A positive affection of human health by nutrition is of high interest, especially for bioactive compounds which are consumed daily in high amounts. This is the case for chlorogenic acids (CGA) ingested by coffee. This molecule class is associated with several possible beneficial health effects observed in vitro that strongly depend on their bioavailability. So far factors influencing bioavailability of CGA such as dose, molecule structure and site of absorption haven´t been investigated sufficiently.
Therefore we performed an in vivo dose-response study with ileostomists, who consumed three different nutritional doses of CGA ingested as instant coffee (4,525 (HIGH); 2,219 (MEDIUM); 1,053 (LOW) μmol CGA). CGA concentrations were determined in ileal fluid, urine and plasma. Furthermore, we conducted an ex vivo study with pig jejunal mucosa using the Ussing chamber model to confirm the in vivo observations. Individual transfer rates of CGA from coffee were investigated, namely: caffeoylquinic acid (CQA), feruloylquinic acid (FQA), caffeic acid (CA), dicaffeoylquinic acid (diCQA) and QA at physiological concentrations (0.2–3.5 mM). Samples were analyzed by HPLC-DAD, -ESI-MS and -ESI-MS/MS.
About ⅔ of the ingested CGA by coffee consumption were available in the colon dose independent. Nevertheless, the results showed that the consumption of higher CGA doses leads to a faster ileal excretion. This corresponds to a plasma AUC0-8h for CGA and metabolites of 4,412 ± 751 nM*h0-8-1 (HIGH), 2,394 ± 637 nM*h0-8-1 (MEDIUM) and 1,782 ± 731 nM*h0-8-1 (LOW) respectively, and a renal excretion of 8.0 ± 4.9% (HIGH), 12.1 ± 6.7% (MEDIUM) and 14.6 ± 6.8% (LOW). Moreover interindividual differences in gastrointestinal transit times were related to differences in total CGA absorption. Thus the variety of patient´s physiology is a decisive bioavailability factor for CGA uptake. This is corroborated ex vivo by a direct proportional relationship of incubation time with absorbed CGA amount.
The consumption of high CGA doses influences the metabolism pattern as an increasing glucuronidation was observed with consumption of increasing CGA doses. However, the different CGA doses have only minor effects on the overall bioavailability which was confirmed ex vivo by a non-saturable passive diffusion of 5-CQA. Furthermore, we identified in the Ussing chamber an active efflux secretion for 5-CQA that decreases its bioavailability and the physicochemical properties of the CGA subgroups as an important bioavailability factor. Transferred amount in increasing order: diCQA, trace amounts; CQA ≈ 1%; CA ≈ 1.5%; FQA ≈ 2%; and QA ≈ 4%.
Altogether, the consumption of increasing CGA doses by coffee had a minor effect on oral bioavailability in ileostomists, such as a slightly increased glucuronidation. Thus, the consumption of high amounts of CGA from coffee in the daily diet is not limiting the CGA concentrations at the site of possible health effects in the human body. However, according to the patient´s physiology the interindividual gastrointestinal transit time which is possibly influenced by dose is influencing CGA bioavailability. Moreover, ex vivo CGA absorption is governed by diffusion as an absorption mechanism corroborating an unsaturable uptake in vivo and by the individual physicochemical properties of CGA.
C-H activations(C-H bond weakening effects) under impact of transition metal atoms
are theoretically investigated,
two model systems are used, one is CH3MX, the other is
n-ButMX, (X=F,Cl,Br,I,H,CN, M include all transition metal
atoms from group 4 to group 10).
C-H activations(C-H bond weakening effects) under impact of transition metal atoms are theoretically investigated, two model systems are used, one is CH3MX, the other is n-ButMX, (X=F,Cl,Br,I,H,CN, M include all transition metal atoms from group 4 to group 10).
