Diffusion-ordered NMR Spectroscopy Research Articles

Study of the preparation, characterization, and solubility of lidocaine complexed with 5-sulfosalicylic acid dihydrate

Objective: This study was to prepare solid dispersions of lidocaine (Lid) with 5-sulfosalicylic acid dihydrate (SSA) by freeze-drying (freeze-dried [FD] Lid/SSA = 1/1) and to evaluate their physical properties. Methods: Here, we evaluated the physicochemical properties and solubility of solid dispersions of lidocaine (Lid) and 5-sulfosalicylic acid dihydrate (SSA) prepared by freeze-drying (freeze-dried [FD] Lid/SSA = 1/1). Results: Differential scanning calorimetry measurements showed that after freeze-drying, the endothermic peak due to Lid melting, the dehydration peak, and the endothermic peak due to SSA melting disappeared. Powder X-ray diffraction results showed that the characteristic Lid and SSA peaks disappeared after freeze-drying, indicating a halo pattern. The near-infrared spectroscopy results suggested that Lid-derived -NH and -CH groups and the Lid-derived -OH and -CH groups from the SSA peak shifted and broadened after freeze-drying, suggesting their involvement in complex formation through Lid/SSA intermolecular interactions. Nuclear Overhauser effect spectroscopy-nuclear magnetic resonance (NMR) measurements showed a cross-peak due to the interaction between the Lid-derived -CH group and the SSA-derived -OH group, suggesting hydrogen bonding. Diffusion-ordered spectroscopy NMR measurements showed that the diffusion coefficients of Lid and SSA aggregated in FD Lid/SSA, suggesting a change in Lid dispersibility in the solvent owing to the formation of a complex with SSA. The solubility of FD Lid/SSA was approximately 88 mg/mL (∼20-fold higher than that of Lid). Conclusions: These findings suggest that complex formation occurred in FD Lid/SSA; this enhanced the solubility of this dispersion.

Water Confinement on Polymer Coatings Dictates Proton-Electron Transfer on Metal-Catalyzed Hydrogenation of Nitrite.

Enzymes can precisely control the speed and selectivity of chemical reactions by modifying locally the solvent-reactant interactions. To extrapolate these attributes to heterogeneous catalysts, we have employed thermoresponsive poly n-isopropylacrylamide (p-NIPAM) brushes bonded to silica spheres containing palladium. These polymers can form hydrogen bonds with water molecules at low temperatures (<32 °C) allowing the polymer to stay swollen. Detailed reaction kinetics of nitrite hydrogenation showed that p-NIPAM decreases the apparent activation barrier by a factor of 3 at low temperatures. Diffusion-ordered spectroscopy nuclear magnetic resonance and ab initio molecular dynamics simulations showed that when p-NIPAM is present, water molecules near the surface are less mobile. This confinement perturbs the water interaction with the metal, reducing the barrier for the proton-electron transfer reduction of nitrite. Notably, this enhancement vanishes at high temperature as the polymer collapses on itself exposing the Pd to unconfined water. The fully reversible nature of this process opens the door for creating homeostatic catalysts with controlled water-confinement.

Metabolomics analysis using matrix-assisted diffusion-ordered spectroscopy (DOSY) and its application in acrylamide-induced cardiovascular toxicity

Untargeted NMR-based metabolomics strategies are increasingly applied for metabolite screening in a wide variety of medical conditions. However, the identification of metabolites remains a great challenge in NMR metabolomics research. Here, we used matrix-assisted diffusion-ordered spectroscopy (DOSY) to identify new biomarkers in body fluids. DOSY-based metabolomics can exploit the unique strengths of NMR and, thanks to improvements in its resolution, can be successfully used to address a significantly wider range of biological issues. In this study, a metabolite model consisting of ten common metabolites was employed to optimize the matrix-assisted DOSY experimental parameters. Polyvinylpyrrolidone (PVP), a high-molecular-weight polymer, was found to be the most appropriate matrix with which to accomplish the separation of the mixture components. 1H NMR and PVP-assisted DOSY analyses were performed to characterize the metabolic profile of serum samples from acrylamide (ACR)-treated rats and study the mechanism of ACR-induced cardiovascular toxicity. Biomarker identification by 1H NMR and PVP-assisted DOSY yielded similar results. However, multivariate statistical analysis showed that, compared with 1H NMR, PVP-assisted DOSY achieved a better discrimination of the serum samples obtained from rats treated with different doses of ACR. A total of 11 differential metabolites were identified. An enrichment analysis of the metabolic pathways showed that, in rats treated with a high dose of ACR, the glutamate and glutathione metabolic pathways and the energy metabolism pathway of the tricarboxylic acid cycle were mainly affected.

