Production Of Small Molecules Research Articles

Transportome-wide engineering of Saccharomyces cerevisiae

Synthetic biology enables the production of small molecules by recombinant microbes for pharma, food, and materials applications. The secretion of products reduces the cost of separation and purification, but it is challenging to engineer due to the limited understanding of the transporter proteins' functions. Here we describe a method for genome-wide transporter disruption that, in combination with a metabolite biosensor, enables the identification of transporters impacting the production of a given target metabolite in yeast Saccharomyces cerevisiae. We applied the method to study the transport of xenobiotic compounds, cis,cis-muconic acid (CCM), protocatechuic acid (PCA), and betaxanthins. We found 22 transporters that influenced the production of CCM or PCA. The transporter of the 12-spanner drug:H(+) antiporter (DHA1) family Tpo2p was further confirmed to import CCM and PCA in Xenopus expression assays. We also identified three transporter proteins (Qdr1p, Qdr2p, and Apl1p) involved in betaxanthins transport. In summary, the described method enables high-throughput transporter identification for small molecules in cell factories.

Implementation of Quality by Design (QbD) Principles in Regulatory Dossiers of Medicinal Products in the European Union (EU) Between 2014 and 2019

BackgroundQuality by Design (QbD) is a systematic risk-based approach to development, with predefined characteristics and quality risk management throughout the life cycle of a product. International Conference on Harmonization (ICH) guidelines Q8–Q11 give guidance on QbD applications with ICH Q8 (R2)—approved in 2009—describing the principles of QbD in detail. Since its adoption over 10 years ago, more information about QbD usage for the development of medicinal products is expected to be written in regulatory dossiers by companies.MethodsThe present study set out to evaluate the implementation of QbD principles and elements in all EU approved marketing applications (MA) (n = 494), based on information available in the European Public Assessment Reports (EPARs), for a period of six years (2014–2019), starting 5 years after QbD adoption.ResultsOf the 494 MAs, 271 were submitted with a full dossier (article 8(3)). According to EMA (38%), out of the 271 full dossier submissions, only 104 were developed using full QbD. This figure did not increase during this period. Interestingly, between 2014 and 2019, several MAs were not developed via full QbD implementation but used one or more QbD elements during development, including design space. In addition, a higher percentage of small molecule products were developed with QbD as opposed to biotechnology-derived products (78% vs. 22%, respectively).ConclusionOverall, QbD during development of medicinal products is still not commonly described in dossiers. However, more companies started mentioning QbD elements, thus making it a promising step toward QbD as the standard for development in the future.

Research Analysis and Construction of Application of Deep Filtration Technology in Biopharmaceutical Process

This article analyzes the difference between deep filtration technology and surface filtration technology. The author studied the specific application of deep filtration technology in clarification of fermentation broth, pyrogen removal filtration, small molecule purification, product purification and separation, and purification of macromolecules. This article analyzes the common problems during the application of deep filtration technology. The author puts forward suggestions to improve the resource sharing mechanism, standardize the management of filtering research institutions, increase research on new technologies, and do a good job of comprehensive training of personnel. The purpose of this article is to deepen people’s understanding of deep filtration technology and promote the healthy economic development of the biopharmaceutical industry.

Engineered Bacterial Production of Volatile Methyl Salicylate.

The engineering of microbial metabolic pathways over the last two decades has led to numerous examples of cell factories used for the production of small molecules. These molecules have an array of utility in commercial industries and as in situ expressed biomarkers or therapeutics in microbial applications. While most efforts have focused on the production of molecules in the liquid phase, there has been increasing interest in harnessing microbes' inherent ability to generate volatile compounds. Here, we optimized and characterized the production of methyl salicylate, an aromatic compound found mainly in plants, using a common lab strain of E. coli. We utilized genetic components from both microbes and plants to construct the volatile metabolite circuit cassette. In order to maximize production, we explored expression of methyl salicylate precursors, upregulation of expression by increasing ribosomal binding strength and codon optimization of the methyl transferase gene obtained from plant Petunia x hybrida. Last, we validated and quantified the production of methyl salicylate with liquid chromatography or gas chromatography mass spectrometry (LC-MS or GC-MS) and found that the codon optimized strain with precursor supplementation yielded the highest production compared to the other strains. This work characterizes an optimized metabolite producing genetic circuit and sets the stage for creation of an engineered bacteria diagnostic to be used in volatile assays.

