Construction of a Synergy Combination Model for Turmeric (Curcuma longa L.) and Black Pepper (Piper nigrum L.) Extracts: Enhanced Anticancer Activity against A549 and NCI-H292 Human Lung Cancer Cells.

Extensive research on medicinal herbs for bioactive compounds proposes that they could replace synthetic drugs, reducing side effects and economic burdens. Especially, interest in the synergistic benefits of natural products is increasing, implying that their combined use may enhance therapeutic effectiveness. This study aimed to explore the synergetic effects of turmeric (Curcuma longa L.) and black pepper (Piper nigrum L.) extract on lung normal (MRC-5) and cancer (A549 and NCI-H292) cell lines. The turmeric extract (TM) only affected the lung cancer cell lines, but it had no impact on the MRC-5 cell line. On the other hand, the black pepper extract (BP) did not cause any damage to either the lung normal or cancer cell lines, even at concentrations of up to 400 µg/mL. Response surface methodology was used to predict the ideal synergistic concentrations (EC50) of TM and BP, which were found to be 48.5 and 241.7 µg/mL, respectively. Notably, the selected condition resulted in higher cytotoxicity compared to the exposure to TM alone, indicating a potent synergetic effect. The rate of curcumin degradation under this combined treatment was significantly decreased to 49.72 ± 5.00 nmol/h/µg for A549 cells and 47.53 ± 4.78 nmol/h/µg for NCI-H292 cells, respectively, as compared to curcumin alone. Taken together, this study confirmed the potent synergistic effect of TM and BP on lung cancer cell lines. Further research is required to identify their specific synergetic mechanisms. Our findings provide crucial foundational data on the synergistic effects of TM and BP.

Autocrine insulin-like growth factor 2 signaling as a potential target in the associated development of pulmonary emphysema and cancer in smokers.

BACKGROUND: Tobacco smoking causes pulmonary inflammation, resulting in emphysema, an independent risk factor for lung cancer. Induction of insulin-like growth factor 2 (IGF2) in response to lung injury by tobacco carcinogens, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and polycyclic aromatic hydrocarbon benzo[a]pyrene in combination (NB), is critical for the proliferation of alveolar type 2 cells (AT2s) for lung repair. However, persistent IGF2 overexpression during NB-induced severe injury results in hyperproliferation of AT2s without coordinated AT2-to-AT1 differentiation, disrupting alveolar repair, which leads to the concurrent development of emphysema and lung cancer. The current study aims to verify the role of IGF2 signaling in the associated development of emphysema and cancer and develop effective pharmaceuticals for the diseases using animal models that recapitulate the characteristics of these chronic diseases. METHODS: The pathogenesis of pulmonary emphysema and cancer was analyzed by lung function testing, histological evaluation, in situ zymography, dihydroethidium staining, and immunofluorescence and immunohistochemistry analyses utilizing mouse models of emphysema and cancer established by moderate exposure to NB for up to seven months. RESULTS: Moderate NB exposure induced IGF2 expression in AT2s during the development of pulmonary emphysema and lung cancer in mice. Using AT2-specific insulin receptor knockout mice, we verified the causative role of sustained IGF2 signaling activation in AT2s in emphysema development. IGF2-targeting strategies, including voltage-dependent calcium channel blocker (CCB) and a neutralizing antibody, significantly suppressed the NB-induced development of emphysema and lung cancer. A publicly available database revealed an inverse correlation between the use of calcium channel blockers and a COPD diagnosis. CONCLUSIONS: Our work confirms sustained IGF2 signaling activation in AT2s couples impaired lung repair to the concurrent development of emphysema and cancer in mice. Additionally, CCB and IGF2-specific neutralizing antibodies are effective pharmaceuticals for the two diseases.

Effect of Capsule Burst in Cigarette Filters on the Compound Composition of Mainstream Cigarette Smoke.

The number of cigarette capsule users is increasing; however, a comprehensive epidemiological investigation comparing the harmfulness of capsule and non-capsule cigarettes, particularly concerning the composition of flavor components and mainstream smoke, is lacking. The present study aimed to investigate the effect of capsule burst on the compound composition of mainstream smoke by quantifying the Hoffmann list and flavor (geraniol, eugenol, menthofuran, and pulegone) in mainstream smoke with and without crushing the capsules. The findings indicate that while tar values tended to increase when the capsules were burst, there was no significant change observed in the other Hoffmann list components, such as nicotine, benzo[α]pyrene, tobacco-specific nitrosamines, aromatic amines, and phenolics. However, when the capsules burst, menthofuran and pulegone, which were present in the capsule and could cause toxicity, were found in the mainstream smoke via the International Standard Organization (1.5-4.0 µg/cig and 5.13-6.00 µg/cig smoking regime) and Health Canada Intense (12.8-18.2 µg/cig and 22.77-24.67 µg/cig smoking regime). Therefore, understanding the capsule composition is important, as the toxic components of the capsules can be inhaled as smoke, posing a potential health risk.