Photochemical reactions are of great interest due to their importance in chemical and biological processes. Highly sensitive IR/UV double and triple resonance spectroscopy in molecular beam experiments in combination with ab initio and DFT calculations yields information on reaction coordinates and Intersystem Crossing (ISC) processes subsequent to photoexcitation. In general, molecular beam experiments enable the investigation of isolated, cold molecules without any influence of the environment. Furthermore, small aggregates can be analyzed in a supersonic jet by gradually adding solvent molecules like water. Conclusions concerning the interactions in solution can be derived by investigating and fully understanding small systems with a defined amount of solvent molecules. In this work the first applications of combined IR/UV spectroscopy on reactive isolated molecules and triplet states in molecular beams without using any messenger molecules are presented. Special focus was on excited state proton transfer reactions, which can also be described as keto enol tautomerisms. Various molecules such as 3-hydroxyflavone, 2-(2-naphthyl)-3-hydroxychromone and 2,5-dihydroxybenzoic acid have been investigated with regard to this question. In the case of 3-hydroxyflavone and 2-(2-naphthyl)-3-hydroxychromone, the IR spectra have been recorded subsequent to an excited state proton transfer. Furthermore the dihydrate of 3-hydroxyflavone has been analyzed concerning a possible proton transfer in the excited state: The proton transfer reaction along the water molecules (proton wire) has to be induced by raising the excitation energy. However, photoinduced reactions involve not only singlet but also triplet states. As an archetype molecule xanthone has been analysed. After excitation to the S2 state, ISC occurs into the triplet manifold leading to a population of the T1 state. The IR spectrum of the T1 state has been recorded for the first time using the UV/IR/UV technique without using any messenger molecules. Altogether it is shown that IR/UV double and triple resonance techniques are suitable tools to analyze reaction coordinates of photochemical processes.
In this thesis, collision-induced dissociation (CID) studies serve to elucidate relative stabilities and to determine bond strengths within a given structure type of transition metal complexes. The infrared multi photon dissociation (IRMPD) spectroscopy combined with density functional theory (DFT) allow for structural analysis and provide insights into the coordination sphere of transition metal centers. The used combination of CID and IRMPD experiments is a powerful tool to obtain a detailed and comprehensive characterization and understanding of interactions between transition metals and organic ligands. The compounds’ spectrum comprises mono- or oligonuclear transition metal complexes containing iron, palladium, and ruthenium as well as lanthanide containing single molecule magnets (SMM). The presented investigations on the different transition metal complexes reveal manifold effects for each species leading to valuable results. A fundamental understanding of metal to ligand interactions is mandatory for the development of new and better organometallic complexes with catalytic, optical or magnetic properties.
Clusters bridge the gap between single atoms or molecules and the condensed phase and it is the challenge of cluster science to obtain a deeper understanding of the molecular foundation of the observed cluster specific properties/reactivities and their dependence on size. The electronic structure of hydrated magnesium monocations [Mg,nH2O]+, n<20, exhibits a strong cluster size dependency. With increasing number of H2O ligands the SOMO evolves from a quasi-valence state (n=3-5), in which the singly occupied molecular orbital (SOMO) is not yet detached from the metal atom and has distinct sp-hybrid character, to a contact ion pair state. For larger clusters (n=17,19) these ion pair states are best described as solvent separated ion pair states, which are formed by a hydrated dication and a hydrated electron. With growing cluster size the SOMO moves away from the magnesium ion to the cluster surface, where it is localized through mutual attractive interactions between the electron density and dangling H-atoms of H2O ligands forming "molecular tweezers" HO-H (e-) H-OH. In case of the hydrated aluminum monocations [Al,nH2O]+,n=20, different isomers of the formal stoichiometry [Al,20H2O]+ were investigated by using gradient-corrected DFT (BLYP) and three different basic structures for [Al,20H2O]+ were identified: (a) [AlI(H2O)20]+ with a threefold coordinated AlI; (b) [HAlIII(OH)(H2O)19]+ with a fourfold coordinated AlIII; (c) [HAlIII(OH)(H2O)19]+ with a fivefold coordinated AlIII. In ground state [AlI(H2O)20]+ (a) which contains aluminum in oxidation state +1 the 3s2 valence electrons remain located at the aluminium monocation. Different than for open shell magnesium monocations no electron transfer into the hydration shell