 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 106
32. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 107
33. GC/MS chromatogram of a typical tomato whole fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 108
34. GC/MS chromatogram of a typical tomato whole fruit
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 109
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Agriculture Division
35. GC/MS chromatogram of a typical tomato whole fruit 0.5 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 110
36. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 111
37. GC/MS chromatogram of a typical pepper whole fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 112
38. GC/MS chromatogram of a typical pepper whole fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 113
39. GC/MS chromatogram of a typical pepper whole fruit 0.4 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 114
40. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution...................................................... 115
41. GC/MS chromatogram of a typical squash whole fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 116
42. GC/MS chromatogram of a typical squash whole fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 117
43. GC/MS chromatogram of a typical squash whole fruit 0.08 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 118
44. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 119
45. GC/MS chromatogram of a typical cucumber whole fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution...................................................... 120
46. GC/MS chromatogram of a typical cucumber whole fruit
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 121
47. GC/MS chromatogram of a typical cucumber whole fruit 0.07 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 122
48. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 123
49. GC/MS chromatogram of a typical cantaloupe whole fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 124
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50. GC/MS chromatogram of a typical cantaloupe whole fruit
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 125
51. GC/MS chromatogram of a typical cantaloupe whole fruit
0.05 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 126
52. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 127
53. GC/MS chromatogram of a typical carrot root control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 128
54. GC/MS chromatogram of a typical carrot root 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 129
55. GC/MS chromatogram of a typical carrot root 0.05 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 130
56. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 131
57. GC/MS chromatogram of a typical radish root control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 132
58. GC/MS chromatogram of a typical radish root 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 133
59. GC/MS chromatogram of a typical radish root 0.025 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 134
60. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 135
61. GC/MS chromatogram of a typical radish tops control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 136
62. GC/MS chromatogram of a typical radish tops 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 137
63. GC/MS chromatogram of a typical radish tops 5.0 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 138
64. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 139
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65. GC/MS chromatogram of a typical head lettuce heads with wrapper leaves
control / 0.05 ppm [ 2 H6]cyfluthrin standard solution ......................................... 140
66. GC/MS chromatogram of a typical head lettuce heads with wrapper leaves
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 141
67. GC/MS chromatogram of a typical head lettuce heads with wrapper leaves
2.0 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 142
68. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 143
69. GC/MS chromatogram of a typical head lettuce heads without
wrapper leaves / 0.05 ppm [ 2 H6]cyfluthrin standard solution ............................ 144
70. GC/MS chromatogram of a typical head lettuce heads without wrapper leaves
0.01 ppm cyfluthrin standard / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 145
71. GC/MS chromatogram of a typical head lettuce heads without wrapper leaves
0.05 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 146
72. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 147
73. GC/MS chromatogram of a typical leaf lettuce fresh leaves control /
0.05 ppm [ 2 H6]cyfluthrin standard solution...................................................... 148
74. GC/MS chromatogram of a typical leaf lettuce fresh leaves
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 149
75. GC/MS chromatogram of a typical leaf lettuce fresh leaves 2.5 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 150
76. GC/MS chromatogram of a typical leaf lettuce fresh leaves 3.5 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 151
77. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 152
78. GC/MS chromatogram of a typical grape fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 153
79. GC/MS chromatogram of a typical grape fresh fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution...................................... 154
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80. GC/MS chromatogram of a typical grape fresh fruit 0.5 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 155
81. GC/MS chromatogram of a typical grape fresh fruit 1.0 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 156
82. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 157
83. GC/MS chromatogram of a typical dried pea dry pea control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 158
84. GC/MS chromatogram of a typical dried pea dry pea 0.01 ppm
cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ....................... 159
85. GC/MS chromatogram of a typical dried pea dry pea 0.15 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 160
86. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 161
87. GC/MS chromatogram of a typical dried bean dry seed control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 162
88. GC/MS chromatogram of a typical dried bean dry seed 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 163
89. GC/MS chromatogram of a typical dried bean dry seed 0.15 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 164
90. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 165
91. GC/MS chromatogram of a typical mustard green leaves control /
0.05 ppm [ 2 H6]cyfluthrin standard solution...................................................... 166
92. GC/MS chromatogram of a typical mustard green leaves 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 167
93. GC/MS chromatogram of a typical mustard green leaves 4.0 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 168
94. GC/MS chromatogram of a typical mustard green leaves 5.5 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 169
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95. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 170
96. GC/MS chromatogram of a typical broccoli flowering heads and
stems control / 0.05 ppm [ 2 H6]cyfluthrin standard solution ............................... 171
97. GC/MS chromatogram of a typical broccoli flowering heads and stems
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 172
98. GC/MS chromatogram of a typical broccoli flowering heads and stems
0.75 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 173
99. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 174
100. GC/MS chromatogram of a typical cabbage heads with wrapper leaves
control / 0.05 ppm [ 2 H6]cyfluthrin standard solution ......................................... 175
101. GC/MS chromatogram of a typical cabbage heads with wrapper leaves
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution....... 176
102. GC/MS chromatogram of a typical cabbage heads with wrapper leavers
3.5 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 177
103. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 178
104. GC/MS chromatogram of a typical cabbage heads without wrapper
leaves control / 0.05 ppm [ 2 H6]cyfluthrin standard solution .............................. 179
105. GC/MS chromatogram of a typical cabbage heads without wrapper leaves
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 180
106. GC/MS chromatogram of a typical cabbage heads without wrapper leaves
0.02 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 181
107. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 182
108. GC/MS chromatogram of a typical celery untrimmed leaf stalks control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 183
109. GC/MS chromatogram of a typical celery untrimmed leaf stalks
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 184
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110. GC/MS chromatogram of a typical celery untrimmed leaf stalks
3.5 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 185
111. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 186
112. GC/MS chromatogram of a typical celery trimmed leaf stalks control
/ 0.05 ppm [ 2 H6]cyfluthrin standard solution .................................................... 187
113. GC/MS chromatogram of a typical celery trimmed leaf stalks
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 188
114. GC/MS chromatogram of a typical celery trimmed leaf stalks
0.1 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 189
115. GC/MS chromatogram of a typical celery trimmed leaf stalks
0.70 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 190
116. GC/MS chromatogram of a typical celery trimmed leaf stalks
1.0 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 191
117. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 192
118. GC/MS chromatogram of a typical spinach leaves control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 193
119. GC/MS chromatogram of a typical spinach leaves 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 194
120. GC/MS chromatogram of a typical spinach leaves 6.0 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 195
121. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 196
122. GC/MS chromatogram of a typical tobacco whole green leaves control
/ 0.05 ppm [ 2 H6]cyfluthrin standard solution .................................................... 197
123. GC/MS chromatogram of a typical tobacco whole green leaves
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 198
124. GC/MS chromatogram of a typical tobacco whole green leaves
1.5 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 199
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125. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 200
126. GC/MS chromatogram of a typical tobacco flue cured leaves control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 201
127. GC/MS chromatogram of a typical tobacco flue cured leaves
0.01 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution........ 202
128. GC/MS chromatogram of a typical tobacco flue cured leaves
7.5 ppm cyfluthrin recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution.......... 203
129. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 204
130. GC/MS chromatogram of a typical peanut nutmeat control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 205
131. GC/MS chromatogram of a typical peanut nutmeat 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 206
132. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 207
133. GC/MS chromatogram of a typical peanut hay control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 208
134. GC/MS chromatogram of a typical peanut hay 0.01 ppm cyfluthrin recovery /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 209
135. GC/MS chromatogram of a typical peanut hay 6.0 ppm cyfluthrin recovery /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 210
136. GC/MS chromatogram of a typical peanut hay 6.5 ppm cyfluthrin recovery /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 211
137. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 212
138. GC/MS chromatogram of a typical apple fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 213
139. GC/MS chromatogram of a typical apple fresh fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 214
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140. GC/MS chromatogram of a typical apple fresh fruit 0.05 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 215
141. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 216
142. GC/MS chromatogram of a typical pear fresh fruit control /
0.05 ppm [ 2 H6]cyfluthrin standard solution....................................................... 217
143. GC/MS chromatogram of a typical pear fresh fruit 0.01 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 218
144. GC/MS chromatogram of a typical pear fresh fruit 0.05 ppm cyfluthrin
recovery / 0.05 ppm [ 2 H6]cyfluthrin standard solution ...................................... 219
145. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus cherry fresh fruit
(top) and in the presence of solvent plus peach fresh fruit (bottom)................. 220
146. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus plum fresh fruit (top)
and in the presence of solvent plus potato fresh tuber (bottom) ...................... 221
147. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus orange fresh fruit
(top) and in the presence of solvent plus lemon fresh fruit (bottom)................. 222
148. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus grapfruit fresh fruit
(top) and in the presence of solvent plus tomato whole fruit (bottom).............. 223
149. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus pepper whole fruit
(top) and in the presence of solvent plus squash whole fruit (bottom) ............. 224
150. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus cucumber whole
fruit (top) and in the presence of solvent plus cantaloupe whole fruit (bottom). 225
151. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus carrot roots(top)
and in the presence of solvent plus radish roots (bottom)................................ 226
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152. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in
the presence of solvent only and in the presence of solvent plus radish
tops (top) and in the presence of solvent plus head lettuce heads with
wrapper leaves (bottom).................................................................................. 227
153. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus head lettuce
heads without wrapper leaves (top) and in the presence of solvent plus leaf
lettuce fresh leaves (bottom) ........................................................................... 228
154. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus grape fresh fruit
(top) and in the presence of solvent plus dried pea dry pea (bottom) .............. 229
155. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus dried bean dry seed
(top) and in the presence of solvent plus mustard green leaves (bottom)........ 230
156. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus boccoli flowering
heads and stems (top) and in the presence of solvent plus cabbage heads with
wrapper leaves (bottom).................................................................................. 231
157. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus cabbage heads
without wrapper leaves (top) and in the presence of solvent plus celery trimmed
leaf stalks (bottom).......................................................................................... 232
158. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus celery untrimmed
leaf stalks (top) and in the presence of solvent plus spinach leaves leaves
(bottom) .......................................................................................................... 233
159. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus tobacco whole
green leaves (top) and in the presence of solvent plus tobacco flue cured leaves
(bottom) .......................................................................................................... 234
160. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus peanut nutmeat
(top) and in the presence of solvent plus peanut hay (bottom) ........................ 235
161. Linearity curve for the cyfluthrin / [ 2 H6]cyfluthrin standard solutions in the
presence of solvent only and in the presence of solvent plus apple fresh fruit
(top) and in the presence of solvent plus pear fresh fruit (bottom) ................... 236
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Appendix
1. Archive listing of project personnel and notebook numbers............................. 237
2. Sample calculations ........................................................................................ 238
3. Sample calculation of the LOD from cherry fresh fruit ..................................... 240
4. Proposed fragmentation of cyfluthrin to m/z ion 207........................................ 242
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A Modified Analytical Method for the Determination of Cyfluthrin Residues in Various Plant
Matrices by Chemical Ionization – GC/MS
1.0 Summary
An existing analytical method 1 for the measurement of the total toxic residues (TTR) of cyfluthrin
in various crop using gas chromatography/mass spectroscopy (GC/MS) with electron impact
ionization(EI) was updated to GC/MS using chemical ionization (CI).
The new method was developed for apple and pear matrices but was later modified to include
all other matrices due to complications with some of the tougher crop matrices. This method
works for all matrices analyzed except peanut nutmeat which required a modification due to its
oily nature. The initial method used for apple and pear matrices consisted of homogenizing a
5.0 g sample in 45 mL of MeOH/aqueous 1.2 N HCl (4:1, v/v) for 2 min. After filtration into a
100-mL graduated mixing cylinder, the filter cake was returned to the extraction vessel and
homogenized with 45 mL of MeOH for an additional 2 min followed by filtration and combination
with the first extract. The sample was diluted to 100 mL with MeOH/aqueous 1.2 N HCl (4:1).
A 50-mL aliquot of the sample was rotary evaporated to dryness. The dry residue was
reconstituted in 25 mL of hexane in a boiling flask using sonication to dissolve as much of the
solids as possible. A florisil column was prepared by adding 100 mL of hexane and a glass
wool plug into a 250-mL glass bell column. Seven gram of florisil was added to the column
followed by 6 g of sodium sulfate. The hexane was allowed to drain from the column until the
level of hexane reached the top of the column packing. The 25-mL sample extract was added
to the column and allowed to drain dropwise until the solvent level reached the top of the
column packing. The boiling flask that contained the sample extract was washed with 40-mL of
clean hexane. The wash was then added to the florisil column and allowed to drain until the
solvent just reached the top of the column packing. A 125-mL boiling flask was placed under
the florisil column, and the cyfluthrin was eluted from the column with 60-mL of hexane/acetone
(9:1, v/v). The hexane/acetone mixture was rotary evaporated to dryness and reconstituted in
2.5 mL of toluene. The sample extracts were stored at < 10 ºC. The samples were analyzed
using GC/MS equipped with a chemical ionization (CI) source and quantified using a known
amount of deuterated internal standard. The internal standard was added to the samples after
the extraction.
The method used for the analysis of all other crop matrices except peanut nutmeat consisted of
homogenizing a 5.0 g sample in 45 mL of MeOH/aqueous 1.2 N HCl (4:1) for 2 min. After
filtration into a 100-mL graduated mixing cylinder, the filter cake was returned to the extraction
vessel and homogenized with 45 mL of MeOH for an additional 2 min followed by filtration and
combination with the first extract. The sample was diluted to 100 mL with MeOH. A 50-mL
aliquot of the sample was added to a 60-mL vial and evaporated to approximately 4 to 5 mL
using a Turbo Vap LV. Ten milliliters of water was added to the vial and mixed thoroughly. The
sample extract was transferred to a 125-mL separatory funnel. The 60-mL vial was washed
with two 15-mL aliquots of acetone/methylene chloride (1:2) with both aliquots being added to
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the 125-mL separatory funnel. The separatory funnel was shaken for 30 s. The lower organic
phase was drained through a funnel containing a glass wool plug, and 15 g of anhydrous
sodium sulfate into a 60-mL vial. The sodium sulfate was washed with 10 mL of methylene
chloride and collected in the same 60-mL vial. The sample was concentrated to dryness on a
Turbo Vap LV and reconstituted in 25-mL of hexane. A florisil column was prepared by adding
50 mL of hexane and a glass wool plug into a 200-mL glass bell column. Seven gram of florisil
was added to the column followed by 3 g of sodium sulfate. The hexane was allowed to drain
from the column until the level of hexane just reached the top of the column packing. The
25 mL sample extract was added to the column and allowed to drain dropwise until the solvent
level reached the top of the column packing. The 60-mL vial that contained the sample extract
was washed with 40-mL of clean hexane. The wash was then added to the florisil column and
allowed to drain until the solvent just reached the top of the column packing. A 60-mL vial was
placed under the florisil column, and the cyfluthrin was eluted from the column with 40 mL of
hexane/acetone (9:1, v/v). The hexane/acetone mixture was concentrated to dryness on a
Turbo Vap LV and reconstituted in 2.5 mL of toluene. The sample extracts were stored at
< 10 ºC. The samples were analyzed using GC/MS equipped with a CI source and quantified
using a known amount of deuterated internal standard. The internal standard was added to the
samples after the extraction.
The peanut nutmeat samples required a significant change to the extraction scheme and
clean-up due to the oily nature of the nutmeat matrix. In the normal extraction scheme, MeOH
and water would not remove the cyfluthrin from the nutmeat once it was absorbed into the oil.
The nutmeat method consisted of extracting a 5 g sample using 50 mL of hexane on a Soxtec
extractor. The sample was boiled in 50 mL of hexane for 120 min and then rinsed for additional
30 min. The extract was transferred to a 50-mL graduated mixing cylinder. The Soxtec cup that
contained the extract was washed with 5 mL of clean hexane and transferred to the 50-mL
graduated cylinder. The sample was diluted to 50 mL with hexane and mixed thoroughly. A
25-mL aliquot of the sample was added to a 125-mL separatory funnel followed by 25 mL of
ACN presaturated with hexane. The separatory funnel was shaken for 30 s. The lower ACN
phase was transferred to another 125-mL separatory which contained 10 mL of hexane
presaturated with ACN and shaken for 30 s. The lower ACN phase was collected in a 60-mL
vial. Another 25 mL of ACN presaturated with hexane was added to the first separatory funnel
and shaken for 30 s. The lower ACN phase was added to the second separatory funnel
containing 10 mL of hexane presaturated with ACN and shaken for 30 s. The lower ACN phase
was collected in the 60-mL vial with the first ACN phase. The extract was concentrated to
dryness using a Turbo Vap LV. A florisil column was prepared by adding 50 mL of hexane and
a glass wool plug into a 200-mL glass bell column. Seven grams of florisil was added to the
column followed by 3 g of sodium sulfate. The hexane was allowed to drain from the column
until the level of hexane just reached the top of the column packing. The 25-mL sample extract
was added to the column and allowed to drain dropwise until the solvent level reached the top of
the column packing. The 60-mL vial that contained the sample extract was washed with 40 mL
of clean hexane. The wash was added to the florisil column and allowed to drain until the
solvent reached the top of the column packing. A 60-mL vial was placed under the florisil
column and the cyfluthrin was eluted from the column with 40 mL of hexane/acetone (9:1, v/v).
The hexane/acetone (9:1, v/v) mixture was concentrated to dryness on a Turbo Vap LV and
reconstituted in 2.5 mL of toluene. The sample extracts were stored at < 10 ºC. The samples
23