Pyrolysis characteristics and products distribution of petroleum sludges

ABSTRACT Pyrolysis can realise the harmlessness, reduction and resource utilisation of petroleum sludge in a short period. In the present work, a tank bottom sludge (SSOS) and a landing sludge (SLOS) from Shengli Oilfield were used for experimental research. Thermogravimetric testing is used to initially determine the optimal range of pyrolysis temperature. Pyrolysis experiments were performed in a tube furnace reactor. Pyrolysis products were collected and analysed separately. The char yield of SSOS and SLOS were 50% and 70%, respectively. Although there are differences in the oil content of the two types of petroleum sludge, the oil yield remained nearly the same, which were both between 7% and 8%. As the pyrolysis temperature was raised to 500°C, the yield of each product did not change greatly while their composition had obvious changes. High temperature is more conducive to the production of small molecule products. Result showed that pyrolysis treatment of petroleum sludge can effectively recover energy materials in the form of pyrolysis gas and oil. The heating value of char is lower than that of petroleum sludge, which means that char is not suitable for direct use as fuel. Pyrolysis treatment also showed good curing effect on Cr, which reached 85%. However, the solidification effect decreased as pyrolysis temperature increasing. It is necessary to pay attention to the heavy metal contained in char as soil improver. The rich surface structure of char provides evidence to produce high value-added carbon materials.

Gut Microbiota in Intestinal and Liver Disease.

It is known that the gut microbiota, the numerically vast and taxonomically diverse microbial communities that thrive in a symbiotic fashion within our alimentary tract, can affect the normal physiology of the gastrointestinal tract and liver. Further, disturbances of the microbiota community structure from both endogenous and exogenous influences as well as the failure of host responsive mechanisms have been implicated in a variety of disease processes. Mechanistically, alterations in intestinal permeability and dysbiosis of the microbiota can result in inflammation, immune activation, and exposure to xenobiotic influences. Additionally, the gut and liver are continually exposed to small molecule products of the microbiota with proinflammatory, gene regulatory, and oxidative properties. Long-term coevolution has led to tolerance and incorporation of these influences into normal physiology and homeostasis; conversely, changes in this equilibrium from either the host or the microbial side can result in a wide variety of immune, inflammatory, metabolic, and neoplastic intestinal and hepatic disorders.

Cambrex will up capacity at Iowa facility

Citing growing demand for small-molecule drug development and production, Cambrex says it will spend $50 million to expand its capacity to make active pharmaceutical ingredients in Charles City, Io...

A Phytochemical Perspective on Plant Defense Against Nematodes.

Given the large yield losses attributed to plant-parasitic nematodes and the limited availability of sustainable control strategies, new plant-parasitic nematode control strategies are urgently needed. To defend themselves against nematode attack, plants possess sophisticated multi-layered immune systems. One element of plant immunity against nematodes is the production of small molecules with anti-nematode activity, either constitutively or after nematode infection. This review provides an overview of such metabolites that have been identified to date and groups them by chemical class (e.g., terpenoids, flavonoids, glucosinolates, etc.). Furthermore, this review discusses strategies that have been used to identify such metabolites and highlights the ways in which studying anti-nematode metabolites might be of use to agriculture and crop protection. Particular attention is given to emerging, high-throughput approaches for the identification of anti-nematode metabolites, in particular the use of untargeted metabolomics techniques based on nuclear magnetic resonance (NMR) and mass spectrometry (MS).

