A reliable method using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was established to conduct a comprehensive analysis of the chemical constituents of Du-zhi pill (DZP) as well as their metabolites in rat plasma, urine and feces after gastric perfusion. The efficient on-line mass data acquisition modes combined the various off-line mass data mining strategy was applied. A full mass scan was performed, and then accurate MS/MS datasets were obtained through the use of a multiple mass defect filter (MMDF) and dynamic background subtraction (DBS)-dependent data acquisition method. Furthermore, post-acquisition data processing was conducted using various data-mining tools, including extracted ion chromatography (XIC), mass defect filtering (MDF), product ion filtering (PIF), and neutral loss filtering (NLF) (MetabolitePilot™). Finaly, a total of 176 compounds were identified or tentatively characterized in DZP. Moreover, a total of 233 components in vivo, which includes 92 prototype components and 141 metabolites, were unambiguously or tentatively identified in rat plasma, urine and feces. The metabolic pathways, including phase I reactions (hydroxylation, dehydroxylation and hydrogenation) and phase II reactions (acetylation, sulfation, glucuronidation and methylation), for the absorbed constituents, were explored and summarized. This is the first systematic study on the components of DZP and their metabolites in vivo. This study provide a valid analytical strategy for the characterization of chemical compounds and metabolites of TCM formulas. Moreover, an integrative strategy was proposed for the characterization and identification of chemical constituents and metabolites for additional TCM prescriptions.

In this study, four kinds of chemical substances (2,3,5,6-tetramethylpyrazine, β-ionone, citronellal, and paeonol), three kinds of plant essential oils (tea tree essential oil, lavender essential oil, and myrrh essential oil), and their combinations were selected to explore their synergistic effects on tobacco beetle [Lasioderma serricorne (Fabricius) (Coleoptera: Anobiidae)] adults by the behavioral test and laboratory simulation test. Behavioral test results showed that some of the combinations revealed a synergistic effect on tobacco beetle adults, especially the sexual attractant +2,3,5,6-tetramethylpyrazine + β-ionone + citronellal + paeonol (SABCD, one portion of sexual attractant, and 1 mg/L synergistic substances) combination and the food attractant +2,3,5,6-tetramethylpyrazine + paeonol (FAD, 1 ml of food attractant and 1 mg/L synergistic substances) combination showed the best behavioral effect on tobacco beetle adults with average dwell times of 120.97 and 126.74 s, respectively, compared to those of other combinations. Meanwhile, SABCD had the highest selection rate [89.47%, about 1.5 times that of the sexual attractant (S)] on tobacco beetle adults compared with those of other combinations. In addition, laboratory simulation test results showed that the SABCD combination had the highest average selection rate (37.31%, about 2 times that of S) on tobacco beetle adults at 1 mg/L. However, our results showed that there was no significant difference in the indoor simulation results of food attractant synergistic substances. Our results will provide guidance for the development of new pesticides for tobacco beetle adults.

Ginseng, Panax ginseng C. A. Meyer, is an industrial crop in China and Korea. The functional components in ginseng roots and rhizomes are characteristic ginsenosides. This work developed a new high-performance liquid chromatography coupled with electrospray ionization ion trap time-of-flight multistage mass spectrometry (LC-ESI-IT-TOF-MS(n)) method to identify the triterpenoids. Sixty compounds (1-60) including 58 triterpenoids were identified from the ginseng cultivated in China. Substances 1, 2, 7, 15-20, 35, 39, 45-47, 49, 55-57, 59, and 60 were identified for the first time. To evaluate the quality of ginseng cultivated in Northeast China, this paper developed a practical liquid chromatography-diode array detection (LC-DAD) method to simultaneously quantify 14 interesting ginsenosides in ginseng collected from 66 different producing areas for the first time. The results showed the quality of ginseng roots and rhizomes from different sources was different due to growing environment, cultivation technology, and so on. The developed LC-ESI-IT-TOF-MS(n) method can be used to identify many more ginsenosides and the LC-DAD method can be used not only to assess the quality of ginseng, but also to optimize the cultivation conditions for the production of ginsenosides.

The incomplete identification of the chemical components of traditional Chinese medicinal formula has been one of the bottlenecks in the modernization of traditional Chinese medicine. Tandem mass spectrometry has been widely used for the identification of chemical substances. Current automatic tandem mass spectrometry acquisition, where precursor ions were selected according to their signal intensity, encounters a drawback in chemical substances identification when samples contain many overlapping signals. Compounds in minor or trace amounts could not be identified because most tandem mass spectrometry information was lost. Herein, a molecular feature orientated precursor ion selection and tandem mass spectrometry structure elucidation method for complex Chinese medicine chemical constituent analysis was developed. The precursor ions were selected according to their two-dimensional characteristics of retention times and mass-to-charge ratio ranges from herbal compounds, so that all precursor ions from herbal compounds were included and more minor chemical constituents in Chinese medicine were identified. Compared to the conventional automatic tandem mass spectrometry setups, the approach is novel and can overcome the drawback for chemical substances identification. As an example, 276 compounds from the Chinese Medicine of Yi-Xin-Shu capsule were identified.

A new reporter system has been developed for quantifying the activity of potentially DNA-damaging substances in the yeast Saccharomyces cerevisiae. The system relies on two different reporter genes, yEGFP and DsRed-Express2, to screen for DNA-damaging chemicals. The yEGFP gene is fused to the test promoter of RNR2, whose measurable signal has a dose-dependent relationship with DNA damage. The gene encoding DsRed-Express2 is fused to a constitutive promoter of GPD, providing an internal control for normalizing cell numbers in the assay. The dual fluorescent protein assay system is performed by sequentially measuring the yEGFP and DsRed-Express2 fluorescent intensity of the same sample, with the results expressed as the ratio of yEGFP to DsRed-Express2 intensity (yEGFP/DsRed-Express2). The yeast fluorescent protein reporter assay was performed in 96-well microtiter plates in the presence of different concentrations of test substances, which were then characterized. The assay was very efficient, high-throughput, and amenable to full automation. Here, we demonstrate that this system can be used as a biosensor to assess the genotoxic potential of drugs and other chemical substances.

Urban societies are using an increasingly diverse array of chemicals, many of which ultimately end up accumulating in urban harbors, where they can act as contaminants alone or as part of a mixture. In attempt to grasp a more complete picture of anthropogenic chemicals in an urban harbor, we analyzed 940 organic chemicals in sediments in Tokyo Bay, one of the most densely populated and modernized areas in the world. For the chemical analysis, we used targeted analytical methods using a GC-MS-MS and a GC-MS-SIM, and a screening analysis using an automated identification and quantification system with a GC-MS database. We detected 195 organic chemicals in the sediments; the sum of concentrations of compounds detected varied from 6095 to 39140μgkg(-1)dry wt. Since their concentrations increased with proximity to the innermost part of the bay, their sources seem to be mainly sewage treatment plants (STPs) and rivers flowing to this area. Additional confirmation comes from the nature of the identified pollutants, which are characteristic of chemicals used in households as well as fecal matter, business activities and urban run-off. From these results, it was confirmed that sediments in Tokyo Bay are still polluted with a wide range of chemicals, particularly domestic chemicals, despite nearly 100% of wastewater from household and business activities being treated by STPs.