Molecular insights into fragrance release from SDS and SLES micelles

Understanding the interaction between fragrance and surfactant micelles is essential for formulators to develop performing fragrant products. The equilibrium of fragrances in micellar solution of two commonly used surfactants in home and body care industries, namely sodium dodecyl sulphate (SDS) and sodium lauryl ether sulphate (SLES), has been investigated. The difference between SDS and SLES in the release of fragrant compounds is assessed by headspace measurements and explained at a molecular level by the different solubilization sites of the perfume within the micellar systems. Diffusion-Ordered Nuclear Magnetic Resonance Spectroscopy (DOSY NMR) shows that SLES micelles solubilize less fragrance than SDS. Subsequently, a shift from fragrance positioning through the water phase from SDS to SLES system is shown with COSMOmic calculations. This study brings knowledge on fragrance/surfactant interactions and shows the interest of using a computational method to screen and compare fragranced surfactants systems allowing formulators to anticipate the performance of their products.

Using Nuclear Magnetic Resonance to Troubleshoot a Stability Issue in a Real-World Formulation Chassis-Application to Consumer Oral Healthcare.

With direct application to current and future consumer healthcare products, this research sheds light on the importance of packaging and its potential effects on both Active Pharmaceutical Ingredient (API) delivery and stability. Industrially sourced, proprietary experimental formulations (PEFs), specifically oral cleansers, based on salicylic acid and hydrogen peroxide, discolored over time at different rates, depending on packaging type used. This discoloration stemmed from an interplay of two factors, involving both spontaneous formulation degradation and the interaction of both degradants and salicylic acid with the internal surface of the packaging. This manuscript reports on the investigation to uncover the origins of discoloration. To investigate this real-world, industrial pipeline problem, we exploited the high dimensionality and simple sample preparation uniquely afforded by NMR. Using a combination of 1D/2D NMR and diffusion-ordered NMR spectroscopy (DOSY) to leverage molecular mass estimations from, we not only quickly confirmed the identities of these degradants, but also assessed their formation as a function of temperature and pH, providing insight into the mechanisms underlying their formation. We were able to identify catechol as the main source of discoloration over a period of several weeks, being formed at the ppm level. Furthermore, we evaluated the formulation-container interaction, employing NMR, ICP-MS, and ATR-IR. Despite this comprehensive analysis, the root causes of discoloration could only tentatively be assigned to a surface Ti complex of salicylic acid and other hydroxy carboxylic acids. Through the understanding of formulation degradation pathways, we were able to support further toxicology assessment, vital to both consumer safety and the manufacturer. This work underscores the invaluable role of NMR in the analysis of intricate proprietary mixtures with a consumer-centric purpose. Our findings demonstrate that conventional analytical techniques falter in the face of such complexity, requiring extensive preparation and pre-analytical processing, highlighting the novelty and crucial relevance of NMR research to manufacturers and consumers. Such an analysis is of value in the pursuit of materials within the consumer-healthcare space, which meet the requirements for successful recycling or re-use.

Effects of Drug Conjugation on the Biological Activity of Single-Chain Nanoparticles.

