1. TSNA Sub-group Storage sub-committee Report Agronomy/Phytopathology Study Group Meeting - October 2007 Krakow, Poland Selected data from two reports Presented by Dr. Lowell Bush University of Kentucky, USA

2. 2006 Coresta Air-Cured TSNA Storage Study Dwayne Beeson – RJR Steve Terrell – Swedish Match NA

3. Objective To determine TSNA change in burley and dark air- cured tobacco leaf from end-of-curing until initial processing (threshing and re-drying). Background / Test Matrix Three burley producers and one dark air-cured producer participated in study. Burley producers were relatively new to burley production and were first and second year growers located in the Piedmont section of North Carolina and Virginia. The dark air-cured producer was located in Western Kentucky and is well experienced in growing both burley and dark air-cure.

4. Cured Leaf Storage ProducerCured Leaf Storage Package On-Farm Storage Location Final Market Package Poole~125 lbs. per sheetStored inside “pack house” on wooden floor Large Bale (42” x 42” x 40”) (~ 1 cubic meter) Renegar~ 200 lbs. per sheetStored on chicken house floor on spaced lumber, ~ 2 sheets high Large Bale EatonNone – stripped and sent to market within 1 week of stripping NALarge Bale Elliott~ 80 lbs. traditional farmer bale Stacked on pallets on concrete floor in metal building Farmer Bale

5. Warehouse Receiving / Core Sampling

6. Sampling Lamina only – samples were freeze-dried within 24 hours Composite samples were collected via combination of coring tool and “grab” whole leaf – mid rib removed All burley samples and related data are single submissions (N = 1) All Dark Air-Cure (Elliott) samples are in triplicate (N=3) ProducerSheet SampleBale Sample PooleGrabCore RenegarGrabCore EatonNACore ElliottNACore

7. Bale Weights / (%) Moisture / Grade ProducerLow Stalk lbs./Moisture/Grade Mid Stalk lbs./Moisture/Grade Upper Stalk lbs./Moisture/Grade PooleNA454 / NA / B1NA Renegar512 / 24.06% / X1509 / 26.71% / B1536 / 25.39% / BFR1 Eaton606 / NA / X1607 / NA / B1601 / NA / BFR2 ElliottNA Receiving Station moisture equipment was only operable for Reneger delivery Poole Lower & Upper stalk bales identity lost after delivery Poole farm storage moisture ~ 22% by feel Eaton bale moisture ~ 24% by feel X1 – Premium quality tan, thin, over ripe, fluffy B1 – Premium quality tan uniform color and length BFR1 – Premium quality bodied, dark red, uniform color and length BFR2 – Very good quality bodied, dark red, uniform color and length

8. Average Storage Temp (F) / RH (%) On-Farm Storage AmbientTobacco ProducerTemp (F)RH (%)Temp (F)RH (%)Weeks Poole54665772~4 Renegar49785182~10 EatonNA ~1 Elliott48674879~4 Warehouse Storage AmbientTobacco ProducerTemp (F)RH (%)Temp (F)RH (%)Weeks Poole55585659~8 RenegarNA 0 Eaton55585479~6 ElliottNA

9. Renegar Data Sample Date Stalk Position Nitrogen (%) Nitrate (%) Total Alkaloids (%) 2 nd Total Alkaloids (%) Nicotine (%) 2 nd / Total Ratio (%) Percent Convert (%) 11/06Low4.153.134.610.613.9913.327.97 Mid5.091.936.330.445.876.972.36 Upper5.831.846.510.446.076.872.76 1/11Low4.073.703.760.303.467.952.84 Mid5.083.405.020.344.676.742.38 Upper5.511.936.130.425.716.822.78

10. Renegar Secondary Alkaloids / TSNA DateStalk Position Nornicotine (%) Myosmine (%) Anabasine (%) Anatabine (%) NAB (ug/g) NAT (ug/g) NNK (ug/g) NNN (ug/g) TSNA (ug/g) 11/06Low0.34580.00870.02070.2385BDL2.3130.2774.0026.592 Mid0.14240.00460.02740.2663BDL1.5810.3061.5023.389 High0.17210.00370.02710.2442BDL1.3030.4010.9402.644 1/11Low0.10120.00410.01780.1756BDL5.6320.7243.5519.907 Mid0.11410.00460.02280.1972BDL12.5191.3608.72222.601 High0.16300.00410.02660.2243BDL2.2300.2873.0945.611 BDL – Below Detection Limit NAB <0.65 NAT <0.16 NNK <0.16 NNN <.01