Analysis of Chemical Reaction Process after Pentaerythritol Tetranitrate Hot Spot Ignition.

In this paper, ReaxFF force field combined with molecular dynamics method was used to study the ignition, deflagration, and detonation of pentaerythritol tetranitrate (PETN) induced by hot spots. The hot spot is 5.6% of the total volume. When the hot spot temperature is 1000 K, the deflagration and detonation of PETN cannot be observed in the simulation time of 200 ps. When the hot spot temperature is 2000 K, it corresponds to the heating time of 20 to 50 ps, deflation and detonation were observed. During hot spot ignition, the products of decomposition of the condensed phase PETN are dominated by NO2 and HONO. The energy required for the C–C bond and C–ONO2 bond cleavage in PETN is high, resulting in only a small amount of CH2O and NO3 during the reaction. Small nitrogen-containing molecules (such as NO2, NO3, HONO, HNO3, etc.) mainly exist during thermal equilibrium, while the number of N2 increases sharply during the thermal runaway stage, and a small amount of NH3 and NH2 are also produced. H2O molecules are formed before CO2 and N2 are produced, and the number always dominates. During the thermal runaway, the entire system can maintain a spontaneous reaction, resulting in a sharp rise in temperature of about 2500 K in 20 ps. During this phase, the catalytic effect of H2O accelerates the formation of CO2 and N2 due to the near Chapman–Jouguet point in the crystal. PETN is a weak oxygen balance explosive that results in a small amount of CO and H2 production during the thermal instability phase. When the reaction is balanced, the relative molecular mass is close to or exceeds that of PETN. The product is only less than 1% of the total mass fraction, while the small molecule product is as high as 78%, and some relative molecular masses are [75,225]. The intermediates account for about 21%. Rapid and complex reaction events make it difficult to accurately predict the structure of these intermediates by existing experiments and calculations, which will be the focus of future research.

An Ontology to Describe Small Molecule Pharmaceutical Product Development and Methodology for Optimal Activity Scheduling

This contribution is the first example of a small molecule pharmaceutical product development ontology. This ontology allows the portfolio-wide visualization of the small molecule pharmaceutical development business as a network of decisions and activities. This ontology was built not only to digitalize, rapidly access, and gain insights from prior decision-making but also to deliver input data for dynamic resource allocation via optimal scheduling of research activities subject to resource, cost, and time constraints. This ontology can be understood as a generic wiring diagram for small molecule product development which outlines the predecessors and successors of activities and decisions, along with needed or allowed overlap between activities. Each activity has multiple modes with direct dollars, resource consumption, and time taken to complete each mode. This generic wiring diagram was instanced for 17 portfolio assets by creating 32 specific scenarios. These scenarios were stored in an ontology as instances and served as foundational digitalized data for a host of future applications. These specific scenarios across the multiple assets served as input to the optimal resource allocation model. In summary, this work describing pharmaceutical Chemistry Manufacturing and Controls (CMC) development activities and decisions is a foundational transformative project that enables full digitization of pharmaceutical CMC data, not only for optimal scheduling but also to generate heuristics on selection of research activities given development risks, a topic which has received virtually no mention in literature.

Global Regulators to Activate Silent Biosynthetic Gene Clusters

Genome mining has recently emerged as a powerful strategy to discover novel microbial secondary metabolites. However, more than 50% of biosynthetic gene clusters are not transcribed under standardized laboratory culture condition. Several methods have been applied to activate silent biosynthetic gene clusters in the microbes so far. Among the regulatory systems for production of secondary metabolites, global regulators, which affect transcription of genes through regulatory cascades, typically govern the production of small molecules. In this review, global regulators to affect production of microbial secondary metabolites were discussed.

