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Rebecca E. Irwin - One of the best experts on this subject based on the ideXlab platform.

  • context dependent medicinal effects of Anabasine and infection dependent toxicity in bumble bees
    PLOS ONE, 2017
    Co-Authors: Evan C Palmeryoung, Jonathan Andicoechea, Alison Hogeboom, Alexander J Kaye, Dash Donnelly, Sara June Connon, Ian Weston, Kimberly Skyrm, Rebecca E. Irwin
    Abstract:

    Background Floral phytochemicals are ubiquitous in nature, and can function both as antimicrobials and as insecticides. Although many phytochemicals act as toxins and deterrents to consumers, the same chemicals may counteract disease and be preferred by infected individuals. The roles of nectar and pollen phytochemicals in pollinator ecology and conservation are complex, with evidence for both toxicity and medicinal effects against parasites. However, it remains unclear how consistent the effects of phytochemicals are across different parasite lineages and environmental conditions, and whether pollinators actively self-medicate with these compounds when infected. Approach Here, we test effects of the nectar alkaloid Anabasine, found in Nicotiana, on infection intensity, dietary preference, and survival and performance of bumble bees (Bombus impatiens). We examined variation in the effects of Anabasine on infection with different lineages of the intestinal parasite Crithidia under pollen-fed and pollen-starved conditions. Results We found that Anabasine did not reduce infection intensity in individual bees infected with any of four Crithidia lineages that were tested in parallel, nor did Anabasine reduce infection intensity in microcolonies of queenless workers. In addition, neither Anabasine nor its isomer, nicotine, was preferred by infected bees in choice experiments, and infected bees consumed less Anabasine than did uninfected bees under no-choice conditions. Furthermore, Anabasine exacerbated the negative effects of infection on bee survival and microcolony performance. Anabasine reduced infection in only one experiment, in which bees were deprived of pollen and post-pupal contact with nestmates. In this experiment, Anabasine had antiparasitic effects in bees from only two of four colonies, and infected bees exhibited reduced—rather than increased—phytochemical consumption relative to uninfected bees. Conclusions Variation in the effect of Anabasine on infection suggests potential modulation of tritrophic interactions by both host genotype and environmental variables. Overall, our results demonstrate that Bombus impatiens prefer diets without nicotine and Anabasine, and suggest that the medicinal effects and toxicity of Anabasine may be context dependent. Future research should identify the specific environmental and genotypic factors that determine whether nectar phytochemicals have medicinal or deleterious effects on pollinators.

  • RESEARCH ARTICLE Testing Dose-Dependent Effects of the Nectar Alkaloid Anabasine on Trypanosome Parasite Loads in Adult Bumble Bees
    2016
    Co-Authors: Winston E. Anthony¤a, Evan C. Palmer-young, Rebecca E. Irwin, Anne S. Leonard¤b, Lynn S
    Abstract:

    The impact of consuming biologically active compounds is often dose-dependent, where small quantities can be medicinal while larger doses are toxic. The consumption of plant secondary compounds can be toxic to herbivores in large doses, but can also improve sur-vival in parasitized herbivores. In addition, recent studies have found that consuming nectar secondary compounds may decrease parasite loads in pollinators. However, the effect of compound dose on bee survival and parasite loads has not been assessed. To determine how secondary compound consumption affects survival and pathogen load in Bombus impatiens, we manipulated the presence of a common gut parasite, Crithidia bombi, and dietary concentration of Anabasine, a nectar alkaloid produced by Nicotiana spp. using four concentrations naturally observed in floral nectar. We hypothesized that increased con-sumption of secondary compounds at concentrations found in nature would decrease sur-vival of uninfected bees, but improve survival and ameliorate parasite loads in infected bees. We found medicinal effects of Anabasine in infected bees; the high-Anabasine diet decreased parasite loads and increased the probability of clearing the infection entirely

  • variable effects of nicotine Anabasine and their interactions on parasitized bumble bees
    F1000Research, 2015
    Co-Authors: Lukas P Thorburn, Lynn S. Adler, Rebecca E. Irwin, Evan C Palmeryoung
    Abstract:

    Secondary metabolites in floral nectar have been shown to reduce parasite load in two common bumble bee species. Previous studies on the effects of nectar secondary metabolites on parasitized bees have focused on single compounds in isolation; however, in nature, bees are simultaneously exposed to multiple compounds. We tested for interactions between the effects of two alkaloids found in the nectar of Nicotiana spp. plants, nicotine and Anabasine, on parasite load and mortality in bumble bees ( Bombus impatiens) infected with the intestinal parasite Crithidia bombi. Adult worker bees inoculated with C. bombi were fed nicotine and Anabasine diet treatments in a factorial design, resulting in four nectar treatment combinations:  2 ppm nicotine, 5 ppm Anabasine, 2ppm nicotine and 5 ppm Anabasine together, or a control alkaloid-free solution. We conducted the experiment twice: first, with bees incubated under variable environmental conditions ('Variable'; temperatures varied from 10-35°C with ambient lighting); and second, under carefully controlled environmental conditions ('Stable'; 27°C incubator, constant darkness). In 'Variable', each alkaloid alone significantly decreased parasite loads, but this effect was not realized with the alkaloids in combination, suggesting an antagonistic interaction. Nicotine but not Anabasine significantly increased mortality, and the two compounds had no interactive effects on mortality. In 'Stable', nicotine significantly increased parasite loads, the opposite of its effect in 'Variable'. While not significant, the relationship between Anabasine and parasite loads was also positive. Interactive effects between the two alkaloids on parasite load were non-significant, but the pattern of antagonistic interaction was similar to that in the variable experiment. Neither alkaloid, nor their interaction, significantly affected mortality under controlled conditions. Our results do not indicate synergy between Nicotiana nectar alkaloids; however, they do suggest a complex interaction between secondary metabolites, parasites, and environmental variables, in which secondary metabolites can be either toxic or medicinal depending on context.

  • Testing Dose-Dependent Effects of the Nectar Alkaloid Anabasine on Trypanosome Parasite Loads in Adult Bumble Bees.
    PloS one, 2015
    Co-Authors: Winston E Anthony, Evan C. Palmer-young, Rebecca E. Irwin, Anne S Leonard, Lynn S. Adler
    Abstract:

    The impact of consuming biologically active compounds is often dose-dependent, where small quantities can be medicinal while larger doses are toxic. The consumption of plant secondary compounds can be toxic to herbivores in large doses, but can also improve survival in parasitized herbivores. In addition, recent studies have found that consuming nectar secondary compounds may decrease parasite loads in pollinators. However, the effect of compound dose on bee survival and parasite loads has not been assessed. To determine how secondary compound consumption affects survival and pathogen load in Bombus impatiens, we manipulated the presence of a common gut parasite, Crithidia bombi, and dietary concentration of Anabasine, a nectar alkaloid produced by Nicotiana spp. using four concentrations naturally observed in floral nectar. We hypothesized that increased consumption of secondary compounds at concentrations found in nature would decrease survival of uninfected bees, but improve survival and ameliorate parasite loads in infected bees. We found medicinal effects of Anabasine in infected bees; the high-Anabasine diet decreased parasite loads and increased the probability of clearing the infection entirely. However, survival time was not affected by any level of Anabasine concentration, or by interactive effects of Anabasine concentration and infection. Crithidia infection reduced survival time by more than two days, but this effect was not significant. Our results support a medicinal role for Anabasine at the highest concentration; moreover, we found no evidence for a survival-related cost of Anabasine consumption across the concentration range found in nectar. Our results suggest that consuming Anabasine at the higher levels of the natural range could reduce or clear pathogen loads without incurring costs for healthy bees.