The field of single-chain nanoparticles (SCNPs) continues to mature, and an increasing range of reports have emerged that explore the application of these small nanoparticles. A key application for SCNPs is in the field of drug delivery, and recent work suggests that SCNPs can be readily internalized by cells. However, limited attention has been directed to the delivery of small-molecule drugs using SCNPs. Moreover, studies on the physicochemical effects of drug loading on SCNP performance is so far missing, despite the accepted view that such small nanoparticles should be significantly affected by the drug loading content. To address this gap, we prepared a library of SCNPs bearing different amounts of a covalently conjugated therapeutic drug-sulfasalazine (SSZ). We evaluated the impact of the conjugated drug loading on both the synthesis and biological activity of SCNPs on pancreatic cancer cells (AsPC-1). Our results reveal that covalent drug conjugation to the side chains of the SCNP polymer precursor interferes with chain collapse and cross-linking, which demands optimization of reaction conditions to reach high degrees of cross-linking efficiencies. Small-angle neutron scattering and diffusion-ordered spectroscopy nuclear magnetic resonance (DOSY NMR) analyses reveal that SCNPs with a higher drug loading display larger sizes and looser structures, as well as increased hydrophobicity associated with a higher SSZ content. Increased SSZ loading led to reduced cellular uptake when assessed in vitro, whereby SCNP aggregation on the surface of AsPC-1 cells led to reduced toxicity. This work highlights the effects of drug loading on the drug delivery efficiency and biological behavior of SCNPs.

Spatially encoded pure-shift diffusion-ordered NMR spectroscopy yielded by chirp excitation

Spatially-encoded diffusion-ordered NMR spectroscopy (SPEN-DOSY) has emerged as a new time-efficient tool for the analysis of mixtures of small molecules in solution. Time efficiency is achieved using the concept of spatial parallelization of the effective gradient area, instead of the sequential incrementation used in conventional diffusion experiments. The data acquired with such sequences are then usually processed to extract diffusion coefficients, but cases when peak overlap in the 1H spectrum are difficult to address. Such limitation in conventional diffusion experiments is addressed via using the Pure Shift Yielded by CHirp Excitation (PSYCHE)-iDOSY sequence. Here we have adapted the PSYCHE-iDOSY sequence by using echo planar spectroscopic imaging (EPSI) to acquire SPEN-DOSY data. The pure shift mode of PSYCHE separates the overlapped components and a modified Stejskal-Tanner equation is used to extract the corresponding diffusion coefficient. The primary results obtained with the above-mentioned mixtures seem to open the possibility of separating complex mixtures in less time than PSYCHE-iDOSY.

Supramolecular polymerization behavior of a ditopic self-folding biscavitand

Abstract Reported herein is the supramolecular polymerization of a mixture of a feet-to-feet connected biscavitand and a homoditopic quinuclidinium guest that is regulated by cooperativity in the host–guest association. Diffusion-ordered NMR spectroscopy (DOSY) was used to evaluate the supramolecular polymerization in toluene, CHCl3, and tetrahydrofuran (THF). Upon concentrating the solutions of the biscavitand with the quinuclidinium guest in CHCl3 and THF, the diffusion coefficient (D) values were meaningfully decreased, indicating that the host–guest complexation facilitated supramolecular polymerization. In contrast, the slight change of the D value in toluene suggests that supramolecular polymerization was suppressed, although the binding constant (K) between the cavitand and quinuclidinium guest was reported to be 105 L mol−1 in toluene. The viscosity measurements showed both the critical polymerization concentration (CPC) and entangled concentration (Ce) upon concentrating the CHCl3 solution of the mixture. Neither the CPC nor Ce was seen in the toluene solution of the mixture. Accordingly, the strong negative cooperativity in the 1:2 host–guest complexation of the biscavitand discouraged the supramolecular polymerization in toluene. These findings are valuable in deepening the understanding of host–guest association-driven supramolecular polymerization behaviors regulated by a combination of cooperativity and K value in solution.

Monitoring galactolipid digestion and simultaneous changes in lipid-bile salt micellar organization by real-time NMR spectroscopy