Overexpression of the Brassica rapa bZIP Transcription Factor, BrbZIP-S, Increases the Stress Tolerance in Nicotiana benthamiana.

In higher plants, S1-basic region-leucine zipper (S1-bZIP) transcription factors fulfill crucial roles in the physiological homeostasis of carbon and amino acid metabolisms and stress responses. However, very little is known about the physiological role of S1-bZIP in cruciferous vegetables. Here, we analyzed the physiological function of S1-bZIP from Brassica rapa (BrbZIP-S) in modulating proline and sugar metabolism. Overexpression of BrbZIP-S in Nicotiana benthamiana resulted in delayed chlorophyll degradation during the response to dark conditions. Under heat stress or recovery conditions, the transgenic lines exhibited a lower accumulation of H2O2, malondialdehyde, and protein carbonyls compared to the levels in transgenic control plants. These results strongly indicate that BrbZIP-S regulates plant tolerance against dark and heat stress. We propose that BrbZIP-S is a modulator of proline and sugar metabolism, which are required for energy homeostasis in response to environmental stress conditions.

Plant Response to Cold Stress: Cold Stress Changes Antioxidant Metabolism in Heading Type Kimchi Cabbage (Brassica rapa L. ssp. Pekinensis).

Cold stress is known as the important yield-limiting factor of heading type Kimchi cabbage (HtKc, Brassica rapa L. ssp. pekinensis), which is an economically important crop worldwide. However, the biochemical and molecular responses to cold stress in HtKc are largely unknown. In this study, we conducted transcriptome analyses on HtKc grown under normal versus cold conditions to investigate the molecular mechanism underlying HtKc responses to cold stress. A total of 2131 genes (936 up-regulated and 1195 down-regulated) were identified as differentially expressed genes and were significantly annotated in the category of "response to stimulus". In addition, cold stress caused the accumulation of polyphenolic compounds, including p-coumaric, ferulic, and sinapic acids, in HtKc by inducing the phenylpropanoid pathway. The results of the chemical-based antioxidant assay indicated that the cold-induced polyphenolic compounds improved the free-radical scavenging activity and antioxidant capacity, suggesting that the phenylpropanoid pathway induced by cold stress contributes to resistance to cold-induced reactive oxygen species in HtKc. Taken together, our results will serve as an important base to improve the cold tolerance in plants via enhancing the antioxidant machinery.

Safety and biological activity evaluation of Uncaria rhynchophylla ethanolic extract.

Uncaria rhynchophylla (UR) belongs to the Rubiaceae family, and its dried hooks are usually used in traditional medicine. It is effective in treating diseases related to the central nervous system. This study aimed to evaluate the safety of UR extract, investigate its antimutagenic and antioxidative activities, and elucidate its active components. Extraction and fractionation of the UR extract resulted in yields of 6.71%, 0.037%, 0.042%, 0.152%, 0.332%, and 5.132%, for hexane, ether, DCM, EtOAC, and aqueous fractions, respectively. The four indole alkaloids, total phenolic content (TPC), and total flavonoid content (TFC) of UR extract and its subfractions were measured. Alkaloid content was highest in the UR extract. TPC was the highest in the EtOAC fraction (373.7 ± 20.9 mg gallic acid equivalent (GAE)/g), whereas TFC was the highest in the UR extract (33.5 ± 2.4 mg quercetin equivalent (QE)/g). To assess the safety of UR extract mutagenicity, cytotoxicity, and oxidative stress inducibility assays were performed. The UR extract (2000 µg/plate) showed excellent antimutagenic activity (above 90%) against BaP in both TA98 and TA100 strains. The UR extract exhibited efficient DPPH (RC50 239.2 ± 16.5 µg/mL) and ABTS scavenging activity (RC50 458.7 ± 25.0 µg/mL). The UR extract (150 µg/mL) showed cytoprotective activity (65.6% ± 9.2%) against t-BHP. Among the subfractions, the EtOAC fraction possessed the strongest activities, overall. UR generally showed excellent biological activity at nontoxic concentrations (determined in vitro in current work), although the chemical composition of UR requires further investigation prior to its potential future use.