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were analyzed using GC/MS equipped with a CI source and quantified using a known amount of
deuterated internal standard. The internal standard was added to the samples after the
extraction.
All recovery data and LODs (limits of detection) were measured as part of the cyfluthrin field
residue program. 3-17
Recoveries ranges of cyfluthrin were as follows:
Crop / Matrix
Fortification Range
Low High
Recovery
Low High
Cherry / fresh fruit 0.01 0.5 91 103
Peach / fresh fruit 0.01 0.4 88 101
Plum / fresh fruit 0.01 0.1 86 109
Potato / fresh tuber 0.01 NA 90 100
Orange / fresh fruit 0.01 0.1 94 102
Lemon / fresh fruit 0.01 0.15 88 101
Grapefruit / fresh fruit 0.01 0.1 90 99
Tomato / whole fruit 0.01 0.5 91 95
Pepper / whole fruit 0.01 0.4 86 104
Squash / whole fruit 0.01 0.08 96 99
Cucumber / whole fruit 0.01 0.07 91 100
Cantaloupe / whole fruit 0.01 0.05 90 98
Carrots / roots 0.01 0.05 92 102
Radish / tops 0.01 5.0 80 90
Radish / roots 0.01 0.025 91 108
Head lettuce / heads with wrapper leaves 0.01 2.0 95 105
Head lettuce / heads without wrapper 0.01 0.05 93 98
Leaf lettuce / fresh leaves 0.01 3.5 94 99
Grape / fresh fruit 0.01 1.0 93 101
Dried pea / dry pea 0.01 0.15 78 92
Dried beans / dry seed 0.01 0.15 74 98
Mustard green / leaves 0.01 5.5 83 100
Broccoli / flowering heads and stems 0.01 0.75 92 101
Cabbage / heads with wrapper leaves 0.01 3.5 93 105
Cabbage / heads without wrapper leaves 0.01 0.02 99 107
Celery / trimmed leaf stalks 0.01 1.0 93 103
Celery / untrimmed leaf stalks 0.01 3.5 80 103
Spinach / leaves 0.01 6.0 91 102
Tobacco / whole green leaves 0.01 1.5 95 100
Tobacco / flue cured leaves 0.01 7.5 73 91
Peanut / nutmeat 0.01 NA 76 97
Peanut / hay 0.01 6.5 68 87
Apple / fresh fruit 0.01 0.05 88 121
Pear / fresh fruit 0.01 0.05 78 96
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The LOQ (limit of quantitation) was 0.01 ppm in all the matrices. The LODs (limits of detection)
ranged from 0.00031 ppm to 0.0058 ppm.
2.0 Introduction
Cyfluthrin(cyano-(4-fluoro-3-phonoxyphenyl)methyl-3-(2,2-dichloroethyenyl)-2,2dimethylcyclopropanecarboxylate),
CAS # 68359-37-5 was originally analyzed using a gas
chromatograph equipped with a electron capture detector (ECD). 2 The ECD method was
extremely sensitive, but only gave moderate selectivity and suffered from the problems
associated with using a ECD detector such as baseline noise due to contamination of oxygen or
electronegative molecules. The ECD method was adapted to GC/MS using an EI source in
positive mode. Single reaction monitoring (SIM) was used to yield three different ions (m/z 226,
m/z 206, m/z 163) where the m/z 226 ion was used for quantitation and the other two ions were
used for confirmation. Even though greater selectivity was accomplished with the use of
GC/MS, sensitivity was greatly diminished. In addition, points on the matrix curve were often
greater than a 20% the corresponding points on the solvent curve. GC/MS using chemical
ionization (CI) in negative mode and methane as the CI gas was investigated as a possible
replacement for the GC/MS method using the EI source. CI in negative mode was found to be
more sensitive and more selective than EI in positive mode. SIM lead to a fragment of m/z 207,
which was used as the quantitation ion. To further simplify the method, deuterated internal
cyfluthrin standard was added to the initial sample extract to account for any loss during the
method and instrumental variances between injections.
3.0 Experimental
3.1 Location
The method development and validation was conducted at Bayer Research Park located near
Stilwell, KS from October 2000 through December 2001. Project personnel are listed in
Appendix 1.
3.2 Materials
3.2.1 Apparatus
Assorted Laboratory glassware (including, but not limited to)
— Beakers (assorted sized)
— Boiling flask, 125-mL
— Bottle beaker (4 oz Qorpak)
— Buchner funnels, 5.6 cm
— Graduated pouring cylinder, 50 mL, 100-mL
— Graduated mixing cylinders with ground glass joints, 100-mL
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— Pipets
— Pasteur transfer, 5.75-in
— Separatory funnels, 125-mL
— Spatulas, stainless steel
— Volumetric flask, class A, 50-mL
Balances
— Top-loader electronic, accurate to 0.00001 g (AT 20, Mettler Toledo, Columbus, OH) or
equivalent
— Top-loader electronic, accurate to 0.001 g (FX-200, A&D Engineering, INC., Milpitas,
CA) or equivalent
— Top-loader electronic, accurate to 0.01 g (Precisa 1000C-3000D, Precisa, Dietikon,
Switzerland) or equivalent
Chromatography columns, low-medium pressure with reservoir, 200-mL, 250-mL,
(Kimble/Kontes, Vineland, New Jersey, P/N K420280-0213 and P/N K420280-0222) or
equivalent
Evaporators
— Rotary evaporator, Büchi Rotovapor, Model RE 120 with Büchi 461(Brinkman
Instruments Inc., Wesbury, NY) or equivalent.
— Turbo Vap LV with 60-mL ASE vial conversion kit (Zymark, Hopkinton, MA) or equivalent
GC Column
— HP- P /HQJWK PP , ' P ILOP WKLFNQHVV $JLOHQW 7HFKQRORJLHV 3DOR
Alto, CA) or equivalent
— DB- 06 P /HQJWK PP , ' P ILOP WKLFNQHVV $JLOHQW 7HFKQRORJLHV 3DOR
Alto, CA) or equivalent
Mass spectrometer, 5973 mass selective detector with EI/CI source, (Agilent Technologies,
Palo Alto, CA) or equivalent containing the following components
— Auto sampler, 7683 series injector, (Agilent Technologies, Palo Alto, CA)
— Gas chromatograph, 6890, (Agilent Technologies, Palo Alto, CA)
— Software, chemstation G1701BA version B.01.00, (Agilent Technologies, Palo Alto, CA)
Pipetter, Fisherbrand Finnpipettes 0.20 mL to 10 mL range (Fisher, Pittsburgh, PA) or
equivalent
Soxtec system HT 1043, (Foss North America, Praire, MN) or equivalent
Tissumizer, model T-25 Basic with SDT-182EN probe (Tekmar, Cincinnati, OH) or equivalent
Tumbler, Model A-R2, (Tru-Square Metal Products, Auburn, WA) or equivalent
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Vials
— 60 mL, (I-CHEM, New Castle, DE, P/N S236-0060) or equivalent
— Auto sampler, clear glass with inserts and Viton® lined septum caps, 1.8 mL)
— Sample storage, clear glass with Teflon lined caps, 4-mL
3.2.2 Reagents/Supplies
Celite® 545 filter aid, (Fisher Chemicals, Pittsburgh, PA) or equivalent
Compressed gases, helium and methane, purity •
Filters
— GF/A, 5.5-cm (Whatman Inc., Clifton, NJ) or equivalent
— GHP Acrodisc, 13-PP P 3DOO *HOPDQ /DERUDWRULHV $QQ $UERU 0, 3 1 RU
equivalent
Florisil, 60-100 mesh, 2.5% deactivated. Prepared by adding 25 mL of water to 975 g of florisil
that has been dried overnight at 110° C. Florisil was tumbled overnight to equilibrate moisture.
(Fisher, Pittsburg, PA) or equivalent
Sodium sulfate , anhydrous, (Fisher Chemicals, Pittsburgh, PA) or equivalent
Solvents: Methanol (MeOH), acetone, acetonitrile (ACN), methylene chloride (DCM), hexane,
and toluene (Fisher Chemical, Pittsburgh, PA, Optima Grade) or equivalent
Solvent solutions (all solutions are volume to volume)
— aqueous 1.2 N HCl. Prepared by adding 100 mL of concentrated HCl to a 1000-mL
graduated cylinder and diluting to 1000 mL with water.
— MeOH/aqueous 1.2 N HCL (4:1). Prepared by mixing 3200 mL of MeOH with 800 mL of
aqueous 1.2 N HCl in a 4 L glass bottle.
— acetone/DCM (1:2). Prepared by mixing 1300 mL of acetone with 2600 mL of DCM in a
4 L glass bottle.
— acetone/hexane (1:9). Prepared by mixing 400 mL of acetone with 3600 mL of hexane
in a 4 L glass bottle.
— ACN presaturated with hexane. Prepared by shaking equal amounts of ACN and
hexane.
— Hexane presaturated with ACN. Prepared by shaking equal amounts of hexane and
ACN.
Syringes
— Luer lok single use, 3-mL (Becton Dickinson and Co., Franklin Lakes, NJ) or equivalent
— Hamilton 1700 series gas tight, 0.10 mL, 0.25 mL, 0.50 mL, (Hamilton Company, Reno,
NV) or equivalent
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Bayer Corporation 108139-1
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Water (water obtained from a Millipore Q purification system (Millipore corporation, Bedford,
MA)) or equivalent
3.3 Reference Standards
Analytical standards (Figure 1) of cyfluthrin (50.2%, w/w solution in cyclohexanone) and
[ 2 H6]cyfluthrin (15.6%, w/w in ACN) may be obtained from Bayer. The purity of the standard
must be known so solutions of exact concentration can be prepared. The internal standard
must be injected separately to confirm that there is no interference/crossover to the nondeuterated
standard. If interference/crossover is greater than 20% of the LOQ (limit of
quantitation), then the deuterated standard cannot be used. Both neat standards are stored in a
freezer (< -15 °C) and all dilutions are stored in a refrigerator (< 10 °C).
3.3.1 Cyfluthrin Standard Solutions
3.3.1.1 Primary Standard
Using a balance accurate to at least 0.1 mg, weigh the appropriate amount of cyfluthrin neat
standard into a 50-mL volumetric to yield a standard solution containing 2 J P/ 'LOXWH WKH
neat material to volume with toluene. Sonicate if necessary to completely dissolve the neat
material. The exact amount of neat material to weigh may be calculated using the equation
below.
Mass (mg) =
&RQF J P/ [ )LQDO 9RO P/ [
3XULW\ [ J PJ
For example, a 50.2% pure cyfluthrin standard will require a transfer of
PJ WR \LHOG D VROXWLRQ FRQFHQWUDWLRQ RI J P/ LQ D P/ ILQDO YROXPH
3.3.1.2 Secondary Standards in Toluene
Prepare the following secondary standards in toluene from the primary standard in toluene.
6HFRQGDU\ 6WDQGDUG J P/
Add 5.0 mL of the primary standard in toluene to a 50-mL volumetric flask. Dilute
the solution to volume with toluene. Mix the solution thoroughly.
6HFRQGDU\ 6WDQGDUG J P/
Add 1.0 mL of the primary standard in toluene to a 50-mL volumetric flask. Dilute
the solution to volume with toluene. Mix the solution thoroughly.
28

