Browsing by Author "Prenni, Jessica, advisor"
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Item Open Access Characterizing smoke taint in hops (Humulus lupulus) and investigating the impact of defoliation stress on phytocannabinoid content in industrial hemp (Cannabis sativa)(Colorado State University. Libraries, 2024) Sandoval, Brandon, author; Prenni, Jessica, advisor; Rhodes, Davina, committee member; Broeckling, Corey, committee memberThe family Cannabaceae contains at least 10 genera, with Cannabis (hemp) and Humulus (hop) being two of the most economically important. Both genera have long been valued by humans for their chemical constituents and are used today for both medicinal and recreational purposes. However, adverse environmental factors may impact the chemical profile of these important crops, leading them away from a true-to-type quality. This thesis will explore the effects of an abiotic stress on the chemical profile of each crop: smoke-taint in hops and defoliative hail damage of hemp. The Pacific Northwest contains 97.5% of U.S. commercial hop acreage and has also seen an increase in the number and severity of wildfire events in recent years. While there is extensive research from the wine industry on the impact of smoke taint in grapes, our knowledge of smoke taint in hops is limited. Here, we aimed to characterize smoke taint in hops using laboratory simulated wildfires with distinct fuel types and non-targeted gas chromatography-mass spectrometry. Our results reveal an overall variation in the chemical profiles between smoked and control hops and across fuel types and the detection of known and novel smoke taint markers including guaiacol, 4-methylguaiacol, and xylopyranose. This research provides evidence to support the use of established smoke taint markers for hop analysis and lays the groundwork for future studies to investigate various fuel types and their impact on hop quality. The United States has seen an abrupt increase in commercial industrial hemp production since the Agricultural Improvement Act of 2018. However, the historical prohibition of this crop has resulted in a lack of basic physiological research to guide management practices. For example, abiotic stress can stimulate plants to increase production of secondary metabolites such as phytocannabinoids and this is of high importance to farmers as they as they must balance optimization of CBD yield (crop value) with regulatory requirements (THC < 0.3% by mass) that could lead to crop loss (mandated destruction). In this study we evaluated the impact of defoliation stress (to simulate hail damage) at three different growth stages. Our results indicate that defoliation stress during late flowering yielded no significant change in phytocannabinoid production. However, defoliation stress during vegetative and early flowering yielded a significant increase in phytocannabinoids, including total CBD and THC, at harvest.Item Open Access Characterizing the impact of package type on different beer styles using advanced analytical tools(Colorado State University. Libraries, 2022) Fromuth, Kathryn Lenore, author; Prenni, Jessica, advisor; Sedin, Dana, committee member; Van Buiten, Charlene, committee memberIn 2020 there was over 9,000 breweries in the US, increasing the beer market competition and driving the importance of product stability under variable storage conditions. More breweries, specifically craft breweries, than ever before are choosing to package in cans due to ongoing effects of the current pandemic, the growing availability of smaller can line systems, and increased mobile canning options. Foundational beer stability research has focused on light lager styles packaged in bottles. Limited research has been conducted studying flavor stability in styles relevant to the American craft brewing industry, nor any comparisons of how package type (i.e., cans and bottles) affects flavor stability. Industry utilizes trained sensory panels to evaluate flavor stability; a resource that is both time consuming and expensive. Thus, this is a tool that is often inaccessible or inadequate for providing relevant and timely stability data. This research project, a collaboration between New Belgium Brewing and Colorado State University, aims to address the package-type knowledge gap and sensory panel restrictions by utilizing advanced analytical tools to characterize the changes in metabolite profiles over time between cans and bottles. A low-hopped amber ale (AA) and high-hopped India Pale Ale (IPA) were chosen for their distinct style and relevance to the American craft brewing industry. One batch of an IPA and AA was packaged into cans and bottles, then aged for a six-month period. The samples were stored under cold temperatures (4°C) for