Table 1
Parameters for the MS/MS detection.
| Compound | Ion for Q1 | Ion for Q3 | Acquisation time (ms) | Declustering potential (V) | Collision energy (V) | Collision exit potential (V) |
|---|---|---|---|---|---|---|
| Pseudooxynicotine | 179.1 | 148.1 | 100 | 70 | 17 | 13 |
| Nicotine-1′-N-oxide | 180.1 | 133.1 | 100 | 70 | 21 | 13 |
| Nicotine-d3 (I.S.) | 166.1 | 870 | 100 | 30 | 15 | 9 |
Figure 1
Chromatogram generated in positive MRM mode from a Burley tobacco sample indicating the peaks for nicotine-1′-N-oxide, PON and the internal standard nicotine-d3.
Table 2
Parameters a, b, and c for the quantitation of pseudooxynicotine (PON) and nicotine-1′-N-oxide, and the regression coefficient R2 of the calibration line.
| Compound | a | b | c | R2 |
|---|---|---|---|---|
| PON | −1.3581 E-13 | 1.33470 E-06 | −2.1595 E-02 | 0.9996 |
| Nicotine-1′-N-oxide | — | 3.04019 E-07 | −0.039649 | 0.9993 |
Figure 2
Typical chromatogram of PON and PON-d3 in 3R4F filler and in a standard containing 46.2 ng/mL PON.
Table 3
Method recovery with spiked PON-d4 into 3R4F (n = 3).
| Spiked PON-d4 (ng/mL) | Recovery (%) | RSD (%) |
|---|---|---|
| 302.7 | 100 | 10.1 |
| 1210.8 | 103 | 4.0 |
| 2421.6 | 102 | 3.1 |
Table 4
List of tobaccos, moist snuff, and snus samples evaluated in this study.
| No. | Sample | Description |
|---|---|---|
| 1 | FC L (1) | Eastern NC belt, lower stalk (lug) flue-cured (2016) |
| 2 | FC U (1) | Eastern NC belt, upper stalk (leaf & some tips) flue-cured (2016) |
| 3 | FC L (2) | South Carolina belt, lower stalk (lug) flue-cured (2016) |
| 4 | FC U (2) | South Carolina belt, upper stalk (leaf & some tips) flue-cured (2016) |
| 5 | FC off L | Brazil, lower stalk (lugs & primings) flue-cured (2016) |
| 6 | FC off U | Brazil, upper stalk (leaf & tips) flue-cured (2016) |
| 7 | Bu L (1) | Kentucky & Tennessee, lower stalk (flyings & cutters) Burley (2016) |
| 8 | Bu U (1) | Kentucky & Tennessee, upper stalk (leaf) Burley (2016) |
| 9 | Bu L (2) | North Carolina & Virginia, lower stalk (flyings & cutters) Burley (2016) |
| 10 | Bu U (2) | North Carolina & Virginia, upper stalk (leaf) Burley (2016) |
| 11 | Bu off L | Malawi, lower stalk (flyings & cutters) Burley (2016) |
| 12 | Bu off U | Malawi, upper stalk (leaf) Burley (2016) |
| 13 | O SA U | Turkey, good quality middle to upper stalk, Samsun Oriental (2015) |
| 14 | O Iz U | Turkey, good quality middle to upper stalk, Izmir Oriental (2015) |
| 15 | FC L-Prod | Low stalk flue-cured blend used in cigarette production (2018) |
| 16 | FC U-Prod | Leaf flue-cured blend used in cigarette production (2018) |
| 17 | Bu L-Prod | Low stalk Burly blend used in cigarette production (2018) |
| 18 | Bu U-Prod | Leaf Burley blend used in cigarette production (2018) |
| 19 | O1-Prod | Oriental blend used in cigarette production (2018) |
| 20 | O2-Prod | Oriental blend used in cigarette production (2018) |
| 21 | Moist 1 | Red Seal Natural Fine Cut (2019) |
| 22 | Moist 2 | Skoal Long Cut Classic Wintergreen (2019) |
| 23 | Moist 3 | Long Horn Fine Cut Natural (2019) |
| 24 | Moist 4 | Grizzly Fine Cut Premium Wintergreen (2019) |
| 25 | Moist 5 | Grizzly Long Cut Premium Wintergreen (2019) |
| 26 | Moist 6 | Kodiak Premium Wintergreen (2019) |
| 27 | Moist 7 | Copenhagen Snuff Original Fine Cut (2019) |
| 28 | Moist 8 | Copenhagen Long Cut Mint (2019) |
| 29 | RT1a | 1R6F filler |
| 30 | RT2a | Flue-cured ground tobacco |
| 31 | RT3a | Oriental ground tobacco |
| 32 | RT4a | Burley ground tobacco |
| 33 | RT5a | High TSNA dark air-cured ground tobacco |
| 34 | RT6a | Flavored cigar filler |
| 35 | RT7a | 1R5F filler |
| 36 | RT8a | Unflavored cigar filler |
| 37 | RT9a | Dark air-cured ground tobacco |
| 38 | RT10a | Dark fire-cured ground tobacco |
| 39 | 1S4a | Swedish style Snus |
| 40 | 1S5a | Snus |
| 41 | 3S3a | Moist Snuff |
| 42 | 3S1a | Loose-leaf chewing tobacco |
Table 5
Average levels of nicotine, NNK, PON, and nicotine-1′-N-oxide (on “dry weight basis”).
