Get Your Money’s Worth

Determining the Nicotine Content of Cigarettes

Kaitlyn McHugh
TA: Min A Kim
February 3, 2021
CHEM 1255 – Instrumental Analysis Lab, Chemistry Department, University of Pittsburgh

I submit this lab report as an original document. I assert that all ideas and discussion of data
contained herein is my own work unless otherwise referenced.
X Kaitlyn McHugh .
Results
Nicotine standard solutions of known concentration were prepared to have six different standards
varying from concentrations 0.0 mg/mL to 2.0 mg/L. The amount of naphthalene internal
standard solution to dilute the nicotine standard stock solution was determined with a dilution
calculation:

𝑚𝑔 (𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑛𝑖𝑐𝑜𝑡𝑖𝑛𝑒 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑)(10𝑚𝐿)

𝑉𝑜𝑙 𝑜𝑓 2.0 𝑚𝐿 𝑛𝑎𝑝𝑡ℎ𝑎𝑙𝑒𝑛𝑒 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 = (1)
2 𝑚𝑔/𝑚𝐿
Example: 0.2 mg/mL solution.
𝑚𝑔
𝑚𝑔 (0.2 )(10𝑚𝐿)
𝑚𝐿
𝑉𝑜𝑙 𝑜𝑓 2 𝑚𝐿 𝑖𝑛𝑡𝑒𝑟𝑛𝑎𝑙 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 = = 1 𝑚𝐿 𝑛𝑎𝑝ℎ𝑡ℎ𝑎𝑙𝑒𝑛𝑒 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
2 𝑚𝑔/𝑚𝐿

Standard spectra were obtained for the nicotine 0.0 mg/mL to 2.0 mg/mL standards. As seen in
Figure 1, there are 3 peaks, corresponding to the main compounds in the nicotine mixture:
solvent, which had a retention time around 2.5 minutes, nicotine, with a retention time around 7
minutes, and naphthalene, with a retention time around 9 minutes.

Solvent

Nicotine Naphthalene

Figure 1. Standard Chromatogram, 0.2 mg/mL nicotine solution.

Samples were also prepared using four different cigarette brands: Newport, Pall, L&M, and
Marlboro. Spectra were obtained for these samples, which matched the 3 peaks in the standard
solutions, as shown in Figure 2.

Solvent

Nicotine Naphthalene

Figure 2. Standard Chromatogram, Newport sample solution.

Using the peak area ratio of nicotine to naphthalene for the six standard nicotine solutions, a
calibration curve was constructed, as shown in Figure 3, producing a highly reliable R2 value of
0.9988.

4.0
y = 1.7356x - 0.0359
Peak Area Ratio (Nicotine/Napthalene)

3.5 R² = 0.9988

3.0

2.5

2.0

1.5

1.0

0.5

0.0
0 0.5 1 1.5 2 2.5
-0.5
Nicotine Concentration (mg/mL)

Figure 3. Calibration curve showing peak area ratio of nicotine to naphthalene as a function of
the concentration of nicotine.

Because of the high R2 value, this curve was used in the quantification of the amount of nicotine
in various cigarette brands. First, the concentration of the sample solution injected into the
instrument was calculated using the regression equation, as shown in Table 1:

𝑃𝑒𝑎𝑘 𝐴𝑟𝑒𝑎 𝑅𝑎𝑡𝑖𝑜 = 1.7356 ∗ (𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒) − 0.0359 (2)

Example: Newport sample solution.
0.85 = 1.7356𝑥 − 0.0359
𝑥 = 𝟎. 𝟓𝟎𝟗𝟗 𝒎𝒈/𝒎𝑳

Table 1. The concentration of nicotine of each sample of cigarette brand injected into the
instrument.
Brand Sample Concentration (mg/mL)
Newport 0.5099
Pall 0.4035
L&M 0.3432
Marlboro 0.5022

