Purification of Boron Nitride Nanotubes Enhances Biological Application Properties.

Commercially available boron nitride nanotubes (BNNTs) and their purified form (pBNNTs) were dispersed in aqueous solutions with various dispersants, and their cytotoxicity and drug encapsulation capacity were monitored. Our data suggest that pBNNTs showed an average increase in dispersibility of 37.3% in aqueous solution in the presence of 10 different dispersants. In addition, 100 µg of pBNNTs induced an average decrease in cytotoxicity of 27.4% compared to same amount of BNNTs in normal cell lines. The same amount of pBNNTs can encapsulate 10.4-fold more drug (camptothecin) compared to BNNTs. These data suggest that the purification of BNNTs improves several of their properties, which can be applied to biological experiments and are thus essential in the biological application of BNNTs.

Development and validation of a simple method for the extraction and quantification of crisaborole in skin layers.

Crisaborole is a boron compound recently approved by the US Food and Drug Administration as a 2% ointment for the treatment of mild to moderate atopic dermatitis. This work describes a simple method for the quantification of the drug in the skin layers at the end of in-vitro permeation experiments. Chromatographic separation was carried out on a reverse-phase C18 column using a mixture of trifluoroacetic acid 0.05%-acetonitrile (55:45, v/v) as mobile phase, pumped at 1 ml/min. Column temperature was 35°C and UV detection was performed at 250 nm. The method was linear in the range of concentration from 0.06 to 6 µg/ml (R2 = 1) and was selective, precise and accurate. Depending on the solvent used, the LOQ ranged from 0.014 to 0.030 µg/ml and the LOD from 0.005 to 0.010 µg/ml. The extraction from all the skin layers was quantitative. The developed method was successfully tested in an in-vitro permeation study, proving to be an effective tool in the development of new formulations containing crisaborole.

An Electrokinetic Separation Route to Source Dialysate from Excess Fluid in Blood.

To improve the health of patients with end-stage renal disease, there is a clear need for slow, continuous hemodialysis, and the primary barrier to a wearable device is the requirement of a large reservoir of dialysate. We describe an electrokinetic means of producing dialysate from the excess fluid extant in the peripheral blood of patients undergoing therapy. A critical feature of this process is the retention of essential components of blood, especially serum albumin. In progress toward this goal, we demonstrate the separation of charged from neutral species in blood plasma at a branched microchannel junction by ion concentration polarization (ICP). Further, we introduce a method that reduces the opportunity for damage to proteins and prevents electrode biofouling. The present approach results in as high as 99.7% retention of albumin and successful separation of neutral metabolites and excess fluid to be utilized as a precursor to dialysate.

Development and Validation of an HPLC-UV Method to Quantify Tavaborole During in Vitro Transungual Permeation Studies.

A selective and rapid reversed-phase HPLC-UV method was developed and validated to quantify tavaborole (TAV; AN2690) in biological samples, i.e., in receiving phase and in bovine hoof membrane extract derived from in vitro transungual permeation studies. A simple solid-liquid extraction procedure was used to recover the drug from the bovine hoof slices. TAV chromatographic separation was achieved on a Luna PFP column (150 × 4.6 mm, 5 µm) using a mobile phase consisting of a 70% phosphoric acid solution (10 mM, pH 2.0) with 30% acetonitrile. The detection wavelength was set to 220 nm using a UV detector. The method exhibited good linearity in the calibration ranges, which were 0.5-8.0 and 0.03-2.5 µg/mL for the receiving phase and hoof membranes, respectively. The obtained LOD and LOQ values were 0.023 and 0.069 µg/mL, respectively, for the receiving phase and 0.0024 and 0.007 µg/mL for the bovine hoof membrane extracts. In all cases, the CV for intraday and interday precision was widely below the limit of 2%, demonstrating good precision. The analytical method described was sensitive, precise, linear, and accurate and could be applicable for clinical and bioanalytical studies as an alternative to other analytical methods, which are quite expensive and not always available in research laboratories.

Chiral separability of boron cluster species studied by screening approaches utilizing polysaccharide-based chiral stationary phases.