Role of phosphorus (P) additive in the performance of char-supported nickel (Ni) catalyst on tar reforming

Developing high-efficiency and low-cost tar removal catalysts is immediate and critical for gasification technology. In this work, phosphorus (P) was introduced into char-supported nickel (Ni) catalysts for steam reforming of tar. P and Ni were loaded on the catalyst support precursor, Yining coal oxidized by 20% H2O2, using two kinds of loading sequence, Ni, P co-loading and stepwise loading to obtain (P + Ni)/20%O-C and P-Ni/20%O-C catalysts respectively. Sand and Ni/γ-Al2O3 were selected for blank experiment and comparison. The self-designed dual-stage quartz reactor was used as an evaluation device to test tar steam reforming activity of the catalysts. Preparation conditions for catalyst and reaction technology were also optimized. The effect of P on the structure and acidity of catalysts was investigated by XRD, N2 adsorption, SEM, TEM, XPS, and NH3-TPD. It was found that (P + Ni)/20%O-C prepared by Ni, P co-loading method exhibited superior catalytic performance due to a better dispersion of Ni, more acid sites and higher amount of Ni0. Meanwhile, it is verified that pH 11 and 600 °C is the optimal preparation condition and reaction temperature. Compared to Ni/γ-Al2O3, the (P + Ni)/20% O-C catalyst achieved the highest reforming gas yield (39.3 mmol/g) and lowest tar yield (4.9%), which could be attributed to the function of conversion heavy tar to light fraction of (P + Ni)/20%O-C catalyst. That means most polycyclic materials cracked to small molecule products.

Effect of the epichlorohydrin grafting of starch on its pyrolysis behavior and mechanism

Crosslinking is a common method for preparing biomass-based carbon materials. Corn starch grafted with epichlorohydrin was prepared and then stabilized and carbonized to obtain starch-based near-spherical carbon materials. The pyrolysis of corn starch under the influence of epichlorohydrin was analyzed by TG-MS, and the micro-morphology and crystal structure of the near-spherical carbon materials were characterized by SEM and XRD. The surface chemical evolution of the starch before and after pyrolysis was characterized by in-situ FTIR. The modified starch pyrolysis and the cross-linking reactions between epichlorohydrin and starch were investigated. Results indicate that for the modified starch the temperature at which the weight loss rate is a maximum is lowered so that the pyrolysis is relatively mild, and the destruction of the basic skeleton is reduced and the carbon yield greatly increased compared with pristine starch. The cross-linked starch tends to transform to aromatic structures in pyrolysis, the carbonization degree of the hydrolysate is improved, and the yield of the final product is increased compared to the non-crosslinked material. During pyrolysis, epichlorohydrin cross-links with starch to form a network structure, which stabilizes the starch skeleton, thus reducing the production of volatile small molecules.

Bioinformatic Mapping of Opine-Like Zincophore Biosynthesis in Bacteria.