Comparative Study of Bioactivity and Safety Evaluation of Ethanolic Extracts of Zanthoxylum schinifolium Fruit and Pericarp.

The fruit and pericarp of Zanthoxylum schinifolium (ZS) have been used in traditional medicine; however, few studies have characterized ZS fruit and pericarp. Therefore, in the present study, we evaluated the safety of ZS fruit (ZSF) and pericarp (ZSP) extracts and compared their bioactivity. To evaluate the safety of ZSF and ZSP, mutagenicity, cytotoxicity, and oxidative stress assays were performed and nontoxic concentration ranges were obtained. ZSP was found to be superior to ZSF in terms of its antimutagenic, antioxidant, and anti-inflammatory activities. In the S9 mix, the mutation inhibition rate of ZSP was close to 100% at concentrations exceeding 625 µg·plate-1 for both the TA98 and TA100 strains. ZSP exhibited efficient DPPH (IC50 = 75.6 ± 6.1 µg·mL-1) and ABTS (IC50 = 57.4 ± 6 µg·mL-1) scavenging activities. ZSP inhibited the release of cytokines, involved in IL-1ß (IC50 = 134.4 ± 7.8), IL-6 (IC50 = 262.8 ± 11.2), and TNF-α (IC50 = 223.8 ± 5.8). These results indicate that ZSP contains a higher amount of biochemicals than ZSF, or that ZSP contains unique biochemicals. In conclusion, for certain physiological activities, the use of ZSP alone may be more beneficial than the combined use of ZSF and ZSP.

Taxifolin and Sorghum Ethanol Extract Protect against Hepatic Insulin Resistance via the miR-195/IRS1/PI3K/AKT and AMPK Signalling Pathways.

This study aimed to evaluate the effects of taxifolin and sorghum ethanol extract on free fatty acid (FFA)-induced hepatic insulin resistance. FFA treatment decreased glucose uptake by 16.2% compared with that in the control, whereas taxifolin and sorghum ethanol extract increased the glucose uptake. Additionally, taxifolin and sorghum ethanol extract increased the expression of p-PI3K, p-IRS1, p-AKT, p-AMPK, and p-ACC in FFA-induced hepatocytes. Furthermore, FFA treatment increased the expression of miR-195. However, compared with the FFA treatment, treatment with taxifolin and sorghum ethanol extract decreased miR-195 expression in a dose-dependent manner. Taxifolin and sorghum ethanol extract enhanced p-IRS1, p-PI3K, p-AMPK, p-AKT, and p-ACC expression by suppressing miR-195 levels in miR-195 mimic- or inhibitor-transfected cells. These results indicate that taxifolin and sorghum ethanol extract attenuate insulin resistance by regulating miR-195 expression, which suggests that taxifolin and sorghum ethanol extract may be useful antidiabetic agents.

UPLC-ESI-Q-TOF-MS-Based Metabolite Profiling, Antioxidant and Anti-Inflammatory Properties of Different Organ Extracts of Abeliophyllum distichum.

Plant extracts have gained more attention as natural therapeutic agents against inflammation characterized by an overproduction of several inflammatory mediators such as reactive oxygen species and pro-inflammatory cytokines. Although Abeliophyllum distichum Nakai is generally known for its ornamental value, recent pharmacological research has demonstrated its potential therapeutic properties. Thus, to further evaluate the applicability of A. distichum in the food, cosmetic, and medical industries, we identified the phytochemicals in three organ extracts (fruits: AF, branches: AB, leaves: AL) of A. distichum and determined their antioxidant and anti-inflammatory activities. Using UPLC-ESI-Q-TOF-MS, a total of 19 compounds, including dendromoniliside D, forsythoside B, isoacteoside, isomucronulatol 7-O-Glucoside, plantamajoside, and wighteone were identified in the A. distichum organ extracts. AB exhibited a strong reducing power, an oxygen radical antioxidant capacity, and radical scavenging values compared with other samples, whereas AL exhibited the best anti-inflammatory properties. Gene expression, western blot, and molecular docking analyses suggested that the anti-inflammatory effect of AL was mediated by its ability to suppress lipopolysaccharide (LPS)-induced production of reactive oxygen species and/or inhibit LPS-stimulated activation of extracellular signal-regulated protein kinases (ERK1/2) in RAW264.7 cells. Collectively, these results indicate that AL is a potential source of phytochemicals that could be used to treat inflammation-associated diseases.