The standard deviation was calculated using the LINEST function in Excel to return a value of
±0.052. Then, the calculated concentrations in the sample solution were used to calculate the
amount of nicotine in a cigarette for each brand, using the volume of the extraction solution,
weight of tobacco, and weight of the cigarette. These values were also used to calculate the unit
cost of nicotine for each brand, in dollars per milligram, using the cost per pack and the number
of cigarettes in a pack, as shown in Table 2.
Example: Newport cigarette.
0.5099 𝑚𝑔 𝑛𝑖𝑐𝑜𝑡𝑖𝑛𝑒 0.787 𝑔 𝑐𝑖𝑔𝑎𝑟𝑒𝑡𝑡𝑒
2 𝑚𝐿 𝑠𝑎𝑚𝑝𝑙𝑒 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 ∗ 1 𝑚𝐿 𝑠𝑎𝑚𝑝𝑙𝑒 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
∗ 0.052 𝑔 𝑢𝑠𝑒𝑑
= 15.4 𝑚𝑔/𝑁𝑒𝑤𝑝𝑜𝑟𝑡 𝑐𝑖𝑔𝑎𝑟𝑒𝑡𝑡𝑒 (3)

$8.63 1 𝑝𝑎𝑐𝑘 1 𝑐𝑖𝑔𝑎𝑟𝑒𝑡𝑡𝑒

∗ ∗ = $0.0280 /𝑚𝑔 (4)
1 𝑝𝑎𝑐𝑘 20 𝑐𝑖𝑔𝑎𝑟𝑒𝑡𝑡𝑒𝑠 15.4 𝑚𝑔

Table 2. The amount of nicotine in a cigarette and the unit cost of nicotine for Newport, Pall,
L&M, and Marlboro cigarettes.
Brand Mass of nicotine (mg) Cost per Pack ($) Cost per unit Nicotine ($/mg)
Newport 15.4 8.63 0.0280
Pall 12.0 8.01 0.0334
L&M 9.4 8.96 0.0478
Marlboro 13.1 7.78 0.0298

Using the calculated unit price and mass of nicotine per cigarette, a calibration curve was plotted
to determine the correlation between price and nicotine content for the four brands analyzed, as
shown in Figure 4. A regression line was calculated with an equation of y=-0.0033x+0.076 and
an R2 value of 0.8572. Error bars are shown using the standard deviation of the regression of
0.004155. (This value was calculated using the LINEST Excel function).

0.0600
y = -0.0033x + 0.076
R² = 0.8572
Unit Price of Nicotine ($/mg)

0.0500

0.0400

0.0300

0.0200

0.0100
8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 16.0
Amount of Nicotine (mg)

Figure 4. The unit price was plotted as a function of the amount of nicotine.

The limit of detection, which is the smallest amount of analyte (nicotine) that has a signal
significantly larger than the blank signal (naphthalene), was calculated for the calibration method
in Figure 3. The LOD can be calculated using the measurement of the blank and its standard
deviation. The standard deviation was calculated using the LINEST function in Excel to return a
value of 0.052.
 3𝑠𝑏
𝑠𝑖𝑔𝑛𝑎𝑙 𝑑𝑒𝑡𝑒𝑐𝑡𝑖𝑜𝑛 𝑙𝑖𝑚𝑖𝑡 = 𝑥 = (5)
𝑚
3∗0.052
𝐿𝑂𝐷 = = 0.0090 𝑚𝑔/𝑚𝐿
1.7356

