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As a new form of cigarettes, heated tobacco products (HTPs) have been rapidly distributed worldwide. In this study, an improved method for analyzing gaseous and particulate compounds generated from HTPs is described. Smoke is collected using a GF-CX572 sorbent cartridge with 300 mg of carbon molecular sieves, that is, Carboxen 572 (CX572), and a 9 mm glass-fiber filter (GF). After collection, the CX572 particles from the cartridge are transferred along with the GF and deposited into a vial containing two phases of carbon disulfide and methanol. The CX572 particles settle into the lower carbon disulfide phase, while nonpolar compounds are desorbed. After the sample is allowed to stand, the solution is slowly stirred. The two-phase mixture of carbon disulfide and methanol is combined into a homogeneous solution. Polar compounds are then desorbed, while the desorbed nonpolar compounds remain in solution. For the analysis of carbonyl compounds, an enriched 2,4-dinitrophenylhydrazine solution is added to a portion of the combined solution for derivatization and subsequent high-performance liquid chromatography analysis. For the analysis of volatile organic compounds and water, a portion of the combined solution is analyzed by gas chromatography-mass spectrometry or equipped with a thermal conductivity detector. By applying the proposed GF-CX572 one-cartridge method to the analysis of the mainstream smoke generated from HTPs and traditional cigarettes, several chemical compounds are detected, and the chemical composition of smoke is revealed. The GF-CX572 one-cartridge method can analyze gaseous and particulate chemical compounds from the HTP smoke by utilizing not only the entire puff volume but also one puff volume because the GF-CX-572 cartridge can be replaced with a new cartridge within 3 s. An overview of the chemicals generated from HTPs is obtained in detail by one-puff volume sampling. In addition, the generated chemical compounds strongly depend on the temperature of tobacco leaves in HTPs.

Because of the health effects of secondhand smoke, the Japanese government is trying to establish an effective law for total avoidance of secondhand smoke in indoor environments for tobacco-free Tokyo Olympic and Paralympic games 2020, as requested by the International Olympic Committee (IOC) and the World Health Organization (WHO). Meanwhile, Philip Morris International has begun selling a new heat-not-burn tobacco, iQOS, which it claims is designed not to produce secondhand smoke. There is little scientific data, however, of the hazards and toxicity of iQOS. In this study, we evaluated several harmful compounds (nicotine, tar, carbon monoxide (CO) and tobacco-specific nitrosamines (TSNAs)) in the mainstream smoke and fillers of iQOS, and compared their concentrations with those from conventional combustion cigarettes. The concentrations of nicotine in tobacco fillers and the mainstream smoke of iQOS were almost the same as those of conventional combustion cigarettes, while the concentration of TSNAs was one fifth and CO was one hundredth of those of conventional combustion cigarettes. These toxic compounds are not completely removed from the mainstream smoke of iQOS, making it necessary to consider the health effects and regulation of iQOS.

Objective: The World Health Organization (WHO) Framework Convention on Tobacco Control (FCTC) reported that understanding the use and impact of smokeless tobacco (SLT) products is complicated by product diversity. Many different SLT products with different characteristics are used worldwide. ZERO STYLE STIX(TM) (sold by Japan Tobacco Inc.) is a brand of snuff, a type of smokeless tobacco. Our objective was to determine the constituents of the gas from SLT and analyze the ingredients in tobacco fillers., Methods: ZERO STYLE STIX smokeless tobacco was released in the Japanese market in 2010. Nicotine, menthol, and tobacco-specific nitrosamines in the smokeless tobacco fillers were determined by gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-tandem mass spectrometry (LC/MS/MS). The gaseous compounds were collected by a smoking machine using two smoking protocols, i.e., the ISO and Health Canada Intense methods. Nicotine and menthol in the gas were determined by GC/MS., Results: Nicotine, menthol, and the total tobacco-specific nitrosamines were detected in the tobacco fillers. The level of menthol in the snuff was more than ten times that of nicotine. The determined levels of the two components of the gas from the snuff were higher when using the Health Canada Intense protocol than when using the ISO protocol. In addition, flavors other than menthol were emitted from the smokeless tobacco., Conclusion: The new type of snuff introduced in the Japanese market in 2010 contained added flavors, and was attractive smokeless tobacco. Flavors in tobacco products need to regulate on the basis of FCTC 9 and 10 in JAPAN.