the first 30 days, and then at room temperature (20°C) for the subsequent time. Aliquots were collected biweekly for a total of 13 timepoints throughout the six-month aging period and stored at -80°C until chemical analysis. Chemical analysis was conducted by gas chromatography coupled to a mass spectrometer detector (GC-MS) and direct analysis in real time mass spectrometry (DART-MS) to address the research questions. Multivariate (MVA) and univariate (UVA) statistical analysis of the GC-MS data allowed for the characterization of the impacts of package container on the chemical profiles of AA and IPA over time. MVA of the DART-MS data explored the predictive power of the tool for streamlining beer flavor stability analysis. Partial least squares discriminant (PLS-DA) and Multiple Analysis of the Variance (MANOVA) statistical analyses were used to explore data produced by GC-MS and helped define a group of 17 detected metabolites important to explaining the data variation. PLS-DA models of AA samples demonstrated good model fit and package type predictability (R2 = 0.981, Q2 =0.964). This was not observed in IPA which indicates package effects are styles dependent. Differences in AA samples are due, in part, to can and bottle baseline differences in the detected amino acid and ester metabolites. Differences in the physical packaging process of cans, oxidations, and low hop polyphenol concentrations are proposed mechanisms for explaining the observed baseline differences. Analysis of variance (ANOVA) found ten metabolites in AA cans significantly (P ≥ 0.05) changing over time as compared to four metabolites in AA bottles. This indicates higher instability in cans for AA samples. Four detected hop volatiles (humulene, β-myrcene, α-calacorene, pinocarvone), identified by estimated marginal mean of linear models (95% confidence interval) had exhibited significant changes over time that were dependent on package type interactions, but to varying magnitudes and directions depending on the metabolite's polarity and susceptibility to packaging material interactions (e.g., scalping). PLS-DA models of data produced by DART-MS indicated a poor model fit and lack of beer storage time predictability in AA samples (R2 = 0.554, Q2 = -0.151) and IPA samples (R2 = 0.622, Q2 = -0.079). These results lack the evidence that DART-MS is a useful tool for streamlining beer stability analysis. However, results for package type predictability matched GC-MS analysis conclusions in that package type predictability is style dependent. The overall study results demonstrate there is much nuance in the effects of package type on beer flavor stability, and those effects depend on style, packaging material, and the individual metabolite. Targeted analysis is needed to fully understand the mechanisms driving the effects of package type on beer stability.Item Open Access Investigating the relationship between cover crop species diversity, composition and function of the soil microbiome(Colorado State University. Libraries, 2023) Seitz, Valerie, author; Prenni, Jessica, advisor; Wrighton, Kelly, committee member; Schipanski, Meagan, committee member; Nishimura, Marc, committee memberCropping diversification, such as cover cropping, can contribute to sustainable agriculture by enhancing soil health and promoting ecosystem services through interactions with the soil microbial community. One important mechanism through which cover crops impact soil health is via root exudation, the release of organic compounds from plant roots into the soil region surrounding the roots, the rhizosphere. Root exudation varies among cover crop species, growth stages, and edaphic and environmental conditions resulting in a myriad of effects on the rhizosphere. Plant-derived inputs, like root exudates, modulate the soil microbial community, influencing microbial biomass, community structure, and catalyzing biogeochemistry. As a result, cover crops are linked to microbial changes that impact soil nutrient cycling and organic matter decomposition leading to a legacy impact on primary crop yield and health. Understanding the intricate relationship between cover crop root exudation composition and the soil microbiome is crucial for optimizing cover crop selection, management practices, and harnessing cover crops for precision microbiome management in agroecosystems. My dissertation demonstrates that cover crop root exudation differs considerably across cover crop species, and cultivars within species, and reveals cover crop metabolic impacts on soil microbial composition and function, which play a large role in the generation and maintenance of healthy soils to support our agricultural needs.Item Open Access Metabolite fingerprinting of hops (Humulus lupulus) to track chemical