Discussion
In this experiment, which ultimately sought to determine the amount of nicotine in four
different cigarette brands using Gas Chromatography-Flame Ionization Detection (GC-FID), it
was ultimately found that this value could be determined reliably with a small standard
deviation, ranging from ±0.052. Using the calibration curve produced from the six standard
solutions, the nicotine concentration was able to be quantified for Newport, Pall, L&M, and
Marlboro cigarettes. After determining the price for these cigarettes, a relationship between unit
price and the amount of nicotine was shown by plotting a calibration curve.
GC-FID can be a preferred method for analyzing the concentrations of organic
compounds, such as nicotine, because it works best for organic, highly volatile, and thermally
stable compounds. This procedure requires that the analyte be completely vaporized as it moves
through the column and to the detector. FID was used in this experiment, as it does not respond
to water, so it requires less sample preparation than other detectors, and it only responds to
organic compounds. Because GC-FID is not the most accurate form of GC, when compared to
extremely high-resolution methods like Gas Chromatography-Mass Spectrometry (GC-MS), an
internal standard was used in this experiment. The addition of the naphthalene internal standard
increases the precision of results because it limits any external factors that could affect the
results. Any factor influencing the nicotine peaks will be affecting the naphthalene peaks to the
same degree. Therefore, taking the ratio of these two peak areas can decrease the variability
when compared to the nicotine peak area alone.
Despite the use of the internal standard method, there are still many other sources of error
could have impacted the results. One source of error comes from the sample preparation. In this
experiment, potassium hydroxide is used in the solvent stock solution preparation, to ensure that
all the nicotine is in the free base form. The free base form of nicotine is much easier to vaporize,
so it is better at moving through the GC column. If there is still protonated salt form of nicotine
present when the sample is injected into the column, this can affect the results, as less nicotine
will be vaporized and be detected at the column. This would result in an underestimation of the
amount of nicotine in cigarettes. Another source of error comes from the sensitivity of the GC-
FID, which requires that all solutions be filtered before injection. If these solutions are not
filtered, it can result in unwanted solids entering the GC-FID. If the solutions are filtered
improperly, it can result in extra peaks, which is why it is important to first filter a small amount
of solution into a waste beaker before filtering into the sample vials.
The results show that a calibration curve was produced to calculate the concentration of
nicotine in different cigarette brands, plotting the peak area ratio of nicotine as a function of
nicotine concentration. This curve, as shown in Figure 3, has an R2 value of 0.9988, which
indicates good reliability in predicting the concentration of unknown solutions. The calculated
standard deviation was also relatively low, at 0.052. After using this curve to calculate the
amount of nicotine in cigarettes, the values were compared to literature values of nicotine, which
stated that the average value of nicotine in cigarettes was 10 to 12 mg.2 This average value is
close to all the calculated values for the four brands studied, which range from 9.4 to 15.4 mg.
This indicates that our method was very accurate for predicting the nicotine content of various
cigarettes. A second calibration curve was used to predict the relationship between unit price of
nicotine and the amount of nicotine in cigarettes, as shown in Figure 4. This curve had a lower
R2 value of 0.8572. This shows that there is a weaker correlation between unit price and amount
of nicotine. The calculated standard deviation of the calibration curve was 0.004155 and is
shown as error bars on the plot in Figure 4. These error bars show that the regression line could
become even more unreliable, with a smaller R2 value, as the values could be more spread out.
While there were numerous possible sources of error in this experiment and places for
improvement, the data produced appears to be of relatively high reliability. The R2 value of
0.9988 and standard deviation value indicates that GC-FID is a preferred method for determining
the concentration of unknown solutions. However, the lower R2 value in the second calibration
curve indicates that it is less reliable in predicting the relationship between unit price and amount
of nicotine. GC-FID is very useful in a variety of applications due to its ability to yield reliable
results, as shown in numerous industrial examples as well as in the context of the procedure used
in this experiment, making it one of the most important analytical instruments in chemistry.
References
1. Dewar, A., et al. Do You Get More “Bang for Your Buck”? Determination of Nicotine in
Cigarettes. Department of Chemistry, University of Pittsburgh. 2011.
2. Jewell, Tim. How Much Nicotine Is in a Cigarette and Other Tobacco Products? 18
November 2019. https://www.healthline.com/health/how-much-nicotine-is-in-a-
cigarette#nicotine-in-cigarettes.