Objective: Some Japanese smokers imported Japanese cigarette brands from other Asian countries for personal use because of the increase in tobacco tax in 2010, which meant that privately importing the cigarettes was cheaper than purchasing them in Japan. We assumed that the health hazards of locally made cigarettes were different from those of privately imported cigarettes, despite the brands being the same. In this study, we carried out a comparative toxicological analysis of domestic and privately imported cigarettes., Methods: Privately imported cigarettes of five brands and domestic cigarettes of the brand named "MILD SEVEN ORIGINAL" were selected for the study. The level of polycyclic aromatic hydrocarbons (PAHs) in mainstream smoke was determined by high-performance liquid chromatography using fluorescence detection. The mutagenicity of mainstream smoke was assessed using the Salmonella typhimurium strains TA100, TA98, and YG1024 with the metabolic activation system (S9mix) by preincubation assay. The levels of heavy metals in cigarette fillers were determined by inductively coupled plasma-mass spectrometry., Results: The level of PAH in the mainstream smoke from the privately imported cigarettes was higher than that in the smoke from the domestic cigarettes. However, the mutagenic activity of the mainstream smoke from the privately imported cigarettes was equivalent to that from the domestic cigarettes. The levels of nickel and cadmium in the fillers of the privately imported cigarettes were higher than those in the domestic cigarettes., Conclusion: The levels of constituents in the fillers and mainstream smoke from the privately imported cigarettes were higher than those in the fillers and mainstream smoke from the domestic cigarettes. The majority of Japanese smokers are unaware of these findings. The guidelines regulating tobacco products recommend that articles 9 and 10 of the World Health Organization (WHO) Framework Convention on Tobacco Control should be fully implemented in Japan.

We have developed a new analytical method for the determination of nicotine, tar, volatile organic compounds and carbonyls in main-stream cigarette smoke using a sorbent cartridge packed with Carboxen 572 (CX-572) and a Cambridge filter pad (CFP) followed by the two-phase/one-pot elution method. A CX-572 cartridge is installed between the intake of the CFP and the pump of the smoking machine. Gaseous compounds collected with the CX-572 cartridge and total particulate matter (TPM) collected with the CFP are coeluted simultaneously in the same vial and then analyzed by high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC/MS) and gas chromatograph-thermal conductivity detector (GC/TCD). Carbonyl compounds are determined by adding derivatizing reagent (2,4-dinitrophenylhydrazine, DNPH) to the eluate followed by HPLC analysis. VOCs and nicotine are determined by GC/MS, and water is determined by GC/TCD. The same sample eluate solution is used for HPLC, GC/MS and GC/TCD analyses. As a result of measuring main-stream cigarette smoke generated from reference cigarettes, almost all carbonyl compounds and VOCs except formaldehyde were passed through a CFP and trapped in a CX-572 cartridge. 100% of nicotine, tar and TPM were trapped in a CFP. 50% of water and 53% of formaldehyde were trapped in a CFP. The one-pot data is almost equal to the sums of CFP (particulate matter) and CX-572 (gaseous compounds) data. The two-phase/one-pot elution method can simultaneously measure nicotine, tar, volatile organic compounds and carbonyl compounds in cigarette smoke with simple operation and small amounts of reagents., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In Japan, ultralow-nicotine cigarette brands with ventilation holes on the cigarette filters have been widely marketed to smokers. The use of these cigarette brands leads to compensation smoking. Menthol cigarette brands that have a cooling and numbing effect are also sold. In 2013, smokeless tobacco products similar to the Swedish snus, which is banned for sale in the European Union except in Sweden, were released in Japanese markets. These tobacco products have "toxicity," have a "ventilation filter," are "attractive," and promote "dependence." Tobacco smoke and smokeless tobacco are classified into "Group 1: Carcinogenic to humans" by the International Agency for Research on Cancer (IARC). The purpose of Articles 9 and 10 of the World Health Organization Framework Convention on Tobacco Control (WHO FCTC) is to regulate the contents of tobacco products as well as to regulate tobacco product disclosures. The implementation of some other articles has gradually advanced in Japan. However, that of Articles 9 and 10 is late. Japanese governmental authorities are being urged to immediately implement Articles 9 and 10.