is observed for closed shell AlI. However, clusters of type (a) are high energy isomers (DE»+190 kJ mol-1) and the activation barrier for reaction into cluster type (b) or (c) is only approximately 14 kJ mol-1. The performed ab initio calculations reveal that unlike in [Mg,nH2O]+, n=7-17, for which H atom eliminiation is found to be the result of an intracluster redoxreaction, in [Al,nH2O]+,n=20, H2 is formed in an intracluster acid-base reaction. In [Mg,nH2O]+, n>17, the magnesium dication was found to coexist with a hydrated electron in larger cluster sizes. This proves that intermolecular electron delocalization - previously almost exclusively studied in (H2O)n- and (NH3)n- clusters - can also be an important issue for water clusters doped with an open shell metal cation or a metal anion. Structures and stabilities of hydrated magnesium water cluster anions with the formal stoichiometry [Mg,nH2O]-, n=1-11, were investigated by application of various correlated ab initio methods (MP2, CCSD, CCSD(T)). Metal cations surely have high relevance in numerous biological processes, and as most biological processes take place in aqueous solution hydrated metal ions will be involved. However, in biological systems solvent molecules (i.e. water) compete with different solvated chelate ligands for coordination sites at the metal ion and the solvent and chelate ligands are in mutual interactions with each other and the metal ion. These interactions were investigated for the hydration of ZnII/carnosine complexes by application of FT-ICR-MS, gas-phase H/D exchange experiments and supporting ab initio calculations. In the last chapter of this work the Free Electron Laser IR Multi Photon Dissocition (FEL-IR-MPD) spectra of mass selected cationic niobium acetonitrile complexes with the formal stoichiometry [Nb,nCH3CN]+, n=4-5, in the spectral range 780 – 2500 cm-1 are reported. In case of n=4 the recorded vibrational bands are close to those of the free CH3CN molecule and the experimental spectra do not contain any evident indication of a potential reaction beyond complex formation. By comparison with B3LYP calculated IR absorption spectra the recorded spectra are assigned to high spin (quintet, S=2), planar [NbI(NCCH3)4]+. In [Nb,nCH3CN]+, n=5, new vibrational bands shifted away from those of the acetonitrile monomer are observed between 1300 – 1550 cm-1. These bands are evidence of a chemical modification due to an intramolecular reaction. Screening on the basis of B3LYP calculated IR absorption spectra allow for an assignment of the recorded spectra to the metallacyclic species [NbIII(NCCH3)3(N=C(CH3)C(CH3)=N)]+ (triplet, S=1), which has formed in a internal reductive nitrile coupling reaction from [NbI(NCCH3)5]+. Calculated reaction coordinates explain the experimentally observed differences in reactivity between ground state [NbI(NCCH3)4]+ and [NbI(NCCH3)5]+. The reductive nitrile coupling reaction is exothermic and accessible (Ea=49 kJ mol-1) only in [NbI(NCCH3)5]+, whereas in [NbI(NCCH3)4]+ the reaction is found to be endothermic and retarded by significantly higher activation barriers (Ea>116 kJ mol-1).
In the 1st project, successful development of 2nd generation of a palladium catalyst for the selective hydrogenation of carboxylic acids to aldehydes was accomplished. This project was done in cooperation with Dipl. Chem. Thomas Fett from Boeringer Ingelheim, Austria. The new catalyst is highly effective for the conversion of diversely functionalized aromatic, heteroaromatic and aliphatic carboxylic acids to the corresponding aldehydes in the presence of pivalic anhydride at 5 bar hydrogen pressure, which was otherwise achieved either at 30 bar of hydrogen pressure or by using waste intensive hypophosphite bases as reducing agent. Our method has increased the synthetic importance of this valuable transformation. Selective hydrogenation of carboxylic acids to the corresponding aldehydes is now possible with industrial hydrogenation equipment as well as laboratory scale glass autoclaves. It might also convince the synthetic organic chemists to use this transformation for routine aldehyde synthesis in the laboratories.
In the 2nd project, a microwave assisted Cu-catalyzed protodecarboxylation of arenecarboxylic acids to arenes is achieved. This work was done in collaboration with Dipl. Chem. Filipe Manjolinho under the supervision of Dr. Nuria Rodríguez. In the presence of 1-5 mol% of inexpensive CuI/1,10-phenanthroline catalyst generated in situ under microwave radiations, diversely functionalized arenes and heteroarene carboxylic acids have been decarboxylated to the corresponding arenes in good yields at 190 °C in 5-15 min. The loss of volatile arenes with the release of CO2 is controled by the use of sealed high pressure resistant microwave vessels. These reactions are highly beneficial for parallel synthesis in drug discovery due to their short reaction time. Microwave technology will also help in the future to develop more effective catalysts for protodecarboxylation rections.
