Tobacco analysis

Hecht, S.S., R.M. Omaf, andD. Hoffmann Determination of A -nitrosonomicotine in tobacco by high-speed liquid chromatography Anal. Chem. 47 (1975) 2046-2048. Hecht, S.S., R.M. Omaf, and D. Hoffmann Chemical studiesontobaccosmoke.XXXIII.A -Nitrosonornicotine in tobacco Analysis of possible contributing factors and biological implications J. Natl. Cancer Inst. 54 (1974) 1237-1244. 

Vimses are also detectable with imprinted sensor materials thus leading to the first tme rapid on-line analysis for these species that are too small for e.g. light scattering experiments. So we e.g. succeeded in determining the tobacco mosaic vims (TMV) in plant saps as well as the Human Rhinovims (HRV). 

Efforts are still being made to estimate that elusive notion quality in smoking tobacco by chemical analysis it does at least seem to be clearly established that a low content of protein and of nicotine is desirable, and in that connection the isolation by Bucherer of several species of... 

The analysis of combustion products presents problems of complexity similar to that of feedstock and raw fuel analysis. A highly complex matrix of aliphatic material often exists (as unburnt fuel in the combustion exhaust), whilst the species of interest, for example, carcinogens or mutagens are often at very low concentrations. A classic example of multidimensional GC is its use in the analysis of flue-cured tobacco essential oil condensate. 

Figure 3.5 Two-dimensional GC analysis of tobacco essential oil using non-polar primary and polar secondary separ-ations. The top tr-ace indicates the primary separ-ation, with the four resulting heart-cut cliromatograms shown below being obtained on the transfer of approximately 1-2 min fractions of primary eluent. Reproduced from B.M. Gordon et al. J. Chwmatogr. Sci. 1988, 26, 174 (23).

Dinitrophenyl-hydrazine has been successfully employed in the analysis of simple aldehydes, substituted aldehydes, glyoxal and gluteraldehyde (43-45), all the isomers of the C3 to C7 aliphatic ketones (44,45) and in the determination of formaldehyde in tobacco smoke (46). 

C03-0083. Nicotine is an addictive compound found in tobacco leaves. Elemental analysis of nicotine gives these data C 74.0%, H 8.65%, N 17.35%. What is the empirical formula of nicotine The molar mass of nicotine is 162 g/mol. What is the molecular formula of nicotine ... 

The first step in the analysis is extraction of the tobacco with buffer solution (pH 4.5) containing 20 mM ascorbic acid. The nitrosamines are then concentrated by partition with dichloromethane, and a chromatographic clean-up on alumina. In the final step, the concentrate is analyzed by GC-TEA and confirmation of the nitrosamines is obtained by GC-MS (O. If isolated amounts of the nitrosamines are below levels needed for GC-MS confirmation, we employ confirmatory techniques proposed by Krull et a. ( 5). 

NDELA Analysis. Using a conventional analytical method which we developed in 1977, we determined that MH-30 treated tobaccos contain 100-170 ppb of N-nitrosodiethanolamine (NDELA 17). The availability of the TEA detector made it possible to develop an analytical method which permitted routine monitoring of NDELA in tobacco and its smoke. 

This method requires about 40 g of tobacco which are extracted with ethyl acetate in the presence of ascorbic acid. A trace amount of C-NDELA is added as an internal standard for quantitative analytical work. The filtered extract is concentrated and NDELA is enriched by column chromatography of the concentrate on silica gel. The residues of fractions with p-activity are pooled and redissolved in acetonitrile. Initially, we attempted to separate NDELA on a 3% OV-225 Chromosorb W HP column at 210 C using a GC-TEA system with direct interface similar to the technique developed by Edwards a. for the analysis of NDELA in urine (18). We found this method satisfactory for reference compounds however, it was not useful for an optimal separation of NDELA from the crude concentrate of the tobacco extract (Figure 4). Therefore, we silylated the crude concentrate with BSTFA and an aliquot was analyzed by GC-TEA with direct interface. The chromatographic conditions were 6 ft glass column filled with 3% OV-... 

In order to ascertain that the NDELA formation does not occur as a result of trapping of the smoke or during the analysis, we added diethanolamine to tobacco prior to extraction with ethyl acetate in the presence of ascorbic acid. The control value for NDELA was 121 ppb and the experiment with 5.5 iqg diethanolamine addition yielded 113 ppb NDELA. For control of the smoke analysis we added 5.5 mg of DELA in the solvent trap and smoked cigarettes known to be free of DELA. Analysis of the trapped material showed no significant quantities of NDELA, so that artifactual formation of this nitrosamine during smoke collection and analysis can be ruled out. 

