Influences of surface chemistry on the separation behavior of stationary phases for reversed-phase and ion-exchange chromatography: a comparison of coated and grafted supports prepared by ring-opening metathesis polymerization

A series of poly(norborn-2-ene) (poly-NBE), poly(7-oxanorborne-2-ene-5,6-dicarboxylic acid) (poly-ONDCA), as well as poly(norborn-2-ene-co-7-oxanorborne-2-ene-5,6-dicarboxylic acid) (poly-NBE-co-ONDCA) based silica supports were prepared via ring-opening metathesis polymerization (ROMP) using both coating and grafting techniques. Poly-NBE-grafted and poly-NBE-coated supports were used for the reversed-phase separation of phenols; poly-NBE, poly-ONDCA as well as poly-NBE-co-ONDCA-grafted supports were used for comparative studies on the separation of a series of anilines and lutidines. As expected, grafted supports possess superior separation capabilities compared to their coated analogues. Compared to pure poly-NBE- and poly-ONDCA-grafted stationary phases, supports consisting of poly-NBE-co-ONDCA block-copolymers possess both hydrophobic and ion-exchange sites and represent optimum stationary phases for the separation of isomeric basic analytes.     

Metathesis polymerization-derived monolithic membranes for solid-phase extraction coupled with diffuse reflectance spectroscopy

Novel monolithic disks were prepared via ring opening metathesis polymerization (ROMP) from norborn-2-ene (NBE), a crosslinker, i.e., 1,4,5,8,8a-hexahydro-1,4,4,5,8, exo, endo-dimethanonaphthalene (DMN-H6) and tris(norborn-5-ene-2-ylmethylenoxy)methylsilane, respectively, 2-propanol and toluene (25:25:41:9, all %, w/w) using RuCl2(PCy3)2(CHPh) (Cy=cyclohexyl) as initiator and triphenylphosphine (PPh3) as modulator. Disks 1-2 mm thick were prepared by polymerization in a mold, disks thinner than 1mm were prepared by impregnation of nylon or other porous filters prior to the polymerization step. These disks were evaluated for the preconcentration of iodine and selected organic solutes from dilute aqueous samples by solid-phase extraction (SPE). Quantitative measurement of the extracted solutes was achieved by diffuse-reflectance spectroscopy (DRS) directly on the surface of the disk.     

Chiral beta-cyclodextrin-based polymer supports prepared via ring-opening metathesis graft-polymerization

A series of norborn-2-ene-derivatized beta-cyclodextrins (beta-CDs), 6-O-(norborn-2-ene-5-carboxyl)-beta-CD (1), tetrakis(6-O-norborn-2-ene-5-carboxyl)-beta-CD (2), (3), 6-O-(6-norborn-2-ene-5-car-6-O-(7-oxanorborn-2-ene-5-carboxyl)-beta-CD bonylaminohexoyl)-beta-CD (4), 6-O-(norborn-2-ene-5-ylmethoxymethylsilyl)-beta-CD (5), tris(6-O-norborn-2-ene-5-ylmethoxymethylsilyl)-beta-CD (6), tetrakis(6-O-norborn-2-ene-5-ylmethoxymethylsilyl)-beta-CD (7) and hexakis(6-O-norborn-2-ene-5-ylmethoxymethylsilyl)-beta-CD (8), have been synthesized. Compounds 1-3 were prepared via reaction of beta-CD with norborn-2-ene-5-carboxylic chloride and 7-oxanorborn-2-ene-5-carboxylic chloride, respectively; compounds 5-8 were synthesized from norborn-2-ene-5-yl-methyldichlorosilane and beta-CD, respectively. Compound 4 was accessible by reaction of norbom-2-ene-5-carboxylaminohexoyl chloride with beta-CD. Compounds 1-8 were surface grafted onto norborn-2-ene-derivatized silica-based supports using ring-opening metathesis polymerization employing the ruthenium-based initiator bis(tricyclohexylphosphino)benzylideneruthenium dichloride [Cl2Ru(CHC6H5)(PCy3)2, Cy=cyclohexyl, 9]. Generally speaking, the resulting chiral stationary phases (CSPs) I-VIII may be prepared with high reproducibility and may be used within a pH of 2-10. Thus, relative standard deviations (sigman-1) of the mean resolution (Rs) are <7%. The CSPs were used for the enantioselective separation of beta-blockers, N-dansyl-, N-3,5-dinitrobenzoyl- and Fmoc-protected amino acids and were characterized in terms of chemical stability, selectivity (alpha') and resolution (Rs). Additionally, the role of the spacer as well as influences of capacity and the degree of substitution of the beta-CD moiety on the separation characteristics were determined.     