Bayer Corporation 108139-1
Agriculture Division
6HFRQGDU\ 6WDQGDUG J P/
$GG P/ RI WKH J/mL secondary standard to a 50-mL volumetric flask.
Dilute the solution to volume with toluene. Mix the solution thoroughly.
6HFRQGDU\ 6WDQGDUG J P/
$GG P/ RI WKH J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask.
Dilute the solution to volume with toluene. Mix the solution thoroughly.
3.3.2 [ 2 H6]Cyfluthrin Standard Solutions
3.3.2.1 Primary Deuterated Standard
Note: Due to the [ 2 H6]cyfluthrin neat standard concentration of 15.6%, w/w in ACN, the ACN will
evaporate quickly when weighing out a small aliquot of standard. The weight measurement
should be taken quickly. The exact concentration of the primary deuterated standard was not
critical as long as standards containing internal standard from one primary standard were not
compared to standards made from a new primary standard.
Using a balance accurate to at least 0.1 mg, weigh the appropriate amount of [ 2 H6]cyfluthrin
neat standard into a 50-P/ YROXPHWULF WR \LHOG D VWDQGDUG VROXWLRQ FRQWDLQLQJ WR J P/
in parent equivalence. Dilute the neat material to volume with toluene. Sonicate if necessary to
completely dissolve the neat material. The exact amount of neat material to weigh may be
calculated using the equation below and adding a correction for deuterated standards to convert
to cyfluthrin parent molar equivalents.
Mass (mg) =
&RQF J P/ [ )LQDO 9RO P/ [ [ PRO ZW > 2 H2]Standard
3XULW\ [ J PJ [ PRO ZW 3DUHQW 6WDQGDUG
3.3.2.2 Deuterated Secondary Standards in Toluene
Prepare the following deuterated secondary standards in toluene from the primary deuterated
standard in toluene.
'HXWHUDWHG 6HFRQGDU\ 6WDQGDUG J P/ RI > 2 H6]cyfluthrin:
$GG EHWZHHQ WR P/ RI D WR J P/ SULPDU\ GHXWHUDWHG VWDQGDUG
to a 50-mL volumetric flask. Dilute the solution to volume with toluene. Mix the
solution thoroughly.
'HXWHUDWHG 6HFRQGDU\ 6WDQGDUG 6ROXWLRQ J PO
$GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG WR D -mL
volumetric flask. Dilute the solution to volume with toluene. Mix the solution
thoroughly.
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3.3.3 Linearity Standard Solutions
Linearity standards consist of a mixture of the cyfluthrin and [ 2 H6]cyfluthrin. The [ 2 H6]cyfluthrin
concentration was held constant in each linearity standard (same level used in the sample
analysis) while the cyfluthrin concentration was varied from the lowest designated point to the
highest designated point.
3.3.3.1 Linearity Standards
0 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG WR D -mL
volumetric flask. Dilute the solution to volume with toluene. Mix the solution
thoroughly.
0.002 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution
to volume with toluene. Mix the solution thoroughly.
0.004 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGDU\ VWDQGDUG WR a 50-mL volumetric flask. Dilute the solution
to volume with toluene. Mix the solution thoroughly.
0.010 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution
to volume with toluene. Mix the solution thoroughly.
0.020 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution
to volume with toluene. Mix the solution thoroughly.
0.050 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution
to volume with toluene. Mix the solution thoroughly.
0.10 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution
to volume with toluene. Mix the solution thoroughly.
0.20 ppm Add 1.0 mL of the 2.5 J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution to
volume with toluene. Mix the solution thoroughly.
1.0 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG and 10.0 mL of the
J P/ VHFRQGDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution to
volume with toluene. Mix the solution thoroughly.
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5.0 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ VHFRQGary standard to a 50-mL volumetric flask. Dilute the solution
to volume with toluene. Mix the solution thoroughly.
10.0 ppm $GG P/ RI WKH J P/ GHXWHUDWHG VHFRQGDU\ VWDQGDUG DQG P/ RI WKH
J P/ SULPDU\ VWDQGDUG WR D -mL volumetric flask. Dilute the solution to
volume with toluene. Mix the solution thoroughly.
3.3.4 Quantification Standard
4XDQWLILFDWLRQ 6WDQGDUG J P/
$GG P/ RI WKH J P/ VHFRQGDU\ VWDQGDUG DQG P/ RI WKH J P/
deuterated secondary standard solution to a 50-mL volumetric flask. Dilute the
solution to volume with toluene. Mix the solution thoroughly.
3.4 Analytical Method
3.4.1 Matrix Preparation
1. Add approximately 2.5 lbs of the raw agricultural commodity (RAC) to the vertical
batch processor. Add about an equal portion of palletized dry ice to the sample
in the processor. Macerate the combined dry ice and RAC until a homogenous
mixture is obtained.
2. Pour the homogenized mixture into a plastic Tupperware container and store
open at –20 ± 5 °C until the last traces of dry ice have sublimed. Store the
homogenized RAC in a freezer at –20 ± 5 °C.
3.4.2 Procedure (Apples and Pears only)
3.4.2.1 Extraction
1. Weigh 5.0 g of sample into a 120-mL glass bottle. Note: begin recovery samples
at this point by fortifying the control matrix sample with a standard.
2. Add 45 mL of the solvent mixture MeOH/aqueous 1.2 N HCl (4:1, v/v) into the
bottle. Homogenize for 2 min using a tissumizer.
3. Vacuum filter the sample solution using a Buchner funnel into a stoppered
100-mL graduated cylinder.
4. Return the solids including the paper filter back to the glass bottle, and add
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45 mL of MeOH into the bottle. Homogenize for 2 min, then filter a second time
into the same stoppered 100-mL graduated cylinder.
5. 8VLQJ D +DPLOWRQ V\ULQJH WUDQVIHU / RI WKH J P/ LQWHUQDO VWDQGDUG
solution to the filtrate. Adjust the volume to the 100-mL mark with the solvent
mixture and mix.
6. Split the 100-mL filtrate into two 50-mL portions and discard the filter.
7. Rotovap one of the 50-mL sample portions to remove all the solvents as possible
(some matrices may contain oily residues that can not be evaporated). Add
25 mL of hexane to the flask, sonicate in a water bath for 60 s.
8. Proceed to the florisil clean-up step.
3.4.2.2 Florisil Clean-up
1. Place glass wool at the bottom of a Kontes column (15 mm ID x 250 mm length,
250 mL reservoir), add 100 mL of hexane to the column.
2. Add 7 g of 2.5% deactivated florisil into the column and tap the column to remove
the air bubbles.
3. Add 6 g of anhydrous sodium sulfate to the column and drain the excess hexane
into a waste container. Leave enough hexane in the column just to cover the bed
of sodium sulfate.
4. Pour the 25-mL hexane sample extract from step 7 of the extraction into the
column. Let the extract run through the column dropwise into a waste container.
5. Add 40 mL of hexane to the boiling flask, mix and pour into the column. Let the
extract run through the column dropwise into a waste container.
6. Elute the analyte with 60 mL of hexane/acetone (90/10%) dropwise into a
125-mL boiling flask.
7. Rotovap to dryness.
3.4.2.3 Final Extract Preparation for Analysis
1. Pipet 2.5 mL of toluene into the flask and rotate to dissolve all the residue.
2. Filter approximately 1 mL of the sample solution into a sample vial using a
P -mm, acrodisc filter for GC/MS analysis.
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3.4.3 Procedure (All Matrices Except Nutmeat, Apples, and Pears)
3.4.3.1 Extraction
1. Weigh 5.00 g (± 0.02 g) of the sample into a 4 oz Qorpak bottle beaker (Fisher
P/N. 03-320-10C) or equivalent. NOTE: Begin recovery samples at this point by
fortifying the control tissue matrix samples with a standard.
2. Add 45 mL of methanol (MeOH)/aqueous 1.2 N HCl (4:1, v/v).
3. Blend the sample using an ultra-turrax blender or equivalent equipped with
tissumizer probe 18N or equivalent for approximately 2 min.
4. Add approximately 2 g of Celite®.
5. Vacuum filter the contents of the beaker through a Whatman GF/A filter in a
5.6-cm Buchner funnel into a 100-mL graduated mixing cylinder.
6. Return the filter cake to the 4 oz beaker and add 45 mL of MeOH and blend the
sample an additional 2 min.
7. Vacuum filter the contents through the same Buchner funnel containing a new
GF/A filter into the 100-mL graduated mixing cylinder.
8. Add the 2 [H6]cyfluthrin internal standard (0.05 ppm) to the 100-mL graduated
mixing cylinder.
9. Bring the volume in the 100-mL graduated mixing cylinder to 100 mL with MeOH.
Mix thoroughly.
10. Transfer 50 mL of the sample to a 60-mL I-Chem vial.
11. Evaporate to approximately 4 to 5 mL using a Turbo Vap LV
(water bath 55 ± 5 °C).
12. Add 10 mL of water to the I-Chem vial and mix.
13. Transfer the aqueous sample to a 125-mL separatory funnel.
14. Wash the I-Chem vial with 15 mL of acetone/methylene chloride (1:2) and
transfer to the 125-mL separatory funnel.
15. Repeat Step 14.
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16. Shake the separatory funnel for 30 s.
17. Drain the lower organic phase into a 60-mL I-Chem vial through a funnel
containing a glass wool plug and approximately 15 g of anhydrous sodium
sulfate.
18. Wash the sodium sulfate with approximately 10 mL of methylene chloride.
19. Evaporate to dryness on a Turbo Vap LV (water bath 40 ± 5° C).
20. Reconstitute the residue in 25 mL of hexane.
21. Proceed to the Florisil clean-up (Sec 3.4.3.4, Step 1)
3.4.3.2 Extraction (Nutmeat)
1. Weigh 5.00 g (± 0.02 g) of the sample into a paper thimble for the soxtec. Begin
recovery samples at this point by fortifying the control tissue matrix samples with
a standard.
2. Add 50 mL of hexane to the soxtec stainless steel beaker.
3. Place the cup on the soxtec and extract ~ two hours at 140 °C in the boil
position.
4. Turn the soxtec to the rinse position and rinse for 1 h.
5. Allow the cups to cool to room temperature.
6. Add the d6-cyfluthrin internal standard (0.05 ppm) to the soxtec cup.
7. Transfer the sample to a 50-mL graduated cylinder.
8. Wash the soxtec cup with 5 mL of clean hexane and transfer to the graduated
cylinder.
9. Bring the volume in the graduated cylinder to 50 mL with hexane.
10. Proceed to the ACN / hexane partition (Sec 3.4.3.3, Step 1)
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3.4.3.3 ACN/Hexane Partition
1. Transfer 25 mL of the sample from the 50-mL graduated cylinder to a 125-mL
separatory funnel.
2. Add 25 mL of ACN presaturated with hexane to the separatory funnel and shake
for approximately 30 s.
3. Transfer the lower ACN layer to another 125-mL separatory funnel already
containing 10 mL of hexane presaturated with ACN and shake for approximately
30 s.
4. Collect the lower ACN layer into a 60-mL I-Chem vial.
5. Repeat Steps 2 through 4. Note: Both ACN fractions in Step 13 are collected in
the same 60 mL vial.
6. Concentrate the sample in the 60-mL vial to dryness on a Turbo-vap LV (water
bath 45 ± 5 °C) and reconstitute in 25 mL of hexane followed by sonication.
7. Proceed to the Florisil Clean-up (Sec 3.4.3.4, Step 1)
3.4.3.4 Florisil Clean-up
1. Place a plug of glass wool at the bottom of a Kontes column (11 mm ID x 250
length, 200 mL reservoir), and add 50 mL of hexane to the column.
2. Slowly add approximately 7 g of 2.5% deactivated florisil into the column while
tapping the column to remove air bubbles.
3. Add approximately 3 g of anhydrous sodium sulfate to the column.
4. Drain the hexane until it is just above the sodium sulfate layer. Discard the
hexane.
5. Pour the 25-mL sample extract from Step 20 (extraction for all matrices except
apple, pear, and nutmeat) or Step 6 (ACN / hexane partition nutmeat) into the
column. Let the extract run through the column dropwise. Discard the hexane.
6. Wash the I-Chem vial with 40 mL of hexane and transfer to the column. Allow
the hexane to drip until it just reaches the top of the sodium sulfate. Discard the
hexane.
7. Elute the analyte with 40 mL of hexane/acetone (9:1) dropwise into a 60-mL
I-Chem until no more drops are apparent.
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8. Concentrate the sample to dryness using a turbo-vap (water bath 40 ± 5 °C).
9. Add 2.5 mL of toluene to the 60-mL I-Chem vial and mix thoroughly.
10. Proceed to the final extract preparation for analysis
3.4.3.5 Final Extract Preparation for Analysis
1. To complete the preparation for instrumental analysis, the extract is to be added
to a GC vial.
3.4.4 Analysis By Gas Chromatography/Mass Spectrometry (GC/MS)
A. GC conditions:
Column: DB-5MS capillary column, 5% phenyl methyl, 12 m length, 0.2 mm
ID, 0.33- P ILOP WKLFNQHVV DOO PDWULFHV H[FHSW DSSOH DQG SHDU
Column: HP-5 capillary column, 5% phenyl methyl, 12 m length, 0.2 mm
ID, 0.33- P ILOP WKLFNQHVV DSSOH DQG SHDU
Injector port temp: 280 °C (all matrices except apple and pear)
250 °C (apple and pear)
Flow: 1 mL/min
Mode: Splitless then split at 1.0 min
Gas type: Helium
Initial oven temp: 150 °C
Initial time: 4 min
Ramps: 30 °C to a final temp of 300 °C and hold for 11 min
Total run time: 20.0 min
Approx. Retention Time: 9.4 min
B. Mass spectrometer conditions:
Ionization mode: negative chemical ionization (NCI)
Ionization gas: methane
Acquisition mode: Single interaction monitoring (SIM)
Solvent delay: 4 min
Ion masses: cyfluthrin m/z 207
[ 2 H6]cyfluthrin m/z 213
MS quad temp: 106 °C
MS source temp: 150 °C
MSD transfer line temp: 300 °C
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C. Procedure:
1. Use the chemstation software to run the automated autotune for NCI.
2. Add an aliquot of the final extract or an aliquot of the 0.050 ppm quantification
standard (0.050 ppm cyfluthrin and 0.050 ppm [ 2 H6]cyfluthrin) solution in toluene
into a GC auto sampler vial.
3. Inject 1 / IURP WKH ILUVW TXDQWLILFDWLRQ VWDQGDUG
4. ,QMHFW / IURP -10 samples (1-10 in a sample set).
5. ,QMHFW / IURP WKH VHFRQG TXDQWLILFDWLRQ VWDQGDUG
6. Compare the ratio (response factor) of the cyfluthrin / [ 2 H6]cyfluthrin peak area of
each sample extract solution to the average ratio of the respective peak area of
the quantitation standards on either side.
7. Compare the ratio of the cyfluthrin / [ 2 H6]cyfluthrin peak area of the quantitation
standard injected before and after each group of sample extract solution
injections. If the ratios vary by > 20% of the ratio of the first quantitation
standard, prepare new aliquots of the quantitation standard and sample extract
solutions and repeat steps 1 through 6. If there is < 20% variation, proceed to
part C.
C. Calculations
1. Compare the GC retention time of cyfluthrin / [ 2 H6]cyfluthrin in each of the
injections made. The retention time of the cyfluthrin should be within ± 0.03 min
of the [ 2 H6]cyfluthrin. Also, the retention time of the cyfluthrin in the sample
injections should be with 0.10 min of the average of the retention times for the
cyfluthrin in the sample extract’s respective quantitation standard injections. If
these criteria are met, then proceed to Step 2. If not, then samples should be reanalyzed.
2. Calculate the concentration of cyfluthrin in each sample by comparing the
detector response factor (peak area of cyfluthrin/[ 2 H6]cyfluthrin for the sample to
the average response factor for the quantitation standards injected before and
after the sample.
response factor sample target sample weight
ppm = X X standard conc. X dilution factor
Avg standard response factor actual sample weight
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The standard concentration is equal to the ppm relative to matrix weight in an
undiluted sample taking into account the initial sample size, aliquoting of the
original sample, and the final extract volume. The following equation may be
used to calculate the ppm equivalence of the standard solution concentration.
S x F x T
Standard conc. (ppm equivalence) =
W x A
where 6 $QDO\WLFDO VWDQGDUG FRQFHQWUDWLRQ J P/
F = Final extract volume (mL),
T = Total initial extract volume (mL),
W = Initial target sample weight (g),
A = Aliquot taken
An example sample residue calculation showing the use of the above formulae is presented in
Appendix 2.
D. Detector Linearity Curves
1. To demonstrate linearity of the response factor of cyfluthrin versus the
corresponding [ 2 H6]cyfluthrin internal standard in either solvent or matrix extract,
a series of 7 different concentrations were used. More points were sometimes
needed depending on the amount of residue measured.
Transfer 150 / IURP D FRQWURO H[WUDFW FRQWDLQLQJ QR > 2 H6]cyfluthrin standard into
7 different auto sampler vials. Evaporate the solutions to dryness. Then transfer
/ RI WKH DSSURSULDWH VWDQGDUG WKDW FRQWDLQV F\IOXWKULQ DQG > 2 H6]cyfluthrin into
each of the 7 auto sampler vials to yield the solvent + matrix containing curve.
7UDQVIHU / RI WKH DSSURSULDWH VWDQGDUG LQWR HDFK RI DQRWKHU DXWR VDPSOHU
vials to yield the solvent only curve. The contents of the vials are shown below:
Vial 1 Blank 0.00000 J P/ F\IOXWKULQ J P/ > 2 H6]cyfluthrin
Vial 2 0.002 ppm 0.0020 J P/ F\IOXWUKLQ J P/ > 2 H6]cyfluthrin
Vial 3 0.004 ppm 0.0040 J P/ F\IOXWUKLQ J P/ > 2 H6]cyfluthrin
Vial 4 0.010 ppm 0.010 J P/ F\IOXWUKLQ J P/ > 2 H6]cyfluthrin
Vial 5 0.020 ppm 0.020 J P/ F\IOXWKULQ J P/ > 2 H6]cyfluthrin
Vial 6 0.050 ppm 0.050 J P/ F\IOXWKULQ J P/ > 2 H6]cyfluthrin
Vial 7 0.100 ppm 0.10 J P/ F\IOXWKULQ J P/ > 2 H6]cyfluthrin
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2. Sequentially analyze the solutions in each of the auto sampler vials and calculate
a response factor for cyfluthrin standards versus [ 2 H6]cyfluthrin standards in each
solution.
3. On a graph comparing response factor versus concentration of cyfluthrin in each
solution (ppm relative to matrix), plot the curves for the fortified solvent and the
fortified matrix samples.
Using the equation: y = mx + b
where m = the coefficient (slope) of x,
x = the concentration (ppm),
b = the constant or y intercept,
Calculate the linear regression using Lotus 123 or equivalent spreadsheet
program of the data points for each set of solvent and matrix samples and
determine the correlation coefficient for the regression analysis of the data.
Plot the curve for the values of the regressed data points for solvent and matrix
data. Compare the slopes of the regressed curves. The regressed matrix curve
should be parallel to the regressed curve, and the response factors at each
concentration should indicate values differing by less than 20% between the
solvent curve and the matrix curve to show that any matrix effect was within
acceptable limits.
E. Confirmatory Procedure
Note: For cyfluthrin, use the peak area from the m/z 207.
1. For all standard solutions and samples, individually integrate all the ionchromatograms.
2. Examine each sample chromatogram. Inspect the analyte ion-chromatogram for
a peak eluting at the same retention time as the sample internal standard
(± 0.03 min). Any peak falling within this window was considered to be the
sample analyte. If a sample analyte peak was found, continue with the
confirmatory data analysis.
3. Further confirmation could be obtained by analyzing the questionable sample by
Bayer method 108139. 1
3.4.5 Disposal of Solutions
All solvent wastes should be disposed of in approved hazardous waste containers.
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3.5 Method Validation
3.5.1 Requirements
1. Analyze three unfortified control samples and three fortified control samples at
the LOQ, using three different control samples when possible.
2. Recovery values must fall between 70% and 120%.
3. The control ppm value must be less than 20% of the fortification ppm level.
4. (DFK *& 06 PHDVXUHPHQW RI D UHFRYHU\ ZDV FRPSDUHG WR D J P/
(0.050 ppm) quantitation standard containing cyfluthrin and [ 2 H6]cyfluthrin. All
recovery samples must fall within the linearity curve.
5. The correlation coefficient from the linear regression of the response factors of
the linearity curve points, as described in 3.4.4 D, must be greater than 0.99.
6. Calculate recoveries by the following equation:
3.5.2 Recovery Procedures
ppm found – control ppm
Recovery = X 100%
ppm fortification level
1. Fortify aliquots of control matrix (in the 4 oz glass bottle beaker that was used for
the 5 gram sample extraction) with the appropriate secondary standard using a
/ JDV WLJKW V\ULQJH WR DSSO\ WKH DSSURSULDWH VWDQGDUG VROXWLRQ IRU WKH
recovery level desired. An example is given below.
SSP / RI WKH J P/ VHFRQGDU\ VWDQGDUG VROXWLRQ
2. Proceed to sample extraction (Sec 3.4.2.1, Step 2, Sec 3.4.3.1 Step 2, or
Sec 3.4.3.2 Step 2).
4.0 Results and Discussion
A flow diagram of the residue analytical method is presented in Figures 2 thru 4.
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4.1 Sample Extraction (Except Nutmeat), Liquid/Liquid Partition,
and Florisil Clean-up
The extraction described in the original method 1 was retained except for a reduction in sample
weight with corresponding decrease in solvent volumes and the addition of HCl. HCl was used
in corn fodder in the original method due to the basic nature of the matrix. The extraction using
MeOH / aqueous 1.2 N HCl (4:1, v/v) followed by MeOH was considered adequate for the
release of cyfluthrin residues.
The method continued with a liquid/liquid partition. The aqueous phase containing the sample
was partitioned versus acetone / DCM (1:2, v/v). The cyfluthrin was transferred to the organic
phase. Many polar compounds extracted using the polar extraction solvents were left behind in
the aqueous phase. The method ends using normal phase absorption chromatography
(2.5% deactivated florisil) where the cyfluthrin was loaded onto the florisil in hexane, washed
with hexane and selectively removed with hexane/acetone (9:1, v/v). Any compounds that have
a stronger retention than cyfluthrin was left behind on the florisil packing.
4.1.1 Sample Extraction (Apple and Pear Only), and Florisil Clean-up
The extraction described in the original method 1 was retained except for a reduction in sample
weight with corresponding decrease in solvent volumes and the addition of HCl. HCl was used
in corn fodder in the original method due to the basic nature of the matrix. The extraction using
MeOH/aqueous 1.2 N HCl (4:1, v/v) followed by MeOH was considered adequate for the
release of cyfluthrin residues.
The method continued using normal phase absorption chromatography (2.5% deactivated
florisil) where the cyfluthrin was loaded onto the florisil in hexane, washed with hexane and
selectively removed with hexane/acetone (9:1, v/v). Any compounds that have a stronger
retention than cyfluthrin were left behind on the florisil packing.
4.1.2 Sample Extraction (Nutmeat Only), ACN / hexane Partition, and
Florisil Clean-up
Due to the oily nature of the nutmeat, the original extraction procedure was not yielding
acceptable recoveries. The nutmeat sample was placed on a Soxtec and boiled for 2 h in
50 mL of hexane followed by a 1 h rinse. The method continued with a ACN / hexane partition
where the cyfluthrin was transferred to the ACN phase. The oily compounds were left behind in
the hexane phase. The ACN phase was partitioned once more with clean hexane and
concentrated to dryness. The sample continued through the florisil clean-up as described in
Section 4.1.
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4.2 Addition of Internal Standard Solution
A standard containing the deuterated standard was added immediately after the extraction to
account for any loss during sample clean-up. The addition was made using the same accurate
pipette for each addition. The accuracy of the pipette was important since the response factors
calculated in the sample extracts were compared to the response factor of the quantification
standard, which was made using an accurate pipette.
4.3 GC/MS of Cyfluthrin
In the current method, the gas chromatography procedure in combination with the mass
spectrometer using negative chemical ionization was selected to replace the gas
chromatographic/mass spectrometry procedure using positive electron impact ionization. This
modification was performed to increase sensitivity and selectivity thus reducing the amount of
clean-up. In addition, the use of an internal standard compensated for loss during sample
clean-up. Due to the increase in sensitivity, less sample was required to maintain an LOQ of
0.01 ppm.
Figures 5 through 144 show typical negative chemical ionization selected ion monitoring (SIM)
ion-chromatograms of cyfluthrin, and [ 2 H6]cyfluthrin standard solutions and matrix extract
solutions. Cyfluthrin is represented by ion m/z 207. [ 2 H6]Cyfluthrin is represented by ion
m/z 213. Proposed fragmentation of Cyfluthrin is shown in Appendix 4.
The presence of eight diastereomers (four pairs) in cyfluthrin creates three peaks that are not
base line resolved. All three peaks are integrated as one peak area and used to quantitate
cyfluthrin.
4.4 Recovery of Cyfluthrin
Tables 1 through 28 list data for recoveries, control values, standard deviations, and average
recoveries for cyfluthrin from various plant matrices. Figures 5 through 144 show typical
chromatograms of standards and control samples, and recovery samples for various plant
matrices. All recoveries cyfluthrin at all fortification levels fell within the 70% to 120% range
except for the 6.5 ppm fortifications in peanut hay which were just below 70%.
4.5 Limits of Quantitation and Detection (Except Apple and Pears)
The limits of quantitation (LOQ) based on the lowest ppm levels fortified of were 0.01 ppm for all
the plant matrices analyzed.
The limit of detection (LOD) is defined as the lowest concentration that can be determined to be
statistically different from a blank or negative control. The LOD was calculated by taking the
standard deviation of the residue values from the analysis of the recovery samples at the LOQ
and using the equation shown below:
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LOD = Standard deviation X t 0.99
Where t 0.99 = the one tailed t-statistic at the 99%
Confidence level for n-1 replicates
Values for t and an example LOD calculation based on the recovery samples measured as part
of the apple field study is shown in Appendix 3. Table 29 shows the LODs for the different crop
matrices.
4.6 Linearity of Detector Response
Detector response factor curves for cyfluthrin are shown for standards in solvent solutions and
in the presence of crop matrix extracts (Figures 145 thru 161). The correlation coefficient for the
detector response factor curves were all > 0.99, indicating that the detector response factor was
linear over the range of concentrations tested.
4.7 Time Interval of the Analytical Method
If a set of 12 samples were started in the morning, and an analytical instrument was utilized
which had been previously set-up for the analysis of cyfluthrin, the GC/MS data was usually
available by the morning of the next day.
4.8 Conclusions
A modified method for measuring the residue of cyfluthrin in a wide variety of plant matrices
has been revised and validated. Limits of detection (LODs) are shown in Table 29. Recovery
ranges in each crop matrix are shown in Table 30.
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5.0 Bibliography
1. F.E. Sandie and R.R. Gronberg. 1998. An Analytical Method for the Determination of
&\IOXWKULQ DQG -Cyflutrhin Residues in Various Crops. Bayer Ag Div Report No.
108139.
2. A.M. Harbin, R. G. Minor, P.L. Freeseman, and L.K. Pfankuche. 1983. A gas
chromatographic method for BAYTHROID TM residues in crops. Bayer Ag Div Report No.
85823. MRID 40301501.
3. D. R. Fischer. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
on Broccoli and Cabbage (Crop Subgroup 5A - Head and Stem Brassica). Bayer Ag Div
Report No. 110339.
4. D. R. Fischer. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
on Cantaloupe, Cucumber, and Summer Squash (Crop Subgroup 9 - Cucurbit
Vegetables). Bayer Ag Div Report No. 110894.
5. D. R. Fischer. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
on Mustard Greens (Crop Subgroup 5B – Leafy Brassica Greens). Bayer Ag Div Report
No. 110895.
6. D. R. Fischer. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
on Potatoes. Bayer Ag Div Report No. 110988.
7. D. R. Fischer. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
on Tobacco. Bayer Ag Div Report No. 110340.
8. D. R. Fischer. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
on Grapes. Bayer Ag Div Report No. 110989.
9. E. C. Beedle. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
in Dried Peas and Dried Beans (Crop Subgroup 6C – Dried, Shelled Pea an Bean –
Except Soybean). Bayer Ag Div Report 110479.
10. E. C. Beedle. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
in Celery and Spinach Crop Group 4 . Bayer Ag Div. Report 110338.
11. E C. Beedle. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
in Carrots and Radishes (Bridging Trials). Bayer Ag Div Report 110958
12. E. C. Beedle. BAYTHROID 20 WP and BAYTHROID 2 EC - Magnitude of the Residue
in Leaf Lettuce and Head Lettuce (Bridging Trials). Bayer Ag Div Report 110628
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13. A. M. Harbin. BAYTHROID 20 WP and BAYTHROID 2 - Magnitude of the Residue in
Apples and Pears (Crop Group 11 - Pome Fruit). Bayer Ag Div Report No. 110031.
14. Lemke, V. J. 2002. BAYTHROID 2 EC and BAYTHROID 20 WP – Magnitude of the
Residue on Peppers and Tomatoes (Bridging Trials). Bayer Ag Div Report No. 110981.
15. Duah, F. K. 2002. BAYTHROID 2 EC and BAYTHROID 20 WP – Magnitude of the
Residue on Sweet and Tart Cherries, Peaches, and Plums (Crop Group 12 – Stone
Fruits). Bayer Ag Div Report No. 110314.
16. Duah, F. K. and V. J. Lemke. 2002. BAYTHROID 2 EC and BAYTHROID 20 WP –
Magnitude of the Residue on Grapefruits, Lemons, and Oranges (Crop Group 10 –
Citrus Fruits, Bridging Trials). Bayer Ag Div Report No. 111025.
17. Duah, F. K. 2002. BAYTHROID 2 EC and BAYTHROID 20 WP – Magnitude of the
Residue in/on Peanuts. Bayer Ag Div Report No. 111036.
45

