### J.R. Simplot Company Compositional Analyses 1 

### Appendix 9

Compositional Analyses

INDEX

Section Page

Index 1

Introduction 2

Results 3

Materials and M ethods 70

Conclusions 7 9

References 8 3

Certification 8 7

Annex A: Combined Lit erature Range Supplement 8 8

### J.R. Simplot Company Compositional Analyses 2 

### INTRODUCTION

The field

- grown tubers of both events and untransformed control varieties were analyzed to determine their composition al equivalence. S ignificant differences in the amount of a compound between an event and control were considered important only if the mean values were outside the normal range

. In order to determine if values fell within the normal range

, m ean values were compared with tolerance intervals that were calculated from commercial control varietie s and with values found in the scientific literature.

### J.R. Simplot Company Compositional Analyses 3 

### RESULTS

Compositional A nalysis of Ranger Russet Events and Controls

Data on the biochemical composition of Ranger Russet events and controls, which represent the mean,

range, p

- value s and /or adjus ted p

- values , are shown in Tables 1 (key proximates, vitamins, and minerals), 2 (free amino acids), 3 (total amino acids), 4 (glycoalkaloids)

, 5 ( sugars), and 6 (acrylamide in fried product)

.

### J.R. Simplot Company Compositional Analyses 4 

### Table 1 : Ranger Russet Proximates, Vitamins, and Minerals 1

Co mpound TI CLR 2

Control Ranger N= 33 F10

N= 33 F37

N= 30 Protein (%) Mean 2.384 2.382 2.429 1.258

- 3.594 0.70

- 4.60

Range 1.67

- 3.2 2 1.78

- 3.52 1.68

- 3. 30

APV 0. 4416 0.8202

Fat (%) Mean 0.122 4 0.126 3 0.134 1 0.000

- 0.341 0.02

- 0.20

Range 0.03

- 0.3 0 0.08

- 0.3 0 0.06

- 0.3 0

APV 0. 4667 0.758 5

Ash (%) Mean 1.103 1.140 1.115 0.391

- 1.888 0.44

- 1.90

Range 0.14

- 1.54 0 .00

- 1.67 0.33

- 1.5 2

APV 0. 9861 1.0000

Crude Fiber (%) Mean 0.528 2 0.536 1 0.548 3 0.142

- 0.690 0.17

- 3.50

Range 0.27

- 0.74 0.33

- 0.7 6 0.40

- 0.73

APV 0.6080 0.7904
Carbohydrates (%) Mean 20.24 20.75 21.00 12.29

- 25.92 13.30

- 30.53

Range 14.8 0

- 23.8 0 16.83

- 24 .00 16.94

- 25.6 0

APV 0. 0812 0.211 2

Calories (kcal/ 100g) Mean 91.41 93.4 8 94.7 6 59.09

- 114. 9 70.00

- 110.2

Range 67. 4

- 104 .0 76.8 0

- 104. 3 80.4 0

- 111 .0

APV 0. 0637 0.2070

Moisture (%) Mean 76.17 75.6 3 75.34 70.28

- 83.9 1 63.20

- 86.90

Range 74 .00

- 81.7 0 72.55

- 79.4 0 71.9 0

- 78.73

APV 0. 0632 0.27 40

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adj usted P

- value , B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper. Adjusted P

- values indicating significant differences with controls are underlined.
2 Literature range s are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992), Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 5 

### Table 1 (Continued) : Ranger Russet Proximates, Vitamins, and Minerals 1
Compound TI CLR 2

Control Ranger N=33 F10

N=33 F37

N= 30
B 3 (mg/ 100g) Mean 2.41 5 2. 500 2.698 0.922

- 3.123 0.09

- 3.10

Range 1.43

- 3.43 1.90

- 3.43 2 .00

- 3. 70

APV 0. 0006 0.0077

B 6 (mg/ 100g) Mean 0.1348 0.135 2 0.1299 0.059

- 0.192 0.13

- 0.41

Range 0.11

- 0.16 0.10

- 0.22 0.10

- 0.18

APV 0.2953 0.6397

Vitamin C (mg/ 100g) Mean 25.87 30.6 8 29.9 5 0.000

- 129. 3 1.00

- 54.00

Range 11.36

- 51.4 0 16.16

- 56.5 0 16.15

- 59 .00

APV 0.0 039 0.0254

Cu (ppm) Mean 1.1637 1.12 10 1.1182 0.111

- 2.236 0.15

- 7.00

Range 0.75

- 1.85 0.64

- 1.6 1 0.73

- 1.6 0

APV 0. 4992 0.721 1

Mg (ppm) Mean 284. 8 285. 7 287.8 101. 5

- 371.5 112.5

- 550.0

Range 204. 6

- 405. 6 207.6

- 408.2 204. 4

- 420. 1

APV 0.6687 0.7903

K (ppm) Mean 498 2 508 8 5069 271 1

- 6882 3500.

- 6250

Range 3 670

- 688 5 3900

- 7432 3770

- 7049

APV 0. 9992 0.9992

1 TI = Toler ance Interval, CLR = Combined Literature Range, APV = adjusted P

- value , B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper , ppm = parts per million

. Adjusted P

- values indicating significant differences with controls are underlined.

2 Literature ranges are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992 ), Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 6 

### Table 2 : Ran ger Russet Free Amino Acids ( parts pe r million) 1
Free Amino Acid (ppm) TI 2 CLR 3
Control Ranger N=32 F10

N=33 F37

N=33
ALA Mean 108. 9 134. 4 154. 8 100.0

- 208. 2 12 .00

- 236 .0

Range 100 .0

- 182 .0 100.0

- 248.0 100 .0

- 305 .0

APV NA NA
ARG Mean 707. 3 94 8.0 1011 100.0

- 106 8 120 .0

- 1472

Ran ge 352 .0

- 1218 332.5

- 1740 381 .0

- 1885

APV 0.1692 0.0470
ASN Mean 2351 567. 2 642.5 500.0

- 4952 3 12 .0

- 6980

Range 1209

- 3530 381. 8

- 872 .0 355 .0

- 1100

APV 0.0 266 0.0 206

ASP Mean 558. 5 600. 2 624. 9 100.0

- 1411 64 .00

- 752 .0

Range 266 .0

- 119 7 242 .0

- 122 5 303 .0

- 127 9

APV 0.1490 0.0077
GLN Mean 958.6 166 8 171 5 100.0

- 344 9 440 .0

- 5396

Range 399. 1

- 1648 409 .0

- 2860 718 .0

- 2610

APV <0.0001 <0.0001

GLU Mean 557. 4 652. 4 657.0 100.0

- 119 2 450 .0

- 742 .0

Range 252. 7

- 877.9 302.9

- 883. 4 301.5

- 966 .0

APV 0.0 896 0. 0632

GLY Mean 100 .0 100.5 100. 1 100.0

- 116. 1 2 .00

- 104 .0

Range 100 .0

- 100 .0 100 .0

- 114.4 100.0

- 103. 4

APV NA NA
HIS Mean 101. 1 101. 8 103. 1 100.0

- 136. 9 34 .00

- 656 .0

Range 100 .0

- 126 .0 100 .0

- 125 .0 100 .0

- 138 .0

APV NA NA
ILE Mean 129

. 4 120.3 142.0 100.0

- 217.8 28 .00

- 330 .0

Range 100 .0

- 223 .0 100 .0

- 172. 4 100 .0

- 226. 8

APV NA NA
LEU Mean 100. 7 100.7 100.2 100.0

- 173.9 16 .00

- 262 .0

Range 100 .0

- 120. 9 100 .0

- 123.8 100 .0

- 104 .0

APV NA NA

1 Free amino acids included to show efficac y of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit ( 100 ppm) for statistical analysis

. NA = Adjusted p

- value not determined because observations near the detection limits ar e too l ow to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.
2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

. GLN high level reported as 5396 ppm from the mean of 4 sites ( Davies 1977 ) omitting a level of 18,244 ppm from a single site, which appeared to be an outlier.

### J.R. Simplot Company Compositional Analyses 7 

### Table 2 (Continued) : Ran ger Russet Free Amino Acids ( parts per million) 1
Free Amino Acid (ppm) TI 2 CLR 3
Control Ranger N =32 F10

N=33 F37

N=33
LYS Mean 175. 3 180.1 195.3 100.0

- 557. 3 18 .00

- 638 .0

Range 100 .0

- 435. 3 100 .0

- 372.8 100 .0

- 400. 1

APV 0.8 18 6 0.2167

MET Mean 114. 5 114. 5 118.9 100.0

- 191. 6 18 .00

- 216 .0

Range 100 .0

- 222 .0 100 .0

- 178 .0 100 .0

- 208 .0

APV NA NA

PHE Mean 168. 6 158. 2 174. 1 100.0

- 294. 9 0 .00

- 408 .0

Range 100 .0

- 248 .0 100 .0

- 294.7 100 .0

- 250 .0

APV 0.5366 0.7848

PRO Mean 169. 8 204. 8 277.3 100.0

- 630.5 0 .0 0

- 968 .0

Range 100 .0

- 458.6 100 .0

- 747. 8 100 .0

- 762 .0

APV NA NA
SER Mean 113.4 122.9 137. 7 100.0

- 198. 1 30 .00

- 256 .0

Range 100 .0

- 178 .0 100 .0

- 218 .0 100 .0

- 237 .0

APV NA NA
THR Mean 120. 2 135.5 144. 8 100.0

- 219.8 28 .00

- 286 .0

Range 100 .0

- 186.0 10 0 .0

- 244. 7 100 .0

- 239. 1

APV NA NA
TRP Mean 100 .0 100 .0 100 .0 100.0

- 113. 1 14 .00

- 348 .0

Range 100 .0

- 100 .0 100 .0

- 100 .0 100 .0

- 100 .0

APV NA NA
TYR Mean 103. 1 110.6 115. 2 100.0

- 225.2 18 .00

- 638 .0

Range 100 .0

- 140 .0 100 .0

- 198 .0 100 .0

- 188 .0

APV 0.5223 0.8052

VAL Mean 236. 1 236. 4 281.3 100.0

- 460. 6 30.00

- 740.0

Range 125. 6

- 476 .0 117.5

- 416 .0 100 .0

- 497 .0

APV 0.9486 0. 0681

1 Free amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell bel ow detection limits and were adjusted to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- va lues indicating significant differences with controls are underlined.
2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

.

### J.R. Simplot Company Compositional Analyses 8 

### Table 3 : Rang er Russet Total Amino Acids ( parts per milllion) 1

Total Amino Acid (ppm) TI CLR 2 Control Ranger N=32 F10

N=33 F37

N=33
ALA Mean 661.3 715. 1 729.3 100.0

- 1335 392 .0

- 952 .0

Range 448.3

- 1090 389. 8

- 1179 473. 3

- 956.3
APV 0.5063 0.4379

ARG M ean 117 4 143 9 149 5 367.6

- 192 2 700 .0

- 1383

Range 765.4

- 234 4 71 1.0

- 229 7 955. 9

- 276 7
APV 0.1291 0.0719

ASP+ASN Mean 551 8 304 4 312 1 100.0

- 13340 3385

- 7380

Range 2080

- 848 6 1960

- 5572 2010

- 457 1
APV 0.0165 0.0182

GLU+GLN Mean 314 4 43 6 8 43 80 568.4

- 7011 2915

- 6035

Range 237 8

- 422 3 2600

- 6070 2760

- 5690
APV <0.0001 <0.0001

GLY Mean 146 8 1607 147 2 100.0

- 2865 500 .0

- 1990

Range 462 .0

- 308 8 484 .0

- 3284 463 .0

- 345 7
APV 0.7382 0.9446

HIS Mean 424.0 420. 3 466. 5 100.0

- 7 60. 7 133 .0

- 469 .0

Range 100 .0

- 887. 8 208.9

- 851.5 145. 2

- 1034
APV 0.9038 0.1934

ILE Mean 717.5 771.7 755. 7 144.8

- 1346 525 .0

- 953 .0

Range 380.0

- 961.2 431.8

- 1186 50 7.0

- 1016
APV 0.3851 0.5198

LEU Mean 138 6 1460 1455 476.9

- 217 4 68 5 .0

- 1383

Range 911 .0

- 2103 939 .0

- 2119 920 .0

- 222 5
APV 0.4719 0.4808

LYS Mean 992.4 1137 1125 100.0

- 3363 687 .0

- 1368

Range 745. 5

- 1177 809. 4

- 1808 868 .0

- 1616
APV 0.1320 0.1763

MET Mean 371.0 395. 2 402. 2 100.0

- 593. 3 300 .0

- 500 .0

Range 140. 2

- 836.4 218.5

- 744.7 214.6

- 682. 1
APV 0.2895 0.2189

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined.
2 Literature ran ges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 9 

### Table 3 (Continued) : Rang er Russet Total Amino Acids ( parts per milllion) 1
Total Amino Acid (ppm) TI CLR 2 Control Ranger N=32 F10

N=33 F37

N=33
PHE Mean 857.9 850.3 874.3 397.1

- 1395 552 .0

- 1087

Range 597. 7

- 120 8 64 6.0

- 1262 637. 6

- 158 9
APV 0.8401 0.6817

PRO Mean 809.4 886.1 919.9 100.0

- 1492 355 .0

- 1464

Range 564 .0

- 1473 567. 7

- 1747 574 .0

- 1861
APV 0.4608 0.2788

SER Mean 66 9. 0 727.5 693. 4 100.0

- 136 2 500 .0

- 1022

Range 318. 7

- 140 3 293.8

- 1205 345.3

- 154 2
APV 0.0996 0.5270

THR Mean 682.5 755. 5 739.6 226.0

- 1315 436 .0

- 855 .0

Range 279.4

- 1070 379.0

- 146 5 445. 4

- 1287
APV 0.0848 0.1662

TRP Mean 216.8 217.9 2 14. 3 114.7

- 434.7 14.00

- 348.0

Range 192 .0

- 243 .0 174 .0

- 272 .0 179 .0

- 261 .0
APV 0.8646 0.7703

TYR Mean 590. 5 642. 4 649. 9 100.0

- 152 3 457 .0

- 942 .0

Range 317.3

- 760 .0 469.3

- 933 .0 458.5

- 91 5.0
APV 0.0887 0.0542

VAL Mean 100 7 103 1 101 4 1 00.0

- 1860 752 .0

- 1450

Range 559.9

- 151 2 656.2

- 1657 639.4

- 131 9
APV 0.8817 0.9906

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underli ned. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.
2 Literature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 10 

### Table 4 : R anger Russet Glycoalkaloids ( mg/100 g) 1
Glycoalkaloids (mg/100g) TI CLR 2

Control Ranger N=32 F10

N=33 F37

N=33
Mean 6.70 4 6.97 9 6.805 0.000

- 33.0 5 3.2 0

– 210.4
Range 3.06

- 10.23 2.84

- 9.51 4.28

- 9.42

P

- value 0.8285 0.9812

1 TI = Toleran ce Interval, CLR = Combined Literature Range

, P

- values indicating significant differences with controls are underlined.

2 Literature ranges from Kozukue et al. (2008).

### J.R. Simplot Company Compositional Analyses 11 

### Table 5: R anger Russet Sugars (percentage fresh weight) 1

Fructose + Glucose (%)
Ranger Control F10 F37 TI 1 CLR 2
Fructose + Glucose Fresh Mean 0.185 0.18 2 0.178

0.000

- 0.307 0.00

- 0.80

Range 0.047

- 0.362 0.040

- 0.437 0.034

- 0.374

P

- Value 0.9924 0.9613

N 12 12 12 Fructose + Glucose Month 1 Mean 0.086 0. 0 69 0.0 57

0.000

- 0.307 0.00

- 0.80

Range 0.043

- 0.141 0.036

- 0.110 0.038

- 0.075

P

- Value 0.0243 0.0009

N 21 21 17 Fructose + Glucose Month 3 Mean 0.270 0.245 0.286

0.000

- 0.307 0.00

- 0.80

Range 0.073

- 0.450 0.068

- 0.550 0.082

- 0.552

P

- Value 0.6579 0.8986

N 27 27 24 Fructose + Glucose Month 5 Mean 0.190 0.178 0.224

0.000

- 0.307 0.00

- 0.80

Range 0.083

- 0.412 0.073

- 0.358 0.069

- 0.602

P

- Value 0.9696 0.9571

N 21 21 18 Sucrose (%)
Ranger Control F10 F37 TI CLR

Sucrose Fresh Mean 0.188 0.174 0.188

0.000

- 0.315 0.05

- 0.32

Range 0.082

- 0.293 0.080

- 0.252 0.043

- 0.284

P

- Value 0.7892 0.9999

N 12 12 12
Sucrose Month 1 Mean 0.137 0.136 0.138

0.000

- 0.315 0.05

- 0.32

Range 0.094

- 0.179 0.073

- 0.186 0.077

- 0.183

P

- Value 0.9327 0.9826

N 21 21 17
Sucrose Month 3 Mean 0.061 0.063 0.056

0.000

- 0.315 0.05

- 0.32

Range 0.009

- 0.127 0.018

- 0.121 0.014

- 0.122

P

- Value 0.7420 0.9032

N 27 27 24
Sucrose Month 5 Mean 0.068 0.067 0.055

0.000

- 0.315 0.05

- 0.32

Range 0.008

- 0.129 0.009

- 0.135 0.008

- 0.150

P

- Value 0.9367 0.7556

N 21 21 18

1 TI = Tolerance Interval, CLR = Combined Literature Range

, P

- values indicating significant differenc es with controls are underlined.
2 

### Literature ranges from Blenkinsop et al. ( 2002 ) ; Matsuura

- Endo et al. ( 2006 ); Menendez et al. ( 2002 ).

### J.R. Simplot Company Compositional Analyses 12 

### Table 6 : Ranger Russet, Acrylamide L evels in Processed Material ( parts per billion) 1

Acrylamide (ppb) Ranger Control F10 F37

Fresh Mean 37 1.9 15 0

. 6 127

. 3

2010 & 2011 Range 175

- 513 98

- 236 62

- 242

P

- value

— <0.0001 <0.0001

N 36 36 36

Month 2 Mean 588.4 207.7 210.3

2009 Range 436

- 688 135

- 254 164

- 254

P

- value

— <0.0001 <0.0001

N 9 9 6

Month 3 Mean 630.6 237.9 221. 8

2010 & 201 1 Range 366

- 1010 132

- 370 120

- 374

P

- value

— <0.0001 <0.0001

N 23 24 24

Month 5 Mean 148.0 70.8 81.7

2009 Range 124

- 196 54

- 83 78

- 88

P

- value

— <0.0001 <0.0001

N 9 9 6

Month 6 Mean 460.4 144.3 180.8

2010 Range 171

- 714 77

- 195 112

- 299

P

- value

— <0.0001 <0.0001

N 12 12 12

Month 7 Mean 443.0 145.7 161.3

2009 Range 359

- 497 130

- 173 146

- 190

P

- value

— <0.0001 <0.0001

N 9 9 6

1 P

- values indicating significant differences with controls are underlined.

### J.R. Simplot Company Compositional Analyses 13 

### Compositional Analysis of Event F10

Tubers of event F10 were equivalent to those of untransformed Ranger Russet except for the following expected changes :

 F10 tubers contained, on average, 5 67 parts per million (ppm) free ASN as compared to controls, which contained 2351 ppm ( APV = 0. 026 6 )

. This change was expected and is linked to a reduced acrylamide

- forming potential.

 F10 tubers contained, on average

, 1 668 ppm free GLN as compared to controls, which contained
959 ppm ( APV = <0.0001 )

. The increase in this amide amino acid counter

- balan ces, in part, the
losses in the other amide amino acid ASN. The new levels are still normal (which means, within the normal range) for potato.

 F10 tubers contained, on average, 3044 ppm total ASN+ASP as compared to controls, which
contained 5518 ppm ( APV = 0.0 165 ). The new level fits within the tolerance interval and is still normal for potato.

 F10 tubers contained, on average, 43 68 ppm total GLN+GLU as compared to controls, which
contained 31 44 ppm ( APV <0.0001 )

. The new levels are still normal for potato.

