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Accession number 1309 Lorillard Research Center Greensboro WATER SOLUBLE ADDITIVES USED FOR PREVENTION OF NITROSATION DURING LABORATORY FERMENTATION OF PF2-1977 TOBACCO Submitted by: R. A. Shoffner, F. R. Perini and J. H. Bell Report number: H-173-S-3 Qate: 3/9/79 Summary or Abstract: Propyl gallate, a water soluble antioxidant, has been tested as an additive on tobacco. It was found to be 76% effective in controlling nitrosation at the 2% w/w level. RAS/sac Xc: Dr. H. J. Minnemeyer Dr. F. J. Schultz Library

INTRODUCTION This report describes work that has been done using water soluble additives on tobacco to control the formation of N-nitrosonornicotine. EXPERIMENTAL Fermentation experiments were carried out in the laboratory fermentation chamber. T4e tobacco system used for study was PF2-1977, ground and sieved. All experiments were run at ti70% moisture. The data were collected from two series of 4-day runs and one series of 8-day runs. RESULTS AND DISCUSSION Previous experiments have shown that hydroquinone was 95% efficient* at a 1% w/w level and 99% efficient at a 1.5% w/w level in controlling NNN formation. Although hydroquinone used topically in many cosmetic preparations, it has hazardous side effects when ingested. Because of the questionable health aspects of hydroquinone, other water soluble antioxidants were investigated. These included: Vitamin A Palmitate, L-Cysteine Hydrochloride, Tannic Acid, Propyl Gallate, and Sodium Bisulfite. Vitamin A palmitate and L-cysteine hydrochloride showed either an increase in ivNN formation or had little effect during an eight day run. Preliminary tests with sodium bisulfite and propyl gallate at 1% levels showed ti50% effectiveness for an eight-day run. Tannic acid at the same concentration was 38% effective. These results are shown in Table 1. Table 2 contains data collected from two four-day runs testing the last three above mentioned additives at higher levels. Sodium bisulfite at 2% and 5% levels interfered with both fermentation and nitrosation. Tannic acid at the 2% level was still only 39% effective. Propyl gallate at the 2% level showed an increase in efficiency to 76%.

CONCLUSIONS Propyl gallate is currently used as an antioxidant for foods. There is no specific data on human toxicity and animal experiments show very low toxicity. Its solubility is only 0.35g/100 ml H20 and therefore heat is necessary to increase the solubility. Economic figures on use of propyl gallate indicate the cost for treatinq 100 lbs of tobacco to be approx. $ .36/lb. this figure is based on cost of a small quantity of additive and would decrease if larger quantities were purchased. ** * A formula expressing the % efficiency of additives in preventing nitrosation on tobacco is explained in a current report. ** Price obtained from ICN Biochemicals Division and is based on a cost of $20.00 for 500g.

Table 1 Additive Concentration # Days % Nit. % TVB % Nic. COD pH NNN % Effectiveness (% w w (dry) (ppm) Untreated - - 4.23 1.07 3.21 145 6.4 4.2 PF2-1977 H20 (ti70$ moisture) - 8 3.86 0.56 3.17 127 8.5 24.3 - Vit. A Palmitate 1% 8 3.94 0.54 3.20 125 8.4 50.0 0 Vit. A Palmitate 5% 8 3.86 0.54 3.00 131 8.5 - 0 L-Cysteine Hydrochloride 1% 8 4.10 0.62 3.27 122 8.2 25.5 0 L-Cysteine Hydrochloride 5% 8 4.09 0.83 3.25 131 7.3 22.3 10.0 Sodium Bisulfite 1% 8 4.19 0.55 3.08 135 8.4 13.5 53.7 Propyl Gallate 1% 8 3.95 0.55 3.05 126 8.4 13.7 52.7 Tannic Acid 1% 8 3.99 0.53 2.94 126 8.4 16.6 38.3 TEV9II00

4; Table 2 Additive Concentration # Days % Nit. % TVB % Nic. COD pH NNN % Effectiveness (% w/w) (dry) (ppm) Untreated - - 4.23 1.07 3.21 145 6.4 4.2 PF2-1977 H20 (ti70$ moisture) - 4 3.93 0.62 3.29 127 8.5 30.6 Sodium Bisulfite 2% 4 3.75 0.64 3.33 138 8.1 it It 5% 4 3.99 1.08 3.32 143 6.9 11.7 71.6% Tannic Acid 2% 4 4.10 0.56 3.01 126 8.3 20.2 39.4% Propyl Gallate 2% 4 3.75 0.57 3.05 130 8.0 10.6 75.8% zEVstioo