Bayer Corporation 108139-1
Agriculture Division
Table 1. Recovery of cyfluthrin from cherry fresh fruit measured as part of the field residue
Program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD175-00D-019-3 0.00089 BD175-00D-019-4 0.0100 91 98 3.5
0.01 BD173-00HA-013-1 0.00040 BD173-00HA-013-2 0.00985 94
BD174-00H-013-3 0.00017 BD174-00H-013-4 0.00977 96
BD177-00H-013-1 0.00022 BD177-00H-013-2 0.00987 96
BD173-00HA-013-3 0.00005 BD173-00HA-013-4 0.00979 97
BD174-00H-013-1 0.00020 BD174-00H-013-2 0.00994 97
BD172-00H-013-3 0.00033 BD172-00H-013-4 0.0101 98
BD174-00H-013-5 0.00032 BD174-00H-013-6 0.0101 98
BD172-00H-013-1 0.00012 BD172-00H-013-2 0.0102 100
BD176-00H-013-3 0.00034 BD176-00H-013-4 0.0104 101
BD175-00D-019-1 0.00043 BD175-00D-019-2 0.0105 101
BD176-00H-013-1 0.00024 BD176-00H-013-2 0.0105 103
0.50 BD177-00H-013-4 0.00061 BD177-00H-013-5 0.475 95 96 1.2
BD177-00H-013-6 0.477 95
BD177-00H-013-7 0.486 97
1 Bayer Ag Div Report No. 110314. Percent recovery values were corrected for control
interferences.
46

