Waterpipe smoke: source of toxic and carcinogenic VOCs, phenols and heavy metals?

The use of the waterpipe, a traditional aid for the consumption of tobacco, has spread worldwide and is steadily increasing especially among the youth. On the other hand, there is a lack of knowledge regarding the composition of mainstream waterpipe smoke and the toxicological risks associated with this kind of smoking habit. Using a standardized machine smoking protocol, mainstream waterpipe smoke was generated and further analyzed for twelve volatile organic compounds (VOCs) and eight phenolic compounds by applying gas chromatography-mass spectrometry and reverse-phase high-performance liquid chromatography-fluorescence detection, respectively. Additionally, seventeen elements were analyzed in waterpipe tobacco and charcoal prior to and after smoking, applying inductively coupled plasma-mass spectrometry to assess the maximum exposure of these elements. For the first time ever, we have been able to show that waterpipe mainstream smoke contains high levels of the human carcinogen benzene. Compared with cigarette smoke yields, the levels were 6.2-fold higher, thus representing a significant health hazard for the waterpipe smoker. Furthermore, we found that waterpipe mainstream smoke contains considerable amounts of catechol, hydroquinone and phenol, each of which causing some health concern at least. The analysis of waterpipe tobacco and charcoal revealed that both matrices contained considerable amounts of the toxic elements nickel, cadmium, lead and chromium. Altogether, the data on VOCs, phenols and elements presented in this study clearly point to the health hazards associated with the consumption of tobacco using waterpipes.

Waterpipe smoking: the role of humectants in the release of toxic carbonyls.

In recent years, the number of waterpipe smokers has increased substantially worldwide. Here, we present a study on the identification and quantification of seven carbonylic compounds including formaldehyde, acetaldehyde and acrolein in the mainstream smoke of the waterpipe. Smoking was conducted with a smoking machine, and carbonyls were scavenged from the smoke with two impingers containing an acidic solution of 2,4-dinitrophenylhydrazine. The derivatives were then analyzed by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). For instance, during one waterpipe smoking session, up to 111 ± 12 µg formaldehyde could be detected. This value is about 5 times higher when compared to one 2R4F reference cigarette. We also found a distinct filter effect of the bowl water for all carbonyls investigated. Our data further demonstrate that increasing amounts of humectants in the unburned tobacco lowers the temperature in the waterpipe head during smoking, thereby resulting in decreasing levels of carbonyls in the smoke produced. Altogether, considerable amounts of toxic carbonyls are present in the waterpipe smoke, thus conferring a health risk to waterpipe smokers.

Exposure to allergens of different cattle breeds and their relevance in occupational allergy.

INTRODUCTION: Cattle are an important source of allergens in the working area of farmers. Asthma caused by cow allergens is a significant occupational problem. Yet in allergological testing, the results of in vivo and in vitro diagnostic tests are often inconsistent even in cases with clearly cattle-related symptoms. OBJECTIVES AND METHODS: The aim of this study was to investigate four different commercial cow allergen extracts and to compare them with self prepared extracts of different cattle breeds by means of SDS-PAGE and immunoblotting using the sera of 42 German farmers with asthma and rhino-conjunctivitis caused by cattle contact. RESULTS: The commercial extracts investigated in this study showed only minor differences in protein pattern. Using sera in immunoblotting experiments distinct bands were found for all symptomatic farmers, even in 13 farmers with a negative result in commercially available serological allergy tests. Bands with molecular weights in the range between about 11 and 67 kDa were observed; reactivity with the major allergen Bos d 2 at about 20 kDa was detected in all farmers, although it was not the strongest band in all cases. CONCLUSIONS: We demonstrate for the first time the allergenic relevance of additional proteins with molecular weights of 14, 30, 55 and approx. 67-97 kDa in more than 50% of farmers with cattle related symptoms. One of our most striking results was that 32% of the investigated farmers with cattle related symptoms showed negative results with commercial serological tests but distinct reactions with cow allergen in immunoblotting experiments. The Bos d 2 content in hair showed differences between certain breeds whereas German Brown and Simmental had particularly higher quantities of Bos d 2 in their hair than breeds such as Holstein-Friesian. These results strongly support the following recommendation: test results with commercial extracts that are contradictory to the clinical symptoms should be supplemented by skin tests using extracts of the hair of the farmers' own cattle.

Waterpipe smoking: analysis of the aroma profile of flavored waterpipe tobaccos.

In the last years the habit of smoking waterpipes has spread worldwide, especially among young people and emerged as global health issue. Although research is now under way for no less than 40 years in the field of waterpipe smoking, in comparison to cigarette smoking there is still insufficient knowledge on the real composition and the toxicity of the smoke inhaled and the resulting levels of exposure against particular hazardous ingredients. In most cases for waterpipe smoking a highly flavored tobacco called "moassel" is used. However, the number, quantity and toxicity of the added flavorings are widely unknown. In this study the static headspace gas chromatography-mass spectrometry (SHS-GC-MS) was used to identify 79 volatile flavor compounds present in waterpipe tobacco. Among these eleven compounds were analyzed quantitatively. The results show that waterpipe tobacco contains high amounts of the fragrance benzyl alcohol as well as considerable levels of limonene, linalool and eugenol, all of which are known as being allergenic in human skin. The proposed SHS-GC-MS method has been validated and found to be accurate, simple and characterized by low limits of detection (LOD) in the range of 0.016 to 4.3 µg/g tobacco for benzaldehyde and benzyl alcohol, respectively. The identification and characterization of waterpipe tobacco ingredients indeed reveals crucial for the assessment of potential health risks that may be posed by these additives in smokers.