Microwave-assisted solid-phase synthesis of 2,5-diketopiperazines: solvent and resin dependence

Solid-phase synthesis of diketopiperazines (DKPs) was preformed using various combinations of resins (polystyrene, TentaGel, ArgoGel, and PEGA) and solvents (toluene, tert-butyl alcohol, water, and toluene/2-butanol (1:4, v/v). The DKPs were synthesized from solid-phase bound dipeptides via intramolecular aminolysis. Both thermal and microwave-assisted solid-phase synthesis of DKPs gave high yields of products independently of resin and organic solvent used; however, only the PEGA resin resulted in high yields of DKPs in water independent of heating method. The short reaction times, high yields, and the possibility to run reactions in water when an appropriate resin is used makes the microwave-assisted solid-phase synthesis the method of choice. The method should be suitable for solid-phase synthesis of diketopiperazine-based libraries.     

固相合成及组合化学用的高聚物载体

综述了用于固相合成及组合化学的高聚物载体的制备方法、特征、应用和最新进展,重点介绍了目前广泛用作固相载体的交联聚苯乙烯树脂、聚酰胺树脂和TentaGel树脂,并对几类新型载体如聚乙二醇、聚四氢呋喃衍生物交联剂改性的聚苯乙烯树脂、非芳环体系的POEPOP,树脂和SPOCC树脂作了简要的概述。     

Synthesis of monofunctionalized gold nanoparticles by fmoc solid-phase reactions

In this communication, solid-phase reactions for the synthesis of Lys-monofunctionalized gold nanoparticles are described. A controlled and selective fabrication of linear nanoparticle arrays can be achieved through peptide linkage systems, and therefore it is essential to prepare Fmoc amino acid nanoparticle building blocks susceptible to Fmoc solid-phase peptide synthesis. Gold nanoparticles containing carboxylic acids (2) in the organic shell were covalently ligated to Lys on solid supports through amide bond coupling reactions. We employed Fmoc-Lys-substituted polymer resins such as Fmoc-Lys-Wang or Fmoc-Lys-HMPA-PEGA. The low density of Lys on the matrix enabled 2 nm-sized gold nanoparticles to react with Lys in a 1:1 ratio. Subsequent cleavage reactions using 60% TFA reagent resulted in Lys transfer from the solid matrix to gold nanoparticles, and the Fmoc-Lys-monofunctionalized gold nanoparticles (5) were obtained with 3-15% yield. Synthesis using HMPA-PEGA resin increased productivity due to the superior swelling properties of PEGA resin in DMF. Monofunctionalization of nanoparticles was microscopically characterized using TEM for the ethylenediamine-bridged nanoparticle dimers (6). By counting the number of 6, we found that at least 60% of cleaved nanoparticles were monofunctionalized by Lys. This method is highly selective and efficient for the preparation of monofunctionalized nanoparticles.     

Solid-phase organic tagging resins for labeling biomolecules by 1,3-dipolar cycloaddition: application to the synthesis of a fluorescent non-peptidic vasopressin receptor ligand

Two novel solid-phase organic tagging (SPOrT) resins were synthesized to facilitate the labeling of peptides and small organic compounds with a fluorescent probe. Both resins were obtained from the commercially available backbone amide linker (BAL) resin. Following the solid-phase synthesis of model compounds, a tripeptide and benzazepine, the fluorescent probe derived from Lissamine Rhodamine B was incorporated through CuI-catalyzed 1,3-dipolar cycloaddition. Final cleavage in acidic media enabled access to both types of molecules in good yield with high purity. The SPOrT resin was successfully applied to the preparation of the first non-peptidic fluorescent compound with a nanomolar affinity for the human vasopressin V2 receptor (V2R) subtype. This molecule will find application in binding assays that use polarization or fluorescence resonance energy-transfer (FRET) techniques. The SPOrT resins are also well suited for other tags and the parallel synthesis of a fluorescently tagged library for protein screening.     