variations(Colorado State University. Libraries, 2022) Nasiatka, Katie, author; Prenni, Jessica, advisor; Rhodes, Davina, committee member; Van Buiten, Charlene, committee memberIn the brewing industry, identification of quality crops that provide unique organoleptic properties to beer flavor (aroma, taste) are of critical importance. Hops represent a key ingredient in beer and are utilized to impart specific flavors. India Pale Ales (IPAs) are a popular style of "hoppy beers" in the U.S. and customer expectations for consistency, quality, and unique organoleptic properties of hops are growing. While the contribution of chemical compounds in hops (Humulus lupulus) such as alpha-acids (e.g. humulone) is well-understood, the influence of the hop metabolome (e.g. composition of hop chemical compounds) is still in the early stages of discovery. There is a gap in the knowledge regarding our understanding of chemistry variations in hops among cultivars and growing locations that impact the sensory quality. Traditional sensory evaluation relies on the ability to organize a group of unbiased and trained panelists, who are also subject to sensory fatigue, which can add to the challenge of this method. An alternative approach, ambient mass spectrometry (AMS) is an objective, intuitive, analytical tool capable of rapid chemical fingerprinting. The overall goal of this research is to develop a robust, high-throughput assay using AMS technology to evaluate hop quality that is reflective of both cultivar and environmental variations impacting sensory. To address this goal, twelve hop samples were sourced from three different suppliers across four different farms located in Washington and Oregon over two growing seasons. The samples included three commercial cultivars, Cascade, Centennial, and Strata. The hop samples were extracted using an 80% ethanol solution and fingerprints were acquired by Direct Analysis in Real Time Mass Spectrometry (DART-MS). The resulting data were used to train predictive models and validation was performed to evaluate classification accuracy. Additionally, authentic standards of important hop compounds (hop alpha-acids, terpenes) were used to putatively annotate DART-MS signals reflective of sensory attributes. This study demonstrates the potential of this approach for rapid evaluation of hops quality and lays groundwork for further method optimization. Ultimately, implementation of this tool could have applications for quality assurance programs and for phenotyping of hops for producers and craft brewers.Item Open Access Metabolomic profiles associated with physiological resistance to Sclerotinia sclerotiorum (Lib.) de Bary in common bean(Colorado State University. Libraries, 2015) Robison, Faith M., author; Brick, Mark, advisor; Prenni, Jessica, advisor; Schwartz, Howard, committee member; Byrne, Pat, committee member; Heuberger, Adam, committee memberCommon bean (Phaseolus vulgaris L.) is an important global food crop with a recently sequenced and annotated genome. Plant metabolic and hormone processes are being increasingly recognized as central to disease resistance. For common bean, the molecular and metabolic processes that mediate resistance to white mold disease (caused by Sclerotinia sclerotiorum, (Lib.) de Bary) are largely unknown. Identifying metabolites associated with Sclerotinia infection may provide novel targets to breed for enhanced resistance. The metabolic changes that occur during S. sclerotiorum infection of a detached leaf were characterized using a non-targeted metabolomics workflow spanning primary and secondary metabolism, and a targeted panel of 13 hormones. Partial resistant (A195, beige seed coat color) and susceptible (Sacramento, light red kidney market class) Andean bean lines were inoculated with isolate S20 for non-targeted metabolite profiling at 16, 24, and 48 hours post inoculation (hpi) and at 8 and 16 hpi for hormones. Metabolites from healthy tissue adjacent to the necrotic lesion were extracted with the solvent methanol:water (80:20) and detected using non-targeted UPLC-TOF-MS and GC-MS workflows, and hormones were profiled using UPLC-MS/MS. The analysis detected 140 metabolites that varied between A195 and Sacramento, with the greatest metabolite variation occurring at 16 hpi. The metabolites that varied included amines, amino acids, saccharides, organic acids, phytoalexins, hormones, ureides, and molecules involved in cell wall and membrane composition. The diversity in observed metabolic changes points towards a multi-faceted response associated with plant resistance to S. sclerotiorum in common bean. The integration of metabolomics and genomic data discover functional markers of metabolic resistance to white mold.Item Open Access Peeling back the potato: at