Establishment of a UPLC-PDA/ESI-Q-TOF/MS-Based Approach for the Simultaneous Analysis of Multiple Phenolic Compounds in Amaranth (A. cruentus and A. tricolor).

We used ultraperformance liquid chromatography coupled with a photodiode-array detector and electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-PDA/ESI-Q-TOF/MS) to rapidly and accurately quantify 17 phenolic compounds. Then, we applied this method to the seed and leaf extracts of two Amaranthus species to identify and quantify phenolic compounds other than the 17 compounds mentioned above. Compounds were eluted within 30 min on a C18 column using a mobile phase (water and acetonitrile) containing 0.1% formic acid, and the specific wavelength and ion information of the compounds obtained by PDA and ESI-Q-TOF/MS were confirmed. The proposed method showed good linearity (r2 > 0.990). Limits of detection and quantification were less than 0.1 and 0.1 µg/mL, respectively. Intra- and interday precision were less than 2.4% and 1.8%, respectively. Analysis of amaranth seed and leaf extracts using the established method showed that the seeds contained high amounts of 2,4-dihydroxybenzoic acid and kaempferol, and leaves contained diverse phenolic compounds. In addition, six tentatively new phenolic compounds were identified. Moreover, seeds potentially contained 2,3-dihydroxybenzaldehyde, a beneficial bioactive compound. Thus, our method was an efficient approach for the qualitative and quantitative analysis of phenolic compounds, and could be used to investigate phenolic compounds in plants.

Ethyl Acetate Fraction of Aqueous Extract of Lentinula edodes Inhibits Osteoclastogenesis by Suppressing NFATc1 Expression.

Bone tissue is continuously remodeled by the coordinated action of osteoclasts and osteoblasts. Nuclear factor-activated T cells c1 (NFATc1) is a well-known transcription factor for osteoclastogenesis and transcriptionally activated by the c-Fos and nuclear factor-kappa B (NF-κB) signaling pathways in response to receptor activation of NF-κB ligand (RANKL). Since excessive RANKL signaling causes an increase of osteoclast formation and bone resorption, inhibition of RANKL or its signaling pathway is an attractive therapeutic approach to the treatment of pathologic bone loss. In this study, we show that an ethyl acetate fraction (LEA) from the shiitake mushroom, Lentinula edodes, inhibited RANKL-induced osteoclast differentiation by blocking the NFATc1 signaling pathway. We found that the water extract and its subsequent ethyl acetate fraction of L. edodes significantly suppressed osteoclast formation. Comparative transcriptome analysis revealed that LEA specifically downregulated a set of RANKL target genes, including Nfatc1. Next, we found that LEA suppresses Nfatc1 expression mainly through the inhibition of the transactivity of p65 and NFATc1. Moreover, treatment of LEA rescued an osteoporotic phenotype in a zebrafish model of glucocorticoid-induced osteoporosis. Collectively, our findings define an undocumented role of the shiitake mushroom extract in regulating bone development.

In vitro toxicological activity of particulate matter generated by coal combustion.

Herein, the toxicity of particles generated from the complete combustion of 1 g coal at 500, 700, and 900 °C were compared, and combustion at 700 °C generated the most toxins. Chemical analyses revealed that all components except catechol, resorcinol, and aromatic amines were most abundant at 700 °C. Toxicity results confirmed that the relative mutagenicity, cytotoxicity, redox cycling, and production of reactive oxygen species was highest for particles generated at 700 °C. Particles generated during combustion at 700 °C exhibited higher toxicity toward biological systems due to a higher content of toxic compounds.

Correction to: Effects of roasting conditions on physicochemical properties and antioxidant activities in Ginkgo biloba seeds.

[This corrects the article DOI: 10.1007/s10068-018-0348-7.].

Effects of roasting conditions on physicochemical properties and antioxidant activities in Ginkgo biloba seeds.