Zinc is an essential nutrient in biological systems due to its structural or catalytic requirement in proteins involved in diverse cellular processes. To meet this cellular demand, microbes must acquire sufficient zinc from their environment. However, many environments have low zinc availability. One of the mechanisms used by bacteria to acquire zinc is through the production of small molecules known as zincophores. Similar to bacterial siderophores used for iron uptake, zincophores are synthesized by the bacterium and exported and then reimported as zincophore-zinc complexes. Thus far, only four zincophores have been described, including two from the human pathogens Staphylococcus aureus and Pseudomonas aeruginosa, in which they play a critical role in zinc acquisition during infection, and one in a soil bacterium. To determine what other microbes may produce zincophores, we used bioinformatic analyses to identify new zincophore biosynthetic gene clusters (BGCs) and predict the diversity of molecules synthesized. Genome neighborhood network analysis identified approximately 250 unique zincophore-producing species from actinobacteria, firmicutes, proteobacteria, and fusobacteria. This indicates that zincophores are produced by diverse bacteria that inhabit a broad range of ecological niches. Many of the BGCs likely produce characterized zincophores, based on similarity to the characterized systems. However, this analysis also identified numerous BGCs that, based on the colocalization of additional modifying enzymes and sequence divergence of the biosynthetic enzymes, are likely to produce unique zincophores. Collectively, these findings provide a comprehensive understanding of the zincophore biosynthetic landscape that will be invaluable for future research on these important small molecules.IMPORTANCE Bacteria must acquire essential nutrients, including zinc, from their environment. For bacterial pathogens, this necessitates overcoming the host metal-withholding response known as nutritional immunity. A novel type of zinc uptake mechanism that involves the bacterial production of a small zinc-scavenging molecule was recently described in the human pathogens Staphylococcus aureus, Pseudomonas aeruginosa, and Yersinia pestis, as well as the soil-associated bacterium Paenibacillus mucilaginosus This suggests that zincophores may be important for zinc acquisition in diverse environments. In this study, we sought to identify other zincophore-producing bacteria using bioinformatics. We identified almost 250 unique zincophore-producing species, including human and animal pathogens, as well as isolates from soil, rhizosphere, plant, and marine habitats. Crucially, we observed diversity at the amino acid and gene organization levels, suggesting that many of these species are producing unique zincophores. Together, our findings highlight the importance of zincophores for a broad array of bacteria living in diverse environments.

Catalytic oxidation of dimethyl phthalate over titania-supported noble metal catalysts

Semi-volatile organic compounds (SVOCs) are organic compounds with the boiling point ranging between 240/260 ℃ and 380/400 ℃. Detailed knowledge regarding catalytic removal of SVOCs from indoor environment is very limited as it remains challenge to explore such reaction due to the viscosity nature of target contaminants. Here, we established a facile methodology to explore the heterogeneous catalytic oxidation reaction of dimethyl phthalate (DMP), a model SVOC, over the surface of supported catalyst. DMP was found to be gradually oxidized over the surface of titania supported catalysts including palladium (Pd), platinum and ruthenium with increasing temperature. The cleavage of side chain of DMP occurs at 75 ℃ over the surface of Pd/TiO2, which is significantly lower than that of the other two catalysts. Carbon dioxide was observed as the main product of the catalytic oxidation reaction. However, aromatic products and small molecule products were still observed as side-product in different temperature range. Density functional theory calculations further show that DMP can react with reactive oxygen species to form phthalic acid. While the cleavage of the DMP side chain occurs to form products such as methyl benzoate. This work thus provides basic knowledge about indoor SVOCs catalytic oxidation removal.

ReaxFF MD simulations of thermolysis mechanism of 2, 6-diamino-3, 5-dinitropyrazine-1-oxidated

As one of the important high energetic insensitive explosives, LLM-105 has attracted considerably attention. In this work, the thermal decomposition of LLM-105 was analyzed at 2000, 2250, 2500, 2750, 3000, 3250, and 3500 K. Reaction times were controlled to be 200 ps. The results showed that the main small molecule products in this reaction stage were H2O, N2, CO2, and H2. At low temperatures, water molecules form earlier than nitrogen ones, while the opposite is found at high temperatures. In the initial stage of the reaction, LLM-105 often undergoes a large amount of intermolecular H-transfer and dimerization, and this phenomenon is significantly reduced as the temperature rises. On the other hand, LLM-105 itself is very easy to polymerize to form clusters, and its cyclic structure is more likely to form a macromolecular cluster by diverse linkages (e.g. CNNC). As the reaction progresses, the atoms of H, O, and N gradually escape from the cluster structure, but most of the oxygen atoms stay stable. The activation energy of the initial decomposition reaction of LLM-105 is 167.9 kJ/mol, which is in a reasonable range.

Decomposition and Energy-Enhancement Mechanism of the Energetic Binder Glycidyl Azide Polymer at Explosive Detonation Temperatures.

