Analysis and electroanalysis of perchlorate in water and food samples: a critical review.

Perchlorate, ClO4-, with diverse applications, has become one of the major contaminants in surface and groundwater sources. This highly soluble and stable anion poses a considerable threat to human health given that it contaminates drinking water, vegetables, milk, and other contaminated food products. ClO4- can impair the thyroid function; thus, drinking water with high levels of this anion is a severe problem worldwide. However, due to the high solubility, stability, and mobility of ClO4-, its remediation and monitoring remain a major challenge. Considering the various analytical methods, including electrochemistry, each method has advantages and disadvantages in terms of detection sensitivity, selectivity, analysis time, and cost. Also, sample preconcentration and clean-up must be performed for the analysis of more complex matrices such as food and biological samples to ensure a low detection limit and selectivity. Both ion chromatography (IC) and capillary electrophoresis (CE) coupled with electrochemical detection, in addition to liquid chromatography (LC)-mass spectrometry (MS), are expected to play key roles due to their lower detection limit with excellent sensitivities and selectivity. Herein, we also discuss the perspective on various electrode materials for the detection of ClO4- regarding whether ClO4- can be measured at the lowest levels with the highest selectivity.

Evaluation of hydrophilic interaction chromatography versus reversed-phase chromatography for fast aqueous species distribution analysis of Nickel(II)-Histidine complex species.

Nickel (Ni) is a trace heavy metal of importance in biological and environmental systems, with well documented allergy and carcinogenic effects in humans. With Ni(II) as the dominant oxidation state, the elucidation of the coordination mechanisms and labile complex species responsible for its transportation, toxicity, allergy, and bioavailability is key to understanding its biological effects and location in living systems. Histidine (His) is an essential amino acid that contributes to protein structure and activity and in the coordination of Cu(II) and Ni(II) ions. The aqueous low molecular weight Ni(II)-Histidine complex consists primarily of two stepwise complex species Ni(II)(His)1 and Ni(II)(His)2 in the pH range of 4 to 12. Four chromatographic columns, including the superficially porous Poro-shell EC-C18, Halo RP-amide and Poro-shell bare silica-HILIC columns, alongside a Zic-cHILIC fully porous column, were evaluated for the fast separation of the individual Ni(II)-Histidine species. Of these the Zic-cHILIC exhibited high efficiency and selectivity to distinguish between the two stepwise species Ni(II)His1 and Ni(II)His2 as well as free Histidine, with a fast separation within 120 s at a flow rate of 1 ml/min. This HILIC method utilizing the Zic-cHILIC column was initially optimized for the simultaneous analysis of Ni(II)-His-species using UV detection with a mobile phase consisting of 70% ACN and sodium acetate buffer at wwpH 6. Furthermore, the aqueous metal complex species distribution analysis for the low molecular weight Ni(II)-histidine system was chromatographically determined at various metal-ligand ratios and as a function of pH. The identities of Ni(II)His1 and Ni(II)-His2 species were confirmed using HILIC electrospray ionization- mass spectrometry (HILIC-ESI-MS) at negative mode.

Rapid and sensitive simultaneous separation and electrochemical detection of tetracaine hydrochloride and oxymetazoline hydrochloride in pharmaceutical formulations via core-shell reversed-phase liquid chromatography.

Tetracaine hydrochloride (TCH) is a nasal anesthetic and oxymetazoline hydrochloride (OZH) is a nasal decongestant. A moderate to acute overdosage of OZH and TCH can lead to mydriasis, nausea, cyanosis, tachycardia, dyspnoea, cardiovascular failure, disorientation, seizures, and even death. Liquid chromatography (LC) has been mainly utilized for the individual determination of either TCH or OZH; however, there is a need for rapid and efficient methods for simultaneous analysis in pharmaceutical formulations and aqueous samples. This study highlights the use of the fast and efficient separation capabilities of core-shell silica particles in liquid chromatography (LC) for the simultaneous determination of TCH and OZH using UV detection and the enhanced selectivity afforded by electrochemical detection at a boron-doped diamond (BDD) electrode. Rapid reversed-phase (RP) separation and detection of OZH and TCH in nasal spray and eye drops was achieved within 45 s using a poroshell 120 EC-C18 column, by adjusting the ratio of organic solvent, mobile phase pH, detection potential and mobile phase flow rate. Sensitivity was compared using ultraviolet (UV) detection at 280 nm, and ECD at + 1.3 V with detection limits of 40 and 70 nM for TCH and OZH, respectively. The developed rapid method was utilized successfully in the analysis of pharmaceutical formulations, where the estimated levels of TCH and OZH in these formulations are in agreement with the specified values outlined by the manufacturers.