Nonvolatile Nitrosamines In Tobacco. A method which we developed several years ago for the analysis of tobacco-specific nitrosamines (TSNA 31) involves extraction of tobacco with buffered ascorbic acid TpH 4.5) followed by partition with ethyl acetate, chromatographic clean-up on silica gel, and analysis by HPLC-TEA (Figure 9). Results obtained with this method for a large spectrum of tobacco products (Table IV), strongly support the concept that the levels of nitrate and alkaloids, and especially the methods for curing and fermentation, determine the yields of TSNA in tobacco products. Recent and as yet preliminary data from snuff analyses indicate that aerobic bacteria play a role in the formation of TSNA during air curing and fermentation. 

The smoke analysis of cigarettes made from the same tobacco blend, but with and without filter tips revealed that cellulose acetate retains TSNA selectively (Table VIIl). This phenomenon is clearly established for a large number of filter cigarettes. 

Shen, J. and Shao, X., Comparison of accelerated solvent extraction, Soxhlet extraction, and ultrasonic-assisted extraction for analysis of terpenoids and sterols in tobacco. Ana/. Bioanal. Chem., 383, 1003, 2005. 

Busch, M., Seuter, A., and Hain, R., Functional analysis of the early steps of carotenoid biosynthesis in tobacco. Plant Physiol. 128, 439, 2002. 

Dybczynski R, Polkowska-Motrenko H, Samczynski Z and Szopa Z (1998) Virginia tobacco leaves (CTA-VTL-z) - new Polish CRM for inorganic trace analysis including microanalysis. Fresenius J Anal Chem 360 384-387. 

Dybczynski R, Poikowska-Motrenko H, Samczynski Z, and Szopa Z (1996) Preparation and Certification of the Polish Reference Material Oriental Tobacco Leaves (CTA-OTL-i) for Inorganic Trace Analysis. Raporty IChTJ Seria A nr 1/96. Institute of Nuclear Chemistry and Technology, Warsawa. 

Dominguez, L. M. and Dunn, R. S., Analysis of OPA-derived amino sugars in tobacco by high-performance liquid chromatography with fluorimetric detection, /. Chromatogr. Sci., 25, 468, 1987. 

A deeper understanding of the molecular mechanisms underlying tobacco addiction will lead to the identification of different types of smokers. Classifying smokers according to the underlying biological processes involved in their addiction will lead to new treatments for tobacco dependence. Patient-specific therapy with both choice of treatment and dose of drug informed by DNA analysis seems likely to be more effective than conventional therapy with fewer unwanted effects. 

Figure 11.4 Analysis of in vitro synthesized RNAs. 32P-Radiolabeled RNAs (48 nucleotides) capped with m7Gp3G (A and C) or m27,3 °Gp3G (B and D) were digested with either RNase T2 (A and C) or RNase T2 plus tobacco acid pyrophosphatase (TAP) (B and D) followed by anion-exchange HPLC on a Partisil 10SAX/25 column as described in the text. Fractions of 1 ml were collected, and the Cerenkov radiation was determined. The elution times of the following standard compounds, detected by ultraviolet (UV) absorption, are indicated with arrows 3,-CMP (Cp), S UMP (Up), 37-AMP (Ap), 3 -GMP (Gp), 3, 5 -m7GDP (pm7Gp), 3, 5 -GDP (pGp), 5 -GDP (p2G), 5 -GTP (p3G), and guanosine-SCtetraphosphate (P4G).

Perham, R.N., and Thomas, J.O. (1971) Reaction of tobacco mosaic virus with a thiol-containing imi-doester and a possible application to X-ray diffraction analysis./. Mol. Biol. 62, 415—418. 

Fig. 11.3 Purification ofSOl-lOOxELP-proteins from transgenic tobacco plants by inverse transition cycling and analysis by SDS-PAGE. 1 15 pg of total soluble leaf protein extracted in raw extract buffer 2 cleared supernatant of original 15 pg total soluble leaf protein after heat treatment (60 min, 95 °C) 3 cleared supernatant of original 300 pg leaf protein after heat treatment 4 cleared supernatant of original 300 pg leaf protein after heat treatment (60 min, 60 °C) with 2 M NaCI 5 redissolved spider silk-elastin protein pellet from original 300 pg of total soluble leaf protein after heat treatment (60 min, 60 °C) with 2 M NaCI.

---