Replacing existing inert binders with energetic ones in composite explosives is a novel way to improve the explosive performance, on the proviso that energetic binders are capable of releasing chemical energy rapidly in the detonation environment. Known to be a promising candidate, the reaction mechanism of glycidyl azide polymer (GAP) at typical detonation temperatures higher than 3000 K has been theoretically studied in this work at the atomistic level. By analyzing and tracking the cleavage of characteristic chemical bonds, it was found that at the detonation temperature, GAP was able to release a large amount of energy and small molecule products at a speed comparable to commonly used explosives in the early reaction stage, which was mainly attributed to the decomposition of azide groups into N2 and the main chain breakage into small fragments. Moreover, N2 generation was found to be accelerated by H atom transfer at an earlier reaction step. The dissociation energy of the main chain was lowered with structure deformation so as to facilitate the fragmentation of the GAP chain. Based on this analytical study of reaction kinetics, GAP was found to have higher reactivity at the detonation temperature than at lower temperatures. The small molecules' yield rate is of the same order of magnitude as an explosive detonation reaction, indicating that GAP has the potential to improve the performance of composite explosives. Our study reveals the chemical decomposition mechanism of a typical energetic binder, which would aid in the future design and synthesis of energetic binders so as to achieve both sensitivity-reducing and energy-enhancing performance goals simultaneously.

Roles of bromine radicals, HOBr and Br2 in the transformation of flumequine by the UV/chlorine process in the presence of bromide

In the presence of bromide (Br−), new reactive species, such as bromine and reactive bromine species (RBS), are formed by the UV/chlorine process. In this study, the effects of Br− on the kinetics, products, and pathways of flumequine (FLU) transformation by the UV/chlorine process were investigated. The pseudo first-order rate constant (k′) for FLU degradation in the UV/chlorine process was enhanced by 2.5 times by 50 μM Br− at pH 7, as HOBr, Br2, RBS and HO• were formed, and they contributed 29.2%, 7.8%, 57.4% and 4.8% to k′, respectively. The second-order rate constants (k) of FLU reacting with HOBr, OBr−, Br2 and Br• were determined to be 115.6 M−1 s−1, negligible, 1.5 × 106 M−1 s−1 and 2.4 × 108 M−1 s−1, respectively. The contribution of Br2 was enhanced at a higher Br− level and a lower pH, which accounted for 48.2% of k′ at pH 6. The overall k′ value decreased by 97.9% from pH 6 to 9, during which k′ by HOBr, Br2 and RBS decreased significantly. The formation of brominated products was mainly attributed to HOBr and Br2, while RBS and HO• primarily promoted hydroxylation and ketonization during UV/chlorine treatment with Br−. The formation of total organic bromine (TOBr) by the UV/chlorine process with Br− was comparable to that during dark chlorination with Br−, while the yield of brominated disinfection byproducts in the former was much higher, further indicating that RBS and HO• promoted the formation of small-molecule products. In addition, the acute toxicity of FLU was significantly reduced during the UV/chlorine process with Br−. This is the first study to reveal the significant roles of Br2 and Br• in the degradation of micropollutants by the UV/chlorine process in Br−-containing water.

Soft Textured Implants: Roughness, Friction, and the Complications

Friction and shear between soft medical implants and the contacting cells and tissues can lead to the production and secretion of pro-inflammatory proteins and small molecules: cytokines, chemokines, and damage-associated molecular patterns (DAMPs). Biotribological considerations should be included in biocompatibility studies and the design of soft medical implants. Clinical experience with textured breast implants has revealed a number of complications linked to chronic inflammation. Long-term exposure to elevated levels of pro-inflammatory cytokines, proteins, and DAMPs may lead to chronic inflammation, which can have numerous negative biological consequences: pain, swelling, seromas, DNA damage, and even cancer. Contact pressure, surface roughness, surface elastic modulus, and friction all contribute to the dynamics and shear stresses reacted across these interfaces during use; in many soft implant applications, controlled mobility (without migration) is desirable, and slip is essentially unavoidable. Depending on the magnitude of shear stress, cells will undergo a wide range of responses from proliferation and remodeling, to cytokine and chemokine production, and even cell death (apoptosis or necrosis). In soft medical implants, friction coefficients and shear stresses increase with increasing roughness. Models of breast implant contact pressure as a function of geometry and relative size reveal that the shear stresses of the aggressively textured implants may exceed 100 Pa, while the smooth implants are likely well below 100 Pa. At shear stresses of around 100 Pa, epithelial cells have been shown to produce pro-inflammatory cytokines and undergo programmed cell death (apoptosis). Increasing roughness and texturing of soft medical implants should be considered a risk factor for implant biocompatibility, particularly for larger, high-projection designs.