Isoquinolinequinone N-oxides as anticancer agents effective against drug resistant cell lines.

The isoquinolinequinone (IQQ) pharmacophore is a privileged framework in known cytotoxic natural product families, caulibugulones and mansouramycins. Exploiting both families as a chemical starting point, we report on the structured development of an IQQ N-oxide anticancer framework which exhibits growth inhibition in the nM range across melanoma, ovarian and leukaemia cancer cell lines. A new lead compound (16, R6 = benzyl, R7 = H) exhibits nM GI50 values against 31/57 human tumour cell lines screened as part of the NCI60 panel and shows activity against doxorubicin resistant tumour cell lines. An electrochemical study highlights a correlation between electropositivity of the IQQ N-oxide framework and cytotoxicity. Adduct binding to sulfur based biological nucleophiles glutathione and cysteine was observed in vitro. This new framework possesses significant anticancer potential.

Profiling of phenolic flavorings using core-shell reversed-phase liquid chromatography with electrochemical detection at a boron-doped diamond electrode.

A high-performance liquid chromatography (HPLC) method equipped with a boron-doped diamond (BDD) electrode was established for the simultaneous determination of phenol, 4-ethylphenol (4-EP), guaiacol, 4-ethylguaiacol (4-EG), 4-vinylguaiacol (4-VG), eugenol, and o-, m- and p-cresol. The separation was performed on a reversed-phase HALO C18 core-shell column (3.0 × 50 mm, 2.7 µm) with a mobile phase comprising 10 mM formate, pH 3, and 15% acetonitrile (ACN) (v/v), a flow rate of 1.5 mL/min, corresponding to a total run time of 9 min. The electrochemical detection (ECD) was set at +1.5 V vs. Pd/H2 in oxidative mode. Under optimized operating conditions, good linearity was obtained for the nine phenolics with corresponding coefficients of determination (R2) above 0.998. The limits of detection (LODs, S/N = 3) were 10 nM-1 µM, with an 80-fold increase in sensitivity for guaiacol achieved with ECD over ultraviolet (UV) detection. The sensitive and selective HPLC-ECD method was successfully applied for the identification and quantification of the nine phenolics in Islay, Irish, Scotch, and Highland whiskey samples, with significantly higher concentrations of the flavorings determined in Islay whiskey.

Biotin interference in immunoassays based on biotin-strept(avidin) chemistry: An emerging threat.

Biotinylated antibodies/antigens are currently used in many immunoassay formats in clinical settings for diversified analytes and biomarkers to offer high detection selectivity and sensitivity. Biotin cannot be synthesized by mammals and must be taken as an essential supplement. Normal intake of biotin from various foods and milk causes no effect on the streptavidin/biotin-based immunoassays. However, overconsumption of biotin (daily doses 100-300 mg) poses a significant problem for immunoassays using the biotin-strept(avidin) pair. Biotin interferences are noted in immunoassays of thyroid markers, drugs, hormones, cancer markers, the biomarker for cardiac function (ß-human chorionic gonadotropin), etc. The biotin level required for serious interference in test results varies significantly from test to test and cannot easily be predicted. Immunoassay manufacturers with technologies based on strept(avidin)-biotin binding must investigate the interference from biotin (up to at least 1200 ng/mL or 4.9 µM of biotin) in various formats. There is no concrete solution to circumvent the biotin interference encountered in blood samples, short of biotin removal. Considering the short half-life of biotin in the human body, patients must stop taking biotin supplements for >48 h before the test. However, this scenario is not considered for patients in emergency situations or those with biotinidase deficiency, mitochondrial metabolic disorders or multiple sclerosis. Apparently, a rapid analytical procedure for biotin is urgently needed to quantify for its interference in immunoassays using strep(avidin)-biotin chemistry. To date, there is no quick and reliable procedure for the detection of biotin at below nanomolar levels in blood and biological samples. Traditional lab-based techniques including HPLC/MS-MS cannot process an enormous number of public samples. Biosensors with high detection sensitivity, miniaturization, low cost, and multiplexing have the potential to address this issue.