Bayer Corporation 108139-1
Agriculture Division
Table 2. Recovery of cyfluthrin from peach fresh fruit measured as part of the field residue
Program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD158-00H-013-1 0.00010 BD158-00H-013-2 0.00887 88 95 3.2
0.01 BD159-00H-013-1 0.00000 BD159-00H-013-2 0.00898 90
BD163-00D-019-5 0.00056 BD163-00D-019-6 0.00969 91
BD157-00H-013-1 0.00000 BD157-00H-013-2 0.00923 92
BD157-00H-013-3 0.00006 BD157-00H-013-4 0.00937 93
BD163-00D-019-1 0.00000 BD163-00D-019-2 0.00953 95
BD162-00H-013-1 0.00018 BD162-00H-013-2 0.00964 95
BD165-00HA-013-1 0.00050 BD165-00HA-013-2 0.00997 95
BD163-00D-019-3 0.00000 BD163-00D-019-4 0.00964 96
BD158-00H-013-3 0.00022 BD158-00H-013-4 0.00977 96
BD158-00H-013-5 0.00024 BD158-00H-013-6 0.00988 96
BD165-00HA-013-3 0.00025 BD165-00HA-013-4 0.00988 96
BD159-00H-013-3 0.00000 BD159-00H-013-4 0.00966 97
BD161-00H-013-1 0.00000 BD161-00H-013-2 0.00966 97
BD160-00H-013-1 0.00000 BD160-00H-013-2 0.00974 97
BD160-00H-013-3 0.00000 BD160-00H-013-4 0.00974 97
BD159-00H-013-5 0.00006 BD159-00H-013-6 0.00992 99
BD164-00HA-013-1 0.00010 BD164-00HA-013-2 0.0102 101
0.40 BD165-00HA-013-6 0.00037 BD165-00HA-013-8 0.389 97 98 1.0
BD165-00HA-013-9 0.391 98
BD165-00HA-013-7 0.396 99
1 Bayer Ag Div Report No. 110314. Percent recovery values were corrected for control
interferences.
47

Bayer Corporation 108139-1
Agriculture Division
Table 3. Recovery of cyfluthrin from plum fresh fruit measured as part of the field residue
Program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD170-00H-013-1 0.00000 BD170-00H-013-2 0.00863 86 97 6.1
0.01 BD169-00H-013-5 0.00013 BD169-00H-013-6 0.00899 89
BD-168-00D-019-3 0.00014 BD-168-00D-019-4 0.00940 93
BD170-00H-013-3 0.00000 BD170-00H-013-4 0.00935 94
BD168-00D-019-1 0.00000 BD168-00D-019-2 0.00949 95
BD167-00H-013-5 0.00000 BD167-00H-013-6 0.00964 96
BD166-00H-013-3 0.00014 BD166-00H-013-4 0.00977 96
BD167-00H-013-3 0.00014 BD167-00H-013-4 0.00977 96
BD169-00H-013-3 0.00012 BD169-00H-013-4 0.00978 97
BD166-00H-013-1 0.00029 BD166-00H-013-2 0.0102 99
BD171-00H-013-1 0.00010 BD171-00H-013-2 0.0102 101
BD167-00H-013-1 0.00013 BD167-00H-013-2 0.0106 105
BD169-00H-013-1 0.00000 BD169-00H-013-2 0.0109 109
0.10 BD168-00D-019-6 0.00000 BD168-00D-019-9 0.0955 96 97 0.58
BD168-00D-019-7 0.0966 97
BD168-00D-019-8 0.0966 97
1 Bayer Ag Div Report No. 110314. Percent recovery values were corrected for control
interferences.
48

Bayer Corporation 108139-1
Agriculture Division
Table 4. Recovery of cyfluthrin from potato fresh tubers measured as part of the field
residue Program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Tubers BD107-00D-017-3 0.00010 BD107-00D-017-4 0.00908 90 95 3.4
0.01 BD105-00H-005-3 0.00016 BD105-00H-005-4 0.00957 94
BD107-00D-017-5 0.00060 BD107-00D-017-6 0.00996 94
BD107-00D-017-8 0.00000 BD107-00D-017-9 0.00968 97
BD106-00D-017-3 0.00002 BD106-00D-017-4 0.00971 97
BD106-00D-017-7 0.00004 BD106-00D-017-8 0.0100 100
1 Bayer Ag Div Report No. 110988. Percent recovery values were corrected for control
interferences.
49

Bayer Corporation 108139-1
Agriculture Division
Table 5. Recovery of cyfluthrin from orange fresh fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD132-00D-015-1 0.00006 BD132-00D-015-2 0.00944 94 99 2.7
0.01 BD134-00H-005-1 0.00000 BD134-00H-005-2 0.00980 98
BD132-00D-015-7 0.00035 BD132-00D-015-8 0.0101 98
BD133-00H-005-3 0.00000 BD133-00H-005-4 0.00988 99
BD132-00D-015-5 0.00031 BD132-00D-015-6 0.0102 99
BD132-00D-015-3 0.00000 BD132-00D-015-4 0.0102 102
BD133-00H-005-1 0.00025 BD133-00H-005-2 0.0105 102
0.10 BD134-00H-005-3 0.00022 BD134-00H-005-6 0.0970 97 97 0.58
BD134-00H-005-5 0.0972 97
BD134-00H-005-7 0.0980 98
1 Bayer Ag Div Report No. 111025. Percent recovery values were corrected for control
interferences.
50