The use of Nuclear Magnetic Resonance spectroscopy for studying lipid digestion in vitro most often consists of quantifying lipolysis products after they have been extracted from the reaction medium using organic solvents. However, the current sensitivity level of NMR spectrometers makes possible to avoid the extraction step and continuously quantify the lipids directly in the reaction medium. We used real-time 1H NMR spectroscopy and guinea pig pancreatic lipase-related protein 2 (GPLRP2) as biocatalyst to monitor in situ the lipolysis of monogalactosyl diacylglycerol (MGDG) in the form of mixed micelles with the bile salt sodium taurodeoxycholate (NaTDC). Residual substrate and lipolysis products (monogalactosyl monoacylglycerol (MGMG); monogalactosylglycerol (MGG) and octanoic acid (OA) were simultaneously quantified throughout the reaction thanks to specific proton resonances. Lipolysis was complete with the release of all MGDG fatty acids. These results were confirmed by thin layer chromatography (TLC) and densitometry after lipid extraction at different reaction times. Using diffusion-ordered NMR spectroscopy (DOSY), we could also estimate the diffusion coefficients of all the reaction compounds and deduce the hydrodynamic radius of the lipid aggregates in which they were present. It was shown that MGDG-NaTDC mixed micelles with an initial hydrodynamic radius rH of 7.3 ± 0.5 nm were changed into smaller micelles of NaTDC-MGDG-MGMG of 2.3 ± 0.5 nm in the course of the lipolysis reaction, and finally into NaTDC-OA mixed micelles (rH of 2.9 ± 0.5 nm) and water soluble MGG. These results provide a better understanding of the digestion of galactolipids by PLRP2, a process that leads to the complete micellar solubilisation of their fatty acids and renders their intestinal absorption possible.

Review on NMR as a tool to analyse natural products extract directly: Molecular structure elucidation and biological activity analysis.

Natural products, the small organic molecules produced by plants, microbes and invertebrates, often present in the form of a mixture, this leads to the structural characterisation of natural extracts often requiring time-consuming multistep purification procedures. Nuclear magnetic resonance (NMR) technology is traditionally utilised as a tool for the structural elucidation of pure compounds. Contemporarily, an up-to-date trend in the application of NMR in natural product research is shifting to the direct NMR analysis of crude mixtures, to obtain molecular structure and biological activity information without performing cumbersome separation. To review works of literature on the evolution, principle and progress of NMR technologies for analysing mixtures, we focus on the successful application of NMR technologies in direct analysis of natural product extracts. Based on our research experience, academic tracking and extensive literature search, which involved, but not limited to, the use of various databases, like Web of Knowledge and PubMed. The keywords used, in various combinations, to retrieve recent literature on the successful application of NMR technologies to sheer natural product extracts, and excluded artificially natural product mixture and biofluid. NMR technologies for direct natural extracts analysis, including two-dimensional J-resolved spectroscopy (2D-JRES), pure shift NMR, diffusion-ordered NMR spectroscopy (DOSY), statistical correlation spectroscopy (STOCSY), concentration-ordered NMR spectroscopy (CORDY), saturation transfer difference (STD) and water-ligand observed via gradient spectroscopy (WaterLOGSY) were illustrated. By these methods, molecular structure and biological activity information will be directly obtained from NMR analysis of natural products extract, aiming to save experimental time and expenses.

Recycled PLA for 3D Printing: A Comparison of Recycled PLA Filaments from Waste of Different Origins after Repeated Cycles of Extrusion.

The objective of this work is to evaluate the reprocessing of PLA 3D printing waste from different origins, into filaments and films, and without the addition of any additive. Two types of waste were considered: a blend of different printing wastes (masks, visors, other components) of personal protective equipment coming from an association of Spanish coronamakers, and PLA waste from a single known commercial source. Both types of materials were subjected to repeated extrusion cycles and processed into films by compression molding. Samples were characterized after each cycle and their mechanical and viscosity properties evaluated. Diffusion-ordered NMR spectroscopy (DOSY) experiments were also carried out to estimate molecular weights. The results show a better performance for the PLA waste from the known origin, capable of withstanding up to three re-extrusion cycles per two for the waste blending, without significant degradation. Additionally, a model to address collection and mechanical recycling cycles under two different scenarios (full traceability and not full traceability) was proposed.

Using surface plasmon resonance, capillary electrophoresis and diffusion-ordered NMR spectroscopy to study drug release kinetics

Nanomedicines, including polymer nanocarriers with controlled drug release, are considered next-generation therapeutics with advanced therapeutic properties and reduced side effects. To develop safe and efficient nanomedicines, it is crucial to precisely determine the drug release kinetics. Herein, we present application of analytical methods, i.e., surface plasmon resonance biosensor technology (SPR), capillary electrophoresis, and 1H diffusion-ordered nuclear magnetic resonance spectroscopy, which were innovatively applied for drug release determination. The methods were optimised to quantify the pH-triggered release of three structurally different drugs from a polymer carrier. The suitability of these methods for drug release characterisation was evaluated and compared using several parameters including applicability for diverse samples, the biological relevance of the experimental setup, method complexity, and the analysis outcome. The SPR method was the most universal method for the evaluation of diverse drug molecule release allowing continuous observation in the flow-through setting and requiring a small amount of sample.