Quantitative Determination of Resin Loading in Solid-Phase Organic Synthesis Using (13)C MAS NMR

The use of quantitative carbon nuclear magnetic resonance spectroscopy ((13)C NMR) for the determination of resin loadings has been investigated. Magic angle spinning (MAS) NMR spectra have been obtained for solvent-swollen resins on a conventional 7 mm CP/MAS probe using the two pulse phase modulation (TPPM) proton decoupling sequence. Loadings of resin-bound organic compounds were evaluated via addition of tetrakis(trimethylsilyl)silane as reference or using the carbon resonances of the polymeric resin material as an internal standard. Results for several functionalized Wang and trityl resins are consistent with those obtained using well-established analytical methods. The (13)C NMR method has interesting applications in the field of solid-phase organic synthesis (SPOS), since no functional group acting as a support for the attachment of a quantifiable chromophore must be available in the material of interest.     

Influence of chemical modification of polymeric resin on retention of polar compounds in solid-phase extraction

Summary Three polymeric adsorbents, two of which had been chemically modified with different hydrophilic functional moieties and the third, which was the corresponding unmodified polystyrene-divinylbenzene (PSDVB) resin, were compared for solid-phase extraction (SPE) of several polar pesticides and phenolic compounds from water samples. The SPE system was online coupled to a liquid chromatograph with UV detector. Chemical modification of the PS-DVB resin with either 2-carboxy-3/4-nitrobenzoyl or 2,4-dicarboxybenzoyl, improved the efficiency of the SPE process by increasing polar interactions with the analytes. The adsorbent with the nitro group gave higher recoveries, mainly for the most polar analytes. This adsorbent enabled 100 mL river water to be preconcentrated to determine the target analytes in this matrix.     

Polystyrene resins cross-linked with di- or tri(ethylene glycol) dimethacrylates as supports for solid-phase peptide synthesis

Polystyrene resins cross-linked with di(ethylene glycol) dimethacrylate (DEGDMA) and tri(ethylene glycol) dimethacrylate (TEGDMA), DEGDMA-PS and TEGDMA-PS, were synthesized by suspension copolymerization. Four functionalized resins, chloromethyl resin, 4-hydroxymethylphenoxymethyl resin (Wang resin), 4-methylbenzhydrylamine resin (MBHA resin) and 2-chlorotrityl chloride resin, were prepared from DEGDMA-PS and TEGDMA-PS. DEGDMA-PS and TEGDMA-PS showed high reactivity in the functionalization reactions in comparison with Merrifield resin (polystyrene cross-linked with divinylbenzene, DVB-PS). DEGDMA-PS-Wang resin and TEGDMA-PS-Wang resin were used as the solid-phase support for the synthesis of a difficult sequence, the fragment of acyl carrier protein 65-74. The yields of the crude peptide synthesized using DEGDMA-PS-Wang resin, TEGDMA-PS-Wang resin and DVB-PS-Wang resin were 92.3%, 91.6% and 78.8%, respectively. The purities of the crude peptides were 85.7%, 88.1% and 73.3%, respectively.     

Influence of resin cross-linking on solid-phase chemistry

A range of PS-DVB resins were prepared by suspension polymerization with styrene, p-chloromethyl styrene, and DVB. Yields of polymerization increased (from 40% to almost 80%) with increasing cross-linking. The beads exhibited the expected swelling characteristics, with the 0.3% resin swelling to almost 9 times its dry volume in CHCl3. Kinetics of cleavage of the dye Methyl Red from the range of Rink linked resins showed rate enhancements of up to 500% between the 6.0 and the 0.3% cross-linked resins. Total synthesis of Kawaguchipeptin B was carried out on the resins, and their performance during the syntheses was investigated. Contrary to expectations, the purities of the cyclic peptide product increased with increasing resin cross-linking doubling from the 0.3-6.0% resin. A Suzuki reaction showed the half-lives of reaction increased with increasing resin cross-linking with an 11-fold increase in half-life between the 0.3-2.7% resin. Surprisingly, we observed very little reaction in the case of the 3.0 and 6.0% cross-linked resins.     

Magnetic nanoparticles as an orthogonal support of polymer resins: applications to solid-phase suzuki cross-coupling reactions

[reaction: see text] Most of the reactants immobilized on conventional solid-phase resins are buried inside the interiors of lightly cross-linked polystyrene beads. An orthogonal support of solid-phase resins needs to be small enough to penetrate the interpolymeric chain spaces of a swollen resin to reach reaction sites. In this paper, we report the use of magnetic nanoparticles ( approximately 4 nm) as an orthogonal matrix to assist solid-phase reactions. A magnetic nanoparticle-supported homogeneous Pd catalyst was employed for promoting the Suzuki cross-coupling of an aryl halide on resins and an excessive arylboronic acid in solution. The workup separating three components (the catalyst, product, and remaining arylborate) is a chromatography-free process. The Pd catalyst was magnetically isolated and recycled from the reaction mixture by applying an external magnetic field. Then, a filtration process was followed to recover the excess borate reagent from the resins/product. Our work here presents the first example of an orthogonal matrix of solid-phase resins and shows the promise of employing nanomaterials in organic synthesis.     

