Immobilization of liposomes onto quartz crystal microbalance to detect interaction between liposomes and proteins

To study the interaction between liposomes and proteins, intact liposomes were immobilized on a metal planar support by chemical binding and/or bioaffinity using a quartz crystal microbalance (QCM). A large decrease in the resonance frequency of quartz crystal was observed when the QCM, modified by a self-assembled monolayer (SAM) of carboxythiol, was added to liposome solutions. The stable chemical immobilization of intact liposomes onto SAM was judged according to the degree with which adsorbed mass depended on the prepared size of liposomes, as well as on the activation time of SAMs when amino-coupling was introduced, where the liposome coverage of electrodes was 69+/-8% in optimal conditions. When avidin-biotin binding was used on amino-coupling liposome layers, liposome immobilization finally reached 168% coverage of the electrode surface. Denatured protein was also successfully detected according to the change in the frequency of the liposome-immobilized QCM. The adsorbed mass of denatured carbonic anhydrase from bovine onto immobilized liposomes showed a characteristic peak at a concentration of guanidine hydrochloride that corresponded to a molten globule-like state of the protein, although the mass adsorbed onto deactivated SAM increased monotonously.     

Dynamic interaction between melamine and cyanuric acid in artificial urine investigated by quartz crystal microbalance

The dynamic interaction between melamine (M) and cyanuric acid (CA) in artificial urine was investigated by a flow injection analysis-quartz crystal microbalance (FIA-QCM) system. Melamine was used as the recognition element and immobilized onto the QCM gold surface. The process of M and CA interaction was recorded by FIA-QCM in real-time. The multilayer complex of M and CA was successfully formed on the crystal surface of the QCM when CA and M were introduced into the FIA-QCM system alternately. The influence of pH on the M and CA interaction indicated that the M-CA multilayer complex possesses high stability over a wide range of pH values, especially in physiological urine condition. The association and dissociation rate constants were determined under artificial urine condition and the association constant for M and CA was calculated as 10(2) L mol(-1). The M-CA complex was further characterized by Fourier transform infrared spectroscopy (FT-IR) and atomic force microscopy (AFM). Mechanism study suggested that the formation of the multilayer complex in artificial urine was mainly attributed to the intermolecular hydrogen bonding between M and CA. The work provides direct insight into the dynamic interaction between M and CA under physiological conditions and would be helpful for the research of kidney stones induced by melamine contamination.     

In situ investigation of dye adsorption on TiO2 films using a quartz crystal microbalance with a dissipation technique

Dye adsorption plays a crucial role in dye-sensitized solar cells. Herein, we demonstrate an in situ liquid-phase analytical technique to quantify in real time adsorption of dye and coadsorbates on flat and mesoporous TiO(2) films. For the first time, a molar ratio of co-adsorbed Y123 and chenodeoxycholic acid has been measured.     

In situ kinetic study of zinc sulfide activation using a quartz crystal microbalance with dissipation (QCM-D)

We have studied the activation kinetics of zinc sulfide (ZnS) using silver as an activator by a quartz crystal microbalance with dissipation (QCM-D). The zinc sulfide coating on QCM-D sensor was shown to have similar crystallographic structure, composition, and surface properties as nature sphalerite through the characterization of X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and xanthate adsorption measurement using QCM-D. The activation of ZnS sensor by silver was confirmed by the mass increase in ZnS sensor coupled with subsequent xanthate adsorption during QCM-D measurement, the change of surface wettability, and the presence of Ag(2)S on the surface. Two distinct stages on the silver uptake vs. time curve were identified and fitted well by a logarithmic function for the initial stage and a parabolic law in the later stage, which agrees with the two-stage zinc-silver reaction kinetics reported previously. Argon sputtering followed by XPS measurement on the ZnS surface demonstrated the penetration of silver into the bulk ZnS after activation. The present study is the first of its kind to apply the QCM-D technique to investigate sphalerite activation, which introduces a new in situ approach to investigate surface adsorption and activation in many mineral processes and surface modifications.     

Electrochemical studies on the interaction of heparin with crystal violet and its analytical application

In this paper, crystal violet (CV) was used to determine heparin concentration by linear sweep voltammetry on a dropping mercury electrode (DME). In Britton-Robinson (B-R) buffer solution, pH 3.0, CV had a well-defined second-order derivative linear sweep voltammetric reductive wave at −0.74 V (vs. SCE). After the addition of heparin to the CV solution, the reductive peak current decreased greatly with the positive movement of the peak potential and without appearance of new peaks in the scanning potential range. Based on the decrease in the reductive peak current, a new voltammetric method for the determination of heparin was established. The conditions for the interaction and the electrochemical detection were optimized, and interfering substances were investigated. Under the optimal conditions, the decrease in reductive peak currents of CV was proportional to heparin concentration in the range 0.1–8.0 mg/L with the linear regression equation Δip″(nA) = 400.42 + 1563.11c (mg/L), (n = 14, γ = 0.993). The detection limit was 0.092 mg/L. This new method was further successfully applied to the determination of heparin content in heparin sodium injection samples with satisfactory results. The binding ratio and binding mechanism were also studied by the electrochemical method. The text was submitted by the authors in English.     