Based on the microwave assisted protodecarboxylation strategy, decarboxylative coupling of arenecarboxylic acids with aryl triflates and tosylates was also conducted under microwave radiation which provided higher yields of the corresponding biphenyls from deactivated substrates in short reaction time compared to the conventional heating.
In the 3rd project, crystalline, potassium (trifluoromethyl)trimethoxyborate was successfully applied for the synthesis of benzotrifluorides under the oxidative conditions. This project was done in cooperation with Dipl. Chem. Annette Buba. In the presence of Cu(OAc)2 and molecular oxygen, arylboronates were coupled with K+[CF3B(OMe)3] in DMSO at 60 °C. A variety of benzotriflurides was synthesized in good yields under the optimized reaction conditions. This protocol for the oxidative trifluoromethylation of arylboronates is the base for the development of decarboxylative trifluoromethylation reaction of arenecarboxylic acids.
The 4th project discloses the simple and straightforward synthesis of trifluoromethylated alcohols by nucleophilic addition of potassium (trifluoromethyl)trimethoxyborate to carbonyl compounds. This project was done in cooperation with Dr. Thomas Knauber and Dipl. Chem. Annette Buba. In the presence of K+[CF3B(OMe)3] in THF at 60 °C, diversely functionalized aldehydes and ketones were successfully converted into the corresponding trifluoromethylated alcohols.
The 3rd and 4th projects demonstrate the successful establishment of crystalline and shelf stable potassium (trifluoromethyl)trimethoxyborate as highly versatile CF3-source in nucleophilic trifluoromethylation reactions. These new protocols are characterized by their user-friendliness and broad applicability under mild reaction conditions, thus they are beneficial for late stage introduction of CF3-group into organic molecules.
Colorectal cancer is the second most prevalent cancer form in both men and women in the Europe. In 2002, alimentary cancer (oesophagus, stomach, intestines) made up 26% of the annual incident cases of cancer amongst males in Europe, whereby about half of those were cancers of the colon and rectum (Eurostat 2002). Epidemiological evidence accumulating over the last decades indicates that besides a genetic disposition, diet plays a strong epigenetic role in the genesis of cancer. It is generally assumed that diet is causal for up to 80% of colorectal cancer (Bingham 2000). With the prospect of an approximated 50% rise in global cancer incidence over the first two decades of the 21st century, the World Health Organisation (WHO) has emphasized the need for an improvement in nutrition. Indeed there is increasing public health awareness with respect to nutrition. Today, living healthily is associated with less consumption of animal fats and red (processed) meat, moderate or no consumption of alcohol coupled with increased physical activity, and frequent intake of fruits, vegetables and whole grains (Bingham 1999; Johnson 2004). This idealogy partly stems from scientific epidemiological evidence supportive of an inverse correlation between the consumption of fruits and vegetables and the development cancer. Besides fibre and essential micro-nutrients like ascobate, folate, and tocopherols, the anti-carcinogenic properties of fruits and vegetables are generally thought to be rooted in the bioactivity of secondary plant components like flavonoids (Johnson 2004; Rice-Evans and Miller 1996; Rice-Evans 1995). Along with the increased public health awareness, has also come a burgeoning and lucrative dietary supplement industry, which markets products based on polyphenols and other potentially healthy compounds, sometimes with questionable promises of better health and increased longevity. These claims are based on accumulating in vitro and in vivo evidence indicating that flavonoids and polyphenols in fruits and vegetables can hinder proliferation, induce apoptosis of cancerous cells (Kern et al. 2005; Kumar et al. 2007; Thangapazham et al. 2007), act as antioxidants (Justino et al. 2006; Rice-Evans 1995) and influence cell signalling pathways (Marko et al. 2004; Joseph et al. 2007; Granado-Serrano et al. 2007), all of which are potential mechanisms proposed for their anti-carcinogenic activity. However, not only is the vast variety of supplements worrisome, but also problematic, is their easy accessibilty (just a click away on the internet) and the amount that can potentially be consumed. Such supplements are usually offered in pharmaceutical form (tablets, capsules, powder, concentrates) containing concentrations well beyond what is normally comsumable from the diet. For example, quercetin’s recommended intake is about 1g daily. However, estimates portend a possible daily increase of upto 1000 fold of the daily intake of quercetin (Hertog et al. 1995). Mindful of the concept of dose coined from the words of swiss scientist Paracelsus “What