 F10 tubers at 1 month after harvest contained, on average, 0.0 69 % of the reducing sugars glucose plus fructose as compared to controls, which contained 0.0 86 % ( P

- value = 0.0243). The decrease in reducing sugars was expected from silencing the promoters as sociated with the R1
and PhL genes. The new levels are still normal for potato.

 French fries from potato strips from fresh F10 potato tubers contained 150.6 ppb acrylamide as compared to the control, which had 37 1.9 ppb ( P

- value <0. 0001). This change was i ntended.

 The impact on nutritional quality would be minimal from the observed differences in free ASN,
free GLN, total ASN + ASP, and total GLN + GLU because as non

- essential amino acids they can be synthesized by the body. The observed decrease in reducin g sugars falls within the normal range for potatoes. However, the combination of lower ASN and reducing sugars contribute to the reduced potential of F10 to form acrylamide and would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in light of recent toxicol ogy studies (NTP 201 2 ).

The following changes were observed :

 F10 tubers contained, on average, 2.5 mg/100 g vitamin B3 as compared to control, which
contained 2.4 mg/100 g (APV = 0. 0006 ). The new levels are still normal for potato.

 F10 tubers contained, on average, 30 .7 mg/ 100 g vitamin C as compared to control, which
contained 25

. 9 mg/100 g ( APV = 0. 0039 ). The new levels are still normal for potato.

### J.R. Simplot Company Compositional Analyses 14



### Th ese change s result in a very small modific ation in the vitamin composition of Ranger Russet F10 that is still standard for potato and will not significantly alter potential dietary intake levels. In a ddition , the levels were within the normal range for potatoes.

It can be concluded that tubers of F10 are substantially equivalent to untransformed Ranger Russet controls except for: (1) lowered free ASN, (2) higher levels of free GLN, (3) lower levels of total ASP + ASN, (4) higher levels of total GLN + GLU, (5) the expected decreased levels of the re ducing sugars glucose plus fructose, (6) reduced acrylamide as expected and intended, and (7) slightly increased levels
of vitamin B3 and vitamin C . In all cases, these compositional changes fall within the normal range for
potatoes.

### J.R. Simplot Company Compositional Analyses 15 

### Compositional Analys is of Event F37

Tubers of event F 37 were equivalent to those of untransformed Ranger Russet except for the following expected changes :

 F 37 tubers contained, on average, 643 parts per million (ppm) free ASN as compared to controls, which contained 2351 ppm ( APV = 0.0 206 ). This change was expected and is linked to a reduced acrylamide

- forming potential.

 F37 tubers contained, on average, 625 parts per million (ppm) free A SP as compared to controls,
which contained 559 ppm (APV = 0.0 077 ). These levels were wit hin the normal range for
potatoes.

 F 37 tubers contained, on average, 1715 ppm free GLN as compared to controls, which contained
959 ppm ( APV <0.0001 ). The increase in this amide amino acid counter

- balances, in part, the
losses in the other amide amino acid ASN. The new levels are still normal for potato.

 F37 tubers contained, on average, 3121 ppm total ASN + ASP as compared to controls, which
contained 5518 ppm ( APV = 0.0 182 ). The new level fits within the tolerance interval and is still normal for potato.

 F37 tubers contained, on average, 4380 ppm total GLN + GLU as compared to controls, which contained 3144 ppm (APV <0.0001). The new levels are still normal for potato.

 F37 tubers at 1 month after harvest contained, on averag e, 0.0 57 % of the reducing sugars glucose plus fructose as compared to controls, which contained 0.0 86 % (P

- value = 0.0009). The decrease in reducing sugars was expected from silencing the promoters associated with the R1
and PhL genes. The new levels are still normal for potato.

 French fr ies from potato strips from fresh F37 potato tubers contained 127.3 ppb acrylamide as compared to the control, which had 37 1.9 ppb (P

- value <0.0001). This change was intended.

 The im pact on nutritional quality would be minimal from the observed differences in free ASN

, ASP, GLN, total ASN + ASP, and total GLN + GLU because as non

- essential amino acids they can be synthesized by the body. The observed de crease in reducing sugars falls within the normal range for potatoes. However, lower ASN and reducing suga rs contributes to the reduced potential of F37 to form acrylamid e and would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in light of recent toxicology studies (NTP 201 2 ).

The following chang es were observed :

 F37 tubers contained, on average, 1011 parts per million (ppm) free A RG as compared to controls, which contained 707 ppm (APV = 0.0 470 ). These levels were within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 16



###  F37 tubers contained, on average, 29.9 mg/100 g vitamin C as compared to controls, which contained 25.9 mg/100 g (APV = 0.0254). The new levels are still normal for potato.

 F37 tubers contained, on average, 2.70 mg/100 g niacin (vitamin B 3 

### ) as compared to controls, which contained 2.42 mg/100 g (APV = 0.0077). The new levels are still normal for potato.

These changes result in very small modifications in the vitamin composition of Ranger Russet F37 that are still standard for potato and will not significantly alter potential dietary intake levels.

It can be concluded that tubers of F37 are substantially equivalent to untransformed Ranger Russet controls except for : (1) lowered free ASN, (2) higher levels of free ASP and free GLN, (3) lower levels of total ASP + ASN, (4) higher levels of total GLU + GLN , (5) de creased levels of the reducing sugars glucose plus fructose, (6) reduced acrylamide as expected and intended, (7) slightly increase d levels of vitamin C and niacin (vitamin B 3 

### )

, and (8) increased levels of free ARG

. In al l cases, these compositiona l changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 17 

### Compositional Analysis of Russet Burbank Events and Controls

Data on the biochemical composition of Russet Burbank events and controls, which represent the mean,

range, p

- value s and /or adjusted p

- valu es , are shown in Tables 7 (key proximates, vitamins, and minerals), 8 (free amino acids ), 9 (total amino acids), 10 (glycoalkaloids)

, 11 ( sugars), and 12 (acrylamide in fried product).

### J.R. Simplot Company Compositional Analyses 18 

### Table 7 : Russet Burbank Proximates, Vitamins, and Minerals 1

Compound TI CLR 2

Control Burbank
N=33 E12

N=33 E24

N=33
Protein (%) Mean 2.317 2.29 8 2.289 1.258

- 3.594 0.70

- 4.60

Range 1.64

- 3.10 1.47

- 3.02 1.56

- 3.04

APV 1.0000 0.7732

Fat (%) Mean 0. 100 0.101 0.10 8 0.000

- 0.341 0.02

- 0.20

Range 0.03

- 0.20 0.05

- 0.20 0.05

- 0.20
APV NA NA

Ash (%) Mean 1.07 9 1.030 1.052 0.391

- 1.888 0.44

- 1.90

Range 0.15

- 2.00 0.00

- 1.46 0.56

- 1.42
APV 1.0000 0.8379

Crude Fiber (%) Mean 0.4 70 0.46 7 0.456 0.142

- 0.690 0.17

- 3.50

Range 0.32

- 0.64 0.33

- 0.70 0.32

- 0.69
APV 1.0000 1.0000

Carbohydrates (%) Mean 18.49 18. 50 18.27 12.2 9

- 25.92 13.30

- 30.53

Range 16.10

- 21.10 14.20

- 22.20 14.80

- 22.00
APV 0.9806 0.7676

Calories (kcal/100g) Mean 84.00 83.90 82.9 8 59.09

- 114. 9 70.00

- 110.2

Range 73.10

- 93.26 64.90

- 96.30 68.4 0

- 95.80
APV 1.0000 0.7774

Moisture (%) Mean 78.02 78.0 9 78.3 1 70.28

- 83.9 1 63.20

- 86.90

Range 75.51

- 80.40 75.50

- 82.50 75.04

- 81.70

APV 1.0000 0.7775

B3 (mg/100g) Mean 1.865 1.90 1 1.84 7 0.922

- 3.123 0.09

- 3.10

Range 1.44

- 2.38 1.43

- 2.53 1.46

- 2 .37
APV 1.0000 0.8855

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value, B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper

. A djusted P

- values indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.
2 Literature ranges are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992 ), Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 19 

### Table 7 (Continued) : Russet Burbank Proximates, Vitamins, and Minerals 1
Compound TI CLR 2

Control Burbank
N=33 E12

N=33 E24

N=33
B6 (mg/ 100g) Mean 0. 143 0.142 0.144 0.059

- 0.192 0.13

- 0.41

Range 0.11

- 0.20 0.11

- 0.23 0.1 0

- 0.22
APV 1.000 0. 8778

Vitamin C (mg/ 100g) Mean 17.14 18.7 6 18.7 5 0.000

- 129. 3 1.00

- 54.00

Range 8.88

- 42.90 11.84

- 32.90 11.06

- 33.80
APV 1.0000 0.9118

Cu (ppm) Mean 0.9 6 5 1.00 2 0.91 8 0.111

- 2.236 0.15

- 7.00

Range 0.50

- 1.34 0.54

- 1.52 0.56

- 1.50
APV 1.0000 1.0000

Mg (ppm) Mean 241.7 246. 4 242.0 101.5

- 371. 5 112.5

- 550.0

Range 165.0

- 350.2 187. 6

- 40 5.0 179.3

- 360.1
APV 1.0000 0.9658

K (ppm) Mean 4681 477 7 4577 2711

- 6882 3500

- 6250

Range 3040

- 633 9 3550

- 6537 3280

- 6537
APV 1.0000 1.0000

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value, B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vi tamin C, Mg = magnesium, K = potassium, Cu = copper , ppm = parts per million

. Adjusted P

- values indicating significant differences with controls are underlined.
2 Literature ranges are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992 ), Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 20 

### Table 8 : Russ et Burbank Free Amino Acids ( parts per million) 1
Free Amino Acid (ppm) TI 2 CLR 3 Control Burbank N=33 E12

N=33 E24

N=33
ALA Mean 156.4 189. 4 157. 7 100.0

- 208. 2 12 .00

- 236 .0

Range 100.0

- 332. 7 100.0

- 354. 7 100.0

- 298. 1

APV NA NA

ARG Mean 519.6 525. 2 484. 8 100.0

- 106 8 120 .0

- 1472

Range 257.1

- 833. 2 242.5

- 912. 9 261.9

- 747.0

APV 0.8671 0.5349

ASN Mean 2421 618 .2 591. 9 500.0

- 4952 3 12 .0

- 6890

Range 822.3

- 4030 425. 2

- 1250 416.0

- 980.0

APV 0.1148 0.1088

ASP Mean 438.9 447.5 451.7 100.0

- 1411 64 .00

- 752 .0

Range 280.3

- 760. 1 293. 4

- 823. 9 280.5

- 743. 8

APV 0.6551 0.5938

GLN Mean 111 1 199 7 1979 100.0

- 344 9 440 .00

- 5396

Range 605. 4

- 1700 865. 2

- 3490 885.0

- 3220

APV 0. 0767 0.0 844

GLU Mean 465. 7 459.9 453.0 100.0

- 119 2 450 .0

- 742 .0

Range 221.1

- 688.9 227.3

- 741 .0 240. 8

- 721 .0

APV 0.9548 0.6847

1 Free amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limit s and were adjusted to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating si gnificant differences with controls are underlined.

2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

. GLN high level reported as 5396 ppm fr om the mean of 4 sites (Davies 1977) omitting a level of 18,244 ppm from a single site, which appeared to be an outlier.

### J.R. Simplot Company Compositional Analyses 21 

### Table 8 (Continued) : Russ et Burbank Free Amino Acids ( parts per million) 1
Free Amino Acid (ppm) TI 2 CLR 3

Control Burbank N=33 E12

N=33 E24

N=33
GLY Mean 100.1 102.2 101.2 100.0

- 116. 1 2 .00

- 104 .0

Range 100.0

- 104.2 100.0

- 12 7.0 100.0

- 120. 7

APV NA NA

HIS Mean 107. 6 107.5 104.9 100.0

- 136. 9 34 .00

- 656 .0

Range 100.0

- 157.0 100.0

- 170.0 100.0

- 143.0

APV NA NA

ILE Mean 152 .0 157. 6 140. 2 100.0

- 217. 8 28 .00

- 330 .0

Range 100.0

- 276.0 100.0

- 303. 3 100.0

- 279.2

APV 0.8093 0. 5534

LEU Mean 145. 7 140.4 124. 7 100.0

- 17 3.9 16 .00

- 262 .0

Range 100 .0

- 267. 1 100 .0

- 302.3 100 .0

- 211.5

APV NA NA

LYS Mean 227. 6 210.0 188. 1 100.0

- 5 57.3 18 .00

- 638 .0

Range 100.0

- 384. 6 100.0

- 407.6 100.0

- 286.0

APV 0. 7610 0. 2799

MET Mean 159. 7 174. 8 161.7 100.0

- 191.6 18 .00

- 216 .0

Range 105.3

- 233.0 103.4

- 251.0 116.9

- 226.0

APV 0. 4689 0.8393

PHE Mean 213. 7 196.1 173.6 100.0

- 294.9 0 .00

- 408

. 0

Range 100 .0

- 359 .0 100 .0

- 354 .0 100 .0

- 283.6

APV 0. 6358 0. 170 8

1 Free amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for st atistical analysis. NA = Adjusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underline d.

2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

.

### J.R. Simplot Company Compositional Analyses 22 

### Table 8 (Continued) : Russ et Burbank Free Amino Acids ( parts per million) 1

Free Amino A cid (ppm) TI 2 CLR 3

Control Burbank N=33 E12

N=33 E24

N=33
PRO Mean 144. 2 159.8 134. 4 100.0

- 630.5 0 .00

- 968 .0

Range 100.0

- 313.7 100.0

- 354.2 100.0

- 258.3

APV NA NA

SER Mean 135. 9 137.0 13 8.0 100.0

- 198.1 30 .00

- 256 .0

Range 100.0

- 277.0 100.0

- 282.0 100.0

- 251.0

APV NA NA

THR Mean 124. 2 14 4.0 136. 9 100.0

- 219.8 28 .00

- 286 .0

Range 100.0

- 194. 7 100.0

- 243. 3 100.0

- 213.4

APV NA NA

TRP Mean 100.5 100.5 100.0 100.0

- 113. 1 14 .00

- 348 .0

Range 100.0

- 111.5 100.0

- 109. 9 100.0

- 100.0

APV N A NA

TYR Mean 152.5 181. 2 167.3 100.0

- 225.2 18 .00

- 638 .0

Range 100.0

- 336.5 100.0

- 290.0 100.0

- 289.9

APV NA NA

VAL Mean 359.5 380. 7 347.0 100.0

- 460. 6 30 .00

- 740 .0

Range 212. 5

- 578.0 186.6

- 624.0 196.3

- 505.0

APV 0. 3722 0. 5272

1 Free amino ac ids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- value not determined because observations near th e detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.
2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

.

### J.R. Simplot Company Compositional Analyses 23 

### Table 9 : Russe t Burbank Total Amino Acids ( parts per million) 1
Total Amino Acid (ppm) TI CLR 2

Control Burbank
N=33 E12

N=33 E24

N=33
ALA Mean 703. 8 718.2 675.4 10 0.0

- 1335 392 .0

- 952 .0

Range 323.0

- 125 8 429. 5

- 105 3 370. 6

- 105 2

APV 0.7852 0.7574

ARG Mean 945.1 10 80 969.4 367.6

- 192 2 700 .0

- 1383

Range 508.0

- 1731 490. 3

- 2159 530.3

- 1498

APV 0.5836 0.7770

ASP+ ASN Mean 5391 2846 2747 100.0

- 1334 0 3385

- 7380

Range 2090

- 822 8 1874

- 5114 1786

- 4708

APV 0.0126 0.010 8

GLU+ GLN Mean 3051 4843 461 5 568.4

- 7011 2915

- 6035

Range 1840

- 398 2 3517

- 737 6 346 2

- 6450

APV 0.0153 0.027 9

GLY Mean 1459 1329 1328 100.0

- 2865 500 .0

- 1990

Range 307.0

- 354 8 489.0

- 288 5 468.0

- 28 60

APV 0.7093 0.7987

HIS Mean 372. 9 425. 9 415. 9 100.0

- 760. 7 133 .0

- 469 .0

Range 100.0

- 761.5 170.4

- 990.5 17 8.0

- 760.2

APV 0.5621 0.9012

ILE Mean 687. 2 705. 2 723. 6 144.8

- 1346 525 .0

- 953 .0

Range 372.0

- 911. 7 405.5

- 126 6 448. 1

- 963. 9

APV 0.8494 0.7007

LEU Mean 122 8 1266 1251 476.9

- 217 4 685 .0

- 1383

Range 530 .0

- 1783 803.7

- 202 4 594. 6

- 195 4

APV 0.7951 0.815 4

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value

. Adjus ted P

- values indicating significant differences with controls are underlined.

2 Literature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 24 

### Table 9 (Continued) : Russe t Burbank Total Amino Acids ( parts per mi llion) 1
Total Amino Acid (ppm) TI CLR 2

Control Burbank
N=33 E12

N=33 E24

N=33
LYS Mean 893. 9 1111 1034 100.0

- 3363 687 .0

- 1368

Range 560.0

- 130 4 763.3

- 179 5 74 1.0

- 135 6

APV 0.3594 0.6609

MET Mean 367.5 418. 5 39 8.0 100.0

- 593. 3 300 .0

- 500 .0

Range 185. 6

- 616.7 228.3

- 826. 4 146.6

- 702. 1

APV 0.6368 0.8296

PHE Mean 787. 1 798.6 763. 5 39 7.1

- 1395 552 .0

- 1087

Range 417.0

- 11 50 541. 5

- 124 7 453. 4

- 1050

APV 0.8665 0.8266

PRO Mean 718. 5 767. 8 71 1.0 100.0

- 1492 355 .0

- 1464

Range 321.0

- 141 5 487.0

- 1425 481.0

- 12 30

APV 0.5091 0.8145

SER Mean 547.6 575.1 587.6 100.0

- 136 2 500.0

- 1022

Range 214.0

- 817.0 219.5

- 984.6 303.7

- 965.0

APV 0.7322 0.7240

THR Mean 646.2 658.5 680.9 226.0

- 1315 500 .0

- 1022

Range 21 9 .8

- 1330.6 2 98.8

- 1346.6 3 30.5

- 1271.2

APV 0. 8247 0. 7064

TRP Mean 199.8 204.4 212.6 114.7

- 434.7 436 .0

- 855 .0

Range 182

- 224 163

- 237 181

- 256

APV 0. 7656 0. 4623

TYR Mean 562.4 620.1 592.9 100.0

- 1523 457 .0

- 942 .0

Range 327.9

- 792.2 266. 3

- 9 89.2 254. 5

- 977.0

APV 0.7383 0.8032

VAL Mean 1176 107 6 104 3 100.0

- 1860 752 .0

- 1450

Range 564.0

- 201 2 612.0

- 151 9 61 9

. 4

- 15 20

APV 0. 7857 0. 9682

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.

2 Literature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 25 

### Table 10 : Ru sset Burbank Glycoalkaloids ( mg/100 g) 1
Glycoalkaloids (mg/100g) TI CLR 2

Control Burbank
N=33 E12

N=33 E24

N=33
Mean 7.40 4 5.67 8 6.845 0.000

- 33.0 5 3.2 0

– 210.4

Range 2. 74

- 18.59 1.69

- 10.53 1.45

- 33.81

P

- value 0.1707 0.8174

1 TI = Tolerance Interval

, CLR = Combined Literature Range

. P

- values indicating significant differences with controls are underlined.
2 Literature ranges from Kozukue et al. (2008).