Physical adsorption of protamine for heparin assay using a quartz crystal microbalance and electrochemical impedance spectroscopy

This study attempted to determine absolute heparin concentration in phosphate buffer solution (PBS, pH 7.4) by using quartz crystal microbalance (QCM) as an affinity biosensor. Electrochemical impedance spectroscopy (EIS) was also used to investigate immobilization of protamine and heparin assay. In addition, the effectiveness of physical adsorption in immobilizing protamine was confirmed by examining the preparation conditions, including the incubation time and protamine concentration. It induced maximum decrease (ca. −100 Hz) in oscillating frequency of QCM by applying 20 mg/ml protamine and 20 min for incubation in PBS. Heparin adsorption onto protamine-modified electrode in PBS revealed an exponential-like binding curve and long duration for reaching the steady state in frequency response of QCM. Moreover, two linear calibration curves were obtained judging from the initial slope (df/dt) and the frequency change (Δf) of QCM obtained after a binding interval (600 s) for heparin concentrations from 0 to 3.0 and 7.0 U/ml, respectively. In EIS analysis, calibration curves with linear concentration range of 0-3.0 U/ml were obtained for heparin in PBS when ferrocyanide was used as an electroactive marker.     

Multilayers of charged polypeptides as studied by in situ ellipsometry and quartz crystal microbalance with dissipation

The buildup of poly(L-glutamic acid) (PGA) and poly(L-lysine) (PLL) multilayers on silica and titanium surfaces, with and without an initial layer of polyethyleneimine (PEI), was investigated and characterized by means of in situ ellipsometry and quartz crystal microbalance with dissipation. A two-regime buildup was found in all systems, where the length of the first slow-growing regime is dependent on the structure of the initial layers. In the second fast-growing regime, the film thickness grows linearly while the mass increases more than linearly (close to exponentially) with the number of deposited layers. The film refractive indices as well as the water contents indicate that the film density changes as the multilayer film builds up. The change in film density was proposed to be due to polypeptides diffusing into the multilayer film as they attach. Furthermore, the use of PEI as the initial layer was found to induce a difference in the thickness increments for PGA and PLL.     

A novel method for in situ measurement of solubility via impedance scanning quartz crystal microbalance studies

We introduce here a novel in situ measurement method for solubility of solids in various liquids. Without any calibration the saturation point can be obtained in a relative manner. We exemplified the new method at four systems including water, organic carbonates and an ionic liquid as the solvents and various salts as dissolved solids.     

The interaction between model biomaterial coatings and nylon microparticles as measured with a quartz crystal microbalance with dissipation monitoring

The interaction between cells and biomaterials has been mimicked using nylon microparticles as pseudo-cells and PLMA and PIBMA as biomaterial model acrylate polymers. The shift of fundamental resonance frequencies was negative for both polymers, indicating mass-coupling to the sensor surface. The shifts of the 3rd, 5th and 7th overtone frequencies were initially positive for both polymers, indicating a particle slip or wobbling on the surface. The QCM technique could discriminate between the two different polymers, showing increased interaction between microparticle and PLMA. The dissipation shift was positive for all overtones on both polymers, but again with faster and more prominent response for PLMA.     

Determination of 4-aminophenol using the quartz crystal microbalance sensor

In this paper we report a method for the determination of 4-aminophenol (4-AP) in solution using a quartz crystal microbalance (QCM) sensor. 4-AP reacts with (para-unsubstituted) phenols to form hydrophobic indophenol dye species that precipitate out and adsorb to the surface of the crystal to produce a shift in the crystal resonant frequency. This frequency change, due to in-situ indophenol mass adsorption, can be related to the initial 4-AP concentration. A range of phenols (namely o-cresol, 1-naphthol, resorcinol, catechol and 8-hydroxyquinoline) and their reaction with 4-AP were tested. Ammonium persulfate (APS) and potassium periodate were used as initiators to improve the speed of the reaction and the rate of formation of the precipitate. APS elicited improved signal in terms of response times and frequency shifts compared with KIO4. Of the phenols studied, resorcinol gave the best response time of 6 min for 4-AP determination. The reaction of resorcinol with 4-AP gave extended response times and signal size with decreasing concentration of 4-AP (in the range 2-5 mM).     