is it that is not poison? All things are poison and nothing is without poison. The right dose differentiates a poison and a remedy.” (“Alle Dinge sind Gift und nichts ist ohn’ Gift; allein die Dosis macht, dass ein Ding kein Gift ist”), it is thus conceivable that such high concentrations may not only reverse the acclaimed positive effects of flavonoids and polyphenols but also have negative effects thereby representing a health risk. The fact that direct evidence of the beneficial effects of flavonoids and polyphenols remains wanting, if not entirely lacking, coupled with the afore-mentioned marketing trend demands for a thorough examination of the possible adverse effects that may arise from increased consumption of flavonoids and polyphenols. The genesis and progression of cancer is usually accompanied by dysfunctional signalling of certain cell signalling pathways. Typical for colon carcinogenesis is the malfunctioning of the Wnt-signalling pathway, a pathway, which is crucial for the growth and development of normal colonocytes. The dysfunction of the Wnt-signalling pathway occurs in a manner that culminates in a proliferation stimulus of colonocytes, while differentiation is increasingly minimized. Hence, tumourigenesis is promoted. Interupting the proliferation stumuli by intervening in the actions of components of the Wnt-signalling pathway is one potential mechanism for the anti-carcinogenic action of flavonoids and polyphenols (Pahlke et al. 2006; Dashwood et al. 2002; Park et al. 2005). However, as previously hinted, the indulgence in the consumption of flavonoids and polyphenols based supplements could instead lead to a proliferation stimulus and provoke or promote carcinogenesis in normal cells or pre-cancerous cells respectively. The aim of this work was to
Polychlorinated dibenzo-p-dioxins, dibenzofurans, and polychlorinated biphenyls are persistent environmental pollutants which ubiquitously occur as complex mixtures and accumulate in the food and feed chain due to their high lipophilic properties. Of the 419 possible congeners, only 29 share a common mechanism of action and cause similar effects, the so called dioxin-like compounds. Dioxin-like compounds evoke a broad spectrum of biochemical and toxic responses, i.e. enzyme induction, dermal toxicity, hepatotoxicity, immunotoxicity, carcinogenicity as well as adverse effects on reproduction, development, and the endocrine system in laboratory animals and in humans. Most, if not all, of the aforementioned responses, are mediated by the aryl hydrocarbon receptor. In the present work, the elicited biochemical effects of a selection of dioxin-like compounds and the non dioxin-like PCB 153 were examined in mouse (in vivo) and in human liver cell models (in vitro). Emphasis was given to the main contributors to the total toxic equivalents in human blood and tissues TCDD, 1-PnCDD, 4-PnCDF, PCB 118, PCB 126, and PCB 156, which likewise contribute about 90 % to the dioxin-like activity in the human food chain.
Three mouse in vivo studies were carried out aiming to characterize the alterations in hepatic gene expression as well as the induction of hepatic xenobiotic metabolizing enzymes after single oral dose. Based on the results obtained from mouse 3-day and 14-day studies, the seven test compounds can be categorized into three classes; the ones which are 'pure' AhR ligands (TCDD, 1-PnCDD, 4-PnCDF, and PCB 126) or solely CAR inducers (PCB 153), and the ones which are AhR/CAR mixed-type inducers (PCB 118, PCB 156). Moreover, the analysis of hepatic gene expression patterns after a single oral dose of either TCDD or PCB 153 revealed that the altered genes fundamentally differed. Profiling of significantly altered genes led to the conclusion that changes in gene expression were associated with different signalling pathways, in fact by AhR and CAR.
For investigating the role of the AhR in mediating biological responses, several experimental approaches were carried out, such as the analysis of blood plasma metabolites in Ahr knockout and wild-type mice. Genotype specifics and similarities were determined by HPLC-MS/MS analysis. Several plasma metabolites could be identified in both genotypes, but also differences were detected. Furthermore, an in vivo experiment was performed aiming to characterize AhR-dependent and -independent effects in female Ahr knockout and wild-type mice. For this purpose, mice received a single oral dose of TCDD and were killed 96 h later. Microarray analysis of mouse livers revealed that although the Ahr gene was knocked out in Ahr-/- mice, the quantity of affected genes were in the same order of magnitude as for Ahr+/+ mice, but the pattern of altered genes distinctly differed. In addition, the relative liver weights of TCDD-treated Ahr+/+ mice were significantly increased which led to the conclusion, that TCDD induced the development of hepatic steatosis in female Ahr wild-type.