The roasting treatment has been used to extend the shelf life of food and improves its quality, and eliminate or reduce toxic. In this study, we investigate the changes in the physicochemical properties and antioxidant activities of Ginkgo biloba seeds (GBS) according to the roasting temperature and duration. As the roasting temperature and duration increased, the pH (from 7.32 to 6.31) and total cyanide content (from 1.49 to 0.70 µg/g) decreased, whereas the titratable acidity (from 0.39 to 0.84%) increased. The antioxidant activities increased rapidly at 210 °C according to the increase in the roasting temperature and duration. The 4'-O-methylpyridoxine (MPN) content in the 210 °C heat treatment group decreased by more than 70% compared to the MPN content in the control group. These results suggest that heat-treated GBS could be used in food materials and medicines for decreasing cyanide and MPN contents as well as for increasing antioxidant compound contents.

Comparison of Mutagenic Activities of Various Ultra-Fine Particles.

Air pollution is increasing, along with consumption of fossil fuels such as coal and diesel gas. Air pollutants are known to be a major cause of respiratory-related illness and death, however, there are few reports on the genotoxic characterization of diverse air pollutants in Korea. In this study, we investigated the mutagenic activity of various particles such as diesel exhaust particles (DEP), combustion of rice straw (RSC), pine stem (PSC), and coal (CC), tunnel dust (TD), and road side dust (RD). Ultra-fine particles (UFPs) were collected by the glass fiber filter pad. Then, we performed a chemical analysis to see each of the component features of each particulate matter. The mutagenicity of various UFPs was determined by the Ames test with four Salmonella typhimurium strains with or without metabolic activation. The optimal concentrations of UFPs were selected based on result of a concentration decision test. Moreover, in order to compare relative mutagenicity among UFPs, we selected and tested DEP as mutation reference. DEP, RSC, and PSC induced concentration-dependent increases in revertant colony numbers with TA98, TA100, and TA1537 strains in the absence and presence of metabolic activation. DEP showed the highest specific activity among the particulate matters. In this study, we conclude that DEP, RSC, PSC, and TD displayed varying degrees of mutagenicity, and these results suggest that the mutagenicity of these air pollutants is associated with the presence of polycyclic aromatic hydrocarbons (PAHs) in these particulate matters.

Effect of heat treatment on 4'-O-methylpyridoxine (MPN) content in Ginkgo biloba seed extract solution.

BACKGROUND: A vitamin B6 derivative, 4'-O-methylpyridoxine (MPN), is responsible for food poisoning by Ginkgo biloba seeds. In this study, we investigate the content of pyridoxine and MPN in MPN standard solution and G. biloba seed extract solution upon heat treatment in order to evaluate the reduction of toxic components in G. biloba seed by such treatment. RESULTS: Heat treatment was conducted at 90-150 °C for 0-60 min, and all samples were adjusted to the same concentration of 1 g L-1 . The MPN content decreased to 994.92-563.69 mg kg-1 for MPN standard solution and to 371.56-76.84 mg kg-1 for G. biloba seed extract solution, and in both cases decreased even further with increasing heat treatment time. However, in all samples, except for the 90 °C heat treatment group, the pyridoxine content in MPN standard solution increased with increasing heat temperature and time; in addition, the extract solution showed a similar tendency. This may be the result of thermal degradation of MPN into pyridoxine. CONCLUSION: We can expect to improve the utilization of functional food materials by applying suitable heat treatment conditions and decreasing the MPN content of the G. biloba seed. © 2018 Society of Chemical Industry.

A UPLC-ESI-Q-TOF method for rapid and reliable identification and quantification of major indole alkaloids in Catharanthus roseus.

We developed a novel ultra performance liquid chromatography-quadrupole time-of-flight (UPLC-Q-TOF) mass spectrometry method that allows sensitive, rapid, and reliable detection and identification of six representative indole alkaloids (vincristine, vinblastine, ajmalicine, catharanthine, serpentine, and vindoline) that exhibit physiological activity in Catharanthus roseus (C. roseus). The alkaloids were eluted on a C18 column with acetonitrile and water containing 0.1% formic acid and 10 mM ammonium acetate, and separated with good resolution within 13 min. Electrospray ionization-Q-TOF (ESI-Q-TOF) analysis was performed to characterize the molecules and their fragment ions, and the characteristic ions and fragmentation patterns were used as to identify the alkaloids. The proposed analytical method was verified in reference to the ICH guidelines and the results showed excellent linearity (R2 > 0.9988), limit of detection (1 ng/mL to 10 ng/mL), limit of quantification (3 ng/mL to 30 ng/mL), intra-day and inter-day precisions, and extraction recovery rates (92.8% to 104.1%) for all components. The validated UPLC-Q-TOF method was applied to the analysis of extracts from the root, stem, and leaves of C. roseus, allowing the identification of six alkaloids by comparison of retention times, molecular ions, and fragmentation patterns with those of reference compounds. Sixteen additional indole alkaloids were tentatively identified by comparison of chromatograms to chemical databases and literature reports. The contents of bis-indole alkaloids (vincristine and vinblastine) were high in the aerial parts, while the contents of mono-indole alkaloids (ajmalicine, catharanthine, serpentine, and vindoline) were high in the roots. The present results demonstrate that the proposed UPLC-Q-TOF method can be useful for the investigation of phytochemical constituents of medicinal plants.