Metabolomics Profiling in Response to a Cold Pressor Test

The cold pressor test (CPT) has been widely used to assess both cardiovascular and hemodynamic responses to stress. Despite the broad utilization of the CPT, there have been few if any studies in which metabolomic profiling was used to study the metabolic response to such a stimulus. Metabolomics allows insight into varying chemical processes, displaying detailed profiles of metabolites, small molecule substrates, intermediates and products of metabolism. The purpose of this study was to investigate the influence of sympathetic activation induced by a 1‐minute cold‐water immersion CPT, upon muscle metabolic responses, through metabolomics profiling of muscle interstitial fluid in adults. Five healthy adults (2 males, 3 females; 29.4 ± 9.4 yrs.; 77.6 ± 19.3 kg) were recruited for this pilot study. Three microdialysis probes (CMA 20, CMA/Microdialysis, Stockholm, Sweden) were percutaneously inserted into the vastus lateralis muscle of the dominant leg using sterile technique. Probes were then perfused with sterile saline containing 5 mM ethanol through the inlet tubing at a rate of 2.0 μL/min, and dialysate from the outlet tubing was collected in 150 μL polyethylene collection vials. One hour (3 × 20‐minute collections) of basal dialysates were collected and pooled. Participants then completed, in a seated position, a CPT in which the right hand was immersed up to the wrist in ice water for 1 minute. A 20‐min dialysate sample was collected during and immediately following the CPT. Microdialysis samples were sent to the West Coast Metabolomics Center (UC Davis Genome Center, Davis, CA, USA) for metabolomic profiling. Enzymatic‐fluorometric ethanol assay was also used to measure ethanol outflow: inflow (O:I) ratio to determine microvascular blood flow. The percent change from pre‐CPT to the CPT sample was calculated for every metabolite found. Data were then sorted on a magnitude basis, from smallest to largest change from basal levels. From the 413 metabolites detected, 207 were found to increase and 206 were found to decrease from baseline to post‐CPT. Nine metabolites were found within the 95th to 100th percentile of positive change from baseline to CPT, including: 1‐monostearin (median: 50.4%; range: 10.0–183.1), 3‐aminoisobutyric acid (median: 52.5%; range: 39.2–95.4), palmitoleic acid (median: 92.2%; range: 35.4–204.5); and 6 “unknown metabolites.” Furthermore, 5 “unknown metabolites” were also detected within the 95th to 100th percentile of negative change (decrease from baseline). Ethanol O:I ratio increased baseline to CPT, indicating decreased blood flow (0.483 ± 0.088 to 0.547 ± 0.027, mean ± SEM). These results indicate that the CPT can elicit both vasoconstriction and a notable metabolic response in muscle tissue, identifiable by the use of ethanol analysis and metabolomics of dialysate from microdialysis of skeletal muscle. Of specific interest, 1‐monostearin and palmitoleic acid were found to be in the group of molecules that increased the most in response to CPT. Although speculative, increased adrenergic stimulation resulting from CPT‐induced sympathetic activation may have resulted in liberation of 1‐monostearin and palmitoleic acid from membranes or other locations, or from increases in desaturase activities.