the intersection of food insecurity and cardiometabolic disorders(Colorado State University. Libraries, 2022) Bagheri Toulabi, Sahar, author; Prenni, Jessica, advisor; Stromberger, Mary, committee member; Gentile, Christophe, committee member; Holm, David G., committee memberAccording to the Centers for Disease Control (CDC), cardiometabolic diseases (CMD) such as cardiovascular disease and diabetes are the leading cause of death in the United States, which in turn, are strongly linked to obesity. Diet composition is known to be a primary contributing factor in the development of obesity, in the end promoting systemic chronic inflammation and CMD. Unfortunately, providing nutritious and affordable foods to Americans is still a challenge. This highlights the urgent need for additional research in both agriculture and nutrition, to provide sustainable, affordable, and high-quality foods which meet basic caloric and micronutrient needs. In recent years, much emphasis has been placed on phytochemicals due to their promising role in alleviating the pathophysiology of CMD. Phenolics have been proven to have direct health impacts such as increasing the antioxidant capacity of serum, reducing inflammatory biomarkers, ACE inhibitory, and improving arterial health. Our research focused on the potato as it is a superior food choice for both health and nutrition per dollar. The explorations in this dissertation were conducted through multidisciplinary groups to evaluate the different aspects of potatoes, potential health benefits, and barriers to entry. Purple flesh genotypes showed the most antioxidant (AOX) activity, and the highest ACE inhibitory potential in-vitro. Importantly the observed variation within other market classes shall provide a great opportunity to improve these cultivars for the different sections of industry, beyond the fresh market. Beyond the bioactivity of the phytochemicals in-vitro, we examined the effect of a whole potato (a high phenolics cultivar) diet on obesity and the subsequence pathophysiology. We observed that the high phenolic potato diet increased satiety in obese, leptin-deficient (ob/ob), mice. The obese mice on the purple potato diet also lost weight when compared to the obese with the control diet. The same trend was observed in the lean animal model (+/ob) who had the purple potato diet for 10 weeks. As a consequence of eating less, adiposity is reduced in both obese and lean animals. These observations were corroborated by a non-targeted metabolomics study of serum and liver in obese and lean animals, who had potato diet or control. Clear segregation was observed between the metabolites fingerprints between potato diet group compared to control diet. Reduction in serum cholesterol and liver triacylglycerol of the obese animal who had the potato diet was very promising. In our last research, we sought the potato growers' decision-making process to adopt new cultivars or not. Each year, growers must decide in which cultivar to grow, and therefore this decision has an impact on the availability of a cultivar to consumers. This decision can be considered vital to public health as these cultivars are demonstrated to vary in traits important to human health. Adopting a new approach (either using new technology or adopting a new crop) brings risk to the system and is therefore associated with complex psychological and economic factors. We develop a multi-factorial model to explain adoption in a potato-growing system. Growers that were more aware of specialty cultivar innovation and associated consumer demand were more open to SCs adoption. Other influencing factors include a grower's experience selling a specialty cultivar in the previous year and access to diverse markets. Our model demonstrates that the current barriers to adoption are access to primary buyers such as warehouses, retailers, and households. Taken together, this research demonstrates how rational expectations stem from economic outcomes, knowledge, and experience in the potato industry. These results are important in helping to consider opportunities for growers to access new, higher-value markets, which may also improve consumer access to nutritious cultivars.Item Open Access Proteomic profiling of the rat renal proximal convoluted tubule in response to chronic metabolic acidosis(Colorado State University. Libraries, 2013) Freund, Dana Marie, author; Curthoys, Norman, advisor; Prenni, Jessica, advisor; Nyborg, Jennifer, committee member; Peersen, Olve, committee member; Dobos, Karen, committee memberThe human kidneys contain more than one million glomeruli which filter nearly 200 liters of plasma per day. The proximal tubule is the segment of the nephron that immediately follows the glomeruli. This portion of the nephron contributes