Bayer Corporation 108139-1
Agriculture Division
Table 6. Recovery of cyfluthrin from lemon fresh fruit measured as part of the field
residue program (1)
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD152-00H-001-3 0.00050 BD152-00H-001-4 0.00934 88 94 5.4
0.01 BD153-00D-015-3 0.00018 BD153-00D-015-4 0.00915 90
BD153-00D-015-5 0.00023 BD153-00D-015-6 0.00944 92
BD153-00D-015-9 0.00036 BD153-00D-015-10 0.0102 99
BD154-00H-001-3 0.00000 BD154-00H-001-4 0.0100 100
0.10 BD154-00H-001-6 0.00189 BD154-00H-001-9 0.0939 92 96 3.8
BD154-00H-001-8 0.0996 98
BD154-00H-001-7 0.101 99
0.15 BD154-00H-001-12 0.00070 BD154-00H-001-14 0.141 94 98 3.5
BD154-00H-001-13 0.148 98
BD154-00H-001-15 0.153 101
1 Bayer Ag Div Report No. 111025. Percent recovery values were corrected for control
interferences.
51

Bayer Corporation 108139-1
Agriculture Division
Table 7. Recovery of cyfluthrin from grapefruit fresh fruit measured as part of the field
residue program (1)
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD148-00H-001-12 0.00040 BD148-00H-001-13 0.00943 90 94 3.7
0.01 BD146-00D-017-3 0.00052 BD146-00D-017-4 0.00955 90
BD148-00H-001-1 0.00087 BD148-00H-001-2 0.0103 94
BD147-00H-001-1 0.00080 BD147-00H-001-2 0.0104 96
BD147-00H-001-3 0.00039 BD147-00H-001-4 0.0101 97
BD146-00D-017-1 0.00050 BD146-00D-017-2 0.0104 99
0.10 BD148-00H-001-4 0.00035 BD148-00H-001-6 0.0981 98 98 0.0
BD148-00H-001-7 0.0983 98
BD148-00H-001-5 0.0985 98
1 Bayer Ag Div Report No. 111025. Percent recovery values were corrected for control
interferences.
52

Bayer Corporation 108139-1
Agriculture Division
Table 8. Recovery of cyfluthrin from tomato whole fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Whole Fruit BD001-00D-013-3 0.00052 BD001-00D-013-4 0.00962 91 93 1.6
0.01 BD002-00H-001-1 0.00043 BD002-00H-001-2 0.00962 92
BD003-00H-001-1 0.00020 BD003-00H-001-2 0.00954 93
BD002-00H-001-3 0.00021 BD002-00H-001-4 0.00963 94
BD001-00D-013-1 0.00028 BD001-00D-013-2 0.00976 95
0.50 BD002-00H-001-5 0.00014 BD002-00H-001-8 0.466 93 94 0.58
BD002-00H-001-9 0.468 94
BD002-00H-001-7 0.471 94
1 Bayer Ag Div Report No. 110981. Percent recovery values were corrected for control
interferences.
53

Bayer Corporation 108139-1
Agriculture Division
Table 9. Recovery of cyfluthrin from pepper whole fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Whole Fruit BD015-00D-013-1 0.00130 BD015-00D-013-2 0.00987 86 91 2.9
0.01 BD015-00D-013-3 0.00059 BD015-00D-013-4 0.00939 88
BD012-00H-001-1(2) 0.00071 BD012-00H-001-2(2) 0.00967 90
BD012-00H-001-1(2) 0.00067 BD012-00H-001-2(2) 0.00975 91
BD013-00H-001-1 0.00000 BD013-00H-001-2 0.00924 92
BD015-00D-013-5 0.00037 BD015-00D-013-6 0.00954 92
BD014-00H-001-1 0.00009 BD014-00H-001-2 0.00955 95
0.40 BD016-00H-001-2 0.00000 BD016-00H-001-4 0.405 101 103 1.7
BD016-00H-001-3 0.415 104
BD016-00H-001-5 0.415 104
1 Bayer Ag Div Report No. 110981. Percent recovery values were corrected for control
interferences.
2 Duplicate injections made to verify instrument performance
54

Bayer Corporation 108139-1
Agriculture Division
Table 10. Recovery of cyfluthrin from squash whole fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Whole Fruit BD040-00H-001-1 0.00007 BD040-00H-001-2 0.00964 96 98 1.3
0.01 BD038-00H-001-1 0.00000 BD038-00H-001-2 0.00969 97
BD037-00D-013-1 0.00000 BD037-00D-013-2 0.00975 97
BD036-00H-001-1 0.00012 BD036-00H-001-2 0.0100 99
BD039-00H-001-1 0.00023 BD039-00H-001-2 0.0101 99
0.080 BD037-00D-013-4 0.00045 BD037-00D-013-5 0.0775 96 98 1.5
BD037-00D-013-7 0.0785 98
BD037-00D-013-6 0.0794 99
1 Bayer Ag Div Report No. 110894. Percent recovery values were corrected for control
interferences.
55

Bayer Corporation 108139-1
Agriculture Division
Table 11. Recovery of cyfluthrin from cucumber whole fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Whole Fruit BD026-00HB-001-1 0.00089 BD026-00HB-001-2 0.00997 91 96 3.2
0.01 BD028-00H-001-1 0.00051 BD028-00H-001-2 0.00989 94
BD024-00H-001-1 0.00052 BD024-00H-001-2 0.0101 96
BD027-00H-001-1 0.00040 BD027-00H-001-2 0.0101 97
BD029-00H-001-1 0.00031 BD029-00H-001-2 0.0101 98
BD025-00D-013-1 0.00030 BD025-00D-013-2 0.0103 100
0.070 BD025-00D-013-4 0.00030 BD025-00D-013-5 0.0690 98 99 1.2
BD025-00D-013-7 0.0700 100
BD025-00D-013-6 0.0704 100
1 Bayer Ag Div Report No. 110894. Percent recovery values were corrected for control
interferences.
56

Bayer Corporation 108139-1
Agriculture Division
Table 12. Recovery of cyfluthrin from cantaloupe whole fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Whole Fruit BD034-00H-001-1 0.00038 BD034-00H-001-2 0.00936 90 95 2.7
0.01 BD032-00H-001-1 0.00041 BD032-00H-001-2 0.00974 93
BD030-00H-001-1 0.00046 BD030-00H-001-2 0.00983 94
BD033-00D-013-3 0.00045 BD033-00D-013-4 0.00988 94
BD031-00H-001-3 0.00039 BD031-00H-001-4 0.0100 96
BD033-00D-013-1 0.00022 BD033-00D-013-2 0.00993 97
BD031-00H-001-1 0.00026 BD031-00H-001-2 0.0100 98
0.050 BD034-00H-001-4 0.00024 BD034-00H-001-7 0.0481 96 97 1.0
BD034-00H-001-5 0.0487 97
BD034-00H-001-6 0.0491 98
1 Bayer Ag Div Report No. 110894. Percent recovery values were corrected for control
interferences.
57

Bayer Corporation 108139-1
Agriculture Division
Table 13. Recovery of cyfluthrin from carrots roots measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Roots BD121-00H-001-1 0.00072 BD121-00H-001-2 0.00994 92 96 3.8
0.01 BD122-00D-013-10 0.00015 BD122-00D-013-11 0.00960 94
BD122-00D-013-3 0.00006 BD122-00D-013-4 0.00968 96
BD122-00D-013-1 0.00016 BD122-00D-013-2 0.00984 97
BD123-00H-001-1 0.00000 BD123-00H-001-2 0.0102 102
0.050 BD122-00D-013-6 0.00004 BD122-00D-013-7 0.0471 94 95 1.5
BD122-00D-013-8 0.0476 95
BD122-00D-013-9 0.0485 97
1 Bayer Ag Div Report No. 110958. Percent recovery values were corrected for control
interferences.
58

Bayer Corporation 108139-1
Agriculture Division
Table 14. Recovery of cyfluthrin from radish roots and tops measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Roots BD129-00H-001-1 0.00090 BD129-00H-001-2 0.0100 91 97 6.0
0.010 BD127-00DA-013-1 0.00057 BD127-00DA-013-2 0.00983 93
BD127-00DA-013-7 0.00078 BD127-00DA-013-8 0.0103 95
BD128-00H-001-1 0.00000 BD128-00H-001-2 0.00972 97
BD127-00DA-013-5 0.00024 BD127-00DA-013-6 0.0100 98
BD127-00DA-013-3 0.00085 BD127-00DA-013-4 0.0116 108
0.025 BD128-00H-001-4 0.00019 BD128-00H-001-5 0.0242 96 98 1.5
BD128-00H-001-6 0.0247 98
BD128-00H-001-7 0.0250 99
Tops BD127-00DA-030-3 0.00288 BD127-00DA-030-4 0.0109 80 85 4.4
0.010 BD129-00H-006-1 0.00333 BD129-00H-006-2 0.0117 83
BD127-00DA-030-1 0.00418 BD127-00DA-030-2 0.0129 87
BD128-00H-006-1 0.00230 BD128-00H-006-2 0.0113 90
5.0 BD128-00H-006-4 0.00021 BD128-00H-006-6 4.22 84 85 0.58
BD128-00H-006-5 4.23 85
BD128-00H-006-7 4.23 85
1 Bayer Ag Div Report No. 110958. Percent recovery values were corrected for control
interferences.
59

Bayer Corporation 108139-1
Agriculture Division
Table 15. Recovery of cyfluthrin from head lettuce heads with wrapper leaves and heads
without wrapper leaves measured as part of the field residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Heads with BD064-00H-001-1 0.00079 BD064-00H-001-2 0.0103 95 99 4.0
wrapper leaves BD065-00D-013-1 0.00036 BD065-00D-013-2 0.0101 97
0.010 BD065-00D-013-3 0.00061 BD065-00D-013-4 0.0103 97
BD066-00H-001-1 0.00038 BD066-00H-001-2 0.0105 101
BD064-00H-001-3 0.00038 BD064-00H-001-4 0.0109 105
2.0 BD065-00D-013-6 0.00007 BD065-00D-013-8 1.95 98 98 0.0
BD065-00D-013-9 1.96 98
BD065-00D-013-7 1.97 98
Heads without BD064-00H-006-1 0.00062 BD064-00H-006-3 0.00971 93 96 2.4
wrapper leaves BD064-00H-006-2 0.00025 BD064-00H-006-4 0.00985 94
0.010 BD066-00H-006-1 0.00012 BD066-00H-006-2 0.00980 97
BD064-00H-006-5 0.00020 BD064-00H-006-6 0.00997 98
0.050 BD066-00H-006-4 0.00000 BD066-00H-006-7 0.0490 98 98 0.0
BD066-00H-006-6 0.0491 98
BD066-00H-006-5 0.0492 98
1 Bayer Ag Div Report No. 110628. Percent recovery values were corrected for control
interferences.
60

Bayer Corporation 108139-1
Agriculture Division
Table 16. Recovery of cyfluthrin from leaf lettuce fresh leaves measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh leaves BD060-00H-001-3 0.00032 BD060-00H-001-4 0.00970 94 95 1.5
0.010 BD060-00H-001-1 0.00046 BD060-00H-001-2 0.00982 94
BD058-00H-001-1 0.00136 BD058-00H-001-2 0.0108 94
BD059-00D-013-1 0.00057 BD059-00D-013-2 0.0103 97
2.5 BD059-00D-013-4 0.00079 BD059-00D-013-7 2.44 98 98 0.0
BD059-00D-013-6 2.45 98
BD059-00D-013-5 2.46 98
3.5 BD059-00D-013-8 0.00203 BD059-00D-013-10 3.41 98 98 0.58
BD059-00D-013-9 3.44 98
BD059-00D-013-11 3.46 99
1 Bayer Ag Div Report No. 110628. Percent recovery values were corrected for control
interferences.
61

Bayer Corporation 108139-1
Agriculture Division
Table 17. Recovery of cyfluthrin from grape fresh fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD010-01D-021-3 0.00052 BD010-01D-021-4 0.00983 93 97 2.0
0.01 BD010-01D-021-1 0.00021 BD010-01D-021-2 0.00965 94
BD002-01H-001-3 0.00016 BD002-01H-001-4 0.00975 96
BD004-01HA-001-1 0.00013 BD004-01HA-001-2 0.00976 96
BD006-01H-001-1 0.00028 BD006-01H-001-2 0.00990 96
BD003-01H-001-1 0.00008 BD003-01H-001-2 0.00978 97
BD011-01H001-11 0.00021 BD011-01H-001-12 0.00987 97
BD001-01H-001-1 0.00000 BD001-01H-001-2 0.00977 98
BD009-01H-001-1 0.00008 BD009-01H-001-2 0.00998 99
BD002-01H-001-1 0.00010 BD002-01H-001-2 0.0100 99
0.50 BD011-01H-001-3 0.00012 BD011-01H-001-4 0.487 97 98 1.0
BD011-01H-001-6 0.488 98
BD011-01H-001-5 0.496 99
1.0 BD011-01H-001-7 0.00013 BD011-01H-001-9 0.982 98 100 1.5
BD011-01H-001-8 0.995 100
BD011-01H-001-10 1.01 101
1 Bayer Ag Div Report No. 110989. Percent recovery values were corrected for control
interferences.
62