In this study, the screening steps of chiral separation strategies with polysaccharide-based chiral stationary phases were applied on boron cluster compounds in normal-phase liquid chromatography (NPLC) and polar organic solvents chromatography (POSC). Since the screening steps were initially developed to analyze organic compounds, their applicability for boron clusters was investigated. Overall, the screening steps in NPLC were applicable for the separation of zwitterions, while for anions mostly no elution was observed. A hypothesis for the latter behavior is precipitation of anions in the nonpolar mobile phases. Ten out of 11 compounds could be partially or baseline separated on the NPLC screening systems. In POSC, all zwitterions were separated on at least one of the screening systems, with an overall lower retention as in NPLC. Anions were detected but not separated in the majority of the experiments. Also their retention on the chiral stationary phases was very limited. This study showed that the chiral discrimination potential of chemically modified polysaccharides is meaningful for chiral separations of structurally chiral boron cluster species, but needs further systematic research, in which recognition mechanisms should be further explored. In addition, some unusual peaks also indicated that conditions with a high separation efficiency must first be searched for some of the tested systems.

Density gradient separation of carborane-containing liposome from low density lipoprotein and detection by inductively coupled plasma spectrometry.

Inductively coupled plasma atomic emission spectroscopy (ICP-AES) was used for analyzing the new cholesterol-based compounds (BCH, BCH-Da, BCH-Db and BCH-Dc) in liposomal formulations. Not only the boron compounds but also the phospholipid compositions of the liposome formulation were quantitatively analyzed. Reasonable limit of detection for boron (0.5 µg/ml) and phosphorous (0.09 µg/ml), respectively, was observed. ICP-MS was also utilized for analyzing BCH in a brain distribution study. The detection limit of boron analysis by ICP-MS is at least three orders of magnitude lower than of that of ICP-AES (1 ng B/ml). The method was linear in the range of 500-1 ng B/ml and the linearity correlation coefficient was 1. In addition, an ultracentrifugation method was developed to separate liposomes from low-density lipoprotein (LDL). Factors such as density gradient and size of liposomes were adjusted to optimize separation and it was observed that in conjunction to time, speed and density gradient, size of the liposome also had impact on the separation using centrifugation method. These findings show the importance of ICP-AES as an analytical method for the analysis of element-based compounds encapsulated in phospholipid vesicles.

Flow-injection MS/MS for gas-phase chiral recognition and enantiomeric quantitation of a novel boron-containing antibiotic (GSK2251052A) by the mass spectrometric kinetic method.

The present work demonstrates, for the first time, the application of the mass spectrometric kinetic method for quantitative chiral purity determination by automatic flow-injection MS/MS. The particular compound analyzed is GSK2251052A, a novel boron-containing systemic antibiotic for the treatment of multidrug-resistant Gram-negative bacterial infections. Chiral recognition and quantitation of GSK2251052A was achieved based on the competitive dissociation kinetics of the Cu(II)-bound trimeric complex [Cu(II)(A)(ref*)2-H](+) (A = GSK2251052A or its R-enantiomer, ref* = L-tryptophan) that gives rise to Cu(II)-bound dimeric complexes. The sensitive nature of the methodology and the linear relationship between the logarithm of the fragment ion abundance ratio and the optical purity, characteristic of the kinetic method, allow chiral purity determination of pharmaceutical compounds during enantioselective synthesis. By using flow-injection MS/MS, enantiomeric quantitation of GSK2251052A by the kinetic method proved to be fast (2 min for analysis of each sample) and to have accuracy comparable to chiral LC-MS/MS and LC-UV methods as well as the method using chiral derivatization followed by LC-MS/MS analysis. This flow-injection MS/MS method represents an alternative approach to commonly used chromatographic techniques as a means of chiral purity determination and is particularly useful for rapid screening of chiral drugs during pharmaceutical development.

Synthesis and characterization of B-heterocyclic π-radical and its reactivity as a boryl radical.

