Determination of Nicotine, pH, and Moisture Content of Six U.S. Commercial Moist Snuff Products -- Florida, January-February 1999

The use of smokeless tobacco (moist snuff and chewing tobacco) can cause oral cancer and precancerous oral lesions (leukoplakia) and is a risk factor for cardiovascular diseases and nicotine addiction (1). Despite these adverse effects, smokeless tobacco is used commonly in the United States by young people, especially male high school students (2). Officials in Florida requested CDC assistance in analyzing six moist snuff products to measure three factors that affect their nicotine dose: pH, nicotine content, and moisture content. This report summarizes the results of the analysis, which indicate that the pH, amount of nicotine, and moisture vary widely among brands.

During January 5-February 7, 1999, University of Miami staff and affiliated persons bought six smokeless tobacco products from stores in Daytona Beach, Fort Myers, Miami, Orlando, Tallahassee, and Tampa/St. Petersburg, Florida. These products were Copenhagen Snuff, Skoal Bandits Straight, Skoal Bandits Wintergreen, Skoal Long Cut Wintergreen, Kodiak Wintergreen, and Hawken Wintergreen,* and were chosen to reflect a cross-section of products from the five leading U.S. moist snuff brands sold in the United States during 1997 (3).

The pH, nicotine, and total moisture content in samples of the six products were analyzed at CDC using a federal standard protocol** (4). Samples were stored in their original containers at -95.8 F (-71 C) until tested. The pH was obtained by suspending 2 g of moist snuff in 10 mL distilled water. Total moisture content (water and tobacco constituents that are volatile at 211.1 F [99.5 C]) was obtained by calculating the weight difference in 5 g of tobacco before and after 3 hours of oven drying at 211.1 F (99.5 C). Nicotine was extracted from moist snuff by using methyl tertbutyl ether, and tobacco extracts were analyzed by gas chromatography to determine the nicotine content. The nicotine extraction and pH measurements were conducted at room temperature. The percentage of free (unprotonated) nicotine, which is dependent on the pH, was calculated according to the Henderson-Hasselbalch equation and by using a pKa value of 8.02 for nicotine (5). Free nicotine content then was calculated by multiplying the percentage of free nicotine by the total nicotine content (percentage of free nicotine x nicotine content). The tests were not blinded to the brands being tested, and all analyses were done in triplicate. Statistical analyses were performed using Statistical Analysis System (SAS) software.

The mean total moisture content ranged from 48.9% to 54.1%, except Hawken Wintergreen, which had a mean total moisture content of 24.7%; the mean nicotine content varied from 7.11 mg/g to 11.04 mg/g, except Hawken Wintergreen, which had a mean nicotine content of 3.37 mg/g; the mean pH varied from 5.24 (Hawken Wintergreen) to 8.35 (Kodiak Wintergreen). The mean amount of nicotine per dry tobacco weight ranged from 0.45% (Hawken Wintergreen) to 2.41% (Skoal Long Cut Wintergreen). Mean free nicotine levels varied from 0.01 mg/g (Hawken Wintergreen) to 6.23 mg/g (Copenhagen Snuff). The percentage of free nicotine varied from a mean value of 0.23% (Hawken Wintergreen) to 68.14% (Kodiak Wintergreen) ( Table 1).

Reported by: Univ of Miami; Florida Office of Tobacco Control, Florida Dept of Health. Air Toxicants Br, Div of Laboratory Sciences, National Center for Environmental Health; Office on Smoking and Health, National Center for Chronic Disease Prevention and Health Promotion, CDC.

Editorial Note:

Editorial Note: The findings in this report indicate that substantial differences exist in the pH, the amount of moisture and nicotine, and the percentage of free nicotine among six commonly used U. S. smokeless tobacco products bought at several locations in Florida. The nicotine dose smokeless tobacco users receive may be controlled by adjusting the concentration of nicotine, varying the size of tobacco cuttings, and altering the pH (6). The pH in tobacco strongly affects nicotine absorption through the nose and mouth, especially free nicotine, the chemical form most readily absorbed across the buccal mucosa into the bloodstream (1). Although pH is a determinant of nicotine absorption, other factors can modulate the absorption rate (e.g., amount of moist snuff used and behavioral and physiologic factors unique to each user); however, these factors probably have little effect on the nicotine absorption rate (7). Among the 562 compounds reported on the smokeless tobacco ingredient list (8), several salts (e.g., ammonium, sodium, and potassium) may alter the pH of smokeless tobacco. The findings in this report confirm that products with high nicotine content and high pH have a high percentage of free nicotine.