### J.R. Simplot Company Compositional Analyses 26 

### Table 11: Rus set Burbank Sugars (percent fresh weight) 1

Fructose + Glucose (%)

Burbank Control E12 E24 TI CLR 2

Fructose + Glucose Fresh Mean 0.195 0.160 0.160

0.000

- 0.307 0.00

- 0.80 Range 0.053

- 0.357 0.069

- 0.339 0.048

- 0.297

P

- Value 0.1326 0.2298

N 12 12 12
Fructose + Glucose Month
1 Mean 0.162 0.107 0.115

0.000

- 0.307 0.00

- 0.80 Range 0.053

- 0.393 0.034

- 0.251 0.022

- 0.275

P

- Value 0.0006 0.0019

N 21 21 21
Fructose + Glucose Month 3 Mean 0.290 0.249 0.276

0.000

- 0.307 0.00

- 0.80 Rang e 0.117

- 0.660 0.081

- 0.580 0.090

- 0.653

P

- Value 0.4164 0.7803

N 27 27 27
Fructose + Glucose Month
5 Mean 0.190 0.145 0.184

0.000

- 0.307 0.00

- 0.80 Range 0.092

- 0.630 0.085

- 0.489 0.069

- 0.556

P

- Value 0.3090 0.4307

N 21 21 21

Sucros e (%)

Burbank Control E12 E24 TI CLR

Sucrose Fresh Mean 0.179 0.213 0.178

0.000

- 0.315 0.05

- 0.32 Range 0.113

- 0.288 0.134

- 0.299 0.116

- 0.259

P

- Value 0.1284 0.9641

N 12 12 12
Sucrose Month 1 Mean 0.168 0.132 0.133

0.000

- 0.315 0.05

- 0.32 Range 0.103

- 0.252 0.060

- 0.258 0.065

- 0.2461

P

- Value 0.0024 0.0028

N 21 21 21
Sucrose Month 3 Mean 0.079 0.073 0.067

0.000

- 0.315 0.05

- 0.32 Range 0.047

- 0.121 0.034

- 0.126 0.040

- 0.112

P

- V alue 0.5819 0.1565

N 27 27 27
Sucrose Month 5 Mean 0.079 0.088 0.076

0.000

- 0.315 0.05

- 0.32 Range 0.035

- 0.127 0.051

- 0.141 0.036

- 0.143

P

- Value 0.6869 0.9047

N 21 21 21

1 TI = Tolerance Interval, CLR = Combined Litera ture Range. P

- values indicating significant differences with controls are underlined.

2 Literature ranges from Blenkinsop et al. (2002); Matsuura

- Endo et al. (2006); Menendez et al. (2002).

### J.R. Simplot Company Compositional Analyses 27



### Table 1 2 : Russet Burbank Acrylamide Levels in Processed Product 1

Acrylamide (ppb) Burbank Control E12 E24

Fresh Mean 49 3.8 1 62.4 1 58.9

2010 & 201 1 Range 286

- 770 95

- 298 63

- 333

P

- value

— <0.0001 <0.0001

N 36 36 36

Month 2 Mean 778.9 237.2 280.4

2009 Range 619

- 950 209

- 251 219

- 389

P

- value

— <0.0001 <0.0001

N 9 9 9

Month 3 Mean 814.3 314.5 329.8

2010 & 201 1 Range 502

- 1240 169

- 524 171

- 495

P

- value

— <0.0001 <0.0001

N 24 24 24

Month 5 Mean 230.4 82.0 93.8

2009 Range 207

- 260 67

- 97 79

- 109

P

- value

— <0.0001 <0.0001

N 9 9 9

Month 6 Mean 401.3 191.5 220.3

20 10 Range 234

- 776 105

- 272 150

- 321

P

- value

— <0.0001 <0.0001

N 12 12 12

Month 7 Mean 661.0 192 .0 202.7

2009 Range 598

- 693 165

- 217 187

- 221

P

- value

— <0.0001 <0.0001

N 9 9 9

1 P

- values indicating significant differences with controls are underlined.

### J.R. Simplot Company Compositional Analyses 28 

### Compositional Analysis of Event E12

Tubers of the event E12 were equivalent to those of untransformed Russet Burbank except for the following expected changes :

 E12 tubers contained, on average, 618 parts per million (ppm) free ASN as compared to
control s, which contained 2421 ppm ( APV = 0. 1148 ). This change was not statistically significant, although linked to a reduced acrylamide

- forming potential.

 E12 tubers contained, on average, 2846 ppm total ASN + ASP as compared to controls, which
contained 5391 p pm ( APV = 0.0 126 ). The new level fits within the tolerance interval and is still normal for potato.

 E12 tubers contained, on average, 4 843 ppm total GLN + GLU as compared to controls, which
contained 3051 ppm ( APV = 0.0 153 ). The new levels are still normal for potato.

 E12 tubers at 1 month after harvest contained, on average, 0.107% of the reducing sugars glucose plus fructose as compared to controls, which contained 0.162% (P

- value = 0.0006). The decrease in reducing sugars was expected from silencing the promoters associated with the R1
and PhL genes. The new levels are still normal for potato.

 French fries from potato strips from fresh E12 potato tubers contained 162.4 ppb acrylamide as compared to control, which had 493.8 ppb ( P

- value <0.0001). This chan ge was intended.

 The impact on nutritional quality would be minimal from the observed differences in free ASN

, total ASN + ASP, and total GLN + GLU because as non

- essential amino acids they can be synthesized by the body. The observed decrease in reducing sugars falls within the normal range for potatoes. However, the combination of lower ASN and reducing sugars contribute to the reduced potential of E12 to form acrylamide and would benefit consumers by addressing a potentially critical food safety issue fo r the potato industry , particularly in light of recent toxicology studies (NTP 201 2 ).

The following change was observed :

 E12 tubers at 1 month after harvest contained, on average, 0.132% of sucrose as compared to controls, which contained 0.168% (P

- value = 0.0024). The new levels are still normal for potato.

This change results in a very small modification in the composition of Russet Burbank E12 that is still standard for potato.

It can be concluded that tubers of E12 are substantially equivalent to untra nsformed Russet Burbank except for: (1) lowered free ASN

, (2) lower levels of total ASP + ASN, (3) higher levels of total GLU + GLN, (4) the expected decreased levels of the reducing sugars glucose plus fructose, (5) reduced

sucrose, and ( 6 ) reduced acryla mid e as expected and intended. In all cases, these compositional changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 29 

### Compositional Analysis of Russet Burbank and Event E24

Tubers of event E24 were equivalent to those of untransformed Russet Burbank excep t for the following expected changes :

 E24 tubers contained, on average, 592 parts per million (ppm) free ASN as compared to controls, which contained 2421 ppm ( APV = 0. 1088 ). This change was not statistically significant

, although linked to a reduced acryl amide

- forming potential

.

 E24 tubers contained, on average, 2747 ppm total ASN + ASP as compared to controls, which
contained 5391 ppm ( APV = 0.01 08 ). The new level fits within the tolerance interval and is still normal for potato.

 E24 tubers contained, on average, 46 1 5 ppm total GLN + GLU as compared to controls, which
contained 3051 ppm ( APV = 0.0 279 ). The new levels are still normal for potato.

 E24 tubers at 1 month after harvest contained, on average, 0.115% of the reducing sugars glucose plus fructose a s compared to controls, which contained 0.162% (P

- value = 0.0019). The decrease in reducing sugars was expected from silencing the promoters associated with the R1
and PhL genes. The new levels are still normal for potato.

 French fries from potato strips f rom fresh E24 potato tubers contained 158.9 ppb acrylamide as compared to controls, which had 493.8 ppb (P

- value <0.0001). This change was intended.

 The impact on nutritional quality would be minimal from the observed differences in free ASN

, total ASN + A SP, and total GLN + GLU because as non

- essential amino acids they can be synthesized by the body. The observed decrease in reducing sugars falls within the normal range for potatoes. However, the combination of lower ASN and reducing sugars contribute to t he reduced potential of E24 to for acrylamide and would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in light of recent toxicology studies (NTP 201 2 ).

The following change was observed :

 E24 tubers at 1 month after harvest contained, on average, 0.133% of sucrose as compared to controls, which contained 0.168% (P

- value = 0.0028). The new levels are still normal for potato.

This change results in a very small modification in the composition of Russet Burbank E24 that is still standard for potato.

It can be concluded that tubers of E24 are substantially equivalent to untransformed Russet Burbank except for: (1) lowered free ASN , (2) lower levels of total ASP + ASN, (3) higher levels of total GLU + GLN, (4) the expected decreased levels of the reducing sugars glucose plus fructose, (5) reduced sucrose, and ( 6 ) reduced acrylamide as expected and intended. In all cases, these compositional changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 30 

### Composi tional Analysis of Atlantic Events and Controls

Data on the biochemical composition of Atlantic events and controls, which represent the mean, range,

p

- value and adjusted p

- values, are shown in Tables 1 3 (key proximates, vitamins, and minerals), 1 4 (free

a mino acids), 1 5 (total amino acids), 1 6 (glycoalkaloids)

, 17 (sugars), and 1 8 (acrylamide in fried p otato chips )

.

### J.R. Simplot Company Compositional Analyses 31 

### Table 1 3 : Atlantic Proximates, Vitamins, and Minerals 1
Compound TI CLR 2

Control Atlantic
N=47 J3

N=47 J55

N=47 J78

N=47 Protein (%) Mean 2.313 2.300 2.259 2.391 1.258

- 3.594 0.70

- 4.60

Range 1.58

- 2.93 1.56

- 3.32 1.41

- 2.86 1.6 0

- 3.14

APV 0.9028 0.5659 0.6000

Fat (%) Mean 0.131 0.122 0.116 0.129 0.000

- 0.341 0.02

- 0.20

Range 0.04

- 0.6 0 0.02

- 0.5 0 0.02

- 0.4 0 0.02

- 0.4 0

APV NA NA NA

Ash(%) Mean 1.216 1.227 1.257 1.268 0.391

- 1.888 0.44

- 1.90

Range 0.49

- 1.59 0.1 0

- 2.04 0.63

- 2.12 0.48

- 1.92

APV 0.7664 0.5720 0.7144

Crude Fiber (%) Mean 0.344 0.342 0.342 0.339 0.142

- 0.690 0.17

- 3.50

Range 0.216

- 0.436 0.215

- 0.457 0. 212

- 0.46 0.157

- 0.447

APV 0.8415 0.8608 0.7465

Carbohydrates (%) Mean 20.3 9 20.6 6 20.50 20.1 4 12.29

- 25.92 13.30

- 30.53

Range 17.1 0

- 24.3 0 17.4 0

- 25.5 0 17.4 0

- 25.6 0 16.2 0

- 25.4 0

APV 1.0000 0.9540 0.623 0

Calories (kcal/ 100g) Mean 91.71 92.7 2 91. 90 91.1 1 59.09

- 114. 9 70.00

- 110.2

Range 80.2 0

- 107.7 78.3 0

- 112.1 78.5 0

- 110.8 75.9 0

- 110.4

APV 1.0000 0.9012 0.8409

Moisture (%) Mean 75.9 9 75.7 3 75.90 76.10 70.28

- 83.9 1 63.20

- 86.90

Range 72 .00

- 79.4 0 70.9 0

- 79.5 0 71.1 0

- 79.3 0 71.4 0

- 80.2 0

APV 1.0 000 0.9237 0.7296

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value, B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper. Adjusted P

- values indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.
2 Literature ranges are from , Lisinska and Leszczynsk i (1989), Rogan et al. (2000)

, Horton and Anderson (1992), Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 32 

### Table 1 3 (Continued) : Atlantic Proximates, Vitamins, and Minerals 1
Compound TI CLR 2

Control Atlantic

N=47 J3

N=47 J55

N=47 J78

N=47
B 3 (mg/ 100g) Mean 1.935 1.959 2.002 2.020 0.922

- 3.123 0.09

- 3.10

Range 1.4 0

- 2.65 1.39

- 2.81 1.5 0

- 2.82 1.25

- 3.38

APV 0.8217 0.5292 0.6738

B 6 (mg/ 100g) Mean 0.124 0.117 0.117 0.127 0.059

- 0.192 0.13

- 0.41

Range 0.09

- 0.16 0.09

- 0.16 0.09

- 0.1 6 0.09

- 0.17

APV 0.0024 0.001 2 1.0000

Vitamin C (mg/ 100g) Mean 20.66 21.31 21.9 1 22.16 0.000

- 129. 3 1.00

- 54.00

Range 9.22

- 38 8.11

- 35.3 10.94

- 34.2 8.65

- 36.1

APV 0.9353 0.5178 1.0000

Cu (ppm) Mean 1.128 1.064 1.012 1.081 0.111

- 2.236 0.15

- 7.00

Range 0.67

- 1.78 0.51

- 1.8 2 0 .50

- 1.91 0.66

- 1.76

APV 0. 8378 0. 3012 0.8204

Mg (ppm) Mean 220.0 216.7 212.8 221.8 101.5

- 371. 5 112.50

- 550.00

Range 158.0

- 291.0 105.0

- 292.0 122.0

- 284.0 152.0

- 306.0

APV 0.8033 0.4692 0.7984

K (ppm) Mean 4870 4819 490 3 4962 2711

- 6882 3500 .00

- 6250 .00

Range 3680

- 6650 2442

- 6951 2766

- 6515 3710

- 6680

APV 0.8675 0.9839 0.8779

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value, B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic a cid = vitamin C, Mg = magnesium, K = potassium, Cu = copper. Adjusted P

- values indicating significant differences with controls are underlined.

2 Literature ranges are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992)

, Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 33 

### Table 1 4 : Atlantic Free Amino Acids ( parts per million) 1

Free Amino Acid (ppm) TI 2 CLR 3

Control Atlantic
N= 47 J3

N= 47 J55

N= 47 J78

N= 47

ALA Mean 10 3.0 103.5 102.7 103.7 100.0

- 208. 2 12 .00

- 236 .0

Range 100.0

- 21 4.0 100.0

- 2 64. 8 100.0

- 202. 2 100.0

- 244. 6

APV NA NA NA

ARG Mean 439.2 489.6 454. 5 523.4 100.0

- 106 8 120 .0

- 1472

Range 157. 8

- 798. 3 157. 2

- 817. 0 100.0

- 771.5 100.0

- 907.0

APV 1.0000 1.0000 0.7597

ASN Mean 226 8 516. 2 502.4 551.1 500.0

- 4952 3 12 .0

- 6980

Range 300.0

- 3710 203.0

- 992.0 187.0

- 802.0 234.0

- 1870

APV 0.0260 0.0499 0.0548

ASP Mean 55 4.0 525. 3 517.8 561.3 100.0

- 1411 64 .00

- 752 .0

Range 235.0

- 197 4 239.0

- 192 8 224.0

- 2002 252.0

- 2206

APV 0.7536 0.6056 1.0000

GLN Mean 1194 1879 193 4 193 2 100.0

- 344 9 440 .0

- 5396

Range 425.0

- 350 4 605.0

- 3686 565.0

- 412 8 582.0

- 426 8

APV 0.0772 0.0654 0.0969

GLU Mean 728. 2 762. 3 748.8 754. 6 100.0

- 119 2 450 .0

- 752 .0

Range 344 .0

- 1357 317.2

- 123 8 334.4

- 1428 331. 2

- 1486

APV 1.0000 1.0000 1.0000

GLY Mean 100.9 100. 7 100.5 100. 9 100.0

- 116. 1 2 .00

- 140 .0

Range 100.0

- 157. 9 100.0

- 138.9 100.0

- 128. 9 100.0

- 139. 8

APV NA NA NA

HIS Mean 102. 4 100. 8 102. 4 100. 5 1 00.0

- 136. 9 34 .0 0

- 656 .0

Range 100 .0

- 153 .0 100 .0

- 130 .0 100 .0

- 152 .0 100 .0

- 120 .0

APV NA NA NA

1 Fr ee amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- value not determined because observati ons near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.

2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

. GLN high level reported as 5396 ppm from the mean of 4 sites (Davies 1977) omitting a level of 18,244 ppm from a single site, which appeared to be an outlier.

### J.R. Simplot Company Compositional Analyses 34 

### Table 1 4 ( Continued ) : Atlantic Free Amino Acids ( parts per million) 1

Free Amino Acid (ppm) TI 2 CLR 3

Control Atlantic
N= 47 J3

N= 47 J55

N= 47 J78

N= 47

ILE Mean 103. 4 100.1 101. 1 101. 4 100.0

- 217. 8 28 .00

- 330 .0

Range 100.0

- 164. 6 100.0

- 108. 5 100.0

- 125.9 1 00.0

- 169. 1

APV NA NA NA

LEU Mean 100.0 100.0 100.0 100.0 100.0

- 173.9 16 .00

- 262 .0

Range 100.0

- 100.0 100.0

- 100.0 100.0

- 100.0 100.0

- 100.0

APV NA NA NA

LYS Mean 115.5 107.3 109.6 110.7 100.0

- 557. 3 18 .00

- 638 .0

Range 100.0

- 295.9 100.0

- 189.1 1 00.0

- 235. 4 100.0

- 297. 7

APV NA NA NA

MET Mean 113.4 109. 5 110.7 109. 1 100.0

- 191. 6 18 .00

- 216 .0

Range 100. 0

- 170.0 100.0

- 163.0 100.0

- 175.0 100.0

- 156.0

APV NA NA NA

PHE Mean 103. 4 100. 4 100. 5 100.4 100.0

- 294. 9 0 .00

- 408 .0

Range 100.0

- 151. 7 10 0 .0

- 133. 2 100 .0

- 112 .0 100 .0

- 121. 5

APV NA NA NA

PRO Mean 240.3 248.5 250.3 256.0 100.0

- 630.5 0 .00

- 968 .0

Range 100.0

- 1090.0 100.0

- 904.0 100.0

- 974.0 100.0

- 1220

APV NA NA NA

SER Mean 106.0 101. 4 103.6 105.0 100.0

- 198. 1 30 .00

- 256 .0

Range 100 .0

- 166.0 100 .0

- 123 .0 100 .0

- 198 .0 100 .0

- 179.0

APV NA NA NA

1 Free amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for statisti cal analysis. NA = Adjusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.

2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

.

### J.R. Simplot Company Compositional Analyses 35 

### Table 1 4 ( Continued ) : Atlantic Free Amino Acids ( parts per million) 1

Free Amino Acid (ppm) TI 2 CLR 3

Control Atlantic
N= 47 J3

N= 47 J55

N= 47 J78

N= 47
THR Mean 106. 5 106. 6 107. 5 108.8 100.0

- 219.8 28 .00

- 286 .00

Range 100.0

- 241. 7 100.0

- 233. 5 100.0

- 323. 7 100.0

- 339.0

APV NA NA NA

TRP Mean 100.7 100.6 100.4 100.3 100.0

- 113. 1 14 .00

- 348 .00

Range 100.0

- 171.0 100.0

- 135.0 100.0

- 124.0 100.0

- 131.0

APV NA NA NA

TYR Mean 101. 9 100.0 100.9 100.7 100.0

- 225.2 18 .00

- 638 .00

Range 100.0

- 145.0 100.0

- 100.0 100.0

- 139.0 100.0

- 150. 2

APV NA NA NA

VAL Mean 14 7.0 134. 5 137.0 143.1 100.0

- 460. 6 30 .00

- 740 .00

Range 100.0

- 278.0 100.0

- 326. 2 100.0

- 258.0 100.0

- 251.0

APV 0.0118 0.0373 1.0000

1 Free amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjust ed to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differe nces with controls are underlined.

2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

.

### J.R. Simplot Company Compositional Analyses 36 

### Table 1 5 : Atlantic Total Amino Acids ( parts per million) 1

Total Amino Acid (ppm) TI CLR 2

Control Atlantic
N=47 J3

N=47 J55

N=47 J78

N=47

ALA Mean 505. 2 550.7 510. 3 557.8 100.0

- 1335 392 .0

- 952 .0

Range 100.0

- 873.0 100.0

- 922.0 100.0

- 904.0 100.0

- 960.0

APV 0.2927 1.0000 0.3308

ARG Mean 1109 119 8 1130 1233 367.6

- 192 2 700 .0

- 1383

Range 70 2.0

- 1464 659. 8

- 164 5 521. 3

- 163 4 690. 5

- 1710

APV 0.3598 1.0000 0.2451

ASP+ ASN Mean 598 6 3721 35 90 378 9 100.0

- 13340 3385

- 7380

Range 296.0

- 1221 5 2110

- 7946 1980

- 7178 2250

- 7814

APV 0 .1591 0.1358 0.1714

GLU+ GLN Mean 3729 4595 462 8 462 4 568.4

- 7011 2915

- 6035

Range 1580

- 647 4 1763

- 796 6 184 2

- 78 20 164 1

- 7386

APV 0.2140 0.1931 0.1977

GLY Mean 986. 7 1083 956.3 1061 100.0

- 2865 500 .0

- 1990

Range 474.0

- 2383 426.1

- 316 3 487.5

- 2 290 598.8

- 270 1

APV 0.3847 1.0000 0.3933

HIS Mean 296. 3 283.4 297.5 306. 4 100.0

- 760.7 133 .0

- 469 .0

Range 100.0

- 683. 5 100.0

- 459.0 100.0

- 490.0 100.0

- 471.0

APV 0.6403 1.0000 0.7345

ILE Mean 780. 2 828. 7 771.4 813. 6 144.8

- 1346 525 .0

- 953 .0

Ran ge 212. 6

- 1374 244. 3

- 1674 24 9.0

- 1156 248.3

- 15 80

APV 0.3589 1.0000 0.3853

LEU Mean 135 3 1422 1343 1435 476.9

- 217 4 685 .0

- 1383

Range 843. 5

- 1867 771. 6

- 218 1 607.8

- 1880 766.1

- 2042

APV 0.4487 1.0000 0.4321

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined.