Objective: The tobacco tax in Japan was increased in October, 2010. Subsequently, the quantity of sales of domestic cigarette products decreased temporarily. However, smokers could import cigarettes directly through the internet. Because of the high value of the yen and the low price of foreign cigarettes, an individual could import cigarettes easily; however, the contents and emissions from privately imported cigarettes have not been investigated yet. In this study, we carried out a comparative analysis of domestic and privately imported cigarettes., Methods: Privately imported cigarettes from five brands and domestic cigarettes from the brand "Mild Seven" were selected for the study. For the fillers of the cigarettes, the levels of nicotine and tobacco-specific nitrosamines (TSNAs) were determined by methods that are usually used for cigarette products. For the mainstream smoke, the levels of tar, nicotine, carbon monoxide, and TSNAs were also determined by the methods used for cigarettes., Results: The average nicotine and TSNA levels in the fillers of the domestic cigarettes were 15.1 ± 0.19 mg/g and 1,920 ± 85.1 ng/g, respectively. The amounts of nicotine and tar in the mainstream smoke from the privately imported cigarettes were lower than those from the domestic cigarettes. However, the levels of TSNAs in the mainstream smoke from the privately imported cigarettes were higher than those from the domestic cigarettes., Conclusions: The amounts of TSNAs in the fillers of and mainstream smoke from the privately imported cigarettes were higher than those from the domestic cigarettes. Japanese smokers should be careful about consuming privately imported cigarettes because there is a variation in the amount of hazardous constituents in these cigarettes, even when they are from the same brand.

Objective: To determine constituents of fillers and mainstream smoke from Neo Cedar., Methods: Neo Cedar is a second-class over-the-counter (OTC) drug and similar to cigarettes in a number of ways. In particular, the design and usage are very similar to those of cigarettes. For the fillers of the drug, the levels of nicotine, tobacco-specific nitrosamines (TSNA), and heavy metals, and mutagenicity were determined using the methods for cigarette products. For the mainstream smoke, the levels of tar, nicotine, carbon monoxide (CO), TSNA, polycyclic aromatic hydrocarbons (PAH), and carbonyl compounds were also determined using the methods for cigarettes. The mainstream smoke from the drug were collected with a smoking machine using two smoking protocols (ISO and Health Canada Intense methods)., Results: The nicotine and total TSNA levels in the fillers of the drug averaged 2.86 mg/g and 185 ng/g, respectively. The nine species of heavy metals were also detected in the fillers of the drug. The levels of nicotine, tar, CO, TSNA, PAH, and carbonyl compounds of mainstream smoke from the drug were higher when determined using the HCI regime than when using the ISO regime. The mutagenicity of the mainstream smoke determined using the HCI regime was also higher than that determined using the ISO regime., Conclusion: In this study, all constituents of Neo Cedar were determined by methods for cigarette products. The drug had a ventilation hole on its filter. Thus, its constituents are different from those determined by the smoking protocols. Neo Cedar users should be careful of higher exposure to the hazardous gases owing to smoking patterns.

We developed a simple method for the simultaneous determination of volatile organic compounds (VOCs) and carbonyls in the mainstream cigarette smoke using a sorbent cartridge at ambient temperature without the traditional cryogenic impinger. A sorbent cartridge is installed between intake filter and the pump of the smoking machine. Collection of cigarette mainstream smoke is performed according to the Canadian Intense regime or the ISO regime. As adsorbent, Carboxen 572 (CX-572) is the most suitable for collection of VOCs and carbonyls in the mainstream cigarette smoke. Elution of VOCs and carbonyls from CX-572 is performed by the two-step elution with carbon disulfide and methanol. VOCs are eluted by first elution with carbon disulfide and carbonyls are eluted by second elution with methanol. For VOCs, a portion of eluate is analyzed by gas chromatography-mass spectrometry. For carbonyls, a portion of eluate is derivatized with enriched 2,4-dinitrophenylhydrazine solution and analyzed by high-performance liquid chromatography. Measurement values by CX-572 cartridge method are very close to those obtained by traditional impinger method except for 2-butanone. Impinger methods use 2,4-dinitrophenylhydrazine solution containing 50% water and 2-butanone-DNPhydrazone may be hydrolyzed with water. In the CX-572 method, the hydrolysis of 2-butanone is prevented because the eluate solution contains no water. CX-572 method can measure cigarette smoke resulting from not only one whole cigarette but also from one puff volume because of its high sensitivity and simple operation., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objective: To measure the nicotine and tobacco-specific nitrosamine (TSNA) contents in whole tobacco from a variety of Japanese domestic cigarette brands and to determine mutagencity., Methods: The test cigarettes were the top ten best selling cigarette brands in Japan in 2006. The nicotine content in whole tobacco was measured by a modified version of the CORESTA Recommended method. The level of TSNA was measured by a modified version of the Health Canada method. Mutagenicity was assessed using the Salmonella typhimurium strains TA100, TA98 and YG1024 with the metabolic activation system (S9mix) by preincubation assays., Results: The nicotine content in the test cigarettes averaged 15.7±1.2 mg/g, and was in the range between 13.7 and 17.2 mg/g. The level of TSNA averaged 1,750 ng/g, and was in the range between 931 and 2,490 ng/g. Mutagenicity was pseudopositive in several samples of the YG1024 tester strain with and without S9mix., Conclusion: The cigarettes brands were categorized into four groups (Ultra-low, Low, Medium, and High) based on the nominal nicotine yield figures printed on the cigarette packets. The nicotine content in whole tobacco of the High group was the highest. However, the level of TSNA of the high group was the lowest. The analyses of hazardous chemical compounds in whole tobacco can contribute to the reduction and regulation of the toxicity of tobacco products.