The first isolation and full characterization of the stable, persistent diazaboracyclic neutral radical 3 is reported. Reduction of base-stabilized difluororoborane 2 provided radical 3 as a neutral molecule having a planar sp(2) boron atom attached to one fluorine and two nitrogen atoms. ESR spectroscopy and DFT calculations indicated that the unpaired electron is delocalized over the six-membered ring. Because of an electronic transition related to the singly occupied molecular orbital, 3 has a characteristic red color, as UV-vis spectroscopy showed an absorption maximum at 498 nm. Although DFT calculations suggested that radical 3 has relatively low spin density on the boron atom in comparison with the nitrogen and carbon atoms in the six-membered ring, 3 reacted as a base-stabilized boryl radical when treated with benzoquinone or benzoyl peroxide.

Isolation of individual boron nitride nanotubes via peptide wrapping.

The isolation of individual boron nitride nanotubes (BNNTs) in aqueous phases has been achieved for the first time from raw materials based on the combination of peptide wrapping with a sonication procedure. Atomic force microscopic observations revealed the representative height and length of individual BNNTs. Fluorescence and infrared absorption spectra suggested the strong pi-pi interactions between BNNTs and the peptide. The absorption maxima of BNNTs were significantly blue-shifted from 200 nm for the original BNNTs to 193 nm. The modulation of the BNNT band gap with peptide wrapping promises potential applications of the peptide/BNNT complexes to various nanotechnologies.

Purification of single-walled boron nitride nanotubes and boron nitride cages.

Continuous laser vaporization of a BN target under N2 atmosphere is up to now the unique route to single-walled boron nitride nanotubes (BN-SWNTs). Although grams of product can be obtained by this technique, the raw material contains in addition to the BN-SWNTs, different by-products made of boron and nitrogen. Since these materials are undesirable for the studying of the intrinsic properties of the nanotubes, we have undertaken a purification process using chemical and physical methods to separate the different components. We show here that most impurities can be removed by successive cycles of washing, sonication, and centrifugation. Furthermore, the two different types of boron nitride nanostructures i.e., BN-SWNTs and BN-cages can be isolated. Efficiency of the separation was monitored by transmission electron microscopy (TEM) at the different steps of the process. Finally, we envisage the further purification of the nanotubes-enriched fraction by functionalizing the nanotubes in a non covalent manner by specific polymers as for carbon nanotubes and BN multi-walled nanotubes.

Boron carbonitride nanotubes.

A comprehensive understanding of the design, synthesis, characterization, and properties of boron carbonitride nanotubes (BCN) is presented in this review. Distinctive structural and electronic properties are revealed in theoretical studies of the BCN nanotubes and compared with the properties of carbon nanotubes. In the experimental studies, BCN nanotubes have been synthesized by various techniques. For different purposes, controllable growth processes have been used to fabricate BCN nanotubes with novel structures, such as nanojunctions and filled nanotubes. Some interesting phenomena originating from the substitution of B and N atoms, such as the phase segregation, are considered theoretically and experimentally. Mainly the physical properties--field electron emission and photoluminescence--are discussed, which turn out to have potential applications in the industry.

An integrated solid-phase extraction system for sub-picomolar detection.

A microchip structure etched on a glass substrate for packed column solid-phase extraction (SPE) and capillary electrochromatography (CEC) is described. A 200 microm long, octadecylsilane (ODS) packed column was secured using two different approaches: solvent lock or polymer entrapment. The former method was utilized for SPE while the latter approach was applied for CEC. In SPE, the ODS packed chamber gave a detection limit of 70 fM for a nonpolar BODIPY (493/503) dye when concentrated for 3 min at an electroosmotic flow rate of 4.14 nL/min, compared to 30 pM for this detector without the SPE step. SPE beds showed reproducible, linear calibration curves (R(2) = 0.9989) between 1 and 100 pM BODIPY at fixed preconcentration times. Breakthrough curves for the 330 pL (ODS-packed) bed indicated a capacity for BODIPY dye of 8.1 x 10(-14) mmol, or 0.25 mmol dye per liter of bed. The ODS-chamber could also be used to analyze dilute amino acid and peptide solutions. In the CEC format, two neutral dyes (BODIPY and acridine orange) were baseline-separated in an isocratic run with a theoretical plate count of 84 (420 000 plates/m) and a reduced plate height of about 1. A labeled peptide was also analyzed by CEC, using the acidic eluent (84% acetonitrile, and 26% aqueous trifluoroacetic acid (0.05%)) preferred for peptide separations on ODS-coated silica particles.