Bayer Corporation 108139-1
Agriculture Division
Table 18. Recovery of cyfluthrin from dried pea dry pea measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Dry pea BD085-00D-013-3(2) 0.00080 BD085-00D-013-4(2) 0.00865 78 85 5.3
BD085-00D-013-3(2) 0.00093 BD085-00D-013-4(2) 0.00875 78
0.01 BD087-00H-001-4 0.00047 BD087-00H-001-5 0.00919 87
BD088-00H-001-1(2) 0.00075 BD088-00H-001-2(2) 0.00950 88
BD085-00D-013-1(2) 0.00100 BD085-00D-013-2(2) 0.00969 87
BD085-00D-013-1(2) 0.00081 BD085-00D-013-2 0.00959 88
BD087-00H-001-1 0.00052 BD087-00H-001-2 0.00968 92
0.15 BD088-00H-001-3 0.00044 BD088-00H-001-4 0.132 88 89 0.58
BD088-00H-001-5 0.134 89
BD088-00H-001-6 0.134 89
1 Bayer Ag Div Report No. 110479. Percent recovery values were corrected for control
interferences.
2 Duplicate injections made to verify instrument performance
63

Bayer Corporation 108139-1
Agriculture Division
Table 19. Recovery of cyfluthrin from dried bean dry seed measured as part of the
field residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Dry seed BD018-00H-005-5 0.00187 BD018-00H-005-6 0.00923 74 84 5.9
0.01 BD019-00H-001-7 0.00129 BD019-00H-001-8 0.00952 82
BD020-00H-001-1 0.00138 BD020-00H-001-2 0.00959 82
BD019-00H-001-5 0.00113 BD019-00H-001-6 0.00945 83
BD019-00H-001-13 0.00079 BD019-00H-001-14 0.00972 89
BD019-00H-001-9 0.00086 BD019-00H-001-10 0.00976 89
BD019-00H-001-11 0.00058 BD019-00H-001-12 0.00968 91
0.15 BD019-00H-001-16 0.00062 BD019-00H-001-17 0.140 93 95 2.5
BD019-00H-001-19 0.143 95
BD019-00H-001-18 0.148 98
1 Bayer Ag Div Report No. 110479. Percent recovery values were corrected for control
interferences.
64

Bayer Corporation 108139-1
Agriculture Division
Table 20. Recovery of cyfluthrin from mustard green leaves measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Leaves BD055-00D-017-3 0.00156 BD055-00D-017-4 0.00981 82 93 6.4
0.01 BD054-00H-005-1 0.00066 BD054-00H-005-2 0.00979 91
BD201-00H-001-1 0.00056 BD201-00H-001-2 0.00995 94
BD053-00H-005-5 0.00082 BD053-00H-005-6 0.0110 98
BD055-00D-017-1 0.00052 BD055-00D-017-2 0.0104 99
4.0 BD055-00D-017-6 0.00116 BD055-00D-017-9 3.95 99 99 0.58
BD055-00D-017-7 3.97 99
BD055-00D-017-8 3.99 100
5.5 BD201-00H-001-4 0.00040 BD201-00H-001-7 5.27 96 96 0.0
BD201-00H-001-5 5.29 96
BD201-00H-001-6 5.30 96
1 Bayer Ag Div Report No. 110895. Percent recovery values were corrected for control
interferences.
65

Bayer Corporation 108139-1
Agriculture Division
Table 21. Recovery of cyfluthrin from broccoli flowering heads and stems measured as part
of the field residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Flowering heads BD043-00H-001-1 0.00071 BD043-00H-001-2 0.00986 92 95 3.4
and stems BD042-00D-013-1 0.00016 BD042-00D-013-2 0.00945 93
0.010 BD041-00H-001-2 BD041-00H-001-5 0.00992 94
BD041-00H-001-3 0.00029(2) BD041-00H-001-4 0.0103 98
BD043-00H-001-3 0.00000 BD043-00H-001-4 0.00997 100
0.75 BD042-00D-013-3 0.00015 BD042-00D-013-5 0.748 100 100 0.58
BD042-00D-013-4 0.750 100
BD042-00D-013-6 0.754 101
1 Bayer Ag Div Report No. 110339. Percent recovery values were corrected for control
interferences.
2 Average residue of two different extractions of control sample BD041-00H-001.
66

Bayer Corporation 108139-1
Agriculture Division
Table 22. Recovery of cyfluthrin from cabbage heads with wrapper leaves and heads
without wrapper leaves measured as part of the field residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Heads with BD047-00H-001-1 0.00079 BD047-00H-001-2 0.0101 93 99 4.7
wrapper leaves BD048-00D-013-1 0.00073 BD048-00D-013-2 0.0104 97
0.010 BD048-00D-013-3 0.00112 BD048-00D-013-4 0.0108 97
BD048-00D-013-5 0.00022 BD048-00D-013-6 0.0104 102
BD049-00H-001-1 0.00046 BD049-00H-001-2 0.0110 105
3.5 BD049-00H-001-4 0.00069 BD049-00H-001-6 3.53 101 101 0.58
BD049-00H-001-7 3.55 101
BD049-00H-001-5 3.56 102
Heads without BD047-00H-006-2 0.00089(2) BD047-00H-006-4 0.0107 99 99 0.58
Wrapper leaves BD047-00H-006-1 BD047-00H-006-3 0.0108 99
0.010 BD049-00H-006-1 0.00085 BD049-00H-006-2 0.0108 100
0.020 BD049-00H-006-4 0.00016 BD049-00H-006-7 0.0207 102 104 2.6
BD049-00H-006-6 0.0208 103
BD049-00H-006-5 0.215 107
1 Bayer Ag Div Report No. 110339. Percent recovery values were corrected for control
interferences.
67

Bayer Corporation 108139-1
Agriculture Division
Table 23. Recovery of cyfluthrin from celery trimmed leaf stalks and untrimmed leaf stalks
measured as part of the field residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Trimmed leaf BD070-00H-001-1 0.00149 BD070-00H-001-2 0.0108 93 98 3.1
stalks BD073-00H-001-1 0.00000 BD073-00H-001-2 0.00983 98
0.010 BD074-00H-001-1 0.00032 BD074-00H-001-2 0.0102 99
BD071-00H-001-1 0.00000 BD071-00H-001-2 0.0100 100
0.10 BD075-00H-001-7 0.00014 BD075-00H-001-10 0.0995 99 100 0.58
BD075-00H-001-8 0.0996 100
BD075-00H-001-9 0.0997 100
0.70 BD075-00H-001-2 0.00061 BD075-00H-001-4 0.711 101 102 1.0
BD075-00H-001-5 0.717 102
BD075-00H-001-3 0.724 103
1.0 BD075-00H-001-11 0.00000 BD075-00H-001-12 1.01 101 101 0.0
BD075-00H-001-13 1.01 101
BD075-00H-001-14 1.01 101
Untrimmed BD073-00H-006-1 0.00000 BD073-00H-006-2 0.00805 80 97 8.2
leaf stalks BD071-00H-006-1 0.00063 BD071-00H-006-2 0.00985 92
0.010 BD070-00H-006-1 0.00061 BD070-00H-006-2 0.0104 98
BD074-00H-006-1 0.00000 BD074-00H-006-2 0.0100 100
BD075-00H-006-1 0.00023 BD075-00H-006-2 0.0103 101
BD072-00D-013-1 0.00013 BD072-00D-013-2 0.0103 102
BD074-00H-006-3 0.00000 BD074-00H-006-4 0.0103 103
3.5 BD072-00D-013-6 0.00065 BD072-00D-013-8 3.51 100 101 0.58
BD072-00D-013-9 3.52 101
BD072-00D-013-7 3.54 101
1 Bayer Ag Div Report No. 110338. Percent recovery values were corrected for control
interferences.
68

Bayer Corporation 108139-1
Agriculture Division
Table 24. Recovery of cyfluthrin from spinach leaves measured as part of the field residue
Program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Leaves BD080-00D-013-1 0.00165 BD080-00D-013-2 0.0109 93 98 3.6
0.01 BD079-00H-001-1 0.00038 BD079-00H-001-2 0.00984 95
BD080-00D-013-4 0.00221 BD080-00D-013-5 0.0117 95
BD077-00H-001-1 0.00075 BD077-00H-001-2 0.0105 98
BD076-00H-001-1 0.00000 BD076-00H-001-2 0.0101 101
BD081-00H-001-1 0.00150 BD081-00H-001-2 0.0116 101
BD078-00H-001-1 0.00053 BD078-00H-001-2 0.0107 102
6.0 BD079-00H-001-3 0.00048 BD079-00H-001-6 5.45 91 91 0.58
BD079-00H-001-4 5.47 91
BD079-00H-001-5 5.50 92
1 Bayer Ag Div Report No. 110338. Percent recovery values were corrected for control
interferences.
69

Bayer Corporation 108139-1
Agriculture Division
Table 25. Recovery of cyfluthrin from tobacco whole green leaves and flue cured leaves
measured as part of the field residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Whole green BD196-00H-004-1 0.00055 BD196-00H-004-2 0.0101 96 98 2.1
leaves BD198-00H-004-1 0.00015 BD198-00H-004-2 0.0100 99
0.010 BD197-00H-004-1 0.00033 BD197-00H-004-2 0.0103 100
1.5 BD196-00H-004-4 0.00044 BD196-00H-004-5 1.43 95 96 0.58
BD196-00H-004-6 1.44 96
BD196-00H-004-7 1.44 96
Flue cured BD198-00H-001-1 0.00056 BD198-00H-001-2 0.00867 81 86 5.0
leaves BD196-00H-001-1 0.00037 BD196-00H-001-2 0.00885 85
0.010 BD197-00H-001-1 0.00020 BD197-00H-001-2 0.00926 91
7.5 BD196-00H-001-4 0.00017 BD196-00H-001-5 5.45 73 74 0.58
BD196-00H-001-6 5.53 74
BD196-00H-001-7 5.55 74
1 Bayer Ag Div Report No. 110340. Percent recovery values were corrected for control
interferences.
70

Bayer Corporation 108139-1
Agriculture Division
Table 26. Recovery of cyfluthrin from peanut nutmeat and hay measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Nutmeat BD091-00D-016-5 0.00022 BD091-00D-016-6 0.00777 76 89 8.4
0.010 BD092-00H-004-5 0.00073 BD092-00H-004-6 0.00837 76
BD092-00H-004-9 0.00055 BD092-00H-004-10 0.00954 90
BD099-00H-004-1 0.00024 BD099-00H-004-2 0.00940 92
BD093-00H-004-5 0.00027 BD093-00H-004-6 0.00954 93
BD102-00D-016-2 0.00000 BD102-00D-016-3 0.00942 94
BD092-00H-004-7 0.00061 BD092-00H-004-8 0.0101 95
BD096-00H-004-1 0.00010 BD096-00H-004-2 0.00978 97
Hay BD099-00H-001-1 0.00072 BD099-00H-001-2 0.00787 72 79 6.4
0.010 BD092-00H-001-5 0.00084 BD092-00H-001-6 0.00806 72
BD091-00D-007-3 0.00225 BD091-00D-007-4 0.0101 78
BD093-00H-001-3 0.00071 BD093-00H-001-4 0.00893 82
BD092-00H-001-3 0.00063 BD092-00H-001-4 0.00911 85
BD096-00H-001-3 0.00132 BD096-00H-001-4 0.00998 87
6.0 BD102-00D-007-7 0.00373 BD102-00D-007-8 4.32 72 73 0.58
BD102-00D-007-9 4.37 73
BD102-00D-007-10 4.37 73
6.5 BD102-00D-007-12 0.00314 BD102-00D-007-14 4.45 68 69 0.58
BD102-00D-007-13 4.46 69
BD102-00D-007-15 4.46 69
1 Bayer Ag Div Report No. 111036. Percent recovery values were corrected for control
interferences.
71