Objectives: Mainstream smoke from cigarettes contains tobacco-specific N'-nitrosamines (TSNAs) listed as Group 1 and 3 carcinogens by the International Agency for Research on Cancer (IARC). Herein, we report on a method of measuring the concentrations of TSNAs in mainstream smoke from the ten top-selling Japanese cigarette brands using an ISO regime by International Organization for Standardization (ISO) and HCI regime of Health Canada., Methods: Tar in mainstream smoke was collected on a Cambridge filter pad using a smoking machine. The filter pad was immersed in 40 mL of ammonium acetate (pH 6.8) and shaken for 30 min. The extract was then loaded into a C18 column. After washing with 5 mL of 10% methanol and eluting with 5 mL of 70% methanol, the eluate was concentrated to 1 mL for LC-MS/MS analysis., Results: The concentrations of TSNAs in all cigarette brands were higher when determined using the HCI regime than when determined using the ISO regime. Furthermore, the concentrations of TSNAs measured using both the ISO and HCI regimes showed negligible correlation to the tar and nicotine concentrations indicated on package labels. The cigarette samples used in the study were categorized into four classes: ultralow-, low-, medium-, and high-yield brands, which corresponded to 1, 3-6, 8-10, and 14 mg tar/cigarette, respectively. The concentration of TSNAs in ultralow-yield cigarettes was 210 ng/cigarette, as measured using the HCI regime, which was nearly equal to that in high-yield cigarettes (180 ng/cigarette)., Conclusions: Exposure to TSNAs from mainstream smoke from ultralow-yield cigarettes is comparable to that from high-yield cigarettes. To properly evaluate the risk of smoking, not only the concentrations of tar and nicotine but also those of other chemicals, including TSNAs, should be printed on package labels.

A new method for the determination of acrolein and other carbonyls in cigarette smoke using a dual cartridge system has been developed. Each cartridge consists of reagent-impregnated silica particles. The first contains hydroquinone (HQ) for the inhibition of acrolein polymerization, while the second contains 2,4-dinitrophenylhydrazine (DNPH) for the derivatization of carbonyls. Smoke samples were firstly drawn through the cartridge containing HQ-impregnated silica (HQ-silica) and then through the DNPH-impregnated silica (DNPH-silica). Acrolein in the sample was completely trapped in the first HQ-silica cartridge. Some other airborne carbonyls were also trapped by the HQ-silica, and those that pass through were trapped in the second DNPH-silica cartridge. Extraction was performed in the reverse direction to air sampling. When solvent was eluted through the dual-cartridges, excess DNPH was washed into the HQ bed where it reacted with acrolein and other trapped carbonyls to form the corresponding hydrazone derivatives. All of the hydrazones derived from airborne carbonyls were completely separated and measured using high-performance liquid chromatography. This HQ-DNPH-method can be applied for the determination of acrolein and other alpha,beta-unsaturated aldehydes, such as crotonaldehyde, in cigarette smoke., (Copyright 2010 Elsevier B.V. All rights reserved.)