Increase of separation resolution through field enhancement in microchips.

An enhanced ability to separate charged species from neutral compounds in a microfluidic chip is demonstrated using a chip design with low-resistance electrode channels operating with a multiport pressure/voltage controller. A factor of 2.7 improvement in resolution was obtained from chips made using identical mask designs but different etch depth protocols. Greater separation power allows one to cover a wider dynamic range for compounds with different electrophoretic mobilities.

Synthesis and separation of diastereomers of uridine 2',3'-cyclic boranophosphate.

The first boron-containing 2',3'-cyclic phosphate-modified analogue, uridine 2',3'-cyclic boranophosphate (2',3'-cyclic-UMPB), was synthesized. 5'-O-Protected uridine was cyclophosphorylated by diphenyl H-phosphonate to yield uridine 2',3'-cyclic H-phosphonate, which upon silylation followed by boronation and subsequent acid treatment gave 2',3'-cyclic-UMPB in high yield. The two diastereomers of 2',3'-cyclic-UMPB were separated by HPLC. An alternative method for synthesis of uridine 2',3'-cyclic phosphorothioate (2',3'-cyclic-UMPS) via H-phosphonate was also described.

Nanotubes of boron nitride filled with molybdenum clusters.

Carbon nanotubes are known to be metallic or semiconducting, depending on their helicity and diameter. However, boron nitride (BN) nanotubes are the only nanotubular product known to date that are predicted to have stable insulating properties that are independent of their atomic structure and morphology. Thus, the BN tube has attracted prime attention as an advanced nanoinsulating shield for all types of encapsulated conducting material, i.e., metal wires, clusters, etc. However, so far there have been no successes in controlled one-dimensional filling of BN nanotubes with conductive material. We report the first experimental results on the synthesis, high-resolution transmission electron microscopy, energy dispersion X-ray analysis, and electron energy loss spectroscopy of BN nanotubes that are filled with Mo clusters over their entire length. This was accomplished by means of two-step thermochemical treatment of chemically vapor-deposited C nanotubes with B2O3, CuO, and MoO3 oxides in a flowing N2 atmosphere. The first examples of BN nanotubes filled with molybdenum clusters are reported and the formation of the first nanocable (approximately 10 nm in length), consisting of a conductive metal core and an insulating BN nanotubular shield is demonstrated.

Boron carbonitride nanofibers: synthesis, characterization, and photoluminescence properties.

Large-scale highly aligned boron carbonitride (BCN) nanofibers with controllable orientations and chemical compositions were synthesized directly on nickel substrates from a gas mixture of N2, H2, CH4, and B2H6 by plasma-enhanced hot filament chemical vapor deposition. The morphology of the BCN nanofibers was examined by scanning electron microscopy, the microstructures were studied by high-resolution transmission electron microscopy, and the bonding states as well as chemical compositions were determined by electron energy loss spectroscopy. To our knowledge, this is the first report on the photoluminescent properties of BCN nanofibers that shows that they are interesting blue- and violet-light-emitting materials with tunable wavelengths. Further studies on field electron emission suggest that BCN nanofibers are also promising candidates for field emission sources.

Synthesis, characterization, and magnetic properties of carbon- and boron-oxide-encapsulated iron nanocapsules.

Carbon- and boron-oxide-encapsulated iron nanocapsules have been synthesized by arc discharge in methane (CH4) and diborane (B2H6) atmospheres respectively. The characterization and magnetic properties of carbon- and boron-oxide-encapsulated iron nanocapsules [abbreviated as Fe(C) and Fe(B) respectively] were investigated and compared. The structure of the Fe(B) nanocapsules is different from that of the Fe(C) nanocapsules. The Fe(C) nanocapsules consist of a crystalline graphite shell and a core of alpha-Fe and/or Fe3C. The Fe(B) nanocapsules consist of an amorphous boron-oxide layer and a core of Fe(B) solid solution, alpha-Fe, gamma-Fe, FeB, and/or Fe3B phases. The saturation magnetizations of both the Fe(C) and the Fe(B) nanocapsules below 300 K decrease monotonically with increasing temperature. The coercivities of the Fe(C) and Fe(B) nanocapsules are almost 2 orders of magnitude higher than that of bulk Fe. The temperature dependence of magnetization at high temperatures indicates the existence of some phase transformations.