Captavidin as a regenerable biorecognition element on boron-doped diamond for biotin sensing.

Captavidin, a nitrated avidin with moderate affinity with biotin, irreversibly adsorbs on carboxymethylcellulose to form a regenerable biorecognition element for biotin. This layer was retained and stabilized on a boron-doped diamond electrode by a Nafion film for repeated impedance analyses of biotin down to below 1 nM. The labeless electrochemical sensing scheme was then demonstrated for the analysis of biotin in blood plasma. The incorporation of captavidin confers detection specificity and regenerability, whereas the Nafion and carboxymethylcellulose layers circumvent the diffusion of endogenous electroactive species. The biosensing layer is simply regenerated by applying oxidation of +2 V for 1 min instead of its submersion in carbonate buffer for 10 min.

Electrochemical Sensing of Biotin Using Nafion-Modified Boron-Doped Diamond Electrode.

Nafion formed on the surface of a boron-doped diamond electrode allows for a chemosensing system for biotin. The modified electrode is capable of oxidizing biotin and offers a detection limit of 5 nM, the average normal level of biotin in blood plasma. The developed method was successfully applied to determine biotin in human plasma samples and a popular health product as two popular models.

Rapid Electrochemical Detection of Pseudomonas aeruginosa Signaling Molecules by Boron-Doped Diamond Electrode.

As the leading cause of morbidity and mortality of cystic fibrosis (CF) patients, early detection of Pseudomonas aeruginosa (PA) is critical in the clinical management of this pathogen. Herein, we describe rapid and sensitive electroanalytical methods using differential pulse voltammetry (DPV) at a boron-doped diamond (BDD) electrode for the detection of PA signaling biomolecules. Monitoring the production of key signaling molecules in bacterial cultures of P. aeruginosa PA14 over 8 h is described, involving sample pretreatment by liquid-liquid and solid-phase extraction. In addition, direct electrochemical detection approach of PA signaling molecules is also reported in conjunction with hexadecyltrimethylammonium bromide (CTAB) to disrupt the bacterial membrane.

Supercritical carbon dioxide versus toluene as reaction media in silica functionalisation: Synthesis and characterisation of bonded aminopropyl silica intermediate.

This research reports supercritical carbon dioxide versus toluene as reaction media in silica functionalisation for use in liquid chromatography. Bonded aminopropyl silica (APS) intermediates were prepared when porous silica particles (Exsil-pure, 3µm) were reacted with 3-aminopropyltriethoxysilane (3-APTES) or N,N-dimethylaminopropyltrimethoxysilane (DMAPTMS) using supercritical carbon dioxide (sc-CO2) and toluene as reaction media. Covalent bonding to silica was confirmed using elemental microanalysis (CHN), thermogravimetric analysis (TGA), zeta potential (ξ), diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy, scanning electron microscopy (SEM) and solid-state nuclear magnetic resonance (CP/MAS NMR) spectroscopy. The results demonstrate that under sc-CO2 conditions of 100°C/414bar in a substantial reduced time of 3h, the surface coverage of APS (evaluated from%C obtained from elemental analysis) prepared with APTES (%C: 8.03, 5.26µmol/m-2) or DMAPTES (%C: 5.12, 4.58µmol/m2) is somewhat higher when compared to organic based reactions under reflux in toluene at a temperature of 110°C in 24h with APTES (%C: 7.33, 4.71µmol/m2) and DMAPTMS (%C: 4.93, 4.38µmol/m2). Zeta potential measurements revealed a change in electrostatic surface charge from negative values for bare Exsil-pure silica to positive for functionalised APS materials indicating successful immobilization of the aminosilane onto the surface of silica.