Development of a sensor for calcium based on quartz crystal microbalance

A new sensor for calcium based on a piezoelectric quartz crystal is presented. The selectivity depends on the ratio lipophilic salt/ionophore of the composition of the coating of the quartz crystal. A crystal coated with a THF solution of PVC (34.5% w/w), DOS (62.1% w/w) and 10,19-bis[(octadecylcarbamoyl) methoxyacetyl]-1,4,7,13,16-pentaoxa-10,19-diaza cycloheneicosane (3.4% w/w) and a salt/ionophore molar proportion of 60%, corresponding to a frequency decrease of the dry crystal of 6.0 kHz, showed a detection limit to calcium of 2.2 mg/L. Both a standard calcium chloride solution and a commercially balanced salt solution for tissue culture were analyzed subsequent to ion chromatographic separation. The results obtained with the sensor developed were compared with those obtained with a conductivity detector. Although the results from both detectors agreed for the standard solution, only the quartz crystal sensor was able to give reliable results for the tissue culture solution.     

Development of a sensor for calcium based on quartz crystal microbalance

A new sensor for calcium based on a piezoelectric quartz crystal is presented. The selectivity depends on the ratio lipophilic salt/ionophore of the composition of the coating of the quartz crystal. A crystal coated with a THF solution of PVC (34.5% w/w), DOS (62.1% w/w) and 10,19-bis[(octadecylcarbamoyl) methoxyacetyl]-1,4,7,13,16-pentaoxa-10,19-diaza cycloheneicosane (3.4% w/w) and a salt/ionophore molar proportion of 60%, corresponding to a frequency decrease of the dry crystal of 6.0 kHz, showed a detection limit to calcium of 2.2 mg/L. Both a standard calcium chloride solution and a commercially balanced salt solution for tissue culture were analyzed subsequent to ion chromatographic separation. The results obtained with the sensor developed were compared with those obtained with a conductivity detector. Although the results from both detectors agreed for the standard solution, only the quartz crystal sensor was able to give reliable results for the tissue culture solution.     

Decoupling of the liquid response of a superhydrophobic quartz crystal microbalance

Recent reports using particle image velocimetry and cone-and-plate rheometers have suggested that a simple Newtonian liquid flowing across a superhydrophobic surface demonstrates a finite slip length. Slippage on a superhydrophobic surface indicates that the combination of topography and hydrophobicity may have consequences for the coupling at the solid--liquid interface observed using the high-frequency shear-mode oscillation of a quartz crystal microbalance (QCM). In this work, we report on the response of a 5 MHz QCM possessing a superhydrophobic surface to immersion in water--glycerol mixtures. QCM surfaces were prepared with a layer of SU-8 photoresist and lithographically patterned to produce square arrays of 5 mum diameter circular cross-section posts spaced 10 microm center-to-center and with heights of 5, 10, 15, and 18 microm. Non-patterned layers were also created for comparison, and both non-hydrophobized and chemically hydrophobized surfaces were investigated. Contact angle measurements confirmed that the hydrophobized post surfaces were superhydrophobic. QCM measurements in water before and after applying pressure to force a Cassie-Baxter (non-penetrating) to Wenzel (penetrating) conversion of state showed a larger frequency decrease and higher dissipation in the Wenzel state. QCM resonance spectra were fitted to a Butterworth-van Dyke model for the full range of water-glycerol mixtures from pure water to (nominally) pure glycerol, thus providing data on both energy storage and dissipation. The data obtained for the post surfaces show a variety of types of behavior, indicating the importance of the surface chemistry in determining the response of the quartz crystal resonance, particularly on topographically structured surfaces; data for hydrophobized post surfaces imply a decoupling of the surface oscillation from the mixtures. In the case of the 15 microm tall hydrophobized post surfaces, crystal resonance spectra become narrower as the viscosity-density product increases, which is contrary to the usual behavior. In the most extreme case of the 18 microm tall hydrophobized post surfaces, both the frequency decrease and bandwidth increase of the resonance spectra are significantly lower than that predicted by the Kanazawa and Gordon model, thus implying a decoupling of the oscillating surface from the liquid, which can be interpreted as interfacial slip.     