Functionalized cross-linked poly(vinyl alcohol) resins as reaction scavengers and as supports for solid-phase organic synthesis

New hydrophilic poly(vinyl alcohol) (PVA-OH) resins were prepared by an inverse suspension polymerization using epichlorohydrin as a cross-linker. These novel resins swell in a variety of solvents commonly used in solid-phase organic synthesis, such as dicholomethane, dioxane, methanol, tetrahydrofuran, and dimethylformamide. In addition, PVA-OH shows excellent swelling in water. The cross-linked PVA-OH beads were functionalized with an aldehyde group and were tested as scavengers for primary amines in three different reactions: amide bond formation, reductive amination reaction, and urea formation. With 1-2 equiv of the PVA aldehyde resin, all the excess primary amines were successfully scavenged. The utility of PVA-OH resins as solid supports in mono- and dipeptide synthesis was also investigated using symmetrical anhydride and MSNT/MeIm (2,4,6-mesitylenesulfonyl-3-nitro-1,2,4-triazolide in the presence of 1-methylimidazol) methods.     

Microwave-Assisted Solid Phase Organic Synthesis.Application to Indole Library Construction

Microwave-assisted organic synthesis (MAOS) has attained increasing popularity due to recent advancement in the instrumentation of microwave technology. Now, MAOS can be performed under controlled temperature and pressure to yield reproducible results. For combinatorial chemistry,the dramatically increased reaction rate under microwave irradiation at high temperature provides an ideal solution to those sluggish reactions, in particular the combinatorial reactions carried out on solid supports. In this presentation, we describe our results on microwave-assisted solid-phase organic synthesis (MASPOS) applied to the construction of indole libraries such as 5. Compounds 4 were synthesized on the Rink amide resins using IRORI MicroKanTM reactors encoded with a radio-frequency (Rf) tag. The resin-bound terminal alkynes 2, prepared via the amide bond, were cross-coupled with the nitroaryl triflate under the conditions adopted from the solution reactions developed by us1,2. The nitro group of 3 was then reduced and sulfonylated to give 4. Ring closure reactions within 4 with Cu(OAc)2 were examined initially in refluxing DCE for 24 h, but no indole product was detected after cleavage from the resin. Therefore, the same reactions were carried out under microwave irradiation at 200 ℃ for 10 min on a Personal Chemistry Emrys Creator, the desired indoles 5 were obtained in 60-95% overall yields calculated from 1 and in ＞90% purities in most cases3. It is necessary to mention that the IRORI microreactors cannot tolerate the high temperature and the resin-bound 4 must be transferred to the reaction vials for the microwave-assisted ring closure reactions. A traceless synthesis of an indole library via MASPOS will be discussed as well.4     

Synthesis and evaluation of poly(oxyethylene glycol) polymer (POP) supports

Mono- and alpha,omega-bis-styryl-oligo(oxyethylene glycol) ethers have been constructed in an efficient two-step synthesis. From these precursors, poly(oxyethylene glycol) polymer (POP) supports of varying monomer and cross-linker composition have been produced. The swelling properties and mass-solvent uptake of these novel materials have been evaluated in a variety of solvents, demonstrating that POP supports exhibit enhanced solvent compatibilities over the commercial resins TENTA-GEL, ARGO-GEL, and Merrifield's resin. The utility of POP supports in solid-phase organic chemistry has also been demonstrated successfully. It is anticipated that these high-loading polymeric supports will have generic application in the solid-phase synthesis of combinatorial libraries and the in situ screening of these libraries in the aqueous environment of a bioassay.     