Synthesis and characterisation of non-bonded 1.7µm thin-shell (TS1.7-100nm) silica particles for the rapid separation and analysis of uric acid and creatinine in human urine by hydrophilic interaction chromatography.

A rapid chromatographic method for the simultaneous determination of uric acid (UA) and creatinine (Cr) in human urine is described, using a non-bonded 1.7µm thin-shell (TS1.7-100nm) silica particle prepared by the seeded-growth approach. The new shell particle was characterised by scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS) and BET analysis. TEM reveals 1.5µm solid core and 100nm shell thickness. DLS shows polydispersity index <0.2. Expanded pore size of 88Å and specific surface area of 78m2/g were determined by BET. Chromatographic results demonstrate that UA, Cr and hypoxanthine (Hyp, as internal standard) can be separated in less than 1min on the in-house packed TS1.7-100 column (4.6 ID x 100mm), using chromatographic conditions with mobile phase 70% acetonitrile, 10mM ammonium acetate buffer, pH 6.78, flow rate 1.25ml/min and UV detection at 254nm. A linear relationship between the ratio of the peak area of the standard UA and Cr to that of the internal standard (Hyp) and the concentration of standards was obtained for both UA and Cr with the square of the correlation coefficients, R2=0.998 for both renal biomarkers. The calculated detection limits were 0.03µg/ml and 0.05µg/ml for UA and Cr respectively. Urine samples tested were found to contain UA and Cr in the concentration range of 782-1206µg/ml and 535-862µg/ml respectively. The recovery ranges for spiked urine standards were 85.7-93.2% for UA and 91.9-94.6% for Cr and the relative standard deviations (RSD) for both biomarkers were 3.05% and 0.88% respectively. The developed rapid HILIC method can have application in determining the concentrations of UA and Cr for early prediction in patients with developing disease conditions, including acute kidney injury (AKI).

A review of oxyhalide disinfection by-products determination in water by ion chromatography and ion chromatography-mass spectrometry.

This paper is a review of ion chromatographic (IC) separations of inorganic oxyhalide disinfection by-products (DBPs) in water and beverages. The review outlines the chemical mechanisms of formation, regulation of maximum allowable levels, chromatographic column selection and speciation. In addition, this review highlights the application of IC coupled to mass spectrometry (MS) for trace and elemental composition analysis of oxyhalides, along with the analytical considerations associated to enable sensitive analysis. Furthermore, a review of literature concerning IC determination of inorganic oxyhalide DBPs in environmental matrices, including water, published since 2005 is presented, with a focus on MS detection, and a discussion on the relative performance of the methods. Finally some prospective areas for future research, including fast, selective, multi-analyte analysis, for this application are highlighted and discussed.

Molecular Signature of Pseudomonas aeruginosa with Simultaneous Nanomolar Detection of Quorum Sensing Signaling Molecules at a Boron-Doped Diamond Electrode.

Electroanalysis was performed using a boron-doped diamond (BDD) electrode for the simultaneous detection of 2-heptyl-3-hydroxy-4-quinolone (PQS), 2-heptyl-4-hydroxyquinoline (HHQ) and pyocyanin (PYO). PQS and its precursor HHQ are two important signal molecules produced by Pseudomonas aeruginosa, while PYO is a redox active toxin involved in virulence and pathogenesis. This Gram-negative and opportunistic human pathogen is associated with a hospital-acquired infection particularly in patients with compromised immunity and is the primary cause of morbidity and mortality in cystic fibrosis (CF) patients. Early detection is crucial in the clinical management of this pathogen, with established infections entering a biofilm lifestyle that is refractory to conventional antibiotic therapies. Herein, a detection procedure was optimized and proven for the simultaneous detection of PYO, HHQ and PQS in standard mixtures, biological samples, and P. aeruginosa spiked CF sputum samples with remarkable sensitivity, down to nanomolar levels. Differential pulse voltammetry (DPV) scans were also applicable for monitoring the production of PYO, HHQ and PQS in P. aeruginosa PA14 over 8 h of cultivation. The simultaneous detection of these three compounds represents a molecular signature specific to this pathogen.