11. Poole Data Sample Date Stalk Positio n Nitrogen (%) Nitrate (%) Total Alkaloids (%) 2 nd Total Alkaloids (%) Nicotine (%) 2 nd / Total Ratio (%) Percent Convert (%) 10/18Atstripping Mid4.871.634.560.394.188.463.66 Atreceivingstation 11/30Mid3.891.243.670.403.2810.805.85 Afterwarehousestorage 1/11Mid5.082.275.140.464.688.983.55

12. Poole --TSNA DateStalk Position NAB (ug/g)NAT (ug/g) NNK (ug/g) NNN (ug/g) TSNA (ug/g) 10/18Atstrippping MidBDL0.315BDL0.2250.540 Atreceivingstation 11/30MidBDL0.190BDL0.2920.482 Afterwarehousestorage 1/11MidBDL0.491BDL0.4460.937 BDL – Below Detection Limit

13. Elliott Data Sample Date Stalk Position Nitrogen (%) Nitrate (%) Total Alkaloids (%) 2 nd Total Alkaloids (%) Nicotine (%) 2 nd / Total Ratio (%) Percent Convert (%) 11/5Low4.223.653.7730.2923.4817.852.23 Mid4.212.613.8590.2713.5887.022.36 Upper4.411.505.0420.3514.6917.062.03 12/07Low4.193.963.4540.2663.1877.862.61 Mid4.202.714.0590.2873.7727.192.03 Upper3.971.344.5420.2824.2596.231.78 Average Results / N = 3

14. Elliott Secondary Alkaloids / TSNA DateStalk Position Nornicotine (%) Myosmine (%) Anabasine (%) Anatabine (%) NAB (ug/g) NAT (ug/g) NNK (ug/g) NNN (ug/g) TSNA (ug/g) 11/5Low0.0780.0020.0180.194BDL1.2940.3460.5562.195 Mid0.0870.0020.0170.165BDL1.4500.3810.7532.457 High0.0950.0030.0240.229BDL0.826BDL0.4011.227 4 weeks on farmstorage 12/07Low0.0830.0030.0160.164BDL1.4770.1890.8312.434 Mid0.0770.0030.02300.187BDL0.779BDL0.3791.158 High0.0770.0020.0210.182BDL1.5690.2980.7512.519 Average Results / N = 3 BDL – Below Detection Limit

15. Poole Temp/RH Data – Storage Shed Temp / RH recorder placed in center of sheet

16. Poole Temp / RH Receiving Station

17. Result Summary (TSNA ug/gm) ProducerStalk Pos.Initial Sample Final Sample Time (Weeks) (%) Change Renegar On-Farm Storage Only High Density Sheets Low6.599.911050% Mid3.3922.60“567% Upper2.645.61“112% Avg.4.2112.71202% Poole On-Farm Storage Low Density Sheets (4 weeks) Warehouse Storage (8 weeks) Mid0.540.486-11% Mid0.480.958+48% Avg. BDL – Below Detection Limit NA – Sample Not Available

18. Result Summary (TSNA ug/gm) ProducerStalk Pos.Initial Sample Final Sample Time (Weeks) (%) Change Eaton Warehouse Storage Only Low8.9710.77620% Mid6.903.48“-50% Upper2.363.46“46% Avg.6.085.90-3% Elliott On-Farm Storage Only Low2.202.43411% Mid2.461.16“-53% Upper1.232.52“105% Avg.1.962.044%

19. Post Curing Study 2005-2006 Altadis - Swedish Match

20. Trial I: Effect of storage mode Variety: ITB 501 (Burley)

21. Trial I (Lamina) * *

22. Trial I (Midrib) * * ** *

23. Trial II: Effect of pressure and moisture during 3 months of bale storage Experiments performed on cutters and tips Baling pressure: normal and low pressure (-50 %) Baling moisture: normal and low humidity - Normal = 22 % for tips and 18-20 % for cutters - Low = 15 % Variety: ITB 501 (Burley)

24. Trial II (Cutters-Lamina)