Waterpipe smoke: a considerable source of human exposure against furanic compounds.

Smoking of waterpipes became increasingly popular in the Western hemisphere in recent years. Yet, up to now only little is known about the health hazards and on the composition of waterpipe smoke. To obtain more information on the ingredients present in waterpipe smoke we utilized two different approaches. Based on headspace-solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) instrumentation we identified new compounds present in the waterpipe smoke. Additional reversed-phase-high performance liquid chromatography-diode array detection (RP-HPLC-DAD) then led us to perform reliable quantification of the newly detected chemical species. Upon identification of a range of different furanic compounds such as 5-(hydroxymethyl)-2-furaldehyde (HMF), 2-furaldehyde, and others, we developed an easy-to-perform and fast RP-HPLC-DAD method to quantify these compounds in the complex matrix of waterpipe smoke. The detection limits range from 0.04 µg for HMF to 7.1 µg for 3-furan methanol per smoking session. Linearity, intra- and inter-day precision and recovery were determined and proved excellent. We analyzed 5 waterpipe tobacco brands and found up to 62.3±11 mg of HMF generated during one waterpipe smoking session. The applied smoking protocol comprised 171 puffs of 530 mL each and 2.6s duration every 20s. Our results reveal that waterpipe smoking constitutes a major source of HMF exposure. Furthermore, we found a distinct filter effect of the bowl water for all furanic compounds investigated except HMF.

Analysis of primary aromatic amines in the mainstream waterpipe smoke using liquid chromatography-electrospray ionization tandem mass spectrometry.

In recent years waterpipe smoking has spread worldwide and emerged as global health issue. Yet only little is known on the composition of waterpipe smoke. Here, we present a study on the identification and quantification of primary aromatic amines (PAAs) in this complex environmental matrix. Smoking of the waterpipe was conducted with a smoking machine and particulate matter was collected on glass fiber pads. We developed a fast, simple and specific liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) approach to simultaneously detect 31 different PAAs in this matrix. The detection limits comprised a range of 0.45-4.50 ng per smoking session, represented by 2-aminobiphenyl and 3,4,5-trichloroaniline, respectively. Intra- and inter-day precision were determined and proved excellent. We detected 31.3 ± 2.2 ng aniline and 28.0 ± 1.6 ng 4,4'-oxydianiline in the smoke of one waterpipe session. The water in the bowl exerted a small but considerable filter effect on PAAs. The method worked-out showed excellent sensitivity and specificity and is thus highly suited for the determination of PAAs in mainstream waterpipe smoke.

Mainstream smoke of the waterpipe: does this environmental matrix reveal as significant source of toxic compounds?

In recent years the number of waterpipe smokers has increased substantially worldwide. Here we report on the concentrations of tobacco-specific nitrosamines (TSNAs) and polycyclic aromatic hydrocarbons (PAHs) in waterpipe smoke and the analysis of selected biomarkers indicative for the body burden in waterpipe users. We further identify high amounts of unburned humectants (glycerol and propylene glycol) in the waterpipe smoke as main part of the so-called "tar" fraction. These results give cause for serious concern. For standardization we applied a machine smoking protocol. Smoke was collected on glass fiber filters and analyzed for nicotine, water, humectants, TSNAs, and PAHs. In addition, we determined carbon monoxide and found high amounts in the smoke being causative for high levels of carboxyhemoglobin (COHb) in the blood of smokers. In comparison to the reference cigarette 3R4F, the nicotine contents were 10-times higher, but TSNA levels were found lower in waterpipe smoke. This finding explained the low levels of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol detected in the urine of waterpipe smokers. Finally, the levels of benzo[a]pyrene were three times higher in waterpipe smoke compared to the reference cigarette. Altogether, the data presented in this study point to the health hazards associated with the consumption of waterpipes.

Thimerosal induces micronuclei in the cytochalasin B block micronucleus test with human lymphocytes.

Thimerosal is a widely used preservative in health care products, especially in vaccines. Due to possible adverse health effects, investigations on its metabolism and toxicity are urgently needed. An in vivo study on chronic toxicity of thimerosal in rats was inconclusive and reports on genotoxic effects in various in vitro systems were contradictory. Therefore, we reinvestigated thimerosal in the cytochalasin B block micronucleus test. Glutathione S-transferases were proposed to be involved in the detoxification of thimerosal or its decomposition products. Since the outcome of genotoxicity studies can be dependent on the metabolic competence of the cells used, we were additionally interested whether polymorphisms of glutathione S-transferases (GSTM1, GSTT1, or GSTP1) may influence the results of the micronucleus test with primary human lymphocytes. Blood samples of six healthy donors of different glutathione S-transferase genotypes were included in the study. At least two independent experiments were performed for each blood donor. Significant induction of micronuclei was seen at concentrations between 0.05-0.5 micro g/ml in 14 out of 16 experiments. Thus, genotoxic effects were seen even at concentrations which can occur at the injection site. Toxicity and toxicity-related elevation of micronuclei was seen at and above 0.6 micro g/ml thimerosal. Marked individual and intraindividual variations in the in vitro response to thimerosal among the different blood donors occurred. However, there was no association observed with any of the glutathione S-transferase polymorphism investigated. In conclusion, thimerosal is genotoxic in the cytochalasin B block micronucleus test with human lymphocytes. These data raise some concern on the widespread use of thimerosal.