Modern separation techniques for the efficient workup in organic synthesis

The shift of paradigm in combinatorial chemistry, from large compound libraries (of mixtures) on a small scale towards defined compound libraries where each compound is prepared in an individual well, has stimulated the search for alternative separation approaches. The key to a rapid and efficient synthesis is not only the parallel arrangement of reactions, but simple work-up procedures so as to circumvent time-consuming and laborious purification steps. During the initial development stages of combinatorial synthesis it was believed that rational synthesis of individual compounds could only be achieved by solid-phase strategies. However, there are a number of problems in solid-phase chemistry: most notably there is the need for a suitable linker unit, the limitation of the reaction conditions to certain solvents and reagents, and the heterogeneous reaction conditions. Further disadvantages are: the moderate loading capacities of the polymeric support and the limited stability of the solid support. In the last few years several new separation techniques have been developed. Depending on the chemical problem or the class of compounds to be prepared, one can choose from a whole array of different approaches. Most of these modern separation approaches rely on solution-phase chemistry, even though some of them use solid-phase resins as tools (for example, as scavengers). Several of these separation techniques are based on liquid-liquid phase separation, including ionic liquids, fluorous phases, and supercritical solvents. Besides being benign with respect to their environmental aspects, they also show a number of advantages with respect to the work-up procedures of organic reactions as well as simplicity in the isolation of products. Another set of separation strategies involves polymeric supports (for example, as scavengers or for cyclative cleavage), either as solid phases or as soluble polymeric supports. In contrast to solid-phase resins, soluble polymeric supports allow reactions to be performed under homogeneous conditions, which can be an important factor in catalysis. At the same time, a whole set of techniques has been developed for the separation of these soluble polymeric supports from small target molecules. Finally, miscellaneous separation techniques, such as phase-switchable tags for precipitation by chemical modification or magnetic beads, can accelerate the separation of compounds in a parallel format.     

Microwave-assisted solid-phase synthesis of hydantoin derivatives

A microwave-assisted synthesis of 3,5- and 1,3,5-substituted hydantoins starting from various resins for solid-phase combinatorial chemistry has been developed. The hydantoins were synthesized from pre-loaded resins with amino acids via treatment with isocyanate or phenylisocyanate and subsequent intramolecular cyclization. Both reactions were performed under microwave irradiation. We studied the cyclative cleavage leading to hydantoin compounds dependent on the nature of the amino acid and the nucleofuge properties of the resin.     

New cation-exchange resins with high reversed-phase character for solid-phase extraction of phenols

High capacity carboxylic acid functionalized resins, prepared by ring-opening metathesis polymerization, were used for solid-phase extraction (SPE) of phenols. Two resins, based on cross-linked poly-(endo,endo-norborn-2-ene-5,6-dicarboxylic acid) exhibited a capacity of 3.5 and 5.1 mequiv. COOH/g, respectively. Particle-loaded PTFE membranes were prepared from a 3.1 mequiv. COOH/g resin. The extraction behavior of the new materials versus two different EPA priority pollutant phenol standards was investigated. For most compounds, a quantitative recovery was observed. The extraction efficiency of the new resins was compared to those of other, commercially available materials such as silica C₁₈ or carboxypropylsilica. The general advantages of the new materials, the mechanism of extraction, the influence of polarity and acidity of the compounds investigated as well as differences between membranes and columns containing the new particles are discussed. Finally, the extraction efficiency of the new resins for phenols from spiked soil samples using both standard SPE as well as batch-techniques was investigated.     

Functionalized Polymeric Sorbents for Solid-Phase Extraction of Polar Pollutants

This study assesses how the physical characteristics of styrene-divinylbenzene (PS-DVB) resins affect the extent to which they are modified when they are functionalized, and how they affect recoveries in on-line solid-phase extraction (SPE) of some polar phenolic compounds, pesticides, and metabolites from water samples which were analyzed by liquid chromatography and UV detection. For this purpose, three commercial PS-DVB resins with different physical characteristics (Amberchrom GC-161m and two different PLRP-S resins) were chemically modified by placing a hydrophilic group, an o-carboxybenzoyl moiety, on their surface; although the physical characteristics are different, the extent of modification did not vary significantly. The results from the SPE process with each sorbent were related to their physical and chemical properties. The polymers with higher surface area provided better recoveries. Moreover, the recoveries for these analytes were better with the chemically-modified polymers due to the higher polarity of these sorbents. The best recovery values were with the sorbent obtained from the chemical modification of Amberchrom GC-161m, the commercial sorbent with higher surface area. For instance, in the analysis of 50 mL of a sample of 2 μg L^–1, the recovery of phenol was 40% with Amberchrom GC-161m and 65% with the chemically-modified Amberchrom GC-161m.