to fluid, electrolyte and nutrient homeostasis by reabsorbing 60-70% of the filtered water and NaCl and an even greater proportion of NaHCO3. The initial or convoluted portion of the proximal tubule reabsorbs nearly all of the nutrients in the glomerular filtrate and is the site of active secretion and many of the metabolic functions of the kidney. For example, the proximal convoluted tubule is the primary site of renal ammoniagenesis and gluconeogenesis, processes that are significantly activated during metabolic acidosis. Metabolic acidosis is a common clinical condition that is characterized by a decrease in blood pH and bicarbonate concentration. Metabolic acidosis also occurs frequently as a secondary complication, which adversely affects the outcome of patients with various life-threatening conditions. This type of acidosis can occur acutely, lasting for a few hours to a day, or as a chronic condition where acid-base balance is not fully restored. Chronic metabolic acidosis, where the decrease in blood pH and bicarbonate last for 7 days, was the main focus of these studies. Acid-base homeostasis is achieved, in part, by the reabsorption of bicarbonate and excretion of ammonium ions and acids by the proximal convoluted tubule. Metabolic acidosis is partially compensated by an adaptive increase in renal ammoniagenesis and bicarbonate synthesis. During acidosis, there is increased extraction and mitochondrial catabolism of plasma glutamine within the renal proximal convoluted tubule. This process generates ammonium and bicarbonate ions that facilitate the excretion of acid and partially restore acid-base balance. This response is mediated by a pronounced remodeling of the proteome of the proximal convoluted tubule that also produces an extensive hypertrophy. Previous studies identified only a few mitochondrial proteins, including two key enzymes of glutamine metabolism, which are increased during chronic acidosis. Here, a workflow was developed to globally characterize the mitochondrial proteome of the proximal convoluted tubule. Two-dimensional liquid chromatography coupled with mass spectrometry (2D/LC-MS/MS) was utilized to compare mitochondrial enriched samples from control and chronic acidotic rats. Label-free quantitative strategies are commonly used in shot-gun proteomics to detect differences in protein abundance between biological sample groups. In this study we employed a combination of two such approaches, spectral counting (SpC) and average MS/MS total ion current (MS2 TIC). In total, forty nine proteins were observed to be significantly altered in response to metabolic acidosis (p-value < 0.05). Of these, 32 proteins were uniquely observed as significantly different by SpC, 14 by MS2 TIC, and only 3 by both approaches. Western blot analysis was used to validate the fold changes of eight of the proteins that showed an increase upon acidosis. Furthermore, using an antibody specific to acetylated lysine modifications indicated that chronic acidosis causes a 2.5 fold increase in this modification specifically in mitochondria. Western blot analysis established that the observed alterations in both protein abundance and lysine acetylation are not due to the associated hypertrophy. This study represents the first comprehensive analysis of whole mitochondrial proteome of the rat renal proximal convoluted tubule and its response to metabolic acidosis. Additionally, our analysis demonstrates an innovative dual approach for protein quantitation. To further our understanding of the impact of acidosis on the mitochondrial proteome, mitochondrial inner membranes were isolated from control and acidotic rat proximal convoluted tubules. Additional LC-MS/MS analysis was performed, representing the first proteomic characterization of the mitochondrial inner membrane proteome of the rat renal proximal convoluted tubule. Specific sites of lysine acetylation were identified both in the inner membrane and whole mitochondria, the majority of which are novel sites. The results presented here showed successful enrichment of mitochondrial inner membranes and described the proteins and the known biological processes of this compartment of the mitochondria. Previous proteomic analysis was performed on brush-border membrane vesicles isolated from proximal convoluted tubules from control, 1 d and 7 d acidotic rats. To validate the observed protein alterations, western blot analysis was performed on freshly isolated apical membrane. Additionally, the results from three independent proteomic studies focused on the apical membrane, mitochondrial, and soluble cytosolic fractions of the proximal convoluted tubules were compiled. Bioinformatics analysis was performed to describe predominate cellular processes and pathways that respond