| No. | Sample | Nicotine (mg/g) | NNK (ng/g) | PON (µg/g) | Nic-Ox (mg/g) | PON method |
|---|---|---|---|---|---|---|
| 1 | FC L (1) | 22.9 | 452.2 | 56.4 | 103.7 | 1 |
| 2 | FC U (1) | 25.6 | 242.9 | 83.1 | 134.9 | 1 |
| 3 | FC L (2) | 16.6 | 265.3 | 38.6 | 50.4 | 1 |
| 4 | FC U (2) | 24.7 | 49.2 | 76.5 | 62.4 | 1 |
| 5 | FC off L | 20.4 | 109.6 | 62.1 | 146.9 | 1 |
| 6 | FC off U | 29.4 | 72.4 | 71.8 | 183.0 | 1 |
| 7 | Bu L (1) | 26.1 | 274.7 | 150.1 | 224.8 | 1 |
| 8 | Bu U (1) | 33.0 | 317.8 | 155.4 | 254.2 | 1 |
| 9 | Bu L (2) | 21.1 | 1791.9 | 134.5 | 209.4 | 1 |
| 10 | Bu U (2) | 24.6 | 1583.3 | 199.2 | 244.2 | 1 |
| 11 | Bu off L | 7.6 | 136.2 | 76.0 | 123.7 | 1 |
| 12 | Bu off U | 7.3 | 174.0 | 69.4 | 110.2 | 1 |
| 13 | O SA U | 14.0 | 44.4 | 102.0 | 226.5 | 1 |
| 14 | O Iz U | 7.9 | 0.5 | 51.5 | 141.7 | 1 |
| 15 | FC L-Prod | 17.9 | 195.4 | 94.0 | 204.5 | 1 |
| 16 | FC U-Prod | 27.2 | 171.0 | 121.7 | 195.4 | 1 |
| 17 | Bu L-Prod | 22.4 | 885.5 | 205.9 | 221.3 | 1 |
| 18 | Bu U-Prod | 30.9 | 755.2 | 284.2 | 232.2 | 1 |
| 19 | O1-Prod | 10.3 | 7.0 | 67.4 | 150.2 | 1 |
| 20 | O2-Prod | 8.0 | 1.2 | 50.8 | 135.8 | 1 |
| 21 | Moist 1 | 6.0 | 210.7 | 5.1 | 17.8 | 1 |
| 22 | Moist 2 | 5.4 | 177.2 | 3.2 | 23.6 | 1 |
| 23 | Moist 3 | 5.5 | 265.4 | 5.8 | 78.0 | 1 |
| 24 | Moist 4 | 7.8 | 564.9 | 10.6 | 88.7 | 1 |
| 25 | Moist 5 | 4.6 | 331.6 | 5.8 | 92.8 | 1 |
| 26 | Moist 6 | 3.9 | 403.9 | 5.5 | 135.8 | 1 |
| 27 | Moist 7 | 7.1 | 310.4 | 7.8 | 27.6 | 1 |
| 28 | Moist 8 | 5.5 | 211.1 | 3.2 | 28.2 | 1 |
| 29 | RT1 | 21.4 | 683.4 | 62.2 | — | 2 |
| 30 | RT2 | 29.0 | 90.3 | 72.3 | — | 2 |
| 31 | RT3 | 5.9 | <LOQ | 31.9 | — | 2 |
| 32 | RT4 | 38.5 | 672.7 | 105.8 | — | 2 |
| 33 | RT5 | 31.9 | 1075.5 | 149.0 | — | 2 |
| 34 | RT6 | 13.1 | 1126.4 | 33.9 | — | 2 |
| 35 | RT7 | 16.5 | 927.8 | 34.9 | — | 2 |
| 36 | RT8 | 15.5 | 904.8 | 63.1 | — | 2 |
| 37 | RT9 | 50.4 | 2160.0 | 80.8 | — | 2 |
| 38 | RT10 | 40.3 | 526.3 | 61.8 | — | 2 |
| 39 | 1S4 | 15.8 | 168.4 | 28.6 | — | 2 |
| 40 | 1S5 | 15.0 | 333.5 | 51.2 | — | 2 |
| 41 | 3S3 | 25.2 | 381.2 | 15.5 | — | 2 |
| 42 | 3S1 | 8.7 | 72.0 | 25.5 | — | 2 |
Figure 3
The variation of levels of NNK as a function of the levels of PON for all analyzed samples. The numbers next to each point indicate the sample number (for the clarity of the graph not all numbers are shown).
Figure 4
Graph showing the dependence of NNK on nicotine-1′-N-oxide levels for all samples. The numbers next to each point indicate the sample number.
Figure 5
Graph showing the dependence of NNK on nicotine levels for all samples. The numbers next to each point indicate the sample number.