Synthesis and electrochemical detection of a thiazolyl-indole natural product isolated from the nosocomial pathogen Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen, capable of surviving in a broad range of natural environments and quickly acquiring resistance. It is associated with hospital-acquired infections, particularly in patients with compromised immunity, and is the primary cause of morbidity and mortality in cystic fibrosis (CF) patients. P. aeruginosa is also of nosocomial importance on dairy farms and veterinary hospitals, where it is a key morbidity factor in bovine mastitis. P. aeruginosa uses a cell-cell communication system consisting of signalling molecules to coordinate bacterial secondary metabolites, biofilm formation, and virulence. Simple and sensitive methods for the detection of biomolecules as indicators of P. aeruginosa infection would be of great clinical importance. Here, we report the synthesis of the P. aeruginosa natural product, barakacin, which was recently isolated from the bovine ruminal strain ZIO. A simple and sensitive electrochemical method was used for barakacin detection using a boron-doped diamond (BDD) and glassy carbon (GC) electrodes, based on cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The influence of electrolyte pH on the peak potential and peak currents was also investigated. At pH 2.0, the peak current was linearly dependent on barakacin concentration (in the range used, 1-10 µM), with correlation coefficients greater than 0.98 on both electrodes. The detection limit (S/N = 3) on the BDD electrode was 100-fold lower than that obtained on the GC electrode. The optimized method using the BDD electrode was extended to bovine (cow feces) and human (sputum of a CF patient) samples. Spiked barakacin was easily detected in these matrices at a limit of 0.5 and 0.05 µM, respectively. Graphical abstract Electrochemical detection of barakacin.

Sub-2-µm seeded growth mesoporous thin shell particles for high-performance liquid chromatography: Synthesis, functionalisation and characterisation.

Nanometer control over the porous shell thickness of sub-2-µm-shell particles is investigated. Three seeded growth mesoporous thin shell particles for HPLC were prepared, with 0.05µm (or 50nm) porous shell layers: particle sizes 1.5µm (solid core diameters 1.4µm), 1.7µm (solid core diameter 1.6µm), 1.9µm (solid core diameter 1.8µm) and compared with a fourth 1.7µm particle (solid core diameter 1.4µm) surrounded by 0.15µm (or 150nm) porous shell thickness. The thin shell particles were functionalised using a mono-functional octadecyldimethylchlorosilane ligand (C20H43SiCl) under optimised reflux conditions and packed in-house in narrow bore columns (2.1 I.D.×50mm) denoted as TS1.5-50-C18, TS1.7-50-C18, and TS1.9-50-C18 respectively. The synthesised thin shell particles and bonded materials were comprehensively characterised using scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential, BET analysis, elemental analysis (CHN), thermogravimetric analysis (TGA) and diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. Experimental data from inverse size exclusion chromatography (ISEC) was used to measure external, internal and total column porosities. Five probe analytes (uracil, naphthalene, acetophenone, benzene and toluene) were chosen for the chromatographic performance analysis of these columns. Column evaluation and measurements of height equivalent to a theoretical plate (HETP) data were performed on naphthalene using 55% acetonitile in water. The retention coefficients for the thin shell particles (TS1.9-50-C18, TS1.7-50-C18, TS1.5-50-C18) were in the range 1.26-1.35 and 5.6 for the core-shell particle (EiS1.7-150-C18). The minimum reduced plate heights range from 3.89 to 4.26 for the thin shell particles and 2.03 for the core-shell particle.