to chronic metabolic acidosis. The results of these studies demonstrate that the physiological response to the onset of metabolic acidosis requires pronounced changes in the renal proteome. The observed proteomic adaptations within the proximal convoluted tubule support the increased extraction of plasma glutamine and the increased synthesis and transport of glucose and of NH4+ and HCO3- ions. Overall, this dissertation describes the profiling of the proximal convoluted tubule proteome in response to chronic metabolic acidosis and provides the framework for future studies.Item Embargo Survival and persistence of Salmonella enterica in dry bulb onion production practices; a risk assessment approach(Colorado State University. Libraries, 2023) Carpenter, Griffin, author; Gutierrez-Rodriguez, Eduardo, advisor; Prenni, Jessica, advisor; Bunning, Marisa, committee member; Uchanski, Mark, committee memberThe first chapter of this research thesis focuses on onion production practices and the potential risks associated with contamination of dry bulb (DB) onions along the cropping cycle with human enteric pathogens. Dry bulb onions are a widely consumed vegetable globally that has been previously thought to be safe from human pathogen contamination. This literature review summarizes the history of outbreak-related information, the biology of etiologies of interest, the body of literature associated with microbial risks and onions, and pre-and post-harvest production practices with respect to the risk of contamination with human pathogens. The information discussed in this review is useful for portraying the complex interactions of the microorganisms of interest and for industry professionals, producers, and consumers with respect to management and applicable risk mitigation efforts in the future for DB onions. The second chapter of this research thesis focuses on determining the microbial risk factors associated with pre-and post-harvest commercial DB onion production practices. In the past 30 years DB onions have not been involved in foodborne illness outbreaks in the United States. However, two major multi-state foodborne outbreaks linked to Salmonella spp. (Sal) in 2020 and 2021 have altered the perception of producers and consumers about the microbial safety of this crop. Despite significant efforts to identify the source and route of contamination, little knowledge exists regarding the risk factors associated with enteric pathogen contamination along the DB onion cropping cycle. Thus, the goal of this research was to develop risk assessment profiles of DB onion production practices capable of identifying and reducing Sal contamination in pre-and post-harvest activities. DB onion cultivars grown in the state of Colorado were used to determine the ability of these onions to potentially inhibit Sal. This was achieved by testing the minimum inhibitory concentration (MIC) with a cocktail of attenuated and pathogenic Sal at 3,600 mg/L of onion slurry. All evaluations indicated that there was no significant inhibition of Sal irrespective of the type of strain or DB onion cultivar. Pre- and post-harvest risk quantification was determined based on field inoculations of a 2-strain attenuated Salmonella (attSal) cocktail. Survival and persistence of attSal was assessed at multiple production stages including at the 3 leaf stage, lifting, topping, curing, harvest, transport, and packing house environments. From these evaluations, results indicate that attSal is capable of surviving in both DB onions and soil for over a period of 64 days across the entire cropping cycle including harvest. Cultivar, agronomic practice, and UV index had no significant impact in our ability to recover attSal and in the survival of these strains along the cropping cycle in soil or DB onion. At harvest, the population of attSal on DB onion was 3.4 MPN/g at the three-leaf stage development (3LS), log 2.07 cfu/g at topping and log 1.87 cfu/g at lifting irrespective of DB onion cultivar. During interstate transport, the population of attSal further decreased to undetectable levels (< 3.0 MPN/g of DB onion). This scenario was considered a low-risk event for packinghouse purposes. Commercially grown DB onions were also included in all packinghouse evaluations. These onions were free from naturally occurring Sal and were inoculated with chalk containing attSal to mimic soil dust contamination. This chalk had an initial attSal population of log 5.5 cfu/g DB onion and for packinghouse purposes, it was considered a high-risk contamination event. A total of 14 locations within the packing line were selected to test the potential transfer of attSal from inoculated DB onions to control treatments and food contact surfaces. Additionally, DB onions from both high and