Charge transport in protic ionic liquids: Effect of protonation state in 1-methylimidazolium – acetate/trifluoroacetate mixtures

Despite the potential to overcome slow charge transport in ionic liquids by decoupling charge transport from mass transport in protic ionic liquids (PILs), the average proton distribution and the contribution of proton conduction to the overall charge transport in PILs has not been fully established. To elucidate the predominant underlying molecular species in the PILs and the molecular dynamics of the PILs, we study the dipolar response and conductivity using dielectric relaxation (DR) spectroscopy and the nucleis chemical environment and molecular mobility using nuclear magnetic resonance (NMR) spectroscopy of prototypical PILs based on 1–methylimidazolium as Brønsted base. To elucidate the effect of the acidity of the Brønsted acid we gradually vary the composition of the PILs from 1–methylimidazoliumacetate [MimH][AcO] to 1–methylimidazoliumtrifluoroacetate [MimH][TFA]. Both, 15N-NMR chemical shifts and the sample conductivities suggest that in neat [MimH][AcO] electro-neutral molecular species dominate, consistent with earlier findings. The dipolar response of these molecular species as detected via the DR spectra is stronger than what one would expect based on the molecular dipole moments estimated from density functional theory calculations, which points to the formation of an extended hydrogen-bonded network. Upon addition of [MimH][TFA], this dipolar response is reduced as the degree of protonation of 1–methylimidazolium gradually increases, which is apparent from the conductance and 15N chemical shifts. Interestingly, diffusion ordered NMR spectroscopy indicates protons to become more mobile than the constituting acid and base molecules, consistent with Grotthuss-like proton transport. To vary the immediate environment of the PILs, we study solutions of the PIL mixtures in aprotic dimethylformamide (DMF) and protic methanol (MeOH) as two limiting cases. We find that [MimH][AcO] exclusively forms electroneutral species in DMF, as evident from the negligible conductivity, while with increasing TFA content the fraction of charged molecular species increases. Although these ionic species give rise to a finite conductivity, demonstrating the presence of mobile ions, a large fraction of these ions is bound in [MimH][TFA] ion-pairs, as detected in the DR spectra. Yet, temperature dependent 1H-NMR linewidths of the acidic proton signals suggest proton exchange to occur on the NMR timescale, presumably between [MimH]+ and [Mim]. Conversely, in MeOH we find no evidence for the presence of ion-pairs, while the high conductivity of these solutions shows that MeOH can efficiently solvate ions and may participate in proton transfer. Together, our results highlight the sensitivity of protonation equilibria to the immediate environment and of proton dynamics to these protonation equilibria. Our results show that tuning these equilibria using PIL mixtures can provide routes to tailor the contribution of proton transport to charge transport in PILs.

Accounting for gradient non-uniformity in spatially-encoded diffusion-ordered NMR spectroscopy

Diffusion-ordered NMR spectroscopy (DOSY) is a powerful tool for the analysis of mixtures. Spatially-encoded (SPEN) DOSY makes it possible to collect a complete DOSY data set in a single scan, through spatial parallelisation of the gradient dimension. One limitation of current SPEN DOSY experiments is that the data is analysed assuming that the field gradient is uniform over the sample. This is usually not the case for high resolution NMR probes, and even less for triple-axis gradient probes. In this work, we have developed methods to account for gradient non-uniformity in the processing of SPEN DOSY experiment. We have first mapped the field gradient, using a stimulated echo (STE) NMR sequence with a weak readout gradient. We have then modified the calculation of the position-dependent effective gradient pulse area that is used in the analysis of SPEN DOSY data. The resulting model was validated through numerical simulations. A comparison of results obtained with and without inclusion of the effect of non-uniform gradients shows that the proposed approach increases the accuracy of SPEN DOSY experiments.

Visible-Light-Induced Control over Reversible Single-Chain Nanoparticle Folding.