  • variable effects of nicotine and Anabasine on parasitized bumble bees
    F1000Research, 2015
    Co-Authors: Lukas P Thorburn, Lynn S. Adler, Rebecca E. Irwin, Evan C Palmeryoung
    Abstract:

    Secondary metabolites in floral nectar have been shown to reduce parasite load in two common bumble bee species. Previous studies on the effects of nectar secondary metabolites on parasitized bees have focused on single compounds in isolation; however, in nature, bees are simultaneously exposed to multiple compounds. We tested for synergistic effects of two alkaloids found in the nectar of Nicotiana spp. plants, nicotine and Anabasine, on parasite load and mortality in bumble bees ( Bombus impatiens ) infected with the intestinal parasite Crithidia bombi . Adult worker bees inoculated with C. bombi were fed nicotine and Anabasine diet treatments in a factorial design, resulting in four nectar treatment combinations:  2 ppm nicotine, 5 ppm Anabasine, 2ppm nicotine and 5 ppm Anabasine together, or a control alkaloid-free solution. We conducted the experiment twice: first, with bees incubated under variable environmental conditions (‘Variable’; temperatures varied from 10-35°C); and second, under carefully controlled environmental conditions (‘Controlled’; 27°C incubator, constant darkness). In ‘Variable’, each alkaloid alone significantly decreased parasite loads, but this effect was not realized with the alkaloids in combination, suggesting an antagonistic interaction. Nicotine but not Anabasine significantly increased mortality, and the two compounds had no interactive effects on mortality. In ‘Controlled’, nicotine significantly increased parasite loads, the opposite of its effect in ‘Variable’. While not significant, the relationship between Anabasine and parasite loads was also positive. Interactive effects between the two alkaloids on parasite load were non-significant, but the pattern of antagonistic interaction was similar to that in the variable experiment. Neither alkaloid, nor their interaction, significantly affected mortality under controlled conditions. Our results do not indicate synergy between Nicotiana nectar alkaloids; however, they do suggest a complex interaction between secondary metabolites, parasites, and environmental variables, in which secondary metabolites can be either toxic or medicinal depending on context.

Sharon E. Murphy - One of the best experts on this subject based on the ideXlab platform.

  • Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine by High Throughput Liquid Chromatography-Mass Spectrometry.
    Chemical research in toxicology, 2016
    Co-Authors: Linda B. Von Weymarn, Dorothy K. Hatsukami, Nicole M. Thomson, Eric C. Donny, Sharon E. Murphy
    Abstract:

    Nicotine is the most abundant alkaloid in tobacco accounting for 95% of the alkaloid content. There are also several minor tobacco alkaloids; among these are nornicotine, anatabine, and Anabasine. We developed and applied a 96 well plate-based capillary LC-tandem mass spectrometry method for the analysis of nornicotine, anatabine, and Anabasine in urine. The method was validated with regard to accuracy and precision. Anabasine was quantifiable to low levels with a limit of quantitation (LOQ) of 0.2 ng/mL even when nicotine, which is isobaric, is present at concentrations >2500-fold higher than Anabasine. This attribute of the method is important since anatabine and Anabasine in urine have been proposed as biomarkers of tobacco use for individuals using nicotine replacement therapies. In the present study, we analyzed the three minor tobacco alkaloids in urine from 827 smokers with a wide range of tobacco exposures. Nornicotine (LOQ 0.6 ng/mL) was detected in all samples, and anatabine (LOQ, 0.15 ng/mL) an...

  • Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers’ Urine by High Throughput Liquid Chromatography–Mass Spectrometry
    2016
    Co-Authors: Linda B. Von Weymarn, Dorothy K. Hatsukami, Nicole M. Thomson, Eric C. Donny, Sharon E. Murphy
    Abstract:

    Nicotine is the most abundant alkaloid in tobacco accounting for 95% of the alkaloid content. There are also several minor tobacco alkaloids; among these are nornicotine, anatabine, and Anabasine. We developed and applied a 96 well plate-based capillary LC-tandem mass spectrometry method for the analysis of nornicotine, anatabine, and Anabasine in urine. The method was validated with regard to accuracy and precision. Anabasine was quantifiable to low levels with a limit of quantitation (LOQ) of 0.2 ng/mL even when nicotine, which is isobaric, is present at concentrations >2500-fold higher than Anabasine. This attribute of the method is important since anatabine and Anabasine in urine have been proposed as biomarkers of tobacco use for individuals using nicotine replacement therapies. In the present study, we analyzed the three minor tobacco alkaloids in urine from 827 smokers with a wide range of tobacco exposures. Nornicotine (LOQ 0.6 ng/mL) was detected in all samples, and anatabine (LOQ, 0.15 ng/mL) and Anabasine were detected in 97.7% of the samples. The median urinary concentrations of nornicotine, anatabine, and Anabasine were 98.9, 4.02, and 5.53 ng/mL. Total nicotine equivalents (TNE) were well correlated with anatabine (r2 = 0.714) and Anabasine (r2 = 0.760). TNE was most highly correlated with nornicotine, which is also a metabolite of nicotine. Urine samples from a subset of subjects (n = 110) were analyzed for the presence of glucuronide conjugates by quantifying any increase in anatabine and Anabasine concentrations after β-glucuronidase treatment. The median ratio of the glucuronidated to free anatabine was 0.74 (range, 0.1 to 10.9), and the median ratio of glucuronidated to free Anabasine was 0.3 (range, 0.1 to 2.9). To our knowledge, this is the largest population of smokers for whom the urinary concentrations of these three tobacco alkaloids has been reported

Lynn S. Adler - One of the best experts on this subject based on the ideXlab platform.

  • variable effects of nicotine Anabasine and their interactions on parasitized bumble bees
    F1000Research, 2015
    Co-Authors: Lukas P Thorburn, Lynn S. Adler, Rebecca E. Irwin, Evan C Palmeryoung
    Abstract:

    Secondary metabolites in floral nectar have been shown to reduce parasite load in two common bumble bee species. Previous studies on the effects of nectar secondary metabolites on parasitized bees have focused on single compounds in isolation; however, in nature, bees are simultaneously exposed to multiple compounds. We tested for interactions between the effects of two alkaloids found in the nectar of Nicotiana spp. plants, nicotine and Anabasine, on parasite load and mortality in bumble bees ( Bombus impatiens) infected with the intestinal parasite Crithidia bombi. Adult worker bees inoculated with C. bombi were fed nicotine and Anabasine diet treatments in a factorial design, resulting in four nectar treatment combinations:  2 ppm nicotine, 5 ppm Anabasine, 2ppm nicotine and 5 ppm Anabasine together, or a control alkaloid-free solution. We conducted the experiment twice: first, with bees incubated under variable environmental conditions ('Variable'; temperatures varied from 10-35°C with ambient lighting); and second, under carefully controlled environmental conditions ('Stable'; 27°C incubator, constant darkness). In 'Variable', each alkaloid alone significantly decreased parasite loads, but this effect was not realized with the alkaloids in combination, suggesting an antagonistic interaction. Nicotine but not Anabasine significantly increased mortality, and the two compounds had no interactive effects on mortality. In 'Stable', nicotine significantly increased parasite loads, the opposite of its effect in 'Variable'. While not significant, the relationship between Anabasine and parasite loads was also positive. Interactive effects between the two alkaloids on parasite load were non-significant, but the pattern of antagonistic interaction was similar to that in the variable experiment. Neither alkaloid, nor their interaction, significantly affected mortality under controlled conditions. Our results do not indicate synergy between Nicotiana nectar alkaloids; however, they do suggest a complex interaction between secondary metabolites, parasites, and environmental variables, in which secondary metabolites can be either toxic or medicinal depending on context.