low-risk contamination events were collected during sorting and packing. In both high and low -risk packing line contamination events, attSal was not recovered from any food contact surface or DB onions (Totl N= 897) over the course of 4 days of processing. A dry sanitation event was implemented in the packing line to assess whether such approaches could reduce contamination from attSal or any other residues left by the crop or by previous activities at the packing line. Our dry sanitation cleaning protocol involved the cleaning of the crop contact surface with a dry brush-single use paper towel, followed by sanitation with a food grade alcohol wipe, followed by a spray of an ethanol alcohol solution (food grade) at 75%, followed by a final removal of alcohol residues with a dry single-use paper towel. This approach proved to be effective in reducing packing line residues Adenosine triphosphate (ATP) measurements and the population of two indicator organisms Enterobacteriaceae (EB) and Escherichia coli (EC). However, the effectiveness varied with the type of surface. Plastic and camel hair bristles were not cleanable. There was no correlation between the population of EB and EC and the presence of attSal from high and low-risk contamination events. Dry sanitation events clearly indicated that it is a viable and useful practice that could be implemented on DB onion packing lines. The absence of cleaning and sanitation will be conducive to significant accumulation of DB onion debris and for the potential proliferation of indicator and pathogenic organisms. These findings are important to industry professionals, producers, and consumers regarding developing risk profiles and application of risk mitigation strategies to improve the microbial safety of DB onions.Item Open Access Understanding the phytochemistry of high-CBD hemp: efficacy of common row cover materials for pollen exclusion and impact on flower phytochemistry(Colorado State University. Libraries, 2022) Bowen, Janina K., author; Prenni, Jessica, advisor; Cranshaw, Whitney, committee member; Uchanski, Mark, committee memberProduction of high-cannabidiol (high-CBD) hemp (Cannabis sativa L.) is steadily increasing in Colorado and across the United States. However, the impact of management practices on flower phytochemistry in this crop remains relatively unexplored. For example, there is high potential for male hemp plants from fiber and grain cultivars to pollinate exclusively female high-CBD hemp plants grown in close proximity, but it is unknown how the cannabinoid content of high-CBD hemp flowers is affected by pollination. We hypothesized that high seed content resulting from pollination will negatively impact the phytochemical yield of high-CBD crops. In this study, three experimental pollen exclusion treatments were applied to two cultivars of high-CBD hemp, 'Cherry Uno' and 'Wife.' Treatments included non-woven thick row cover (largest pore size of approximately 50 microns), non-woven thin row cover (largest pore size approximately 200 microns), woven insect netting (average pore size 700x240 microns), and uncovered controls. A total of 60 high-CBD plants (clones) were planted in a randomized complete block design at the Colorado State University Agricultural Research, Development and Education Center South (ARDEC South) in Fort Collins, Colorado (lat. 40.611804 N; long. -104.997144 W; elevation 1525 meters). Total biomass and seed weights for 60 whole plants were evaluated. Additionally, 5 cm inflorescence samples were taken from each plant, in concordance with the 2019 Colorado Department of Agriculture (CDA) sampling protocol. Seeds and floral material were weighed separately before samples were homogenized in preparation for cannabinoid analysis. Extracts were analyzed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) to determine the quantitative profiles of 20 cannabinoids. Results indicate that for the cultivar Cherry Uno, thick and thin row cover treatments effectively reduced pollination as compared to uncovered controls. The row cover treatments did not result in a statistically significant reduction of pollination for cultivar Wife, which may be due to later flowering in this cultivar. For cultivar Cherry Uno, a significant reduction in CBD concentration of up to 2.7% was observed in the heavily seeded controls compared to covered plants (Uncovered control = 3.77% CBD, 0.13% Δ9THC; Thin row cover = 6.49% CBD, 0.21% Δ9THC). Taken together, our results suggest that implementation of strategies to minimize pollination and/or remove seeds from high-CBD hemp biomass could improve cannabinoid yield. More research is warranted to evaluate the economic viability of such strategies and the effectiveness across different cultivars and growing climates.