We introduce a class of single-chain nanoparticles (SCNPs) that respond to visible light (λmax =415 nm) with complete unfolding from their compact structure into linear chain analogues. The initial folding is achieved by a simple esterification reaction of the polymer backbone constituted of acrylic acid and polyethylene glycol carrying monomer units, introducing bimane moieties, which allow for the photochemical unfolding, reversing the ester-bond formation. The compaction and the light driven unfolding proceed cleanly and are readily followed by size exclusion chromatography (SEC) and diffusion ordered NMR spectroscopy (DOSY), monitoring the change in the hydrodynamic radius (RH ). Importantly, the folding reaction and the light-induced unfolding are reversible, supported by the high conversion of the photo cleavage. As the unfolding reaction occurs in aqueous systems, the system holds promise for controlling the unfolding of SCNPs in biological environments.

Mechanistic Investigations of Phenoxyimine-Cobalt(II)-Catalyzed C(sp2)-C(sp3) Suzuki-Miyaura Cross-Coupling.

The mechanism of phenoxyimine (FI)-cobalt-catalyzed C(sp2)-C(sp3) Suzuki-Miyaura cross-coupling was studied using a combination of kinetic measurements and catalytic and stoichiometric experiments. A series of dimeric (FI)cobalt(II) bromide complexes, [(4-CF3PhFI)CoBr]2, [(4-OMePhFI)CoBr]2, and [(2,6-diiPrPhFI)CoBr]2, were isolated and characterized by 1H and 19F NMR spectroscopies, solution and solid-state magnetic susceptibility, electron paramagnetic resonance (EPR) spectroscopy, X-ray crystallography, and diffusion-ordered NMR spectroscopy (DOSY). One complex, [(4-CF3PhFI)CoBr]2, was explored as a single-component precatalyst for C(sp2)-C(sp3) Suzuki-Miyaura cross-coupling. Addition of potassium methoxide to [(4-CF3PhFI)CoBr]2 generated the corresponding (FI)cobalt(II) methoxide complex as determined by 1H and 19F NMR and EPR spectroscopies. These spectroscopic signatures were used to identify this compound as the resting state during catalytic C(sp2)-C(sp3) coupling. Variable time normalization analysis (VTNA) of in situ catalytic 19F NMR spectroscopic data was used to establish an experimental rate law that was first-order in a (FI)cobalt(II) precatalyst, zeroth-order in the alkyl halide, and first-order in an activated potassium methoxide-aryl boronate complex. These findings are consistent with turnover-limiting transmetalation that occurs prior to activation of the alkyl bromide electrophile. The involvement of boronate intermediates in transmetalation was corroborated by Hammett studies of electronically differentiated aryl boronic esters. Together, a cobalt(II)/cobalt(III) catalytic cycle was proposed that proceeds through a "boronate"-type mechanism.

NMR Spectroscopy Study of the Effect of the Molecular Mass of Hypromellose Phthalate on Its Solubility

Scientific relevance. Hypromellose phthalate is used in enteric coatings for oral medicinal products. The proportion of phthalate groups in the polymer is standardised because it has a significant effect on solubility. Whereas, the molecular mass of hypromellose phthalate is not controlled, and its impact on solubility in media with different pH values is understudied.Aim. The study aimed to employ NMR spectroscopy to investigate the effect the molecular mass of hypromellose phthalate may have on the dissolution kinetics at the pH value declared by the polymer manufacturer.Materials and methods. The study analysed hypromellose phthalate isolated from proton-pump inhibitor enteric coatings and the hypromellose phthalate reference standard. The molecular mass of the polymer was estimated by diffusion-ordered NMR spectroscopy (DOSY) with polyethylene glycols of known molecular masses for calibration. The authors studied the dissolution profiles of hypromellose phthalates of different molecular masses using 1H NMR spectra.Results. The authors developed a procedure for estimating the average molecular mass of hypromellose phthalate by DOSY. The procedure showed variations in the molecular mass of the polymer in the test samples; the molecular mass scatter amounted to 10 kDa. The dissolution profile of the test samples in an aqueous buffer solution (pH 5.59) was described by a linear function during the first hour. The slope characterising the dissolution rate varied from 10° to 36°.Conclusions. The variation in the molecular mass of hypromellose phthalate significantly affects the dissolution rate of the test samples. The function of the dissolution rate against the molecular mass of hypromellose phthalate is non-linear. The article provides a compelling reason for further research to derive a correlation equation for the dissolution rate of hypromellose phthalate as a function of two variables (molecular mass and proportion of phthalate groups in the polymer).