  • Testing Dose-Dependent Effects of the Nectar Alkaloid Anabasine on Trypanosome Parasite Loads in Adult Bumble Bees.
    PloS one, 2015
    Co-Authors: Winston E Anthony, Evan C. Palmer-young, Rebecca E. Irwin, Anne S Leonard, Lynn S. Adler
    Abstract:

    The impact of consuming biologically active compounds is often dose-dependent, where small quantities can be medicinal while larger doses are toxic. The consumption of plant secondary compounds can be toxic to herbivores in large doses, but can also improve survival in parasitized herbivores. In addition, recent studies have found that consuming nectar secondary compounds may decrease parasite loads in pollinators. However, the effect of compound dose on bee survival and parasite loads has not been assessed. To determine how secondary compound consumption affects survival and pathogen load in Bombus impatiens, we manipulated the presence of a common gut parasite, Crithidia bombi, and dietary concentration of Anabasine, a nectar alkaloid produced by Nicotiana spp. using four concentrations naturally observed in floral nectar. We hypothesized that increased consumption of secondary compounds at concentrations found in nature would decrease survival of uninfected bees, but improve survival and ameliorate parasite loads in infected bees. We found medicinal effects of Anabasine in infected bees; the high-Anabasine diet decreased parasite loads and increased the probability of clearing the infection entirely. However, survival time was not affected by any level of Anabasine concentration, or by interactive effects of Anabasine concentration and infection. Crithidia infection reduced survival time by more than two days, but this effect was not significant. Our results support a medicinal role for Anabasine at the highest concentration; moreover, we found no evidence for a survival-related cost of Anabasine consumption across the concentration range found in nectar. Our results suggest that consuming Anabasine at the higher levels of the natural range could reduce or clear pathogen loads without incurring costs for healthy bees.

  • variable effects of nicotine and Anabasine on parasitized bumble bees
    F1000Research, 2015
    Co-Authors: Lukas P Thorburn, Lynn S. Adler, Rebecca E. Irwin, Evan C Palmeryoung
    Abstract:

    Secondary metabolites in floral nectar have been shown to reduce parasite load in two common bumble bee species. Previous studies on the effects of nectar secondary metabolites on parasitized bees have focused on single compounds in isolation; however, in nature, bees are simultaneously exposed to multiple compounds. We tested for synergistic effects of two alkaloids found in the nectar of Nicotiana spp. plants, nicotine and Anabasine, on parasite load and mortality in bumble bees ( Bombus impatiens ) infected with the intestinal parasite Crithidia bombi . Adult worker bees inoculated with C. bombi were fed nicotine and Anabasine diet treatments in a factorial design, resulting in four nectar treatment combinations:  2 ppm nicotine, 5 ppm Anabasine, 2ppm nicotine and 5 ppm Anabasine together, or a control alkaloid-free solution. We conducted the experiment twice: first, with bees incubated under variable environmental conditions (‘Variable’; temperatures varied from 10-35°C); and second, under carefully controlled environmental conditions (‘Controlled’; 27°C incubator, constant darkness). In ‘Variable’, each alkaloid alone significantly decreased parasite loads, but this effect was not realized with the alkaloids in combination, suggesting an antagonistic interaction. Nicotine but not Anabasine significantly increased mortality, and the two compounds had no interactive effects on mortality. In ‘Controlled’, nicotine significantly increased parasite loads, the opposite of its effect in ‘Variable’. While not significant, the relationship between Anabasine and parasite loads was also positive. Interactive effects between the two alkaloids on parasite load were non-significant, but the pattern of antagonistic interaction was similar to that in the variable experiment. Neither alkaloid, nor their interaction, significantly affected mortality under controlled conditions. Our results do not indicate synergy between Nicotiana nectar alkaloids; however, they do suggest a complex interaction between secondary metabolites, parasites, and environmental variables, in which secondary metabolites can be either toxic or medicinal depending on context.

  • secondary metabolites in floral nectar reduce parasite infections in bumblebees
    Proceedings of The Royal Society B: Biological Sciences, 2015
    Co-Authors: Leif Richardson, Lynn S. Adler, Anne S Leonard, Jonathan Andicoechea, Karly H Regan, Winston E Anthony, Jessamyn S Manson, Rebecca E. Irwin
    Abstract:

    The synthesis of secondary metabolites is a hallmark of plant defence against herbivores. These compounds may be detrimental to consumers, but can also protect herbivores against parasites. Floral nectar commonly contains secondary metabolites, but little is known about the impacts of nectar chemistry on pollinators, including bees. We hypothesized that nectar secondary metabolites could reduce bee parasite infection. We inoculated individual bumblebees with Crithidia bombi, an intestinal parasite, and tested effects of eight naturally occurring nectar chemicals on parasite population growth. Secondary metabolites strongly reduced parasite load, with significant effects of alkaloids, terpenoids and iridoid glycosides ranging from 61 to 81%. Using microcolonies, we also investigated costs and benefits of consuming Anabasine, the compound with the strongest effect on parasites, in infected and uninfected bees. Anabasine increased time to egg laying, and Crithidia reduced bee survival. However, Anabasine consumption did not mitigate the negative effects of Crithidia, and Crithidia infection did not alter Anabasine consumption. Our novel results highlight that although secondary metabolites may not rescue survival in infected bees, they may play a vital role in mediating Crithidia transmission within and between colonies by reducing Crithidia infection intensities.

Linda B. Von Weymarn - One of the best experts on this subject based on the ideXlab platform.

  • Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine by High Throughput Liquid Chromatography-Mass Spectrometry.
    Chemical research in toxicology, 2016
    Co-Authors: Linda B. Von Weymarn, Dorothy K. Hatsukami, Nicole M. Thomson, Eric C. Donny, Sharon E. Murphy
    Abstract:

    Nicotine is the most abundant alkaloid in tobacco accounting for 95% of the alkaloid content. There are also several minor tobacco alkaloids; among these are nornicotine, anatabine, and Anabasine. We developed and applied a 96 well plate-based capillary LC-tandem mass spectrometry method for the analysis of nornicotine, anatabine, and Anabasine in urine. The method was validated with regard to accuracy and precision. Anabasine was quantifiable to low levels with a limit of quantitation (LOQ) of 0.2 ng/mL even when nicotine, which is isobaric, is present at concentrations >2500-fold higher than Anabasine. This attribute of the method is important since anatabine and Anabasine in urine have been proposed as biomarkers of tobacco use for individuals using nicotine replacement therapies. In the present study, we analyzed the three minor tobacco alkaloids in urine from 827 smokers with a wide range of tobacco exposures. Nornicotine (LOQ 0.6 ng/mL) was detected in all samples, and anatabine (LOQ, 0.15 ng/mL) an...

  • Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers’ Urine by High Throughput Liquid Chromatography–Mass Spectrometry
    2016
    Co-Authors: Linda B. Von Weymarn, Dorothy K. Hatsukami, Nicole M. Thomson, Eric C. Donny, Sharon E. Murphy
    Abstract:

    Nicotine is the most abundant alkaloid in tobacco accounting for 95% of the alkaloid content. There are also several minor tobacco alkaloids; among these are nornicotine, anatabine, and Anabasine. We developed and applied a 96 well plate-based capillary LC-tandem mass spectrometry method for the analysis of nornicotine, anatabine, and Anabasine in urine. The method was validated with regard to accuracy and precision. Anabasine was quantifiable to low levels with a limit of quantitation (LOQ) of 0.2 ng/mL even when nicotine, which is isobaric, is present at concentrations >2500-fold higher than Anabasine. This attribute of the method is important since anatabine and Anabasine in urine have been proposed as biomarkers of tobacco use for individuals using nicotine replacement therapies. In the present study, we analyzed the three minor tobacco alkaloids in urine from 827 smokers with a wide range of tobacco exposures. Nornicotine (LOQ 0.6 ng/mL) was detected in all samples, and anatabine (LOQ, 0.15 ng/mL) and Anabasine were detected in 97.7% of the samples. The median urinary concentrations of nornicotine, anatabine, and Anabasine were 98.9, 4.02, and 5.53 ng/mL. Total nicotine equivalents (TNE) were well correlated with anatabine (r2 = 0.714) and Anabasine (r2 = 0.760). TNE was most highly correlated with nornicotine, which is also a metabolite of nicotine. Urine samples from a subset of subjects (n = 110) were analyzed for the presence of glucuronide conjugates by quantifying any increase in anatabine and Anabasine concentrations after β-glucuronidase treatment. The median ratio of the glucuronidated to free anatabine was 0.74 (range, 0.1 to 10.9), and the median ratio of glucuronidated to free Anabasine was 0.3 (range, 0.1 to 2.9). To our knowledge, this is the largest population of smokers for whom the urinary concentrations of these three tobacco alkaloids has been reported

Benjamin J Tscharke - One of the best experts on this subject based on the ideXlab platform.