2 Literature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 37 

### Table 1 5 ( Continued ) : Atlantic Total Amino Acids ( parts per million) 1

Total Amino Acid (ppm) TI CLR 2

Control Atlantic
N=47 J3

N=47 J55

N=47 J78

N=47 LYS Mean 160 1 173 7 1715 1746 100.0

- 3363 687 .0

- 1368

Range 647. 8

- 4034 716.0

- 4407 765.5

- 4581 679.1

- 444 6

APV 0 .2754 0.6618 0.3990

MET Mean 312.6 319. 7 313.7 329.0 100.0

- 593. 3 300 .0

- 500 .0

Range 169. 3

- 511.0 100.0

- 499.0 100.0

- 520.0 100.0

- 530.0

APV 0.7163 0.9970 0.4430

PHE Mean 927.1 974.2 946.5 969.7 397.1

- 1395 552 .0

- 1087

Range 555.1

- 1240 556. 3

- 1290 510. 1

- 1260 523. 2

- 1340

APV 0.4102 1.0000 0.4678

PRO Mean 870. 8 943. 9 914. 7 957.9 100.0

- 1492 355 .0

- 1464

Range 409. 3

- 2090 340.1

- 1850 326.3

- 1960 496. 6

- 2080

APV 0.2876 0.9755 0.2784

SER Mean 670.1 717. 7 704.0 730.8 100.0

- 136 2 500 .0

- 1022

Range 256.1

- 1130 271. 5

- 1190 155. 1

- 1150 252. 3

- 1190

APV 0.3548 0.8755 0.2836

THR Mean 807. 6 903.4 85 8.0 877. 1 226.0

- 1315 436 .0

- 855 .0

Range 356.7

- 1234 399. 4

- 1398 357.3

- 121 9 375. 4

- 1180

APV 0.3212 1.0000 0.3421

TYR Mean 843. 4 872. 5 850. 7 894 .0 100.0

- 1523 457 .0

- 942 .0

Range 375.5

- 145 1 335.4

- 161 3 27 0. 8

- 15 20 318. 1

- 1520

APV 0.6265 1.0000 0.4160

VAL Mean 810.3 855.3 805.5 875.8 100.0

- 1860 752.0

- 1450

Range 275.7

- 1380 175.3

- 1400 221.1

- 1390 269.4

- 1490

APV 0.4321 1.0000 0.3117

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined.

2 Literature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 38 

### Table 1 6 : Atlantic Glycoalkaloids ( mg/100 g) 1
Glycoalkaloids (mg/100g) TI CLR 2

Control Atlantic
N=47 J3

N=47 J55

N=47 J78

N=47
Mean 6.274 5.723 5.864 6.084 0.000

- 33.0 5 3.2 0

– 210 .4 0
Range 2.09

- 11.4 0 1.62

- 11.67 1.83

- 11.12 1.65

- 14.27

P

- value 0.2393 0.4833 0.7198

1 TI = Tolerance Interval

, CLR = Combined Literature Range

. P

- values indicat ing significant differences with controls are underlined.

2 Literature ranges from Kozukue et al. (2008).

### J.R. Simplot Company Compositional Analyses 39 

### Table 17: Atlantic Sugars (percent fresh weight) 1

Fructose + Glucose (%)

Atlantic Control J3 J55 J78 TI CLR 2

Fru ctose + Glucose Fresh Mean 0.062 0.049 0.048 0.048

0.000

- 0.307 0.00

- 0.80 Range 0.003

- 0.23 0.002

- 0.385 0.003

- 0.263 0.002

- 0.237

P

- Value 0.0047 0.0161 0.0454

N 47 47 47 47
Fructose + Glucose Month 1 Mean 0.123 0.095 0.092 0.107

0.000

- 0.30 7 0.00

- 0.80 Range 0.031

- 0.278 0.033

- 0.216 0.031

- 0.267 0.033

- 0.268

P

- Value 0.0028 0.0004 0.1012

N 47 46 46 46
Fructose + Glucose Month 2 Mean 0.070 0.061 0.066 0.077

0.000

- 0.307 0.00

- 0.80 Range 0.028

- 0.182 0.025

- 0.172 0.028

- 0.178 0.029

- 0.229

P

- Value 0.2446 0.6004 0.9998

N 26 26 26 26 Sucrose (%)

Atlantic Control J3 J55 J78 TI CLR

Sucrose Fresh Mean 0.162 0.165 0.166 0.166

0.000

- 0.315 0.05

- 0.32 Range 0.057

- 0.341 0.052

- 0.25 0.075

- 0.366 0.045

- 0.309

P

- Value 0.9772 0.9444 0.9246

N 47 47 47 47
Sucrose Month 1 Mean 0.126 0.132 0.123 0.128

0.000

- 0.315 0.05

- 0.32 Range 0.015

- 0.224 0.016

- 0.247 0.017

- 0.21 0.020

- 0.227

P

- Value 0.9295 0.9995 0.9834

N 47 46 46 46
Sucros e Month 2 Mean 0.132 0.142 0.140 0.131

0.000

- 0.315 0.05

- 0.32 Range 0.082

- 0.202 0.073

- 0.295 0.084

- 0.286 0.022

- 0.245

P

- Value 0.6559 0.8069 0.9994

N 26 26 26 26

1 TI = Tolerance Interval, CLR = Combined Literature Range. P

- va lues indicating significant differences with controls are underlined.

2 Literature ranges from Blenkinsop et al. ( 2002 ) ; Matsuura

- Endo et al. ( 2006 ); Menendez et al. (2002).

### J.R. Simplot Company Compositional Analyses 40 

### Table 1 8 : Atlantic Acrylamide Levels in Processed Product ( parts per billion) 1

Acrylamide (ppb) Atlantic Control J3 J55 J78

Fresh Mean 842.7 278.7 233.4 251.1

2010 & 20 11 Range 113

- 4540 37

- 4120 39

- 1400 36

- 1280

P

- value

— <0.0001 <0.0001 <0.0001

N 47 47 47 47

Month 2 Mean 1681.4 529.8 534.3 558.4

2010 & 2011 Range 181

- 9840 80

- 2110 80

- 2850 82

- 1930

P

- value

— <0.0001 <0.0001 <0.0001

N 47 47 47 47

1 P

- values indicating significant differences with controls are underlined.

### J.R. Simplot Company Compositional Analyses 41 

### Compositional Analysis of Atlantic and Event J3

All discussion for the Atlantic variety and eve nt J3 relates to observations summarized from field trials conducted in 2010 and 2011. Compositional equivalence was tested only in the fresh tubers, although reducing sugars and acrylamide were tested during storage to show trait efficacy.

Tubers of event J3 were equivalent to those of untransformed Atlantic except for the following expected changes :

 J3 tubers contained, on average, 516 parts per million (ppm) free ASN as compared to controls,
which contained 2268 ppm ( APV = 0.0260 ). This change was expect ed and is linked to a reduced acrylamide

- forming potential.

 J3 tubers contained, on average, 0.049% of the reducing sugars glucose plus fructose as compared to controls, which contained 0.062% (P

- value = 0.0047). The decrease in reducing
sugars was expecte d from silencing the promoters associated with the R1 and PhL genes. The new levels are still normal for potato.

 After one month of storage, J3 tubers contained, on average, 0.095% of the reducing sugars glucose plus fructose as compared to controls, which contained 0.123% (P

- value = 0.0028). The decrease in reducing sugars was expected from silencing the promoters associated with the R1
and PhL genes. The new levels are still normal for potato.

 P otato ch ips from fresh J3 tubers contained 278.7 ppb acrylami de as compared to control

, which had 842.7 ppb ( P

- value <0.0001 ). This change was intended.

 The impact on nutritional quality would be minimal from the observed differences in ASN
because as a non

- essential amino acid it can be synthesized by the body. The observed decrease in reducing sugars falls within the normal range for potatoes. However, the combination of lower ASN and reducing sugars contribute to the reduced potential of J3 to form acrylamide and
would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in light of recent toxicology studies (NTP 201 2 ).

The following changes were observed :

 J3 tubers contained, on average, 0.1 17 mg/100 g pyridoxine (vitamin B6) as compared to controls, which contained 0.1 24 mg/100 g ( APV = 0.0 024 ) . The new levels are still normal for
potato.

 J3 tubers contained, on average, 134 ppm free VAL as compared to controls, which contained 147 ppm (APV = 0.0118). The new levels are still normal for potato.

These two ch anges result in very small modifications to the vitamin and amino acid composition of Atlantic J3 that are still standard for potato and will not significantly alter potential dietary intake levels.

### J.R. Simplot Company Compositional Analyses 42



### It can be concluded that tubers of J3 are substantially e quivalent to untransformed Atlantic tubers except for: (1) lowered free ASN

, (2) the expected decreased levels of the reducing sugars glucose plus
fructose

, (3) reduced acrylamide as expected and intended, ( 4 ) small decreases in pyridoxine as compared to c ontrols, and (5) a slight decrease of free VAL as compared to controls. In all cases, these compositional changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 43 

### Compositional Analysis of Atlantic and Event J55

All discussion for the Atlantic variety and even t J55 relates to observations summarized from field trials conducted in 2010 and 2011. Compositional equivalence was tested only in the fresh tubers, although reducing sugars and acrylamide were tested during storage to show trait efficacy.

Tubers of the e vent J55, were equivalent to those of untransformed Atlantic except for the following expected changes :

 J55 tubers contained, on average, 502 parts per million (ppm) free ASN as compared to controls,
which contained 2268 ppm ( APV = 0.0499 ). This change was expected and is linked to a reduced acrylamide

- forming potential.

 J55 tubers contained, on average, 0.048% of the reducing sugars glucose plus fructose as compared to controls, which contained 0.062% (P

- value = 0.0161). The decrease in reducing

sugars was expected from silencing the promoters associated with the R1 and PhL genes. The new levels are still normal for potato.

 After one month of storage, J55 tubers contained, on average, 0.092% of the reducing sugars glucose plus fructose as compared to contro ls, which contained 0.123% (P

- value = 0.0004). The decrease in reducing sugars was expected from silencing the promoters associated with the R1
and PhL genes. The new leve ls are still normal for potato.

 Potato chips from fresh J55 tubers contained 233.4 pp b ac rylamide as compared to control

, which had 842.7 ppb (P

- value <0.0001). This change was intended.

 The impact on nutritional quality would be minimal from the observed differences in free ASN
because as a non

- essential amino acid it can be synthesized b y the body

. The observed decrease in reducing sugars falls within the normal range for potatoes. However, the combination of lower ASN and reducing sugars contribute to the reduced potential of J55 to form acrylamide
and would benefit consumers by addressi ng a potentially critical food safety issue for the potato industry, particularly in light of recent toxicology studies (NTP 201 2 ).

The following changes were observed :

 J55 tubers contained, on average 0.117 mg/100 g pyroxidine (vitamin B 6 

### ) as compared to controls, which contained 0.124 mg/100 g (APV = 0.0012). The new levels are still normal for
potato.

 J55 tubers contained, on average, 137 ppm free VAL as compared to controls, which contained 147 ppm (APV = 0.0373). The new levels are still normal for pot ato.

These two changes result in very small modifications to the vitamin and amino acid composition of
Atlantic J55 that are still standard for potato and will not significantly alter potential dietary intake levels

### J.R. Simplot Company Compositional Analyses 44



### It can be concluded that tubers of Atlan tic J55 are substantially equivalent to untransformed Atlantic tubers except for: (1) lowered free ASN, (2) the expected decreased levels of the reducing sugars glucose plus fructose, (3) reduced acrylamide as expected and intended, (4) small decreases in pyroxidine as compared to controls, and (5) a slight decrease in free VAL as compare to controls. In all cases, these compositional changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 45 

### Compositional Analysis of Atlantic and Event J78

All discussion for the Atlantic variety and event J 78 relates to observations summarized from field trials conducted in 2010 and 2011. Compositional equivalence was tested only in the fresh tubers, although reducing sugars and acrylamide were tested during storage to show trait efficacy.

Tubers of the event J78 , were equivalent to those of untransformed Atlantic except for the following expected changes :

 J78 tubers contained, on average, 551 parts per million (ppm) free ASN as compared to controls,
which contained 2268 ppm ( APV = 0.0548 ). This change was expected and is linked to a reduced acrylamide

- forming potential. Although the APV is slightly above our cutoff for significance of APV < 0.0500, it is worth noting the large difference between J78 and the controls.

 J78 tubers co ntained, on average, 0.048% of the reducing sugars glucose plus fructose as compared to controls, which contained 0.062% (P

- value = 0.0454). The decrease in reducing

sugars was not expected because J78 does not contain the silencing cassette for the promot ers associated with the R1 and PhL genes. The new levels are still normal for potato.

 Potato chips from fresh J78 tubers contained 251.1 ppb acrylamide as compared to control, which had 842.7 ppb (P

- value <0.0001). This change was intended.

 The impact on n utritional quality would be minimal from the observed differences in free ASN
because as a non

- essential amino acid it can be synthesized by the body. The observed decrease in reducing sugars falls within the normal range for potatoes. However, the combina tion of lower ASN and reducing sugars contribute to the reduced potential of J78 to form acrylamide
and would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in light of recent toxicology studi es (NTP 201 2 ).

There were no other changes observed for event J78.

It can be concluded that tubers of J78 are substantially equivalent to untransformed Atlantic tubers except for: (1) lowered free ASN, (2) the decreased levels of the reducing sugars glucos e plus fructose was not expected , and (3) reduced acrylamide as expected and intended. In all cases, these compositional changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 46 

### Compositional Analysis of G Event and Control

Data on the biochemical composition of G control and event G11 are shown in Tables 1 9 (key proximates, vitamins, and minerals), 20 (free amino acids), 21 (total amino acids), 22 (glycoalkaloids)

, and 23 (reducing sugars), and 24 (acrylamide i n fried product) . The analysis was based on 2 tub ers per rep, 4 rep s per location (RCB

- Randomized Com p lete Block field experiment), and 6 locations

, all from
2010 field trials. In addition, some acrylamide data were collected in 2009 and 2011.

### J.R. Simplot Company Compositional Analyses 47 

### Table 1 9 : G Variety Proximates, Vitamins, and Minerals 1
Compound

G Control N=20 G11

N=20 TI CLR 2

P rotein (%) Mean 2.54 2.44 1.258

- 3.594 0.70

- 4.60

Range 1.7 0

- 3.07 1.58

- 3.08

APV 0.361

Fat (%) Mean 0.21 0.18 0.000

- 0.341 0.02

- 0.20

Range 0.10

- 0.60 0.10

- 0.30

APV 0.903 Ash (%) Mean 1.16 1.28 0.391

- 1.888 0.44

- 1.90

Range 0.81

- 1.44 1.03

- 1.54

APV 0.278

Crude Fiber (%) Mean 0. 39 0. 38 0.142

- 0.690 0.17

- 3.50

Range 0.30

- 0.47 0.10

- 0.53

APV 0.935
Carbohydrates (%) Mean 20.62 20.48 12.29

- 25.92 13.30

- 30.53

Range 16.20

- 26.30 16.00

- 25.00

APV 0.829

Calories ( kcal/ 100 g ) Me an 94.32 93.1 59.09

- 114.9 70.00

- 110.2

Range 78.4 0

- 115 .0 78 .00

- 108 .0

APV 0.841 Moisture (%) Mean 75.48 75.64 70.28

- 83.91 63.20

- 86.90

Range 70.1 0

- 79.8 0 71.4 0

- 79.5 0

APV 0.871

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value, B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper. Adjusted P

- values indicating significant differences with controls are underlined. NA = A djuste d p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.

2 Literature ranges are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992) , Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 48 

### Table 1 9 (Continued) : G Variety Proximates, Vitamins, and Minerals 1
Compound

G Control N=20 G11 N=20 TI CLR 2

B3 ( mg/ 100 g ) Mean 1.86 1.93 0.922

- 3.123 0.09

- 3.10

Range 1.61

- 2.18 1.68

- 2.18

APV 0.784

B6 ( mg/ 100 g ) Mean 0.18 0.19 0.059

- 0.1 92 0.13

- 0.41

Range 0.17

- 0.20 0.15

- 0.23

APV 0.359

Vitamin C ( mg/ 100 g ) Mean 26.03 29.14 0.000

- 129.3 1.00

- 54.00

Range 18.50

- 36.10 20.50

- 38.40

APV 0.418 Cu (ppm) Mean 1.14 1.17 0.111

- 2.236 0.15

- 7.00

Range 0.74

- 1.54 0.71

- 1.93

APV 0.603 Mg (ppm) Mean 263.3 268.5 101.5

- 371.5 112.5

- 550.0

Range 198.0

- 319.0 221.0

- 338.0

APV 0.567 K (ppm) Mean 4598 4792 2711

- 6882 3500

- 6250

Range 3710

- 5540 3820

- 5690

APV 0.341

1 TI = Tolerance Interval, CLR = Combined Literature Range , APV = adjusted P

- value, B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper , ppm = parts per million

. Adjusted P

- values indicating significant differences with contro ls are underlined.

2 Literature ranges are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992) , Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 49 

### Table 20 : G Variety Free Amino Acids (parts per million) 1

Free amino acids (ppm) G Control

N= 20 G11 N=20 TI 2 CLR 3

ALA Mean 100 100 100.0

- 208.2 12 .00

- 236 .0
Range 100

- 100 100

- 100

APV NA

ARG Mean 609. 8 626. 2 100.0

- 1068 120 .0

- 1472
Range 350

- 973 344

- 1000

APV 0.597

ASN Mean 3 406 794 500.0

- 4952 3 12.0

- 6980
Range 2190

- 4100 440

- 942

APV 0.003

ASP Mean 304. 2 330. 1 100.0

- 1411 64 .00

- 752 .0
Range 264

- 369 270

- 542

APV 0.408

GLU Mean 434.9 475.3 100.0

- 1192 450 .0

- 742 .0
Range 280

- 563 405

- 615

APV 0.516

GLN Mean 889.4 1666 100.0

- 3449 440 .0

- 5396
Range 557

- 1470 954

- 2490

APV 0.017

HIS Mean 107.4 100 100.0

- 136.9 34 .00

- 656 .0
Range 100

- 147 100

- 100

APV NA

ILE Mean 106. 7 102.1 100.0

- 217.8 28 .00

- 330 .0
Range 100

- 147 100

- 134

APV NA

1 Free amino acids included to show efficacy of reducing free ASN and th us reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- value not determined because observations near the detection limits are too low to allow an estimate of var iance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.

2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

. GLN high level reported as 5396 ppm from the mean of 4 sites (Davies 1977) omitting a level of 18,244 ppm from a single site, which appeared to be an outlier.