The findings in this report are subject to at least two limitations. First, the analysis did not use a sales-weighted or representative sample of all U.S. brands or manufacturers; the moist snuff products tested were six leading products manufactured by the two industry leaders. Second, the findings for any specific brand could have been affected by factors unique to the sample delivered to each city surveyed, such as the retailers' duration and conditions of storage (e.g., humidity and temperature) and manufacturing dates.

This study is a new federal analysis of pH, moisture, and nicotine content of smokeless tobacco that quantifies a wide range of nicotine dosing capabilities in moist snuff products. These findings are consistent with other studies (6,9) that have found a wide variation in the nicotine dosing capabilities of these products. The Food and Drug Administration previously found that smokeless tobacco contains components intended to control the delivery of nicotine to the body (10). Smokeless tobacco users who dip or chew eight to 10 times a day may be exposed to the same amount of nicotine as persons who smoke 30 to 40 cigarettes a day (1). In addition, smokeless tobacco contains known cancer-causing agents: nitrosamines, polycyclic aromatic hydrocarbons, and radioactive polonium (1). These findings underscore the need for intensive efforts to prevent children and adolescents from using any tobacco product, including smokeless tobacco, and to educate young users about the risks associated with smokeless tobacco.

References

1. US Department of Health and Human Services. The health consequences of using smokeless tobacco: a report of the advisory committee to the Surgeon General. Bethesda, Maryland: US Department of Health and Human Services, Public Health Service, 1986.
2. CDC. Youth Risk Behavior Surveillance--United States, 1997. MMWR 1998;47(no. SS-3).
3. Maxwell JC Jr, Fenstermacher SD. The smokeless tobacco industry in 1997. The Maxwell Consumer Report. Richmond, Virginia: Davenport & Company LLC, May 22, 1998.
4. CDC. Protocol to measure the quantity of nicotine contained in smokeless tobacco products manufactured, imported, or packaged in the United States. Federal Register 1997;62:24116-9.
5. Lide DR, ed. CRC Handbook of chemistry and physics. 71st ed. Boca Raton, Florida: CRC Press, 1990.
6. Henningfield JE, Radzius A, Cone EJ. Estimation of available nicotine content of six smokeless tobacco products. Tob Control 1995;4:57-61.
7. Tomar SL, Henningfield JE. Review of the evidence that pH is a determinant of nicotine dosage from oral use of smokeless tobacco. Tob Control 1997;6:219-25.
8. US House of Representatives. Smokeless tobacco ingredient list as of April 4, 1994. US House of Representatives, Report to Subcommittee on Health and the Environment, Committee on Energy and Commerce. Washington, DC: Patton, Boggs and Blow, May 3, 1994.
9. Djordjevic MV, Hoffmann D, Glynn T, Connolly GN. US commercial brands of moist snuff, 1994: I. Assessment of nicotine, moisture, and pH. Tob Control 1995;4:62-6.
10. US Department of Health and Human Services, US Food and Drug Administration. Regulations restricting the sale and distribution of cigarettes and smokeless tobacco to protect children and adolescents. Federal Register 1996;61:44396-45318.

TABLE 1. Mean values of nicotine, total moisture and pH of six moist snuff products* -- Florida, January-February 1999+
=======================================================================================================================================================
                                                                                                       Nicotine dry
                                                          Total moisture                 Nicotine         weight       Free nicotine    Free nicotine
Product                     Place of purchase                   (%)        pH        content (mg/g)&       (%)            (mg/g)&            (%)
-----------------------------------------------------------------------------------------------------------------------------------------------------
Copenhagen Snuff            Daytona Beach                       54.8      8.21            10.76            2.38            6.546            60.81
                            Fort Myers                          53.4      7.99            10.32            2.21            4.982            48.27
                            Miami                               52.7      8.05            10.62            2.25            5.471            51.53
                            Tampa/St. Petersburg                55.1      8.48            10.66            2.37            7.920            74.33