Photoinduced superhydrophilic properties of Ti-B binary oxide thin films and their photocatalytic reactivity for the decomposition of NO.

Characterizations of Ti-B binary oxide thin films by means of various spectroscopic measurements have shown that Ti-B binary oxide thin films are formed by ultra fine TiO2 nanoparticles. A dramatic decrease in the contact angle of water droplets to 0 degree under ultraviolet (UV) light irradiation and a return to only half of its initial value under dark conditions were observed for the Ti-B binary oxide thin films. Moreover, ultraviolet irradiation of these thin film photocatalysts in the presence of NO led to photocatalytic decomposition of NO into N2 and O2. Especially when the Ti content was low, the size of the primary nanoparticles of the thin films was smaller and the photocatalytic reaction proceeded efficiently with a high selectivity for the formation of N2 and O2 from NO.

Hydrazide as a ligand moiety in immobilized metal ion affinity chromatography. Separation of BO-IMI and BODIPY-hydrazide.

BODIPY hydrazide (BO-HZ, a commercially available fluorescent dye) and BO-IMI (obtained by coupling the hydrazide moiety of BODIPY to the carboxyl group of N-acetylhistidine) were separated on three forms of a Sepharose-iminodiacetic acid column: Cu(II), Ni(II) and Zn(II). Whereas BO-IMI eluted first on the Cu(II) and Ni(II) columns (a pH gradient from 7.0 to 2.0 was applied), it eluted last on the Zn(II) column. BO-HZ eluted from the Zn(II) column without displacing this metal. The explanation suggested for these results is that BODIPY hydrazide undergoes strong, bidendate binding only to the Cu(II) and Ni(II) columns.

Conformational studies of dithymidine boranomonophosphate diastereoisomers.

The boranophosphate ester nucleotides are a new class of nucleic acid analogues that are isoelectronic and isostructural to normal phosphodiester nucleic acids and that maintain the anionic charge of the nucleic acid backbone. The two P-diastereoisomers of dithymidine boranomonophosphates were separated using reverse phase HPLC; the faster and slower eluting isomers are designated as d(TpBT)-1 and d(TpBT)-2, respectively. Conformations of the isomers were studied using-circular dichroism (CD) and NMR, and compared to the analogous phosphate diester, d(TpT). This comparison allowed the effects of the borane group and chirality of the boranophosphate linkage on sugar and base conformations to be assessed. The CD spectra of the diastereoisomers are consistent with both having a B-type conformation. Analysis of the 1H-1H and 1H-31P coupling constants showed that these conformations are similar to those of the unmodified parent dimer; specifically, the 2'-deoxyribose rings prefer the S (C2'-endo) conformation, and the C4'-C5' and C5'-O5' rotamers are primarily in the gamma + and beta + conformations, respectively. Conformational differences between the diastereoisomers and between the modified and unmodified dimers are manifested by differences in the preferences of the 3'-residues to adopt S sugar pucker and beta + conformations. There is reduced preference for the S sugar pucker of the 3'-residue in d(TpBT)-1 relative to d(TpBT)-2, which is similar to d(TpT). There is less preference for the beta + conformation of the 3'-residue in d(TpBT)-2 relative to d(TpBT)-1 and d(TpT). Based on the CD results, the temperature dependences of the thymidine H6 chemical shifts, and the derived sugar ring and backbone conformational parameters, we conclude that the borane group exerts a minimal influence on the sugar conformations and base stacking interactions. Preliminary assignment of the absolute configuration of the pair of SP and RP diastereoisomers to d(TpBT)-1 and d(TpBT)-2, respectively, is made on the basis of enzyme selectivity and NOE difference experiments.