### J.R. Simplot Company Compositional Analyses 50 

### Table 20 (Continued) : G Variety Free Amino Acids (parts per million) 1

Free amino acids (ppm) G Control

N=20 G11 N=20 TI 2 CLR 3

LYS Mean 128.6 111.6 100.0

- 557.3 18 .00

- 638 .0 Range 100

- 210 100

- 186

APV NA

MET Mean 107.6 101. 7 100.0

- 191.6 18 .00

- 216 .0 Range 100

- 136 100

- 115
APV NA

PHE Mean 107. 6 100 100.0

- 2 94.9 0 .00

- 408 .0 Range 100

- 139 100

- 100

APV NA

PRO Mean 132. 9 123. 3 100.0

- 630.5 0 .00

- 968 .0 Range 100

- 307 100

- 232

APV NA SER Mean 100 100 100.0

- 198.1 30 .00

- 256 .0 Range 100

- 100 100

- 100

APV NA

THR Mean 100 100 100.0

- 219.8 28 .00

- 286 .0

Range 100

- 100 100

- 100

APV NA

TRP Mean 100 100 100.0

- 113.1 14 .00

- 348 .0 Range 100

- 100 100

- 100

APV NA

TYR Mean 101. 6 100 100.0

- 225.2 18 .00

- 638 .0
Range 100

- 124 100

- 100

APV NA

VAL Mean 132.6 112.8 100.0

- 460.6 30 .00

- 748 .0
Range 100

- 212 100

- 179

APV NA

1 Free amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- val ue not determined because observations near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.

2 TI = Tolerance Interval

. 3 C LR = Com bined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

.

### J.R. Simplot Company Compositional Analyses 51 

### Table 21 : G Variety Total Amino Acids (parts per million) 1
Total amino acids (ppm)

G Control
N=20 G11

N=20 TI CLR 2

ALA Mean 794.3 868.4 100.0

- 1335 392

- 952

Range 671

- 938 658

- 1060

APV 0.06

ARG Mean 1317 1403 367.6

- 1922 700

- 1383

Range 982

- 1750 942

- 1800

APV 0.219

ASP+ASN Mean 5655 317 3 100.0

- 13340 3385

- 7380

Range 3940

- 7710 2070

- 6880

APV 0.008

GLU+GLN Mean 3264 4456 568.4

- 7011 2915

- 6035

R ange 2530

- 4080 3080

- 5640

APV 0.034

GLY Mean 747.1 818.4 100.0

- 2865 500

- 1990

Range 531

- 951 572

- 1080

APV 0.053

HIS Mean 366.7 385.7 100.0

- 760.7 133

- 469

Range 216

- 509 303

- 484

APV 0.445

1 TI = Tolerance Interval, CLR = Combined Liter ature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.
2 Lite rature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 52 

### Table 21 (Continued) : G Variety Total Amino Acids ( pa rts per million) 1
Total amino acids (ppm)

G Control N=20 G11

N=20 TI CLR 2

ILE Mean 874.5 929. 3 144.8

- 1346 525

- 953

Range 706

- 1090 667

- 1120

APV 0.071

LEU Mean 156 8 1715 476.9

- 2174 685

- 1383

Range 1260

- 1880 1270

- 2090

APV 0.059

LYS Mean 1055 1185 100.0

- 3363 687

- 1368

Range 700

- 1510 789

- 1580

APV 0.006

MET Mean 380.2 391.1 10 0.0

- 593.3 300

- 500

Range 271

- 488 247

- 485

APV 0.463

PHE Mean 1029 1099 397.1

- 1395 552

- 1087

Range 794

- 1310 810

- 1340

APV 0.146

PRO Mean 839.3 924 100.0

- 1492 355

- 1464

Range 633

- 1200 665

- 1160

APV 0.035

SER Mean 892.2 963.3 100.0

- 1 362 500

- 1022

Range 722

- 1120 733

- 1210

APV 0.058

THR Mean 911.7 1009 226.0

- 1315 436

- 855

Range 738

- 1100 736

- 1240

APV 0.055

TRP Mean 258.5 246.2 114.7

- 434.7 114

- 282

Range 213

- 366 206

- 299

APV 0.425

TYR Mean 825. 6 937.3 100.0

- 1523 4 57

- 942

Range 604

- 989 684

- 1160

APV 0.053

VAL Mean 108 2 115 4 100.0

- 1860 752

- 1450

Range 876

- 1310 876

- 1350

APV 0.056

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.

2 Literature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000)

, and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 53 

### Table 22 : G Variety G lycoalkaloids ( mg/100 g) 1 Glycoalkaloids (mg/100 g) G Control
N=20 G11 N=20 TI CLR 2

Mean 19.82 20.32 0.000

- 33.05 3.20

- 210.40
Range 6.17

- 44.4 5. 81

- 55.3

P

- value 0. 9153

1 TI = Tolerance Interval

, CLR = Combined Literature Range

. P

- values indicating sig nificant differences with controls are underlined.

2 Literature ranges from Kozukue et al. (2008).



### Table 23: G Variety Sugars (percentage fresh weight) 1

Fructose + Glucose (%)

G Control G11 TI CLR 2
Fructose + Glucose Fresh Mean 0. 08 0.08

0.000

- 0.30 7 0.00

- 0.80

Range 0.019

- 0.165 0.017

- 0.150

P

- Value 0.9800

N 40 39
Sucrose (%)

G Control G11 TI CLR

Sucrose Fresh Mean 0.090 0.100

0.000

- 0.315 0.05

- 0.32

Range 0.046

- 0.119 0.045

- 0.184

P

- Value 0.387

N 20 20

1 TI = Tolerance Interval

, CLR = Combined Literature Range . Data represent the mean levels of reducing sugars and sucrose (% FW) in samples of 3

- 5 tubers, using 3 measurements per event and control per trial site. P

- values indicating significant differenc es with controls are underlined.
2 

### Literature ranges from Blenkinsop et al. ( 2002 ) ; Matsuura

- Endo et al. ( 2006 ); Menendez et al. ( 2002 ).

### J.R. Simplot Company Compositional Analyses 54 Acrylamide (ppb) G Control G11

Fresh Mean 1254.2 421.5

2010 & 20 11 Range 105

- 4320 51

- 1200

P

- value

— <0.0001

N 25 21

Month 1 Mean 1769.0 2050.0

2010 & 2011 Range 215

- 6938 463

- 6210

P

- value

— 0.0136

N 8 4

Month 3 Mean 2370.7 971.2

2010 & 2011 Range 57

- 4890 28

- 2200

P

- value

— <0.0001

N 14 13

Month 5 Mean 1060.7 567.6

2009 & 2010 Range 200

- 2550 55

- 1700

P

- value

— 0.0470

N 7 7

Month 7 Mean 729.3 302.8

2010 Range 296

- 1490 102

- 488

P

- value

— 0.0629

N 4 4

1 P

- values indicating significant differences with controls are underlined.



### Table 2 4 : G Variety Acrylamide Levels in Processed Product ( parts per billion) 1

### J.R. Simplot Company Compositional Analyses 55 

### Compositional Analysis of the G Variety and Event G11

The composition data for G11 represent the summary from 6 replicated field trials conducted in 2010. In addition some acylamide data were collected and summarized in 2009 and 2011. Tubers of event G11 were equivalent to those of the untrans formed G variety except for the following expected changes :

 G11 tubers contained, on average, 794 parts per million (ppm) free ASN compared to the control, which contained 3 406 ppm ( APV = 0.003 ). This change was expected and is linked to a reduced acrylami de

- forming potential.

 G11 tubers contained, on average, 1666 ppm free GLN compared to the control , which
contained 889 ppm ( APV = 0.017 ). The increase in this amide amino acid counter

- balances, in part, losses in the ASN amide amino acid. The new levels ar e normal for potato.

 G11 tubers contained, on average, 317 3 ppm total ASN + ASP compared to the control , which
contained 5655 ppm ( APV = 0.008 ). The new level fits within the tolerance interval and is still normal for potato.

 G11 tubers contained, on avera ge, 4456 ppm GLN + GLU compared to the control , which
contained 3264 ppm ( APV = 0.034 ). The new levels are normal for potato.

 P otato ch ips from G11 tubers contained 421.5 ppb acrylamide compared to the control, which
had 1254.2 ppb ( P

- value <0.0001 ). This change was intended.

 The impact on nutritional quality would be minimal from the observed differences in free ASN and GLN, along with total ASN + ASP, and total GLN + GLU because as non

- essential amino acids they can be synthesized by the body. However, t he reduced potential of G11 to form acrylamide would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in light of recent toxicology studies (NTP 201 2 ).

The following changes were observed :

 G11 t ubers contained, on average, 1185 ppm total LYS as compared to controls, which contained 1055 ppm ( APV = 0.006). The new levels are still normal for potato.

 G11 tubers contained, on average, 924 ppm total PRO as compared t o controls, which contained
839 pp m ( APV = 0.035) . The new levels are still normal for potato.

It can be concluded that tubers of event G11 are substantially equivalent to untransformed G tubers except for: (1) changes in ASN and GLN, (2) lower levels of total ASP + ASN, (3) higher levels of total GLN + GLU, ( 4 ) slightly increased total LYS and PRO, and ( 5 ) reduced acrylamide as expected and intended. In all cases, these compositional changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 56 

### Compositional Analysis of H Events and Controls

Data o n the biochemical composition of the H variety and events H37 and H50 are shown in Tables 2 5 (key proximates, vitamins, and minerals), 2 6 (free amino acids), 2 7 (total amino acids), 2 8 (glycoalkaloids)

, 2 9 (reducing sugars), and 30 (acrylamide in fried pro duct) . The analysis was based on 2 tubers per rep, 4 reps per location (RCB field experiment), and 6 locations

, all from 2010 field trials. In addition, some acrylamide data were collected in 2009 and 2011.

### J.R. Simplot Company Compositional Analyses 57 

### Table 2 5 : H Variety Proximates, Vitamins, and M inerals 1
Compound
H Control
N=20 H37

N=20 H50

N=20 TI CLR 2

Protein (%) Mean 2.49 2.37 2.36 1.258

- 3.594 0.70

- 4.60 Range 1.23

- 3.15 1.67

- 2.94 1.34

- 2.85
APV 0.833 0.752
Fat (%) Mean 0.11 0.10 0.11 0.000

- 0.341 0.02

- 0.20 Range 0.10

- 0.20 0.10

- 0.10 0.10

- 0.30
APV NA NA Ash (%) Mean 1.15 1.16 1.13 0.391

- 1.888 0.44

- 1.90 Range 0.47

- 1.71 0.65

- 1.62 0.87

- 1.42
APV 1 .000 0.847 Crude Fiber (%) Mean 0.42 0.43 0.41 0.142

- 0.690 0.17

- 3.50 Range 0.34

- 0.52 0.34

- 0.62 0.33

- 0.51
APV 0.775 0.8 18 Carbohydrates (%) Mean 18.18 18.81 19.08 12.29

- 25.92 13.30

- 30.53 Range 13.90

- 21.00 16.10

- 21.40 15.50

- 22.30
APV 0.904 0.681 Calories ( kcal/100 g ) Mean 83 .00 85.09 86.22 59.09

- 114.9 70.00

- 110.2 Range 66 .00

- 95.4 0 74.1 0

- 93.3 0 74.3 0

- 98.1 0
APV 0.81 0 0.671 Moisture (%) Mean 78.14 77.62 77.38 70.28

- 83.91 63.20

- 86.90 Range 75.4 0

- 82.5 0 75.5 0

- 80.8 0 74.5 0

- 80.4 0
APV 0.916 0.607

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value, B6 = pyridoxine hydrochl orine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper , ppm = parts per million . Adjusted P

values indicating significant differences with controls are underlined. NA = A djusted p

- value not dete rmined because observations near the detection limits are too low to allow an estimate of variance.

2 Literature ranges are from , Lisinska and Leszczynski (1989), Rogan et al. (2000)

, Horton and Anderson (1992 ), Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 58 

### Table 2 5 (Continued) : H Variety Proximates, Vitamins, and M inerals 1
Compound
H Control
N=20 H37

N=20 H50

N=20 TI CLR 2

B3 (mg/100 g) Mean 1.68 1.73 1.66 0.922

- 3.123 0.09

- 3.10

Range 1.40

- 1.98 1.50

- 2.15 1.42

- 2.16
APV 0.901 0.844
B6 (mg/100 g) Mean 0.16 0.16 0.15 0.059

- 0.192 0.13

- 0.41

Range 0.12

- 0.20 0.13

- 0.19 0.13

- 0.17
APV 1.000 1.000 Vitamin C ( mg/100 g ) Mean 28.76 29.62 29.99 0.000

- 129.3 1.00

- 54.00

Range 19.80

- 37.70 21.10

- 39.30 23.10

- 38.70
APV 0.814 0.710 Cu (ppm) Mean 1.15 0.96 0.97 0.111

- 2.236 0.15

- 7.00

Range 0.57

- 2.96 0.61

- 1. 46 0.66

- 1.47
APV 0.880 0.546 Mg (ppm) Mean 246.1 237.4 247.8 101.5

- 371.5 112.5

- 550.0

Range 194.0

- 369.0 156.0

- 329.0 202.0

- 324.0
APV 0.994 0.860 K (ppm) Mean 4562 4779 4679 2711

- 6882 3500

- 6250

Range 4240

- 5120 3810

- 5460 4200

- 5040
APV 0.855 0.623

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value, B6 = pyridoxine hydrochlorine (vitamin B6), B3 = niacin (vitamin B3), ascorbic acid = vitamin C, Mg = magnesium, K = potassium, Cu = copper. Adjusted P

- valu es indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.

2 Literature ranges are from , Lisinska and Leszczynski (1 989), Rogan et al. (2000)

, Horton and Anderson (1992), Talburt and Smith (1987)

.

### J.R. Simplot Company Compositional Analyses 59 

### Table 2 6 : H Variety Free Amino Acids ( parts per million) 1
Free Amino Acids (ppm)

H Control N=20 H37

N=20 H50

N=20 TI 2 CLR 3

ALA Mean 101. 8 100 100 100.0

- 208.2 12 .00

- 236 .0

Range 100

- 128 100

- 100 100

- 100

APV NA NA

ARG Mean 869. 2 977. 7 930.4 100.0

- 1068 120 .0

- 1472

Range 496

- 1330 654

- 1380 523

- 1230

APV 0.412 0.68

ASN Mean 280 5 877.5 672. 7 500.0

- 4952 3 12.0

- 6980

Range 657

- 4100 429

- 2750 280

- 1070

APV <.0001 <.0001

ASP Mean 391.9 405.5 387. 9 100.0

- 141 1 64 .00

- 752 .0

Range 286

- 490 325

- 494 315

- 533

APV 0.854 0.847

GLU Mean 456. 6 459.8 487. 3 100.0

- 1192 450 .0

- 742 .0

Range 324

- 596 315

- 589 372

- 590

APV 0.966 0.424

GLN Mean 1680 2724 2913 100.0

- 3449 440 .0

- 5396

Range 1100

- 3380 1720

- 3620 1140

- 3 770

APV 0.014 0.006

HIS Mean 121. 6 121.3 117. 2 100.0

- 136.9 34 .00

- 656 .0

Range 100

- 163 100

- 211 100

- 171

APV 0.967 0.618

ILE Mean 140. 9 128.3 117.6 100.0

- 217.8 28 .00

- 330 .0

Range 100

- 185 100

- 254 100

- 172

APV 0.441 0.082

1 Free amino ac ids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for statistical analysis. NA = Adjusted p

- value not determined because observations near th e detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.

2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

. GLN high level reported as 5396 ppm from the mean of 4 sites (Davies 1977) omitting a level of 18,244 ppm from a single site, which appeared to be an outlier.

### J.R. Simplot Company Compositional Analyses 60



### Table 2 6 (Cont inued) : H Variety Free Amino Acids ( parts per million) 1

Free Amino Acids (ppm) H Control N=20 H37

N=20 H50

N=20 TI 2 CLR 3

LYS Mean 263.1 206.8 219.9 100.0

- 557.3 18 .00

- 638 .0

Range 131

- 385 100

- 325 100

- 342

APV 0.035 0.079
MET Mean 135.6 132.4 129 .1 100.0

- 191.6 18 .00

- 216 .0

Range 100

- 185 100

- 179 100

- 170

APV 0.934 0.699

PHE Mean 236. 2 206. 3 191. 8 100.0

- 294.9 0 .00

- 408 .0

Range 126

- 362 100

- 342 100

- 281

APV 0.473 0.227

PRO Mean 130. 4 118.1 157. 9 100.0

- 630.5 0 .00

- 968 .0

Range 100

- 294 1 00

- 275 100

- 545

APV NA NA

SER Mean 108.9 107. 2 104. 3 100.0

- 198.1 30 .00

- 256 .0

Range 100

- 141 100

- 147 100

- 133

APV NA NA

THR Mean 115.3 118. 7 119.4 100.0

- 219.8 28 .00

- 286 .0

Range 100

- 162 100

- 152 100

- 150

APV 0.857 0.641

TRP Mean 100.6 104. 8 110.6 100.0

- 113.1 14 .00

- 348 .0

Range 100

- 110 100

- 176 100

- 190

APV NA NA

TYR Mean 129.5 154. 4 156.5 100.0

- 225.2 18 .00

- 638 .0

Range 100

- 209 100

- 256 100

- 251

APV 0.458 0.319

VAL Mean 273.1 259. 7 239.4 100.0

- 460.6 30 .00

- 740 .0

Range 181

- 400 171

- 467 105

- 375

APV 0.933 0.441

1 Free amino acids included to show efficacy of reducing free ASN and thus reducing acrylamide. Many values fell below detection limits and were adjusted to the limit (100 ppm) for statistical analysis. NA = Ad justed p

- value not determined because observations near the detection limits are too low to allow an estimate of variance. APV = adjusted P

- value. Adjusted
P

- values indicating significant differences with controls are underlined.
2 TI = Tolerance Interval

. 3 C LR = Combined Literature Range. Literature ranges are from Davies (1977), Lisinska and Leszczynski (1989), and Shepherd et al. (2010)

.

### J.R. Simplot Company Compositional Analyses 61 

### Table 2 7 : H Variety Total Amino Acids ( parts per million) 1
Total Amino Acids (ppm)

H Control N=20 H37

N=20 H50

N=20 TI CLR 2

ALA Mean 642. 3 677.1 676. 9 100.0

- 1335 392 .0

- 952 .0

Range 492

- 771 510

- 826 507

- 891

APV 0.462 0.416

ARG Mean 1443 1580 154 7 367.6

- 1922 700 .0

- 1383

Range 930

- 1870 1120

- 1990 970

- 1880

APV 0.437 0.423

ASP+ASN Mean 4748 2860 265 5 100.0

- 13340 3385

- 7380

Range 2680

- 6190 1950

- 4910 1700

- 3320

APV <.0001 <.0001

GLU+GLN Mean 3926 5159 5443 568.4

- 7011 2915

- 6035

Range 3090

- 6130 3670

- 6060 2940

- 6620

APV 0.05 0.018

GLY Mean 636. 6 667. 7 668. 7 100.0

- 2865 500 .0

- 1990

Range 497

- 774 514

- 829 501

- 830

APV 0.606 0.374

HIS Mean 360. 7 372.4 365. 3 100.0

- 760.7 133 .0

- 469 .0

Range 258

- 423 279

- 466 247

- 475

APV 0.607 0.839

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations near the detection limits are too low to allow an estimate of variance.
2 Literature ranges are from Lisinska and Leszczynski (1989)

, Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 62 

### Table 2 7 (Continued) : H Variety Total Amino Acids ( parts per million) 1
Total Amino Acids (ppm)

H Control N=20 H37

N=20 H50

N=20 TI CLR 2

ILE Mean 794. 1 811. 9 808.1 144.8

- 1346 525 .0

- 953 .0

Range 573

- 910 607

- 998 556

- 1010

APV 0.6 0.745

LEU Mean 1226 129 4 129 7 476.9

- 2174 685 .0

- 1383

Range 910

- 1520 973

- 1670 930

- 1660

APV 0.554 0.364

LYS Mean 116 1 118 8 1209 100.0

- 3363 687 .0

- 1368

Range 838

- 1360 878

- 1440 817

- 1450

APV 0.589 0.334

MET Mean 372. 9 388. 2 381. 4 100.0

- 593.3 300 .0

- 500 .0

Range 265

- 435 268

- 478 237

- 494

APV 0.627 0.755

PHE Mean 964. 6 981.3 973. 9 397.1

- 1395 552 .0

- 1087

Range 675

- 1100 681

- 1240 609

- 1270

APV 0.736 0.827

PRO Mean 682.5 717.4 767. 9 100.0

- 1492 355 .0

- 1464

Range 488

- 993 502

- 1010 485

- 1270

APV 0. 604 0.487

SER Mean 728.1 754 751.4 100.0

- 1362 500 .0

- 1022

Range 573

- 903 591

- 934 568

- 925

APV 0.612 0.582

THR Mean 766. 3 814.9 820. 9 226.0

- 1315 436 .0

- 855 .0

Range 573

- 924 602

- 1060 566

- 1090

APV 0.468 0.428

TRP Mean 223.4 227.3 235.3 114.7

- 434.7 114 .0

- 282 .0

Range 170

- 291 166

- 273 171

- 352

APV 0.71 0.453

TYR Mean 714. 1 795.1 830.3 100.0

- 1523 457 .0

- 942 .0

Range 551

- 824 628

- 970 553

- 1090

APV 0.543 0.166

VAL Mean 1053 109 3 10 80 100.0

- 1860 752 .0

- 1450

Range 776

- 1200 813

- 1300 716

- 1310

APV 0.609 0.663

1 TI = Tolerance Interval, CLR = Combined Literature Range, APV = adjusted P

- value. Adjusted P

- values indicating significant differences with controls are underlined. NA = A djusted p

- value not determined because observations n ear the detection limits are too low to allow an estimate of variance.
2 Literature ranges are from Lisinska and Leszczynski (1989), Rogan et al. (2000) , and Talley et al. (1984)

.