One step preparation and electrochemical analysis of IQS, a cell-cell communication signal in the nosocomial pathogen Pseudomonas aeruginosa.

Pseudomonas aeruginosa uses a hierarchical cell-cell communication system consisting of a number of regulatory elements to coordinate the expression of bacterial virulence genes. Sensitive detection of quorum sensing (QS) molecules has the potential for early identification of P. aeruginosa facilitating early medical intervention. A recently isolated cell-cell communication molecule, a thiazole termed IQS, can bypass the las QS system of P. aeruginosa under times of stress, activating a subset of QS-controlled genes. This compound offers a new target for pathogen detection and has been prepared in a one step protocol. A simple electrochemical strategy was employed for its sensitive detection using boron-doped diamond and glassy carbon electrodes by cyclic voltammetry and amperometry.

A new N-hydroxyethyliminodiacetic acid modified core-shell silica phase for chelation ion chromatography of alkaline earth, transition and rare earth elements.

Bare core-shell silica (1.7µm) has been modified with iminodiacetic acid functional groups via standard silane chemistry, forming a new N-hydroxyethyliminodiacetic acid (HEIDA) functionalised core-shell stationary phase. The column was applied in high-performance chelation ion chromatography and evaluated for the retention of alkaline earth, transition and heavy metal cations. The influence of nitric acid eluent concentration, addition of complexing agent dipicolinic acid, eluent pH and column temperature on the column performance was investigated. The efficiencies obtained for transition and heavy metal cations (and resultant separations) were comparable or better than those previously obtained for alternative fully porous silica based chelation stationary phases, and a similarly modified monolithic silica column, ranging from ∼15 to 56µm HETP. Increasing the ionic strength of the eluent with the addition of KNO3 (0.75M) and increasing the column temperature (70°C) facilitated the isocratic separation of a mixture of 14 lanthanides and yttrium in under 12min, with HETP averaging 18µm (7µm for Ce(III)).

Fabrication and characterization of nanotemplated carbon monolithic material.

A novel hierarchical nanotemplated carbon monolithic rod (NTCM) was prepared using a novel facile nanotemplating approach. The NTCM was obtained using C60-fullerene modified silica gels as hard templates, which were embedded in a phenolic resin containing a metal catalyst for localized graphitization, followed by bulk carbonization, and template and catalyst removal. TEM, SEM, and BET measurements revealed that NTCM possessed an integrated open hierarchical porous structure, with a trimodal pore distribution. This porous material also possessed a high mesopore volume and narrow mesopore size distribution. During the course of carbonization, the C60 conjugated to aminated silica was partly decomposed, leading to the formation of micropores. The Raman signature of NTCM was very similar to that of multiwalled carbon nanotubes as exemplified by three major peaks as commonly observed for other carbon materials, i.e., the sp3 and sp2 carbon phases coexisted in the sample. Surface area measurements were obtained using both nitrogen adsorption/desorption isotherms (BET) and with a methylene blue binding assay, with BET results showing the NTCM material possessed an average specific surface area of 435 m2 g(-1), compared to an area of 372 m2 g(-1) obtained using the methylene blue assay. Electrochemical studies using NTCM modified glassy carbon or boron doped diamond (BDD) electrodes displayed quasi-reversible oxidation/reduction with ferricyanide. In addition, the BDD electrode modified with NTCM was able to detect hydrogen peroxide with a detection limit of below 300 nM, whereas the pristine BDD electrode was not responsive to this target compound.

Spectroscopic and chromatographic characterisation of a pentafluorophenylpropyl silica phase end-capped in supercritical carbon dioxide as a reaction solvent.

