Does the Level of NNK in Tobacco and Tobacco Products Depend on the Level of Pseudooxynicotine or of Nicotine-1′-N-Oxide?

Ji, Huihua; Moldoveanu, Serban C.

doi:10.2478/cttr-2022-0003

Figures & Tables

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.

Typical chromatogram of PON and PON-d3 in 3R4F filler and in a standard containing 46.2 ng/mL PON.

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).

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.

Graph showing the dependence of NNK on nicotine levels for all samples. The numbers next to each point indicate the sample number.

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

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

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

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	RT1^a	1R6F filler
30	RT2^a	Flue-cured ground tobacco
31	RT3^a	Oriental ground tobacco
32	RT4^a	Burley ground tobacco
33	RT5^a	High TSNA dark air-cured ground tobacco
34	RT6^a	Flavored cigar filler
35	RT7^a	1R5F filler
36	RT8^a	Unflavored cigar filler
37	RT9^a	Dark air-cured ground tobacco
38	RT10^a	Dark fire-cured ground tobacco
39	1S4^a	Swedish style Snus
40	1S5^a	Snus
41	3S3^a	Moist Snuff
42	3S1^a	Loose-leaf chewing tobacco

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

Does the Level of NNK in Tobacco and Tobacco Products Depend on the Level of Pseudooxynicotine or of Nicotine-1′-N-Oxide?

Figures & Tables

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Average levels of nicotine, NNK, PON, and nicotine-1′-N-oxide (on “dry weight basis”)_

Parameters for the MS/MS detection_

Method recovery with spiked PON-d4 into 3R4F (n = 3)_

List of tobaccos, moist snuff, and snus samples evaluated in this study_

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_

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