### J.R. Simplot Company Compositional Analyses 63 

### Table 2 8 : H Variety Glycoalkaloids ( mg/100 g) 1
Glycoalkaloids (mg/100 g) TI CLR 2

H Control
N=20 H 37

N=20 H 50

N=20
Mean 1 0.65 10.76 11.30 0.000

- 33.05 3.20

- 210.40 Range 5. 96

- 21.07 5.41

- 17.25 6. 23

- 22.81

P

- value 0. 8951 0. 4379

1 TI = Tolerance Interval

, CLR = Combined Literature Range

. P

- values indicating significant differences with controls are underlined.
2 Literatur e ranges from Kozukue (2008).



### Table 29: H Variety Sugars (percentage fresh weight) 1

Fructose + Glucose (%)

H Control H37 H50 TI CLR 2

Fructose + Glucose Fresh Mean 0.0 6 0.0 5 0.0 4

0.000

- 0.307 0.00

- 0.80

Range 0.016

- 0.104 0.015

- 0.071 0.013

- 0.078

P

- v alue 0.0830 0.0550

N 4 0 40 40
Sucrose (%)

H Control H37 H50 TI CLR

Sucrose Fresh Mean 0.11 0.13 0.12

0.000

- 0.315 0.05

- 0.32

Range 0.067

- 0.147 0.071

- 0.183 0.101

- 0.162

P

- v alue 0.0450 0.2250

N 20 20 20

1 TI = Tolerance Interval

, CLR = Combined Literature Range . Data represent the mean levels of reducing sugars and sucrose (% FW) in samples of 3

- 5 tubers, using 3 measurements per event and control per trial site. P

- values indicating significant differences wit h controls are underlined. 2 Literature ranges from Blenkinsop et al. (2002); Matsuura

- Endo et al. (2006); Menendez et al. (2002).

### J.R. Simplot Company Compositional Analyses 64 

### Table 30 : H Variety Acrylamide Levels in Processed Product ( parts per billion) 1

Acrylamide (ppb) H Control H37 H50

Fresh Mean 531.2 223.0 174.7

2010 & 20 11 Range 65

- 1110 81

- 512 59

- 545

P

- value

— <0.0001 <0.0001

N 25 25 25

Month 1 Mean 577.0 253.0 195.6

2010 & 2011 Range 104

- 1600 65

- 727 43

- 4 99

P

- value

— 0.0006 <0.0001

N 8 8 8

Month 3 Mean 791.5 440.2 293.2

2010 & 2011 Range 43

- 2980 43

- 1160 32

- 602

P

- value

— 0.0453 0.0009

N 14 14 13

Month 5 Mean 244.9 147.9 141.9

2009 & 2010 Range 148

- 351 70

- 223 92

- 179

P

- value

— 0.1765 0.1940

N 7 7 7

Month 7 Mean 540.5 317.3 257.3

2010 Range 275

- 930 157

- 544 175

- 318

P

- value

— 0.2359 0.1677

N 4 4 4

1 P

- values indicating signifi cant differences with controls are underlined.

### J.R. Simplot Company Compositional Analyses 65 

### Compositional Analysis of the H Variety and Event H37

The composition data for H37 represent the summary from 6 replicated field trials conducted in 2010.

. In addition some acylamide data were collected an d summarized in 2009 and 2011. Tubers of event H37

were equivalent to those of the untransformed H variety except for the following:

 H37 tubers contained, on average, 877.5 parts per million (ppm) free ASN compared to the
control , which contained 2805 ppm ( APV <0.0001). This change was expected and is linked to a reduced acrylamide

- forming potential.

 H37 tubers contained, on average, 2724 ppm free GLN compared to the control , which contained 1680 ppm ( APV = 0.014). The increase in this amide amino acid coun ter

- balances, in part, losses in the ASN amide amino acid. The new levels are normal for potato.

 H37 tubers contained, on average, 2860 ppm total ASN + ASP compared to the control , which contained 4748 ppm ( APV <0.0001). The new level fits within the toler ance interval and is still normal for potato.

 H37 tubers contained, on average, 5159 ppm total GLN + GLU compared to the control , which contained 3926 ppm ( APV = 0.05). The new levels are normal for potato.

 P otato ch ips from H37 potato tubers contained 223 .0 ppb acrylamide compared to the control,
which had 531.2 ppb ( P

- value <0.0001 ). This change was intended.

 The impact on nutritional quality would be minimal from the observed differences in free ASN and GLN, along with total ASN + ASP, and total GLN + GL U because as non

- essential amino acids they can be synthesi zed by the body. However, the reduced potential of H37 to form acrylamide would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in li ght of recent toxicology studies (NTP 201 2 ).

The following changes were observed :

 H37 tubers contained, on average, 207 ppm free LYS as compared to controls, which contained 263 ppm ( APV = 0.035). The new levels are still normal for potato.

 H37 tubers cont ained, on average, 0.13% of sucrose as compared to controls, which contained 0.11% (P

- value = 0.0450). The increase in sucrose is often related to a decrease in reducing sugars, which was expected from silencing the promoters associated with the R1 and PhL genes, but was not statistically different from controls. The new levels of sucrose are still normal for
potato.

It can be concluded that tubers of event H37 are substantially equivalent to untransformed H tubers except for: (1) changes in ASN and GLN, ( 2) lower levels of total ASP + ASN, (3) higher levels of total GLN + GLU, ( 4 ) slightly decreased amounts of free LYS, ( 5 ) slightly increased levels of sucrose, and ( 6 ) reduced

### J.R. Simplot Company Compositional Analyses 66



### acrylamide as expected and intended. In all cases, these compositional changes f all within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 67 

### Compositional Analysis of the H Variety and Event H50

The composition data for H50 represent the summary from 6 replicated field trials conducted in 2010. In addition some acylamide data were collected and summ arized in 2009 and 2011. Tubers of event H50

were equivalent to those of the untransformed H variety except for the following expected changes :

 H50 tubers contained, on average, 673 parts per million (ppm) free ASN compared to the
control , which contained 2805 ppm ( APV <0.0001). This change was expected and is linked to a reduced acrylamide

- forming potential.

 H50 tubers contained, on average, 2913 ppm free GLN compared to the control , which contained 1680 ppm ( APV = 0.006). The increase in this amide amino acid counter

- balances, in part, losses in the ASN amide amino acid. The new levels are normal for potato.

 H50 tubers contained, on average, 2655 ppm total ASN+ASP compared to the control , which contained 4748 ppm ( APV <0.0001). The new level fits within th e tolerance interval and is still normal for potato.

 H50 tubers contained, on average, 5443 ppm GLN+GLU compared to the control , which contained 3926 ppm ( APV = 0.018). The new levels are normal for potato.

 P otato ch ips from H50 potato tubers contained 174 .7 ppb acrylamide compared to the control,
which had 531.2 ppb ( P

- value <0.0001 ). This change was intended.

 The impact on nutritional quality would be minimal from the observed differences in free ASN and GLN, along with total ASN + ASP, and total GLN + GL U because as non

- essential amino acids they can be synthesized by the body. However, the reduced potential of H50 to form acrylamide would benefit consumers by addressing a potentially critical food safety issue for the potato industry, particularly in li ght of recent toxicology studies (NTP 201 2 ).

It can be concluded that tubers of event H50 are substantially equivalent to untransformed H tubers except for: (1) changes in ASN and GLN, (2) lower levels of total ASP + ASN, (3) higher levels of total GLN
+ G LU, and ( 4 ) reduced acrylamide when compared to controls as expected and intended. In all cases, these compositional changes fall within the normal range for potatoes.

### J.R. Simplot Company Compositional Analyses 68 

### Storage Studies: Reducing Sugars and Acryl a mide

The objective of the composition stud ies was to compare all events with their respective controls mostly at the time of harvest. In addition, since many potatoes are stored for months before processing into fries or chips, we tested some of the potatoes for two important attributes, reducing sugars and acrylamide, following storage.

Reducing Sugars. As was noted in the Results section ( Tables 5, 11, 17, 23, and 29) , many of the events had lowered levels of reducing sugars either at the time of harvest or after storage for 1 month (F10, F37, E12, E24 , J3, and J55, J78)

. However, in most cases we did not observe significant differences after
2

- 5 months of storage. This led us to conclude that silencing of the promoters associated with the PhL/R1 genes effectively lowered reducing sugars near the time of harvest but these differences were not sustained throughout storage for 2

- 5 months.

Acrylamide. Samples of fries or potato chips were tested for up to 7 months after harvesting, depending upon the variety. The Atlantic controls and events J3, J55, and J78 were tested only through 2 months of storage because typically that variety would not be stored. Throughout storage, acrylamide levels were consistently lower (p < .0001) in the events compared with controls ( Tables 6, 12, 18, 24,
and 30 ), a lthough there were exceptions with the G and H varieties. In event G11, we observed a mean value of 2,050 ppb compared with 1,769 ppb for the control after 1 month of storage. This was unusual and throughout the rest of the study, acrylamide levels in ev ent G11 were always much lower than controls (46 to 58% reduction at months 5 and 7). For variety H, acrylamide levels were always lower in events H37 or H50, but not statistically significant in months 5 and 7. Overall, the modifications from
insertion of pSIM 1278 resulted in dramatic reductions in acrylamide that persisted throughout the typical storage periods.

Glycoalkaloids

Glycoalkaloids are toxins commonly found in Solanaceous crops, including potato. The principle glycoalkaloids in potatoes are α

- solanine and α

- chaconine. For food safety purposes, an upper limit for glycoalkaloid content of 20 mg per 100 g of potato is generally accepted (S inden 1987; Crocco 1981 ).
This research confirmed that tubers of the events contained equivalent levels of natural glycoalkaloids compared with untransformed controls.

For the varieties included here, most values for glycoalkaloids were close to or below the accepted level of 20 mg per 100 g of potato ( Table 31 )

. Both untran s formed controls and events for the G variety had
mean values that were about the level of 20 mg/100 g . This has been attributed to excessive handling at storage facilities, resulting in increased exposure to light, and thus, higher levels of glycoalkaloids (Percival 1999). Although the con trol mean values nearly exceeded the accepted levels, the G variety ha s a long history of safe use and the slightly higher than expected results would not be expected in normal commercial operations.

Also, the range of values for E24 exceeded 20 mg/100 g but the mean was less than and not significantly
different from the Russet Burbank control. Although a cause was not identified, the higher value for E24

### J.R. Simplot Company Compositional Analyses 69



### could be from excessive exposure to light (Percival 1999). I n all cases, the events were equivalent to the controls with no observed significant differences. All glycoalkaloid levels were within the normal ranges for potato, when compared with the tolerance intervals or combined literature ranges.

Table 31 : Glycoalkaloids (mg/100 g) 1 

### in Ranger Russet, Russet Burbank , Atlantic , G, and H Varieties.

Variety Total Glycoalkaloids (mg/100 g)

Mean Range P

- Value TI CLR 2

Ranger Control 6.704 3.06

- 10.23

0.000

- 33.05 3.20

- 210.4 F10 6.979 2.84

- 9.51 0.8285

F37 6 .805 4.28

- 9.42 0.9812

Burbank Control 7.404 2.74

- 18.59

0.000

- 33.05 3.20

- 210.4 E12 5.678 1.69

- 10.53 0.1707

E24 6.845 1.45

- 33.81 0.8174

Atlantic Control 6.274 2.09

- 11.40

0.000

- 33.05 3.20

- 210.4 J3 5.723 1.62

- 11.67 0.2393

J55 5.864 1.83

- 11.12 0.4833

J78 6.084 1.65

- 14.27 0.7198

G Co ntrol 19.82 6.17

- 44.40 0.000

- 33.05 3.20

- 210.4 G11 20.3 2 5.81

- 55.3 0. 9153

H Control 10.65 5. 96

- 21.07 0.000

- 33.05 3.20

- 210.4 H37 10.76 5.41

- 17.25 0. 8951

H50 11.30 6.23

- 22.81 0. 4379

1 TI = Tolerance Interval

, CLR = Combined Literature Range

. P

- val ues indicating significant differences with controls are underlined.

2 Literature ranges from Kozukue et al. (2008).

### J.R. Simplot Company Compositional Analyses 70 

### MATERIALS AND METHODS

Field Trial Locations

. All locations and varieties have been summarized in Table 3 2

. Each combination of trial s ite, event or contol, and replication would represent one sample for the compositional
assessment. Some trials were conducted only for agronomic and phenotypic data as indicated in Table 32
, and in addition most were u sed for full composition although some were only tested for acrylamide and sugars. 32

Table 3 2 : Field Trial Locations

USDA Notificat ion # Site

State Site County Material

Tested Trial

Design 3 Rows x

Planted Tubers / Rep Seed

Type Regional Specifics Location ID Code Composition Data



### 2009 Fiel d trials

09

- 049

- 114n ID Canyon Ranger Russet, Russet Burbank, Atlantic RCB, 5

reps/line (1 rep/line for Atlantic) 1x 20 (1x15

for Atlantic) Mini

tubers Typical for Southwest Idaho, an important potato

- growing region in a state in the Northwest that produc es about 120 million cwt/year, mainly for the French fry industry, Careful management is needed to limit or prevent heat

- associated agronomic issues. N ot required 1 Ranger Russet, Russet Burbank

, only

09

- 049

- 114n ID Bingham Ranger Russet, Russet Burbank RC B, 5

reps/line 1x 20 Mini

tubers Typical for Southeast Idaho, an important potato

- growing region in a state that produces about 120 million cwt/year, mainly for the French fry industry, with harvests in Fall. The growing season is relatively short because

o f cooler temperatures. N ot required 1 Ranger Russet, Russet Burbank

09

- 049

- 114n WI Oneida G, H 1 rep /line 1x100 Mini

tubers Typical for Wisconsin, a Midwest state that produces about 20 million cwt/year, for both the chip and fresh potato industry, with ha rvests in Fall. Large areas are dominated by muck soils. N ot required 1 Agronomic and phenotypic data only

08

- 353

- 103n FL St. Johns G, H RCB, 4 reps/line 1x2 5 Mini

tubers Typical for Florida that produces almost 8 million cwt/year, mainly for the chip and fresh potato industry, with harvests in Spring. N ot required 1 Agronomic and phenotypic data only
08

- 353

- 104n MI Montcalm Ranger Russet, Russet Burbank, G, H RCB, 4

reps/line 1x 20 Mini

tubers Typical for Michigan, a Midwest state that produces about 15 mil lion cwt/year, for both the chip and fresh potato industry, with harvests in Fall. The climate is characterized by mild temperatures and ample rain. N ot required 1 Ranger Russet, Russet Burbank

– complete composition G, H

– Acrylamide

### J.R. Simplot Company Compositional Analyses 71 

### Table 3 2 (Continued) : Field Trial Locations

USDA Notificat ion # Site

State Site County Material

Tested Trial

Design 3 Rows x

Planted Tubers / Rep Seed

Type Regional Specifics Location ID Code Composition
Data



### 2010 Field trials

10

- 053

- 132n ID Canyon Ranger Russet, Russet Burb ank, Atlantic RCB, 3

reps/line 3x20 Field seed

(G1) See above for Canyon county UOIPAR2010 Ranger Russet, Russet Burbank, Atlantic
10

- 053

- 132n ID Bingham Ranger Russet, Russet Burbank, Atlantic, G, H RCB, 3

reps/line (4 reps/line for G, H; 5 reps/line fo r Atlantic) 3x20

(1x20 for G, H, Atlantic) Field seed

(G1) (Mini

tubers for Atlantic) See above for Bingham county UOIABE2010 Ranger Russet, Russet Burbank, Atlantic, G, H

10

- 053

- 132n WI Oneida G, H RCB, 4 reps/line 1x 20 Field seed

(G1) See above for On eida county RHI2010 G, H

10

- 053

- 132n WI Adams Atlantic RCB, 5 reps/line 1x20 Mini

tubers Typical for Wisconsin, a Midwest

state that produces about 20 million cwt/year, for both the chip and fresh potato industry, with harvests in Fall. WI2010 Atlantic

10

- 076

- 013n WI Vilas G,H 1 rep Variable 4 Mini

tuber/
G1 5 Typical for Wisconsin, a Midwest

state that produces about 20 million cwt/year, for both the chip and fresh potato industry, with harvests in Fall. VILAS2010 Seed production only

10

- 053

- 132n MI Mont calm Atlantic, G, H RCB, 3

reps/line (4 reps/line for G, H) 3x20

(1x20 for G, H) Mini

tubers See above for Montcalm county MSUMONT2010 Atlantic, G, H

10

- 053

- 132n MI Missaukee G, H RCB, 4 reps/line 1x 20 Field seed

(G1) Typical for Michigan, a Midwest

state that produces about 15 million cwt/year, for both the chip and fresh potato industry, with harvests in Fall. MSULAKE2010 G, H

10

- 053

- 132n NE Cherry Atlantic RCB, 5 reps/line 1x20 Field seed

(G1) (Mini

tubers for Atlantic) Typical for Nebraska, a Midwest

state that produces about 9 million cwt/year, mainly for the seed industry, with harvests in Fall. CSS2010 Atlantic

10

- 053

- 132n ND Grand

Forks 2 Ranger

Russet, Russet Burbank, Atlantic RCB, 3

reps/line (5 reps/line for Atlantic) 3x20

(1x20 for Atlantic) Fi eld seed

(G1) (Mini

tubers for Atlantic) Typical for North Dakota, a

Midwest state that produces about 20 million cwt/year, mainly for the French fry industry, with harvests in Fall. The Re d River
V alley is the 3 rd largest potato growing region in the US. NDSU2010 Ranger Russet, Russet Burbank, Atlantic

10

- 053

- 132n WA Adams Ranger Russet, Russet Burbank, Atlantic RCB, 3

reps/line (5 reps/line for Atlantic) 3x20

(1x20 for Atlantic) Field seed

(G1) (Mini

tubers for Atlantic) Typical for Washington, a state i n

the Northwest that produces about 85 million cwt/year, mainly for the French fry industry, with harvests in Fall. Ideal growing conditions give rise to very high yields per acre. WSU2010 Ranger Russet, Russet Burbank, Atlantic

09

- 336

- 103n FL St. Johns G, H RCB, 4 reps/line 1x 25 Field seed

(G1) See above for St. Johns county U FL2010 G, H

### J.R. Simplot Company Compositional Analyses 72 

### Table 3 2 (Continued) : Field Trial Locations

USDA Notifica tion # Site State Site County Material

Tested Trial

Design 3 Rows x

Planted Tubers / Rep Seed Type Regional Specifics Location ID Code Composition Data



### 2011 Field trials

11

- 063

- 103n ID Canyon Ranger Russet, Russet Burbank, Atlantic RCB, 3

reps/line 3x20 Field seed

(G2) (G1 for Atlantic, Mini

tubers for F37) See above for Canyon county UOIPAR2 011 Ranger Russet
, Russet Burbank, Atlantic
11

- 063

- 103n ID Bingham Ranger Russet, Russet Burbank, Atlantic RCB, 3

reps/line 3x20 Field seed

(G2) (G1 for Atlantic, Mini

tubers for F37) See above for Bingham county UOIABE201 1 Ranger Russet, Russet Burbank, Atlantic

1 1

- 063

- 103n WI Adams Atlantic RCB, 3 reps/line 3x20 Field seed

(G1) See above for Adams county

, Wisconsin WI2011 Atlantic

11

- 063

- 103n MI Montcalm Atlantic RCB, 3 reps/line 3x20 Field seed

(G1) See above for Montcalm county MSUMONT201 1 Atlantic

11

- 063

- 103n ND Grand

Forks Ranger

Russet, Russet Burbank, Atlantic RCB, 3

reps/line 3x20 Field seed

(G2) (G1 for Atlantic, Mini

tubers for F37) See above for Grand Forks

county NDSU201 1 Ranger Russet, Russet Burbank, Atlantic
11

- 063

- 103n WA Adams Ranger Russet, Russet Burbank, Atlantic RCB, 3

reps/line 3x20 Field seed

(G2) (G1 for Atlantic, Mini

tubers for F37) See above for Adams county,

Washington WSU201 1 Ranger Russet, Russet Burbank, Atlantic
10

- 326

- 103n FL St. Johns G, H, Atlantic RCB, 4

rep s/line 1x20 F ield seed

( G1) See above for St. Johns county U FL2011 Atlantic

– complete composition G, H

– Acrylamide
11

- 063

- 103n WI Oneida G, H RCB, 4
rep s/line 1x20 Field seed

( G1) See above for Oneida County RHI 2011 G, H

- Acrylamide
11

- 063

- 103n IN Pulaski Atlantic RCB, 3 reps/line 1x20 Field seed

(G1) Typical for Indiana , a Midw est

state that produces about 2 million cwt/year, for both the chip and fresh potato industry, with harvests in Fall. INDIANA 2011 Atlantic

11

- 094

- 106n WI Oneida G,H RCB, 4 reps/ line 1x13 F ield seed (GI) See above for Oneida County ONEIDA2011 G

- Acrylamide
1 USDA/APHIS did not require a unique location identification code in 2009.