Bayer Corporation 108139-1
Agriculture Division
Table 27. Recovery of cyfluthrin from apple fresh fruit measured as part of the field
residue program (1).
Control Recovery Measured
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD048-99H-001-002 0.00304 BD048-99H-001-003 0.0128 97 100 3.0
0.010 BD050-99H-001-001 0.00261 BD050-99H-001-002 0.0126 100
BD049-99H-001-001 0.00251 BD049-99H-001-002 0.0128 103
0.050 BD050-99H-001-003 0.00139 BD050-99H-001-004 0.0454 88 106 13.9
BD043-99D-001-003 0.00111 BD043-99D-001-004 0.0460 90
BD043-99D-001-001 0.00072 BD043-99D-001-002 0.0461 91
BD049-99H-001-003 0.00083 BD049-99H-001-004 0.0497 98
BD048-99H-001-007 0.00358 BD048-99H-001-008 0.0602 113
BD047-99H-001-001 0.00367 BD047-99H-001-002 0.0617 116
BD051-99D-001-003 0.00429 BD051-99D-001-004(2) 0.0633 118
BD051-99D-001-003 0.00429 BD051-99D-001-004(2) 0.0647 121
BD046-99H-001-001 0.00312 BD046-99H-001-002 0.0624 119
1 Bayer Ag Div Report No. 110031. Percent recovery values were corrected for control
interferences.
2 Duplicate injection to verify instrument performance.
72

Bayer Corporation 108139-1
Agriculture Division
Table 28. Recovery of cyfluthrin from pear fresh fruit measured as part of the field
residue program (1).
Control Recovery
Measure
d
Fortification Sample Control Sample Recov Net % Avg % Std Dev
(ppm) Number (ppm) Number (ppm) Recov Recov % Recov
Fresh Fruit BD037-99D-001-001 0.00132(2) BD037-99D-001-004 0.00909 78 84 7.3
0.010 BD037-99D-001-002 BD037-99D-001-005 0.00948 82
BD037-99D-001-003 BD037-99D-001-006 0.00990 86
BD038-99H-001-001 0.00083 BD038-99H-001-002 0.00878 79
BD040-99H-001-001 0.00000 BD040-99H-001-002 0.00957 96
0.050 BD040-99H-001-003 0.00000 BD040-99H-001-004 0.0430 86 90 3.0
BD037-99D-001-011 0.00095 BD037-99D-001-012 0.0448 88
BD039-99H-001-001 0.00144 BD039-99H-001-002 0.0453 88
BD037-99D-001-009 0.00213 BD037-99D-001-010 0.0468 89
BD041-99H-001-001 0.00250 BD041-99H-001-002 0.0469 89
BD038-99H-001-003 0.00000 BD038-99H-001-004 0.0453 91
BD038-99H-001-003 0.00000 BD038-99H-001-006 0.0458 92
BD037-99D-001-011 0.00095 BD037-99D-001-013 0.0480 94
BD037-99D-001-011 0.00095 BD037-99D-001-014 0.0483 95
1 Bayer Ag Div Report No. 110031. Percent recovery values were corrected for control
interferences.
2 Average residue of three different extractions of control sample BD037-99D-001.
73

Bayer Corporation 108139-1
Agriculture Division
Table 29. LODs of Cyfluthrin from crop matrices.
Crop / Matrix LOD (ppm)
Cherry / fresh fruit 0.00070
Peach / fresh fruit 0.00081
Plum / fresh fruit 0.0017
Potato / fresh tuber 0.0011
Oranges / fresh fruit 0.00095
Lemon / fresh fruit 0.0017
Grapefruit / fresh fruit 0.0016
Tomato / whole fruit 0.00037
Pepper / whole fuit 0.00075
Squash / whole fruit 0.00051
Cucumber / whole fruit 0.00056
Cantaloupe / whole fruit 0.00058
Carrots / roots 0.00099
Radish / tops 0.0040
Radish / roots 0.0020
head lettuce / heads with wrapper leaves 0.0011
head lettuce / heads without wrapper leaves 0.00049
leaf lettuce / fresh leaves 0.0023
Grape / fresh fruit 0.00031
dried pea / dry pea 0.0016
dried beans / dry seed 0.00058
mustard green / leaves 0.0017
Broccoli / flowering heads and stems 0.0011
Cabbage / heads with wrapper leaves 0.0013
Cabbage / heads without wrapper leaves 0.00040
Celery / trimmed leaf stalks 0.0019
Celery / untrimmed leaf stalks 0.0026
Spinach / leaves 0.0022
Tobacco / whole green leaves 0.0011
Tobacco / flue cured leaves 0.0021
Peanut / nutmeat 0.0023
Peanut / hay 0.0031
Apple / fresh fruit 0.00080
Pear / fresh fruit 0.0016
74

Bayer Corporation 108139-1
Agriculture Division
Table 30. Cyfluthrin recovery ranges.
Fortification Recovery
Crop / Matrix
Range(ppm) Range(%)
Low High Lo High
Cherry / fresh fruit 0.01 0.5 91 103
Peach / fresh fruit 0.01 0.4 88 101
Plum / fresh fruit 0.01 0.1 86 109
Potato / fresh tuber 0.01 NA 90 100
Orange / fresh fruit 0.01 0.1 94 102
Lemon / fresh fruit 0.01 0.15 88 101
Grapefruit / fresh fruit 0.01 0.1 90 99
Tomato / whole fruit 0.01 0.5 91 95
Pepper / whole fuit 0.01 0.4 86 104
Squash / whole fruit 0.01 0.08 96 99
Cucumber / whole fruit 0.01 0.07 91 100
Cantaloupe / whole fruit 0.01 0.05 90 98
Carrots / roots 0.01 0.05 92 102
Radish / tops 0.01 5.0 80 90
Radish / roots 0.01 0.025 91 108
head lettuce / heads with wrapper leaves 0.01 2.0 95 105
head lettuce / heads without wrapper 0.01 0.05 93 98
leaf lettuce / fresh leaves 0.01 3.5 94 99
Grape / fresh fruit 0.01 1.0 93 101
dried pea / dry pea 0.01 0.15 78 92
dried beans / dry seed 0.01 0.15 74 98
mustard green / leaves 0.01 5.5 83 100
Broccoli / flowering heads and stems 0.01 0.75 92 101
Cabbage / heads with wrapper leaves 0.01 3.5 93 105
Cabbage / heads without wrapper leaves 0.01 0.02 99 107
Celery / trimmed leaf stalks 0.01 1.0 93 103
Celery / untrimmed leaf stalks 0.01 3.5 80 103
Spinach / leaves 0.01 6.0 91 102
Tobacco / whole green leaves 0.01 1.5 95 100
Tobacco / flue cured leaves 0.01 7.5 73 91
Peanut / nutmeat 0.01 NA 76 97
Peanut / hay 0.01 6.5 68 87
Apple / fresh fruit 0.01 0.05 88 121
Pear / fresh fruit 0.01 0.05 78 96
75

Bayer Corporation 108139-1
Agriculture Division
Cl
Cl
H
H
H
H C
O
3 CH3
Figure 1. Chemical name, acronymns, storage conditions, and structures for cyfluthrin and
[
76
2 H6]Cyfluthrin.
O
Cyfluthrin
Cyano-(4-fluoro-3-phonoxyphenyl)methyl-3-(2,2-dichloroethenyl)-2,2dimethylcyclopropanecarboxylate
Molecular Weight 434.30
Storage condition:frozen
Cl
Cl
H
H
H
CD
O
3 CD3
O
H
H
N
N
[ 2 H6]Cyfluthrin
Molecular Weight 440.33
Storage condition:frozen
F
O
F
O

Bayer Corporation 108139-1
Agriculture Division
- place 5.0 g sample material in a 120-mL glass bottle
- add 45 mL of MeOH/aqueous 1.2 N HCl (4:1)
Figure 2. Flow diagram of the residue analytical method for cyfluthrin in apple and pear
matrices
77
- homogenize with Tekmar Tissumizer
for approx 2 min
- vaccum filter through a 5.5-cm GF/A filter contained in a 5.6-cm Buchner funnel
- return the solid extract including the filter back into the glass bottle and
add 45 mL of MeOH followed by another 2 min of homogenization
- Vacuum filter through another GF/A filter
- add internal standard
- Dilute to 100-mL with MeOH/aqueous 1.2 N HCl (4:1)
- rotary evaporate 50 mL of the extract
to dryness
- reconstitute the residue in 25 mL
hexane followed by sonication
- prepare a florisil column by adding glass wool to the bottom of the column, 100 mL of
hexane, 7 g of 2.5% deactivated florisil and 6 g of anhydrous sodium sulfate
- drain excess hexane, leaving just enough hexane to cover the bed of sodium sulfate
- add the 25-mL sample in hexane to the column and allow to drip through the column into
a waste container
- wash the flask that contained the 25-mL hexane sample with 40 mL of hexane, add the
40 mL of hexane to the column and allow the 40 mL to drip through the column into a
waste container
- Elute the cyfluthrin with 60 mL of hexane/acetone (9:1) into a 125-mL boiling flask
- rotary evaporate to dryness
- reconstitute the sample in 2.5 mL of toluene
- ILOWHU DSSUR[ P/ RI WKH VDPSOH XVLQJ D P PP $FURGLVF ILOWHU
- analyze using GC/MS

Bayer Corporation 108139-1
Agriculture Division
- place 5.0 g sample material in a 4 oz bottle beaker
- add 45 mL of MeOH/aqueous 1.2 N HCl (4:1)
Figure 3. Flow diagram of the residue analytical method for cyfluthrin in all matrices except
apple, pear, and nutmeat.
78
- homogenize with Tekmar Tissumizer
for approx 2 min
- vaccum filter through a 5.5-cm GF/A filter contained in a 5.6-cm Buchner funnel
- return the solid extract including the filter back into the bottle beaker and
add 45 mL of MeOH followed by another 2 min of homogenization
- Vacuum filter through another GF/A filter
- add internal standard
- Dilute to 100-mL with MeOH
- concentrate to approx 4 to 5 mL
using a Turbo Vap LV
- dilute to 10 mL with water
- transfer the aqueous sample to a 125-mL separatory funnel
- wash the vial that contained the aqueous sample twice with 15 mL aliquots of
acetone/DCM (1:2) and add to the 125-mL separatory funnel.
- shake funnel for appprox 30 s and drain the lower phase through a funnel containing a
glasswool plug and approx 15 g of anhydrous sodium sulfate into a 60-mL vial
- wash the sodium sulfate with 10 mL of DCM and collect in the same 60-mL vial
- evaporate to dryness using a Turbo Vap LV and reconstitute the residue in 25-ml
of hexane
- prepare a florisil column by adding glass wool to the bottom of the column, 100 mL of
hexane, 7 g of 2.5% deactivated florisil and 3 g of anhydrous sodium sulfate
- drain excess hexane, leaving just enough hexane to cover the bed of sodium sulfate
- add the 25-mL sample in hexane to the column and allow to drip through the column into
a waste container
- wash the flask that contained the 25-mL hexane sample with 40 mL of hexane, add the
40 mL of hexane to the column and allow the 40 mL to drip through the column into a
waste container
- Elute the cyfluthrin with 40 mL of hexane/acetone (9:1) into a 125-mL boiling flask
- rotary evaporate to dryness
- reconstitute the residue in 2.5 mL of toluene and analyze on GC/MS

Bayer Corporation 108139-1
Agriculture Division
- place 5.0 g sample material into a paper thimble for the soxtec
- add 50 mL of hexane to a soxtec stainless steel beaker
- place the soxtec beaker on the soxtec and extract for 2 h at 140 ° C in the boil position
- move the soxtec to the rinse position for 1 h
- turn off the temp to the soxtec and allow the cups to cool
- add the internal standard to the cup
- transfer the sample to a 50-mL graduated cylinder
- wash the cup with 5 mL of hexane and add to the 50-mL cylinder
- dilute the sample to 50-mL with hexane
- transfer 25-mL of the sample from the 50-mL graduated cylinder into a 125-mL sep funnel
- add 25 mL of ACN presaturated with hexane to the sep funnel and shake for 30 seconds
- Drain the lower ACN phase into another 125-mL sep funnel already containing 10 mL of
hexane presaturated with ACN and shake for 30 s
- collect the lower ACN phase in a 60-mL vial
- repeat the the above procedure by adding another 25 mL of ACN presaturated with hexane
to the 1 st separatory funnel
- Concentrate the ACN phases contained in the 60-mL vial to dryness on a Turbo Vap LV
- prepare a florisil column by adding glass wool to the bottom of the column, 100 mL of
hexane, 7 g of 2.5% deactivated florisil and 3 g of anhydrous sodium sulfate
- drain excess hexane, leaving just enough hexane to cover the bed of sodium sulfate
- add the 25-mL sample in hexane to the column and allow to drip through the column into
a waste container
- wash the flask that contained the 25-mL hexane sample with 40 mL of hexane, add the
40 mL of hexane to the column and allow the 40 mL to drip through the column into a
waste container
- Elute the cyfluthrin with 40 mL of hexane/acetone (9:1) into a 125-mL boiling flask
- rotary evaporate to dryness
- reconstitute the residue in 2.5 mL of toluene and analyze on GC/MS
Figure 4. Flow diagram of the residue analytical method for cyfluthrin in nutmeat
79

Bayer Corporation 108139-1
Agriculture Division
Standard Solution of cyfluthrin and [ 2 H6]Cyfluthrin from cherry fresh fruit
[ 2 H6]Cyfluthrin
(m/z 213)
Cyfluthrin
(m/z 207)
Figure 5. GC/MS chromatogram of a typical 0.05 ppm cyfluthrin standard / 0.05 ppm
[ 2 H6]cyfluthrin standard solution.
80