The study of electrochemical palladium behavior using the quartz crystal microbalance

The electrochemical quartz crystal microbalance (EQCMB) method has been used to evaluate the processes which occur in/on the palladium electrode in basic solutions. Hydrogen electrosorption in palladium is accompanied by an additional frequency shift that can be attributed to the stresses generated inside the Pd metal. A non-linear dependence between the mass change and the charge consumed during hydrogen oxidation in the Pd electrode has been found for hydrogen absorbed in the α- and β-phases. This effect precludes the objective estimation of the amount of hydrogen absorbed inside the Pd electrode. The EQCMB method has been used, however, for studying the surface electrode processes on the Pd electrode, i.e. specific anion adsorption, surface oxidation and dissolution. Also, the structure of the palladium oxide formed on the Pd surface during electrochemical oxidation is discussed in this paper and the effect of the anodic limiting potential on the oxide structure is reported. Received: 10 August 1999 / Accepted: 24 September 1999     

Study of hydrogen electrosorption in Pd-Ni alloys by the quartz crystal microbalance

The electrochemical quartz crystal microbalance (EQCMB) method has been used to study the processes of hydrogen absorption/desorption in Pd-Ni alloy electrodes. It was found that hydrogen electrosorption is accompanied by an additional frequency shift, attributed to the stresses generated inside the alloy. The influence of stresses on the EQCMB response depends on the amount of absorbed hydrogen and the alloy composition. From the comparison of the EQCMB results with Pd-Ni alloy absorption capabilities data, it was concluded that the decrease of the hydrogen sorption capacity at a Ni content of ca. 25–30 at% is due to an excessive generation of stresses in the alloy lattice. Also, a dependency of the rate of hydrogen absorption in Pd-Ni alloys on potential is reported. Electronic Publication     

Measurement of sorption in polymer membranes with a quartz crystal microbalance

An application of the oscillating quartz crystal microbalance (QCM) in the measurements of sorption rates and equilibria in polymer—vapor systems is presented. The sorption isotherms of methanol, ethanol and n-heptane in cellulose acetate and poly(hexamethylene adipamide) were measured with the QCM at 298K. The results agree reasonably with the Flory model except at low absorbate activities where appreciable deviations due to localized absorption were observed, as verified with IR spectroscopy. The diffusion coefficient of methanol in cellulose acetate was also calculated from the sorption rate data measured with the QCM. The values obtained at different methanol concentrations correlate well with the published diffusion coefficients.     

Application of the quartz crystal microbalance for the investigation of nanotribological processes

The electrochemical preparation of copper layers was investigated using a quartz crystal microbalance (QCM) with damping monitoring. The damping increase during the deposition could be separated into two contributions arising from an internal and an external friction process. External friction comes from the coupling of the shear motion of the rough metal surface to the viscous liquid. Internal friction occurred only in the coarse grained layers and can be explained by phonon excitations at the grain boundaries.Presented at the 3rd International Symposium on Electrochemical Processing of Tailored Materials held at the 53rd Annual Meeting of the International Society of Electrochemistry, 15–20 September 2002, Düsseldorf, Germany     

Adsorption of liposomes and emulsions studied with a quartz crystal microbalance

The adsorption from phospholipid liposome solutions (1.2%) and phospholipid stabilized oil-in-water emulsions (20% purified soybean oil) with the same phospholipid liposome concentration, has been followed by means of a quartz crystal microbalance allowing the simultaneous determination of changes in resonance frequency and energy dissipation. Both the fundamental resonance frequency and the third overtone were used for following the interfacial processes. The adsorption from the liposome solution resulted in formation of a phospholipid bilayer with an additional and incomplete outer layer of liposomes. The outer layer was removed by dilution leaving a bilayer of phospholipids on the surface. The adsorption process observed from the concentrated emulsion solution was considerably more complex. A slow spreading process that also resulted in some expulsion of material from the interface followed the rapid initial adsorption of emulsion droplets. After rinsing with water a phospholipid bilayer was retained on the surface.     

Establishing linear solvation energy relationships between VOCs and monolayer-protected gold nanoclusters using quartz crystal microbalance

Linear solvation energy relationships (LSERs) have been recognized as a useful model for investigating the chemical forces behind the partition coefficients between vapor molecules and absorbents. This study is the first to determine the solvation properties of monolayer-protected gold nanoclusters (MPCs) with different surface ligands. The ratio of partition coefficients/MPC density (K/ρ) of 18 volatile organic compounds (VOCs) for four different MPCs obtained through quartz crystal microbalance (QCM) experiments were used for the LSER model calculations. LSER modeling results indicate that all MPC surfaces showed a statistically significant (p < 0.05) preference to hydrogen-bond acidic molecules. Through dipole-dipole attraction, 4-methoxythiophenol-capped MPCs can also interact with polar organics (s = 1.04). Showing a unique preference for the hydrogen bond basicity of vapors (b = 1.11), 2-benzothiazolethiol-capped MPCs provide evidence of an intra-molecular, proton-shift mechanism on surface of nano-gold.