2 Because parts of this field were flooded extensively, Ranger Russet and Russet Burbank tubers were evaluated o nly biochemically.
3 Randomized Complete Block Designs contained the same number of blocks as the number of reps in the table.

4 Several lines were grown

5 Field grown/mini

- tuber

### J.R. Simplot Company Compositional Analyses 73



### Testing Laboratories. Analytical testing for Ranger Russet and Russet Burbank s amples were completed partially by the J.R. Simplot Co.

, Boise, ID in 2009 and 2010, with the remainder done by Covance Laboratories, Inc. , Madison, WI. Atlantic samples were analyzed partially by Simplot and partially by
Covance in 2010. In 2011, all anal yses were completed at Covance Labs. All testing for G and H varieties was conducted by Covance Labs. Clarification of testing locatio n by analysis is provided in Table 3 3

. .In 2011 we made a decision to only use one labarotory for compositional data anal ysis for regulatory data. Altough data were collected in two laboratories, both used standard procedures fo r calibrating their instruments and were treated similarly in the statistical analysis.

Table 3 3 : Laboratory Facilities Used for Compositional Analys is

Analysis 2009 2010 2011

Ranger Russet, Russet Burbank G and H 1 Ranger Russet, Russet Burbank, Atlantic G and H Ranger Russet, Russet Burbank, Atlantic G and H 1 Key Proximates, Vitamins and Minerals 

### Covance NA Covance Covance Covance NA

Free Ami no Acids 

### Simplot NA Simplot Covance Covance NA

Total Amino Acids 

### Simplot NA Simplot Covance Covance NA

Sugars 

### Simplot NA Simplot Covance Covance NA

Acrylamide 

### Covance Covance Covance Covance Covance Covance

Glycoalkaloids 

### Simplot NA Simplot Covance Cov ance NA

Vitamin C 

### Simplot NA Simplot Covance Covance NA

1 Compositional data not completed for G and H in 2009 and 2011 , except for acrylamide



### Sample preparation. For the Ranger Russet, Russet Burbank, and Atlantic varieties, s amples of each
event and con trols were obtained by randomly selecting 5 tubers of about 6 inches in length from each

rep and site. Three to nine longitudinal slices with skin from each tuber were taken from the top, bottom, and center. Samples were immediately frozen in liquid N 2 

### . T hese samples were freeze

- dried, ground and stored at

- 80°C until ready for analysis. For acrylamide testing, samples of Ranger Russet and Russet Burbank potatoes were processed into french fries, using standard practice for the foodservice french fry indu stry. The Atlantic variety and events were processed using standard practice for potato chips.

For the G variety, event G 11, the H variet y and events H37 and H 50, two tubers per replication, 2” to 3” in diameter, were picked from each of 4 replications f rom 6 locations. All compositional analyses for G
and H varieties were performed at Covance Labs according to standard industry protocols. Fresh tubers were used for all analyses except for acrylamide. Samples for acrylamide were prepared by slicing 8

t ubers per replication and combining 2 slices per tuber for a total of 16 slices and then frying in a batch fryer , for the purpose of making potato chips

.

### J.R. Simplot Company Compositional Analyses 74



### Moisture content: Simplot Lab. The percentage moisture content was determined by measuring the

mass o f a tuber before and after the water was removed by lyophilization. The equation used for
calculation was: % Moisture = [ (M Initial 

### - M Dried 

### )/M Initial 

### ] x 100

Moisture content: Covance Labs. Moisture levels were determined by Covance Laboratories using
Cov ance protocol M100T100_S:4 (AOAC 2005c ; AOAC 2005 d ).

Ash : Covance Labs. Ash levels were determined by Covance Laboratories using Covance protocol ASHM_S:5 (AOAC 2005 e )

.
Carbohydrates: Covance Labs. Carbohydrate levels were determined by Covance Laborator ies using Covance protocol CHO:6 (USDA 1973).

Calories: Covance Labs

. Total calories were determined by Covance Laboratories using Covance protocol CALC:4 (USDA 1975).

Crude Fiber : Covance Labs. Crude fiber was determined by Covance Laboratories using Co vance protocol CFIB_S:2 (AOAC 2005 f ).

Elements by ICP Emission Spectrometry : Covance Labs. The minerals Copper (Cu), Magnesium (Mg),and Potassium (K) were determined by Covance Laboratories using Covance protocol (ICP_S:13) (AOAC
2005 g ; AOAC 2005 h ).

Fat: Covance Labs. Fat was determined by Acid Hydrolysis using Covance protocol FAAH_S:7(AOAC

2005 i ; AOAC 2005 j ).

Niacin: Covance Labs. Niacin was determined by Covance Laboratories using Covance protocol NIAP_S:11 ( AOAC 2005 k; AOAC 2005 l ).

Protein : Cova nce Labs. Protein was determined using the Kjeldahl method, approximating protein by multiplying Nitrogen by 6.25, as per Covance protocol PGEN_S:4 (AOAC 2005 m ; AOAC 2005 n ).

Pyridoxine : Pyridoxine was determined by Covance Laboratories using Covance p rotocol B6A_S:11 (AOAC 2005 o , Atkin 1943).

Vitamin C and glycoalkaloids: Simplot Lab

. Vitamin C and total glycoalkaloids (sum of 

- chaconine and



- solanine) were extracted by following a fast extraction method as previously described by Shakya and

Navarr e (2006) with slight modification. In brief, freeze

- dried tuber powder (about 200 mg) was placed
into a 2 mL screw cap tube along with 0.9 mL of extraction buffer (50% Methanol, 3.0% metaphosphoric acid, 1 mM EDTA) and 500 mg of 1.0 mm glass beads. Tubes w ere shaken in a BeadBeater (Biospec, Bartelsville, OK) for 10 min at maximum speed, centrifuged for 5 min at 4 °C and the supernatant was transferred to a clean tube. The remaining pellet was re

- extracted with 0.6 mL of extraction buffer and centrifuged. T he supernatants were combined, centrifuged, and concentrated in a SpeedVac (Thermo Savant, Waltham, MA) prior to HPLC analysis. Gallic acid was added as an internal standard for quantification. Samples were kept chilled and protected from bright light at a ll times.

### J.R. Simplot Company Compositional Analyses 75



### An Agilent 1200 HPLC system (Agilent Technologies) equipped with an on

- line solvent degasser, quaternary pump, refrigerated autosampler, column heater, and variable wavelength diode

- array (DAD) and MS detectors was used for sample analysis. Mon olithic HPLC column Onyx C18, 100 x 4.6 (Phenomenex) was used with optimized parameters. Column temperature was 35°C, flow of 1 mL/min
with a gradient elution of 0−1 min 100% buffer A (10 mM formic acid, pH 3.5, with NH 4 

### OH), 1−9 min 0−30% buffer B (100% methanol with 5 mM ammonium formate), 9−10.5 min 30% buffer B, 10.5−14
min 35−6 5% buffer B, 14

- 16 min 65%

- 100% buffer B, and 16

- 16.5 min 100% buffer B. Data acquisition and instrument parameters were controlled using the Agilent ChemStation . UV Diode

- Array Detector (DAD) detection and quantification were at wavelength 244 nm for vitamin C and 210 nm for glycoalkaloids. The external standard met hod of calibration was used, with each curve prepared from 6
to 8 different concentrations of standard solutions.

Vitamin C and glycoalkaloid levels were further confirmed by LC

- MS analysis with an Agilent 1200 HPLC coupled to 6320 ion trap LC/MS. Experim ents were carried out with an ESI source in negative ion mode for vitamin C and positive ion mode for glycoalkaloids. The source was operated using 350°C drying gas (N2) at 12 L/min, 55 psi nebulizer gas (N 2 

### ),and the source voltage with a scan range of m / z 100

- 1300. Automated MS ( 2 ) analysis was conducted using SmartFrag and Agilent ChemStation software (Agilent) with a ramp range from 1500

- 4500 V.

Vitamin C: Covance Labs. Vitamin C levels were determined by Covance Laboratories using protocol VCF_S:5 (AOA C 2005b).

Glycoalkaloids: Covance Labs. Glycoalkaloid levels were determined by Covance Laboratories using protocol COID_S:2 (AOAC 2006).

Free amino acid s: Simplot Lab. Free amino acids were extracted by homogenizing 250 mg ground
freeze

- dried tubers w ith 5 μmol sarcosine as an internal standard in 3.0 mL 0.03 M triethylamine HCl buffer and adding (a) 150 μ 85 mM K hexacyanoferrate trihydrate (K 4 

### Fe(CN) 6 

### 3H 2 

### O), (b) 150 μL 100 mM zinc sulfate (ZnSO 4 

### .7H 2 

### O), and (c) 250 μL 0.1 N NaOH with 3.0 mL 0.03 M TEA b uffer pH 7.0, vortexing the mixture after each addition. The extract was centrifuged for 15 min at 4°C at 4,000 rpm, and the supernatant was transferred to a new tube. The pellet was resuspended in 5 mL nanopure water and centrifuged. The first and second supernatants were pooled and the final volume was adjusted to 12.5 mL with water. The extracted free amino acids were derivatized using the EZ:faast method according to the user’s manual from the manufacturer (Phenomenex, Torrance, CA). Derivatized samples were analyzed by liquid chromatography

- mass spectrometry (LC

- MS) using an Agilent 1200 series HPLC system that was coupled to an Agilent 6300 series ion trap. Bruker’s quant analysis software was used for quantification. For HPLC, we used a 25.0 x 0.3 cm EZ:faast AAA

- MS column, and the mobile phase was 10 mM ammonium formate in water (A) and 10 mM ammonium formate in methanol 1:2, v/v (B), flow rate 0.25 min/mL with a gradient of 68

- 83% B in 13 min and 13

- 18 min 68% buffer B. MS was run in the positive mod e with ESI and auto MS n 

### . The limit of quantitation for most matrices was 0.1 mg/g dry weight (DW) (or 100 ppm in the fresh weight)

.

Free amino acids: Covance Labs. Free amino acid levels were determined by Covance Laboratories using the Covance protocol F A ALC_S :6 ( Schuster 1988 ; Henderson 2000 ; and Barkholt and Jensen 1989).

### J.R. Simplot Company Compositional Analyses 76



### Tryptophan: Covance Labs. Tryptophan levels were determined by Covance Laboratories using the Covance protocol TRPLC_S:3 (AOAC 2005 a ).

Total amino acid s : Simplot Lab

. Amino acid hydro lysis for total amino acid analys is was performed as previously described (Purcell et al . 1972 ; Miedel et al

. 1989) with optimization. About 50 mg freeze dried tissue was weighed and transferred into the bottom of a vacuum hydrolysis tube (5 mL, 10 mm x
10 0 mm (Wilmad LabGlass, NJ). A 1 mL aliquot of constant boiling, 6N hydrochloric acid (Thermo Scientific/Pierce, IL) in the presence of 4 % thioglycholic acid was added directly to the sample. Vacuum was applied until no air bubbles were seen and the tube w as sealed slowly by screwing the PTFE plug down. The hydrolysis was carried at 110°C for 24 hr on a dry block heating system (Reacti

- therm heating module, Thermo Scientific/Pierce, IL). Following hydrolysis and cooling, pressure in the tube was
released sl owly. The sample was removed by Pasteur pipette and a 20 μL aliquot of the sample was vacuum dried using an acid resistant speedVac (Savant SC250DDA, Thermo Scientific, IL). Samples were

re

- suspended in water to make a 10 fold dilution and centrifuged. TR P, cystine and CYS are unstable during acid hydrolysis and so were not estimated quantitatively. GLN and ASNs are deaminated to form
GLU and ASP. An aliquot of the supernatant was subjected to solid phase extraction (SPE) and derivatization steps using the EZ

- faast: easy

- fast amino acid sample testing kit (Phenomenex, Torrance, CA) as per manual. The derivatized amino acids were analyzed using a LC/MS instrument (Agilent Technologies) as described in free amino acid analysis.

Total amino acids: Covance Lab s. Total amino acid levels were determined by Covance Laboratories using the Covance protocol TAALC_S:6 (Schuster 1988 ; Henderson 2000 ; and Barkholt and Jensen 1989).

Sugars : Simplot Lab

. Sugars were extracted by shaking approximately 150 mg freeze

- dried tissues in 1
mL 60% et hanol at 80 °C for 1 h. The supernatant was transferred into a fresh tube, and the pellet was

re

- extracted with 1 mL 60% ethanol for 30 min at 80 °C. The supernatant volume was reduced in a Speedvac to 60 to 70 μL. A known amount of ribose was added as an internal standard. Sugar analyses were performed on an Agilent 1200 series HPLC system, which consisted of an auto sampler, Zorbax carbohydrate column (4.6 × 150 mm, Agilent Technologies, USA), a solvent system of acetonitrile

- water (75 : 25), and a refrac tive index detector, at a flow rate of 1 mL/min. Sugars were quantified using Agilent ChemStation software with external calibration.

Sugars: Covance Labs. Sugar levels were determined by Covance Laboratories using High Performance Anion Exchange Chromatog raph (HPAEC) equipped with a Pulsed Amperometric Detector (PAD) and following Covance protocol SWET_S:9 (Lilla et al. 2005)

.
Acrylamide: The acrylamide levels were determined by Covance Laboratories in Greenfield, IN using the Covance protocol ACMS_GRN_S:4 ( FDA 2003)

.
Statistical Analysis. All attributes for the Russet Burbank, Ranger Russet, and Atlantic lines were analyzed by com bin ing data from multiple test years and locations using the following linear mixed model:

Y ijkm 

### =  i + 

###  j 

### +  k( j ) 

### +  l(j,k) 

### + (   ) i k 

### +  ijk l m

### J.R. Simplot Company Compositional Analyses 77



### •  = mean of treatment (fixed)

•  = year (random)

•  = effect of location [year] (random)

•  = rep [year, location] (random)

•  = residual random error

 j ~ 

### iidN (0,  2 year 

### ),  k( j ) ~ 

### iidN (0,  2 loc( year ) 

### ),  l(j,k) ~ 

### iidN (0,  2 rep(year, loc ) 

### ), (   ) i k ~ 

### iidN (0,  2 loc*treatment 

### ), and  ijkm ~ 

### iidN (0,  2 

### )

(This means that all error terms are assumed to be normally distributed with a mean of 0 and equa l variances ).

Where  i 

### denotes the mean of the i th 

### treatment (fixed effect),  j 

### denotes the effect of the j th 

### year (random effect),  k(j) 

### are the random location (within year effect )

,  l(j,k) 

### are the rep with in year and location effect, (   ) i k 

### denotes the interaction between the i th 

### treatment and random k th 

### location within year effect

, and  ijklm 

### denotes the residual random error.

We tested all data for normality before conducting the analysis mentioned here. Most data met the requirement, but with some attributes such as free amino acids, many distributions were skewed by being too c lose to the detection limit. In those cases, we simply reported means and ranges and did not test for statistical significance.

In cases where all data were from one trial year

, the statistical model did not include the year term.
D ata from multiple lo cations were analyzed together using the following linear mixed model:

Y ijkm 

### =  i + 

###  j 

### +  k(j ) 

### + (   ) ij 

### +  ijkm



### •  = mean of treatment (fixed effect)

•  = effect of location (random)

•  = rep within location effect (random)

•  = residual random error

 j ~ 

### iid N (0,  2 loc 

### ),  k(i) ~ 

### iidN (0,  2 loc(rep) 

### ), (   ) ij ~ 

### iidN (0,  2 loc*treatment 

### ), and  ijkm ~ 

### iidN (0,  2 

### )

(This means that all error terms are assumed to be normally distributed with a mean of 0 and equa l variances ).

Where  i 

### denotes the mean of the i th 

### treatme nt (fixed effect),  j 

### denotes the effect of the j th 

### location (random effect),  k(j) 

### are the random replicate (within location) effects, (   ) ij 

### denotes the interaction between i th 

### treatment and j th 

### location (random effect), and  ijkm 

### denotes the residual random error.

Following analysis with the mixed linear model s , we calculated the False Discovery Rate to minimize the chance of mistakenly concluding that the transformed events were different from controls. When applying this technique, we organized the data in to logical related groups for c omparison. Those groups

### J.R. Simplot Company Compositional Analyses 78



### were: c ompositio n, free amino acids, and t otal a m ino a cids. Thus, t he difference between least squares ( LS ) means of treatment lines and the LS means of corresponding control line s for traits under

c ompositional, f ree a mino a cids and t otal a mino a cids groups w as first compared using Students pair

wise comparison method. Then

, the P

- values were adjusted following the false discovery rate (FDR) approach (Benjamini and Hochberg 1995) within each group, to control the percentage of false signals. A significant difference was established with a FDR

- A djusted P

- V alue < 0.05.

For traits considered independent of the basic composition, it would not be desirable to apply the FDR
method. Fo r example, potato yield , acrylamide, sugar, and glycoalkaloids would not be closely related to basic composition, so it would be preferred to use only Tukey’s test to compare the events with
controls. Thus, t he difference between LS means of treatment lin es and the LS means of corresponding control lines for yield, a crylamide, s ugar and g lycoalkaloids was compared using Tukey’s method. A significant difference was established with a P

- V alue < 0.05.

The following varieties, all grown in the field trial sit es, were used to calculate tolerance intervals ( Vardeman 1992 ) : Snowden, Chieftain, Red Norland, Ida Rose, Russet Burbank, Ranger Russet, Atlantic, G, and H. Total data points were 317 for proximate analysis, 193 for glycoalkaloids, 337 for both free
amin o acids, 252 total amino acids, and 227 for sugars. The tolerance intervals were calculated to contain, with 95% confidence, 99% of the values contained in the population. This statistical tolerance interval and the combined range of values for each analy te available from the published literature were used to interpret the composition results. In interpreting the data, emphasis was placed on the analyte means; means that fell within the tolerance interval and/or combined literature range for the analyte

we re considered to be within the normal variability of commercial potato varieties.