                            Overall mean                        54.0      8.18            10.59            2.30            6.229            58.74
                            SD@                               +/-1.0   +/-0.20          +/-0.17         +/-0.08         +/-1.178         +/-10.56

Skoal Bandits Straight**    Orlando                             49.4      5.47             8.00            1.58            0.022             0.28
                            Tampa/St. Petersburg                47.3      5.57             8.05            1.53            0.029             0.35
                            Tallahassee                         50.1      5.51             7.71            1.55            0.024             0.31

                            Overall mean                        48.9      5.52             7.92            1.55            0.025             0.31
                            SD                                +/-1.2   +/-0.05          +/-0.16         +/-0.02         +/-0.003         +/- 0.03

Skoal Bandits Wintergreen** Daytona Beach                       50.6      6.91             7.12            1.44            0.515             7.24
                            Orlando                             49.3      6.88             7.42            1.47            0.502             6.77
                            Tampa/St. Petersburg                49.8      6.86             7.05            1.40            0.456             6.47
                            Tallahassee                         49.7      6.74             6.83            1.36            0.341             4.99

                            Overall mean                        49.9      6.85             7.11            1.42            0.454             6.37
                            SD                                +/-0.5   +/-0.07          +/-0.22         +/-0.04         +/-0.072         +/- 0.88

Skoal Long Cut Wintergreen  Daytona Beach                       54.9      7.87            11.10            2.46            4.627            41.68
                            Miami                               54.4      7.80            10.95            2.40            4.121            37.64
                            Orlando                             54.2      7.94            10.79            2.35            4.895            45.36
                            Tampa/St. Petersburg                53.1      7.53            11.33            2.42            2.775            24.48

                            Overall mean                        54.1      7.79            11.04            2.41            4.105            37.29
                            SD                                +/-0.7   +/-0.16          +/-0.21         +/-0.04         +/-0.853         +/- 8.23

Kodiak Wintergreen          Daytona Beach                       53.5      8.34             9.01            1.94            6.078            67.46
                            Orlando                             53.0      8.34             8.46            1.80            5.724            67.67
                            Tallahassee                         53.8      8.47             8.23            1.78            6.058            73.63
                            Tampa/St. Petersburg                52.7      8.27             8.54            1.80            5.448            63.79

                            Overall mean                        53.2      8.35             8.56            1.83            5.827            68.14
                            SD                                +/-0.4   +/-0.08          +/-0.30         +/-0.07         +/-0.272         +/- 3.68

Hawken Wintergreen          Orlando                             28.0      5.45             3.00            0.42            000.8             0.27
                            Tallahassee                         25.1      5.61             3.17            0.42            0.012             0.39
                            Tampa/St. Petersburg                20.9      4.65             3.93            0.50            0.002             0.04

                            Overall mean                        24.7      5.24             3.37            0.45            0.007             0.23
                            SD                                +/-3.1   +/-0.45          +/-0.43         +/-0.04         +/-0.005         +/- 0.15
-----------------------------------------------------------------------------------------------------------------------------------------------------
 * Use of trade names and commercial sources is for identification only and does not imply endorsement by the U.S. Department of Health and Human
   Services or CDC.
 + Mean values for three replicated samples.
 & Units for nicotine and free nicotine content are milligrams of nicotine (or free nicotine) per gram of tobacco (mg/g).
 @ Standard deviation.
** Skoal Bandits come in 0.5 g sachets. Each sachet provides half the nicotine indicated.
=======================================================================================================================================================

* Use of trade names and commercial sources is for identification only and does not imply endorsement by U.S. Department of Health and Human Services or CDC.

** The protocol for determining pH, total moisture, and nicotine content used in this analysis was published as a notice to solicit public comment on the protocol in the Federal Register (62 FR 24116, May 2, 1997). The final version of the protocol was published in the Federal Register on March 23, 1999. The differences between the two protocols are minor and would not affect the results of this study; however, the sampling of the products for this study is different from that required by the protocol.

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