  • determination of Anabasine anatabine and nicotine biomarkers in wastewater by enhanced direct injection lc ms ms and evaluation of their in sewer stability
    Science of The Total Environment, 2020
    Co-Authors: Qiuda Zheng, G Eaglesham, Benjamin J Tscharke, Jake W Obrien, Jack Thompson, Katja Shimko, Timothy Reeks, Cobus Gerber, Kevin V Thomas, Phong K Thai
    Abstract:

    Wastewater-based epidemiology (WBE) has been used to estimate tobacco use in the population. However, the increased use of nicotine replacement therapies and e-cigarettes contributes to the load of nicotine metabolites in wastewater, causing over-estimation of tobacco use if nicotine metabolites were used in WBE back-estimation. This study aims to develop a rapid method for determining the tobacco-specific biomarkers, Anabasine and anatabine, in wastewater and to evaluate their in-sewer stability for better estimation of tobacco use by WBE. An enhanced direct injection LC-MS/MS was developed to quantify Anabasine and anatabine as well as nicotine biomarkers (nicotine, cotinine and hydroxycotinine). The method was optimal when wastewater was filtered through 0.2 μm RC syringe filters and a pre-conditioned SPE cartridge (Oasis HLB 1 cc, 30 mg) before 50 μL was injected into the LC-MS/MS system. Limits of quantification varied between 2.7 and 54.9 ng/L with recoveries from 76% to 103% for all five compounds. In sewer reactors, Anabasine and anatabine were less stable than cotinine and hydroxycotinine. They were more stable in the gravity sewer reactor with <20% loss in 12 h than in the rising main sewer reactor with ~30% loss in the same period. We then applied the new method to 42 daily wastewater influent samples collected from an Australian wastewater treatment plant. The five biomarkers were detected in all samples with concentrations ranging from 9.2 to 7430 ng/L. All five compounds were positively correlated with one another. Our results suggested a high throughput analytical method for feasible application in Anabasine and anatabine as biomarkers of tobacco use in routine wastewater monitoring.

  • Estimates of tobacco use by wastewater analysis of Anabasine and anatabine.
    Drug testing and analysis, 2015
    Co-Authors: Benjamin J Tscharke, Jason M. White, Jacobus P. Gerber
    Abstract:

    Wastewater analysis, the chemical analysis of municipal sewage, is fast becoming the technique of choice to monitor changes in community consumption of a range of compounds over time. Currently wastewater analyses which estimate tobacco consumption focus on the major alkaloid nicotine and its urinary metabolite, cotinine. As nicotine is also present in replacement therapies such as nicotine gum and patches, this analysis is not specific and hence does not truly reflect the harmful consumption of tobacco. Two alkaloids – Anabasine and anatabine – which are specific to dried tobacco, were assessed as biomarkers for tobacco consumption in wastewater, together with nicotine and cotinine. Consequently, solid phase extraction (SPE) and liquid chromatography-mass spectrometry (LC-MS) methods for the detection of Anabasine, anatabine, nicotine, and cotinine in municipal wastewater were validated. All compounds were detected in wastewater extracts and found to have satisfactory recovery, accuracy, precision, and stability in wastewater. Daily flow volume and catchment population of the wastewater facility were used to estimate normalized consumption figures of mg/day/1000 people for composite samples collected over one week, in an application of the method. Anabasine and anatabine were found to be suitable wastewater biomarkers of tobacco and can be used to assess tobacco consumption of communities via wastewater analysis. Application of this methodology can be used to collect temporal consumption data which could be used to determine the efficacy of tobacco reduction strategies. Copyright