This research uses solid-state nuclear magnetic resonance (NMR) spectroscopy to characterise the nature and amount of different surface species, and chromatography to evaluate phase properties of a pentafluorophenylpropyl (PFPP) bonded silica phase prepared and end-capped using supercritical carbon dioxide (sc-CO2) as a reaction solvent. Under sc-CO2 reaction conditions (at temperature of 100 °C and pressure of 414 bar), a PFPP silica phase was prepared using 3-[(pentafluorophenyl)propyldimethylchlorosilane] within 1h. The bonded PFPP phase was subsequently end-capped with bis-N,O-trimethylsilylacetamide (BSA), hexamethyldisilazane (HMDS) and trimethylchlorosilane (TMCS) within 1h under the same sc-CO2 reaction conditions (100 °C/4141 bar). Elemental microanalysis, thermogravimetric analysis (TGA), and scanning electron microscopy (SEM) were used to provide support data to solid-state NMR and chromatographic evaluation. Results revealed a surface coverage of 2.2 µmol/m(2) for the non-end-capped PFPP silica phase while the PFPP phase end-capped with BSA gave a higher surface coverage (3.9 µmol/m(2)) compared to HMDS (2.9 µmol/m(2)) and TMCS (2.8 µmol/m(2)). (29)Si CP/MAS NMR analysis of the PFPP end-capped with BSA shows a significant decrease in the amount of Q(3) (free silanols) and Q(4) (siloxane groups) species, coupled with the absence of the most reactive Q(2) (geminal silanols) in addition to increased amount of a single resonance peak centred at +13 ppm (MH) corresponding to -Si-O-*Si-CH3 bond. (13)C CP/MAS NMR shows the resonance corresponding to the propyl linkage (CH3CH2CH2-) and methyl groups (Si(CH3)n) confirming successful silanisation and endcapping reactions in sc-CO2. Chromatographic evaluation of the BSA end-capped PFPP phase with Neue text mixture revealed improved chromatographic separation as evidenced in the enhanced retention of hydrophobic markers and decreased retention for basic solutes. Moreover, chromatography revealed a change in column selectivity for the BSA end-capped PFPP phase with dipropylphthalate eluting before naphthalene, indicating decreased silanol groups and increased hydrophobicity. The extend of BSA end-capping as measured by the increase in column efficiency (67,260 N/m vs. 60,480 N/m) on a 2.1 i.d.×50 mm column, methylene group selectivity (α(CH(2)) = 2.27 vs. 2.14) and decreased silanophilic interactions (S=3.7 vs. 4.10) indicate that the increase in carbon loading (3.9 µmol/m(2) vs. 2.2 µmol/m(2)) and improvement in chromatography in good peak shape and symmetry is attributed to end-capping with trimethylsilyl groups.

Synthesis, characterisation and chromatographic evaluation of pentafluorophenyl and phenyl bonded silica phases prepared using supercritical carbon dioxide as a reaction solvent.

Pentafluorophenyl and phenyl silica stationary phases offer alternative selectivity compared to alkyl bonded C18 and C8 stationary phases, through other interactions such as π-π interactions, dipole-dipole and hydrogen bond interactions. Pentafluorophenyl and phenyl silica bonded stationary phases were efficiently prepared in sc-CO2 specifically pentafluorophenyl propyl (PFPP), pentafluorophenyl (PFP), phenyl propyl (PP) and phenyl (P) silica stationary phases. The bonded phases were characterised by elemental analysis, thermogravimetric analysis (TGA), BET, and by solid-state NMR spectroscopy. Chromatographic performance of the supercritical fluid generated phases was also investigated using the Neue test. The authors present results which demonstrate that pentafluorophenyl and phenyl stationary phases can be prepared successfully under supercritical conditions of 100 °C, 414 bar in a reaction time of 1h with surface coverage comparable to traditional organic solvent based methods. Chromatographic results reveal that the pentafluorophenyl propyl (PFPP) phase provides superior separation performance for Neue test solutes despite having a lower ligand density (C: 5.67%, 2.2 µmol/m²) compared to the phenyl propyl (PP) analogue having the highest ligand density (C: 6.67%, 2.5 µmol/m²). The difference chromatographic performance is attributed to the polarity of the CF bond in PFPP phase. Moreover, as the alkyl chain length decreases, the hydrophobic interaction also decreases, and the PFPP phase (with a propyl linkage) provides better separation compared to the PFP phase.