Comparison of the in vitro toxicological activity of various particulate matter.

Ultrafine particles (UFPs, < 2.5 µm) in air pollutants have been identified as a major cause of respiratory diseases, since they can affect the lung alveoli through the bronchus. In particular, if toxicants such as heavy metals and polycyclic aromatic hydrocarbons (PAHs) are present in UFPs, they can cause diseases such as asthma, chronic obstructive pulmonary disease, and lung cancer. This study compared in vitro toxicity of various particulate matter including UFPs from combustion particles of diesel (diesel exhaust particles (DEP)), rice straw (RS), pine stem (PS) and coal (CC), and road dust particles from tunnel (TD) and roadside (RD). UFPs from combustion particles and road dust were collected with a glass fiber filter using burning systems and a solid aerosol generator. Cell viability was determined by neutral red uptake assay using Chinese hamster ovary strain K1 cells. Redox cycling activity and intracellular reactive oxygen species were measured using 1,4-dithiothreitol (DTT) and 2',7'-dichlorofluorescin diacetate (DCF-DA) assay, respectively. Our in vitro studies validated that combustion particles had high toxicological activity. PS demonstrated the highest activity in cytotoxicity but DEP had the highest activity in the DTT and DCF-DA assays. Overall, since the toxicological activity of particles generated by various means was different, risk assessment should be conducted through various toxicity evaluations rather than one toxicity evaluation.

Quantitative mapping of elements in basil leaves (Ocimum basilicum) based on cesium concentration and growth period using laser ablation ICP-MS.

Quantitative elemental mapping of metallic pollutants in sweet basil was studied by laser ablation (LA)-ICP-MS. For this, the sweet basil was cultivated in Hoagland nutrient solution spiked with 100 and 1000 ng mL-1 of Cs for 10-60 days. Then, the Cs distribution in collected leaves was determined by LA-ICP-MS using lab-synthesized standard pellets based on NIST 1573a tomato leaves. For comparison, S, Ca, and K were also simultaneously determined in this measurement with a13C+ signal from the leaves as an internal standard. The obtained calibration curves showed linear coefficient of determination (R2) of 0.991 for K and 0.999 for Cs. The concentration of Cs measured in the basil leaves increased with growth period and pollutant concentration, and accumulation followed the order of leaf margin, petiole, midrib, and veins. Although no visible symptom was detected, significant suppression of the growth rate was observed due to the presence of high-concentration Cs. The experimental model demonstrated herein showed potential for studying the influence of radioactive pollutants on plants and other organisms in the food chain.

Comparison of the content of tobacco alkaloids and tobacco-specific nitrosamines in 'heat-not-burn' tobacco products before and after aerosol generation.

Standardized methods for collecting smoke and for measuring smoke components in heat not burn tobacco product (HTP) are yet to be established, and there is a lack of consensus as to whether the content of HTP cigarettes can be assayed in the same manner as for conventional cigarettes. Since HTPcigarettes do not generate ash when smoked, we compared the levels of tobacco alkaloids (TAs) and tobacco-specific nitrosamines (TSNAs) of HTP cigarettes before and after aerosol generation. HTP cigarettes were smoked according to two international standardization methods. The TAs and TSNAs contents of the cigarettes were analyzed by UPLC-Q-TOF and UPLC-MSMS, respectively. Smoking was found to significantly decrease the content of nicotine, nornicotine, anatabine, and anabasine by 53 ∼ 100% in all samples, and the maximum inhalable amounts of these entities were determined to be 4.24 mg/cig, 103.52 µg/cig, 258.72 µg/cig, and 33.03 µg/cig, respectively. By contrast, smoking significantly increased the content of NNK and NAB. we suggested that the reduced nicotine content minus the nicotine content remaining in the filter is an amount that could potentially be inhaled during smoking. The increase of NNK and NAB in HTP cigarette after aerosol generation is expected to be caused by the precursor, but more specific behavioral studies should be performed.