Short-chain polyphosphates: Extraction effects on migration and size estimation of Saccharomyces cerevisiae extracts with polyacrylamide gel electrophoresis.

Polyacrylamide gel electrophoresis is commonly used to characterize the chain length of polyphosphates (polyP), more generally called condensed phosphates. After separation, nonradioactive, optical polyP staining is limited to chain lengths greater than 15 monomers with toluidine blue or 4',6-diamidino-2-phenylindole. PolyP chain lengths longer than 62 monomers were correlated to the shortest DNA ladders. In this study, synthetic linear polyPs (Sigma-Aldrich "Type 45", estimated mean length of 45 monomers), trimetaphosphate (trimetaP: 3 ring), tripolyphosphate (tripolyP), pyrophosphate (PPi ), and inorganic orthophosphate (o-Pi ) were visualized after separation by an in situ hydrolytic degradation process to o-Pi that was subsequently stained with methyl green. Statistically insignificant migration reduction of synthetic short-chain polyP after perchloric acid or phenol-chloroform extraction was confirmed with the Friedman test. 31 P diffusion-ordered NMR spectroscopy confirmed that extraction also reduced PPi diffusivity by <10%. Linear regression between the Rf peak migration value and the logarithm of synthetic polyP molecular weights enabled estimation of extracted polyP chain lengths from 2 to 45 monomers. Linear polyP extracts from Saccharomyces cerevisiae grown in aerobic conditions were generally shorter than extracts cultured in anaerobic conditions. Extractions from both aerobic and anaerobic S. cerevisiae included tripolyP and o-Pi , but no PPi .

Combination of Drug Delivery Properties of PAMAM Dendrimers and Cytotoxicity of Platinum(IV) Complexes-A More Selective Anticancer Treatment?

Based on their drug delivery properties and activity against tumors, we combined PAMAM dendrimers with various platinum(IV) complexes in order to provide an improved approach of anticancer treatment. Platinum(IV) complexes were linked to terminal NH2 moieties of PAMAM dendrimers of generation 2 (G2) and 4 (G4) via amide bonds. Conjugates were characterized by 1H and 195Pt NMR spectroscopy, ICP-MS and in representative cases by pseudo-2D diffusion-ordered NMR spectroscopy. Additionally, the reduction behavior of conjugates in comparison to corresponding platinum(IV) complexes was investigated, showing a faster reduction of conjugates. Cytotoxicity was evaluated via the MTT assay in human cell lines (A549, CH1/PA-1, SW480), revealing IC50 values in the low micromolar to high picomolar range. The synergistic combination of PAMAM dendrimers and platinum(IV) complexes resulted in up to 200 times increased cytotoxic activity of conjugates in consideration of the loaded platinum(IV) units compared to their platinum(IV) counterparts. The lowest IC50 value of 780 ± 260 pM in the CH1/PA-1 cancer cell line was detected for an oxaliplatin-based G4 PAMAM dendrimer conjugate. Finally, in vivo experiments of a cisplatin-based G4 PAMAM dendrimer conjugate were performed based on the best toxicological profile. A maximum tumor growth inhibition effect of 65.6% compared to 47.6% for cisplatin was observed as well as a trend of prolonged animal survival.

Soft Side of Aliphatic Polycarbonates: with PCHC-b-PDMS Copolymers toward Ductile Materials

Aliphatic polycarbonates based on CO2 and epoxides are a promising group of polymers due to the utilization of CO2 as a monomer combined with a versatile epoxide feedstock. To increase the attractiveness of these polymers as a material, the poor mechanical properties have to be improved. Herein, we present the synthesis of block copolymers from poly(cyclohexene carbonate) and poly(dimethyl siloxane) (PDMS), where diffusion-ordered NMR spectroscopy measurements indicate a successful synthesis of block structures. The resulting material shows an elongation at break of up to 24% in tensile testing with an incorporation ratio of only 27% PDMS in the copolymer. At the same time, a high tensile strength (27.9 MPa) and modulus of elasticity (1726 MPa) are observed.