The performed in vitro experiments aimed to characterize the effects elicited by selected DLCs and PCB 153 in human liver cell models by the use of HepG2 cells and primary human hepatocytes. In general, primary human hepatocytes were less responsive than HepG2 cells. This was not only observed in EC values derived from EROD assay, but also regarding microarray analysis in terms of differently regulated genes. In vitro REPs gained from both liver cell models widely confirmed the current TEFs, but some deviations occurred. The comparison of the TCDD-altered genes in both human cell types revealed that only a considerably small number of genes was in common up regulated by both human liver cell models, such as the established AhR-regulated highly inducible cytochrome P450s 1A1, 1A2, and 1B1 as well as other AhR target genes. Although the overlap was rather small, the TCDD-induced genes could be consistently associated with the broad spectrum of established dioxin-related biological responses. The gene expression pattern in primary human hepatocytes after treatment with selected DLCs (TCDD, 1-PnCDD, 4-PnCDF, and PCB 126) and PCB 153 was additionally characterized by microarray analysis. The highest response in terms of significantly altered genes was determined for TCDD, followed by 4-PnCDF, 1-PnCDD, and PCB 126, whereas exposure to PCB 153 did not evoke any significant changes in gene expression. The pattern of significantly altered genes was very homogenous among the four congeners. Genes associated with well-established DLC-related biological responses as well as novel dioxin-inducible target genes were identified, whereby an extensive overlap in terms of up regulated genes by all four DLCs occurred. In conclusion, the results from the in vitro experiments performed in primary human hepatocytes provided fundamental insight into the congeners' potencies and caused alterations in gene expression patterns. The obtained findings implicate that although the extent of enzyme inducibilities varied, the gene expression patterns are coincidental. Microarray analysis identified species-specific (mouse vs. human) as well as model-specific (in vitro vs. in vivo and transformed cells vs. untransformed cells) differences. In order to identify novel biomarkers for AhR activation due to treatment with dioxin-like compounds, five candidates were selected based on the microarray results i.e. ALDH3A1, TIPARP, HSD17B2, CD36, and AhRR. Eventually, ALDH3A1 turned out to be the most reliable and suitable marker for exposure to DLCs in both human liver cell models eliciting the highest mRNA inducibility among the five chosen candidates. In which way these species- and cell type-specific markers are involved in the dioxin-elicited toxic responses should be further characterized in vivo and in vitro.
A number of natural products are known that contain an enamide as a key structural feature. This functionality is a very important subunit in various biologically active products and pharmaceutical drug lead compounds. In addition, enamides serve as highly versatile synthetic intermediates, particularly in the pericyclic reaction, formation of heterocycles, cross-coupling and in asymmetric synthesis. As a result, several protocols have been devised for the preparation of enamides. Traditional syntheses include condensation of aldehydes and ketones with amides or from hydroxylamines and acetic anhydride, require harsh conditions and yield mixtures of E/Z products. Several metal catalyzed approaches have been also investigated, such as isomerization of N-allylamides and catalytic cross-coupling of amides with vinyl halides or pseudohalides. These protocols proceed under milder conditions but suffer from the limited availability of these starting materials. The research described in this dissertation focuses on efficient and atom-economic preparation of enamides and thioenamides, using readily available starting materials. We developed catalyst systems generated in situ from bis(2-methallyl)-cycloocta-1,5-diene-ruthenium(II), phosphines and Lewis acid or base, efficiently catalyze the addition of primary amides and thioamides to terminal alkynes with exclusive formation of the anti-Markovnikov products in high yield and stereoselectivity under mild reaction conditions. The generality of the newly developed methodologies is demonstrated by common functional group tolerance. Furthermore, Markovnikov products were formed via phosphine-catalyzed addition of cyclic amides to phenylacetylene derivatives. The hydroamidation protocol of primary amides was successfully used in the synthesis of naturally occurring compounds, such as alatamide, lansiumamide A, botryllamides C and E, and the key intermediate in the synthesis of aristolactam. In order to investigate the reaction mechanism, the addition of various amides and carboxylic acids to terminal alkynes was performed using deuterium labeled starting materials and followed by in situ NMR and GC-MS studies.