### J.R. Simplot Company Compositional Analyses 79 

### CONCLUSION S

T he compositional assessment studies, evaluating proximates, vit amins, amino acids, mi nerals, sugars, and glycoalkaloids, were conducted on Innate TM po tatoes to 1) show equivalence to the untransformed controls, 2) compare the Innate TM potatoes to literature ranges, where applicable; and 3) show that there were no changes in allergens or potential toxins. Ultimately, results of the composition studies
sh owed food safety equivalence between the events and the untransformed controls.

Results of all significant differences between the events and controls are summarized in Table 34 for Ranger Russet and Russet Burbank and in Table 35 for Atlantic, G, and H va rieties. Most of these differences; reduced free ASN and total ASP + ASN, increased free GLN and total GLU + GLN, reduced acrylamide, and lowered reducing sugars were expected because of the intended gene silencing. Other significant differences included decreased free VAL (J3, J55), decreased free LYS (H37), increased free ARG (F37), increased total LYS (G11), increased total PRO (G11), increased Vitamin C (F10, F37), increased niacin (F10, F37), decreased pyridoxine (J3, J55), decreased sucrose (E12, E2 4), and increased sucrose (H37). All these observed differences fell wi thin the tolerance intervals or combined literature values, and therefore were considered to be within normal ranges for potatoes.

It’s important to note that for events E12 and E24, th e free ASN levels were not statistically different from controls at the adjusted p < .05 level . However, these events showed a 7 4 to 76% reduction in free
ASN , which was significant at the adjusted p = .11 level. Such large differences were sometimes not statistically significant, which was a direct result of using the correction for False Discovery Rate (Benjamini and Hochberg 1995). These reductions were of practical significance and are considered similarly to the other events regarding this trait.

N ote that free amino acids would not normally be part of a composi tional assessment, however, they

were included because the quantity of free asparagine is important to show the efficacy of silencing the
Asn1 gene for the purpose of reducing acrylamide. Ma ny of the free amino acids are found at such low levels in potatoes that values barely exceed the detection limit. We chose to adjust all values that were below detection limits to 100 to complete the statistical analysis, and have reported the mean value s. However, many of these values were too low to allow an estimate of variance, so P

- Values were not reported. It might be just as appropriate to report most of these values as “Not Available”, but we chose to include them. For the purpose of assessing the nutritional equivalence of the events, we recommend that the focus should be on comparing total amino acids to controls.

The most important change between tubers of the events and their untransformed controls relates to
reduction of the amino acid a sparagine (ASN) and reducing sugars

. Reduced amounts of free ASN and reducing sugars in potatoes of the events resulted in lowered acrylamide after heat processing compared to the conventional potatoes

.

### J.R. Simplot Company Compositional Analyses 80



### Table 34 . Statistical Differences Observed in Compo sition Data: Events F10, F37, E12 and E24 Variety Event Attribute Difference Expected? Comments

Ranger Russet F10 Decreased Free ASN 

### Yes Increased Free GLN 

### Yes Decreased Total ASP+ASN 

### Yes Increased Total GLU+GLN 

### Yes Decreased Reducing Suga rs 

### Yes Glucose and Fructose Reduced Acrylamide 

### Yes Increased Niacin (B3) 

### No Within TI and CLR
Increased Vitamin C 

### No Within TI and CLR Ranger Russet F37 Decreased Free ASN 

### Yes Increased free ASP 

### Yes Increased Free GLN 

### Yes Decreased T otal ASP+ASN 

### Yes Increased Total GLU+GLN 

### Yes Decreased Reducing Sugars 

### Yes Glucose and Fructose Reduced Acrylamide 

### Yes Increased Vitamin C 

### No Within TI and CLR Increased Niacin (B3) 

### No Within TI and CLR
Increased free ARG 

### No Within TI and CLR
Russet Burbank E12 Decreased Free ASN 

### Yes 74 % reduced, not significant Decreased Total ASN+ASP 

### Yes Increased Total GLU+GLN 

### Yes Decreased Reducing Sugars 

### Yes Glucose and Fructose Reduced Acrylamide 

### Yes Decreased Sucrose

Reduced A crylamide 

### No

Yes Within TI and CLR
Russet Burbank E24 Decreased Free ASN 

### Yes 76% reduced, not significant Decreased Total ASP+ASN 

### Yes Increased Total GLU+GLN 

### Yes Decreased Reducing Sugars

Decreased Sucrose

Reduced Acrylamide 

### Yes

No

Yes Glucose and Fructose

Within TI and CLR Reduced Acrylamide 

### Yes Decreased Sucrose

Reduced Acrylamide 

### No

Yes Within TI and CLR
TI = tolerance interval CLR = combined literature range

### J.R. Simplot Company Compositional Analyses 81



### Table 35. Statistical Differences Observed in Composition Data: Even ts J3, J55, J78, G11, H37 and H50 Variety Event Attribute Difference Expected? Comments

Atlantic J3 Decreased Free ASN 

### Yes Decreased Reducing Sugars 

### Yes Glucose and Fructose Reduced Acrylamide 

### Yes Decreased Pyridoxine (B 6 

### ) 

### No Within TI and CLR
Decreased Free VAL 

### No Within TI and CLR
Atlantic J55 Decreased Free ASN 

### Yes Decreased Reducing Sugars 

### Yes Glucose and Fructose Reduced Acrylamide 

### Yes Decreased Pyr idox ine (B 6 

### ) 

### No Within TI and CLR
Decreased Free VAL 

### No Within TI and CLR
Atlantic J78 Decreased Free ASN 

### Yes Adjusted P = .0548 Decreased Reducing Sugars 

### No Glucose and Fructose Reduced Acrylamide 

### Yes Variety G G11 Decreased Free ASN 

### Yes Increased Free GLN 

### Yes Decreased Total ASP+ASN 

### Yes Increased Total GL U+GLN 

### Yes Reduced Acrylamide 

### Yes Increased Total LYS 

### No Within TI and CLR
Increased Total PRO 

### No Within TI and CLR Variety H H37 Decreased Free ASN 

### Yes Increased Free GLN 

### Yes Decreased Total ASP+ASN 

### Yes Increased Total GLU+GLN 

### Yes Increased Sucrose 

### No Within TI and CLR Reduced Acrylamide 

### Yes Decreased Free LYS 

### No Within TI and CLR Variety H H50 Decreased Free ASN 

### Yes Increased Free GLN 

### Yes Decreased Total ASP+ASN 

### Yes Increased Total GLU+GLN 

### Y es Reduced Acrylamide 

### Yes
TI = tolerance interval CLR = combined literature range

### J.R. Simplot Company Compositional Analyses 82



### For the compositional assessment, all events were compared with their respective controls mostly at the time of harvest. In addition, the potatoes were tested for reducing sugars and acrylamide, following
storage.

Many of the events had lowered levels of reducing sugars either at the time of harvest or after storage for 1 month (F10, F37, E12, E24, J3, and J55, J78), however, in most cases we did not observe sign ificant
differences after 2

- 5 months of storage. It was not expected that J78 would have lowered levels of reducing sugars because it was one of t wo events, J78 and G11, that did not contain the R1/PhL promoter silencing cassette

. Tubers of event s G11 , H 37, and H50 contain the same amount of reducing sugars as tubers of their untransformed counterpart s

. The inability of the silencing construct to limit glucose/fructose formation in H37 and H50 may be due to the fact that the H variety is naturally low in glucose and fructose. Thus, we concluded that silencing of the promoters associated with the PhL/R1 genes effectively lowered reducing sugars near the time of harvest in most events but these differences

were not su stained throughout storage for 2

- 5 month s.

During storage of up to 7 months, acrylamide levels in most events were consistently lower (p < .0001) than controls, although there were exceptions with the G and H varieties. In event G11, we observed a mean of 2,050 ppb compared with 1,769 ppb for t he control after 1 month of storage. This was unusual and throughout the rest of the study, acrylamide levels in event G11 were always much lower than controls (46 to 58% reduction at months 5 and 7). For variety H, acrylamide levels were always lower i n events H37 or H50, but not statistically significant in months 5 and 7. Overall, the modifications from
insertion of pSIM 1278 resulted in dramatic reductions in acrylamide that persisted throughout the typical storage periods.

This research also confir m ed that the events are as safe as controls and did not contain increased levels of natural glycoalkaloids, which are toxins commonly found in Solanaceous crops

, including potato

. Based on the lack of differences between the event s and control s , there wou ld not be a safety issue as a result of the introduction of the reduced acrylamide potential and low black spot bruise traits in Innate TM potatoes. All glycoalkaloid levels were within the normal ranges for potato, when compared with the
tolerance interval s or combined literature ranges.

Because tubers of the events are substantially equivalent to tubers from the untransformed controls

, except for the intended result of lowered ASN , reducing sugars, and acrylamide

, the potatoes will be as
safe as their untr ansformed controls for use as food and feed. In contrast, the transition from standard
tubers to Innate TM tubers, which have significantly lower acrylamide

- forming potential, will reduce the average total exposure to dietary acrylamide by almost one

- third because about 42% of this acrylamide is associated with the consumption of french fries and potato chips in the United States and Europe (Dybing et al . 2005; Mucci and Wilson 2008).

While some statistically significant differences in analyte levels were f ound and discussed, i n cases where the Innate TM tubers were statistically different from the untransformed control variety, the values remained within the natural variation for potatoes, based on tolerance intervals calculated from the
controls or literatur e references.

### J.R. Simplot Company Compositional Analyses 83 

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### J.R. Simplot Company Compositional Analyses 88 

### Annex A

Proximates, Vitamins, Minerals, Free Amino Acids, and Total Amino Acids

– Combined Literature Ranges (CLR) Used in Tables

Proximates, Vitamins, Minerals CLR 2

Substance Units Value Reference Formula Used 1 Formu la Calculation End Units

Protein low % 0.70 Lisinska and Leszczynski 1989 NA 0.70 % Protein high % 4.60 Lisinska and Leszczynski 1989 NA 4.60 %

Fat low % 0.02 Lisinska and Leszczynski 1989 NA 0.02 % Fat high % 0.20 Lisinska and Leszczynski 1989 NA 0.20 %

Ash low % 0.44 Lisinska and Leszczynski 1989 NA 0.44 % Ash high % 1.90 Lisinska and Leszczynski 1989 NA 1.90 %

Crude fiber low % 0.17 Lisinska and Leszczynski 1989 NA 0.17 % Crude fiber high % 3.50 Lisinska and Leszczynski 1989 NA 3.50 %
Carbohydra tes low % 13.30 Talburt and Smith 1987 NA 13.30 % Carbohydrates high % 30.53 Talburt and Smith 1987 NA 30.53 %

Calories low kcal/100g 70.00 Horton and Anderson 1992 NA 8 0.00 kcal/100g

Calories high kcal/100g

dry 551.00 Horton and Anderson 1992 kcal/100g x 0.2 1 110.20 kcal/100g

Moisture low % 63.20 Talburt and Smith 1987 NA 63.20 % Moisture high % 86.90 Talburt and Smith 1987 NA 86.90 %

B3 low mg/200g 0.18 Rogan et al. 2000 mg/200g x 0.5mg/100g 0.09 mg/100g
B3 high mg/200g 6.20 Rogan et al. 2000 mg/20 0g x 0.5mg/100g 3.10 mg/100g

B6 low mg/200g 0.26 Rogan et al. 2000 mg/200g x 0.5mg/100g 0.13 mg/100g
B6 high mg/200g 0.82 Rogan et al. 2000 mg/200g x 0.5mg/100g 0.41 mg/100g

Vitamin C low mg/100g 1.00 Lisinska and Leszczynski 1989 NA 1.00 mg/100g

Vitam in C high mg/100g 54.00 Lisinska and Leszczynski 1989 NA 54.00 mg/100g

Cu low mg/200g 0.03 Rogan et al. 2000 mg/200g x 5 = mg/kg = ppm 0.15 ppm Cu high mg/200g 1.40 Rogan et al. 2000 mg/200g x 5 = mg/kg = ppm 7.00 ppm Mg low mg/200g 22.50 Rogan et al. 2 000 mg/200g x 5 = mg/kg = ppm 112.50 ppm Mg high mg/200g 110.00 Rogan et al. 2000 mg/200g x 5 = mg/kg = ppm 550.00 ppm
K low mg/200g 700.00 Rogan et al. 2000 mg/200g x 5 = mg/kg = ppm 3500.00 ppm K high mg/200g 1250.00 Rogan et al. 2000 mg/200g x 5 = mg /kg = ppm 6250.00 ppm
1 0.2 = approximate conversion from dry weight to fresh weight (internal communication), NA = value already converted

2 CLR=combined literature range

### J.R. Simplot Company Compositional Analyses 89 1 0.2 is the approximate conversion from dry weight to fresh weight ( internal commun ication)

2 CLR=combined literature range Free A mino A cids CLR 2

Substance Value Units Reference Formula Wt (FW) Formu la Used 1 (ppm)

ALA low 6.0 mg/100g Davies 1977 89.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 12 ALA high 118.0 mg/100g Davies 1977 89.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 236

ARG low 60.0 mg/100g Davies 1977 174.2 mg/100g x 10mg/1 000g x 1000g/kg x 0.2; mg/kg=ppm 120 ARG high 736.0 mg/100g Davies 1977 174.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 1472

ASN low 11.8 mM/kg Shepherd et al. 2010 132.1 mM/kg x FW x 0.2 312 ASN high 3490.0 mg/100g Davies 1977 132.1 mg/100g x 10 mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 6980

ASP low 32.0 mg/100g Davies 1977 133.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 64 ASP high 376.0 mg/100g Davies 1977 133.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 752

GABA low 15.0 mg/100g Davi es 1977 NA mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 30 GABA high 448.0 mg/100g Davies 1977 NA mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 896

GLN low 220.0 mg/100g Davies 1977 146.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 440

GLN h igh 2968 .0 mg/100g Davies 1977 146.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 5396

GLU low 225.0 mg/100g Davies 1977 147.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 450 GLU high 371.0 mg/100g Davies 1977 147.1 mg/100g x 10mg/1000g x 1000g/ kg x 0.2; mg/kg=ppm 742

GLY low 1.0 mg/100g Lisinska and Leszczynski 1989 75.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 2

GLY high 52.0 mg/100g Lisinska and Leszczynski 1989 75.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 104

HIS low 17.0 mg/100g Davies 1977 155.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 34 HIS high 328.0 mg/100g Davies 1977 155.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 656

ILE low 14.0 mg/100g Davies 1977 131.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/k g=ppm 28 ILE high 165.0 mg/100g Davies 1977 131.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 330

LEU low 8.0 mg/100g Davies 1977 131.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 16 LEU high 131.0 mg/100g Davies 1977 131.2 mg/100g x 10mg/1000 g x 1000g/kg x 0.2; mg/kg=ppm 262

LYS low 9.0 mg/100g Davies 1977 146.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 18 LYS high 319.0 mg/100g Davies 1977 146.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 638

MET low 9.0 mg/100g Davies 1977 149 .2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 18 MET high 108.0 mg/100g Davies 1977 149.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 216

ORN low 2.0 mg/100g Davies 1977 NA mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 4 ORN high 244.0 mg/ 100g Davies 1977 NA mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 488

PHE low 0.0 mg/100g Davies 1977 165.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 0 PHE high 204.0 mg/100g Davies 1977 165.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 40 8

PRO low 0.0 mg/100g Davies 1977 115.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 0 PRO high 484.0 mg/100g Davies 1977 115.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 968

SER low 15.0 mg/100g Davies 1977 105.1 mg/100g x 10mg/1000g x 1000g/ kg x 0.2; mg/kg=ppm 30 SER high 128.0 mg/100g Davies 1977 105.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 256

### J.R. Simplot Company Compositional Analyses 90 2 CLR=combin ed literature range Free A mino A cids (Continued) CLR 2

Substance Value Units Reference Formula Wt (FW) Formula Used 1 (ppm)

THR low 14.0 mg/100g Davies 1977 119.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 28 THR high 143.0 mg/100g Davies 1977 1 19.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 286

TRP low 7.0 mg/100g Davies 1977 204.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 14 TRP high 174.0 mg/100g Davies 1977 204.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 348

TYR low 9.0 mg/100g Lisinska and Leszczynski 1989 181.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 18

TYR high 319.0 mg/100g Lisinska and Leszczynski 1989 181.2 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 638

VAL low 15.0 mg/100g Davies 1977 117.1 mg/100 g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 30
VAL high 370.0 mg/100g Davies 1977 117.1 mg/100g x 10mg/1000g x 1000g/kg x 0.2; mg/kg=ppm 740

1 0.2 is the approximate conversion from dry weight to fresh weight (internal communication)

### J.R. Simplot Company Compositional Analyses 91

Total Amino Acids CLR 2

Substance Value Units Reference Formula
Wt (FW) Formula Used 1 Calc (ppm)

THR low 18.3 μM/g Talley et al. 1984 119.1 μM/g x FW x 0.2 436 THR high 35.9 μM/g Talley et al. 1984 119.1 μM/g x FW x 0.2 855

SER low 23.8 μM/g Talley et al. 1984 105.1 μ M/g x FW x 0.2 500 SER high 48.6 μM/g Talley et al. 1984 105.1 μM/g x FW x 0.2 1022

PRO low 15.4 μM/g Talley et al. 1984 115.1 μM/g x FW x 0.2 355 PRO high 63.6 μM/g Talley et al. 1984 115.1 μM/g x FW x 0.2 1464

ALA low 22.0 μM/g Talley et al. 1984 89. 1 μM/g x FW x 0.2 392 ALA high 53.4 μM/g Talley et al. 1984 89.1 μM/g x FW x 0.2 952

VAL low 32.1 μM/g Talley et al. 1984 117.1 μM/g x FW x 0.2 752 VAL high 61.9 μM/g Talley et al. 1984 117.1 μM/g x FW x 0.2 1450

ILE low 20.0 μM/g Talley et al. 1984 131.2 μM/g x FW x 0.2 525 ILE high 36.3 μM/g Talley et al. 1984 131.2 μM/g x FW x 0.2 953

LEU low 26.1 μM/g Talley et al. 1984 131.2 μM/g x FW x 0.2 685 LEU high 52.7 μM/g Talley et al. 1984 131.2 μM/g x FW x 0.2 1383

TYR low 12.6 μM/g Talley et al. 19 84 181.2 μM/g x FW x 0.2 457 TYR high 26.0 μM/g Talley et al. 1984 181.2 μM/g x FW x 0.2 942

PHE low 16.7 μM/g Talley et al. 1984 165.2 μM/g x FW x 0.2 552 PHE high 32.9 μM/g Talley et al. 1984 165.2 μM/g x FW x 0.2 1087

HIS low 4.3 μM/g Talley et al. 1984 155.2 μM/g x FW x 0.2 133 HIS high 15.1 μM/g Talley et al. 1984 155.2 μM/g x FW x 0.2 469

ORN low 0.0 μM/g Talley et al. 1984 136.16 μM/g x FW x 0.2 0 ORN high 1.7 μM/g Talley et al. 1984 136.12 μM/g x FW x 0.2 46

LYS low 23.5 μM/g Talley et al. 1 984 146.2 μM/g x FW x 0.2 687 LYS high 46.8 μM/g Talley et al. 1984 146.2 μM/g x FW x 0.2 1368

ARG low 20.1 μM/g Talley et al. 1984 174.2 μM/g x FW x 0.2 700 ARG high 39.7 μM/g Talley et al. 1984 174.2 μM/g x FW x 0.2 1383

TRP low 2.8 μM/g Talley et al

. 1984 204.2 μM/g x FW x 0.2 114 TRP high 6.9 μM/g Talley et al. 1984 204.2 μM/g x FW x 0.2 282

ASP low 677 Mg/200g Rogan et al. 2000 NA mg/200g x 5 = mg/kg = ppm 3385 ASP high 1476 Mg/200g Rogan et al. 2000 NA mg/200g x 5 = mg/kg = ppm 7380

GLU low 5 83 Mg/200g Rogan et al. 2000 NA mg/200g x 5 = mg/kg = ppm 2915 GLU high 1207 Mg/200g Rogan et al. 2000 NA mg/200g x 5 = mg/kg = ppm 6035

1 0.2 is the approximate conversion from dry weight to fresh weight (internal communication)

2 CLR=combined literatu re range