Acid soap and phase behavior of stearic acid and triethanolamine stearate

Crystals of partially neutralized stearic acid with triethanolamine (TEA) were prepared by mixing these two materials above 80 degrees C and then cooling. The crystalline composition and the structure and melting behavior of the resultant products were characterized with small-angle and wide-angle X-ray diffraction, thermal analysis, microscopy, and infrared spectroscopy. It was discovered that an acid-soap complex of 2:1 fixed stoichiometric ratio exists between stearic acid and TEA stearate. A binary phase diagram of stearic acid and TEA soap is built based on the experimental results; this is the first published record of a binary phase diagram for amine-based soap. Its behavior is significantly different from that of binary systems of fatty acid and alkali soap.     

Fluorimetric determination of pantothenic acid in foods by liquid chromatography with post-column derivatization

A method to determine the content of free pantothenic acid in various foods by reverse phase liquid chromatography-fluorimetry is reported. It includes a purification of the samples by successive passages through anion and cation exchange cartridges and a post-column derivatization of pantothenic acid as the fluorescent 1-alkylthio-2-alkylisoindole (reaction of beta-alanin, formed by hot alkaline hydrolysis of pantothenic acid, with orthophthaldialdehyde in the presence of 3-mercaptopropionic acid). An enzymatic hydrolysis prior to the purification step (pepsin at 50 degrees C for 3 h, then pantetheinase and alkaline phosphatase at 20 degrees C for 18 h) made it possible to release the bound pantothenic acid and thus to obtain the total Vitamin B5 content of these foodstuffs. The method proposed for the determination of free and bound pantothenic acid gives a good recovery rate (96-101%) and a satisfactory repeatability (R.S.D.r less than 8%). Owing to its low detection limit (0.65 microg g(-1)) and the good resolution of the pantothenic acid peak, it could most probably be applied to the determination of this vitamin in any foodstuff.     

Amino acid analysis by high-performance liquid chromatography with methanesulfonic acid hydrolysis and 9-fluorenylmethylchloroformate derivatization

Experiments were undertaken to verify a method for complete amino acid analysis of plant and animal tissues and waste products from a single hydrolysis and high-performance liquid chromatographic run. Using methanesulfonic acid, hydrolysis of cytochrome c at 115 degrees C for 22 h yielded recoveries equal to or higher than hydrolysis at 115 degrees C for 70 h or at 150 degrees C for 22 h. Triple evacuation of the hydrolysis tube alternated with nitrogen flush gave recovery improvements over single evacuation. Refrigerated storage of samples under vacuum for up to 4 days between hydrolysis and further analysis was not different from immediate analysis. However, recoveries of several amino acids were reduced by refrigerated storage in air. Recoveries of individual amino acids were determined by hydrolysis of biological samples with and without added cytochrome c. Although recoveries from biological samples were lower for several amino acids, precision was sufficient to allow quantitation after correction for incomplete recoveries. Derivatization with 9-fluorenylmethylchloroformate (FMOC) was chosen because derivatives are formed with both primary and secondary amino acids, derivatives are quite stable, and detection may be either UV absorbance or fluorescence. Derivative yield is sensitive to the pH of the reaction mixture. A pH of 8.0 gave reproducible derivative yield for all physiological amino acids. Solvent extraction of excess FMOC, when compared to addition of amantadine to react with excess FMOC, gave both higher recoveries and greater precision. Following derivatization, samples could be kept at 4 degrees C for at least 24 h before high-performance liquid chromatographic analysis without loss of response. Derivative yield and detector response were constant across a wide range of molar ratio of FMOC to total amino acids. Gradient elution was required to separate FMOC derivatives on a reversed-phase column. The capability of the pumping system to produce exponential gradients permitted rapid and easy fine-tuning of the gradient.     

The effect of octyl glucoside on the lamellar phase of diluted C12E4 and alcohol systems

A systematic study on phase behavior of the mixture of nonionic surfactants with alcohols at 30.0+/-0.1 degrees C was carried out. The total surfactant concentration was kept to 0.1 M varying the mole ratio of n-octyl beta-d-glucopyranoside (OG) and tetraethylene glycol monododecyl ether. Two uniphasic regions were found, the lamellar phase at low OG mole fraction and micelles at high OG mole fraction. The presence of OG favors the lamellae-micelle transition. Alkanols and benzyl alcohol were used as cosurfactants. The more hydrophobic alcohols (octanol and decanol) increase the OG content in the mixed bilayers. On the contrary, benzyl alcohol is not as favorable to the OG incorporation in the lamellar phase as in the mixed micelles. The L(3) phase has only been found as a uniphasic region with hexanol.     

A method for the determination of the hepatic enzyme activity catalyzing bile acid acyl glucuronide formation by high-performance liquid chromatography with pulsed amperometric detection

A method for the determination of the activity of hepatic glucuronyltransferase catalyzing formation of bile acid 24-glucuronides using high-performance liquid chromatography (HPLC) with pulsed amperometric detection (PAD) has been developed. Bile acid 24-glucuronides were simultaneously separated on a semimicrobore column, Capcell Pak C18UG120, using 20 mM ammonium phosphate (pH 6.0)-acetonitrile (27:10 and 16:10) as the mobile phase in the stepwise gradient elution mode. A 1 M potassium hydroxide solution for the hydrolysis of the 24-glucuronides, which liberates the corresponding bile acids and glucuronic acid, was mixed with the mobile phase in a post-column mode, and the resulting eluant was heated at 90 degrees C, the 24-glucuronides being monitored using a pulsed amperometric detector; the limit of detection was 10 ng. The proposed method was applied to the determination of the hepatic enzyme activity catalyzing bile acid 24-glucuronide formation and the result exhibited the efficient 24-glucuronide formation of the monohydroxylated bile acid, lithocholic acid.     

Self-organization of polybases neutralized with mesogenic wedge-shaped sulfonic acid molecules: an approach toward supramolecular cylinders

Complexes consisting of poly(4-vinylpyridine) and mesogenic wedge-shaped ligands 4'-[3",4",5"-tris(dodecyloxy)benzoyloxy]azobenzene-4-sulfonic acid and 4'-[3",4",5"-tris(octyloxy)benzoyloxy]azobenzene-4-sulfonic acid have been prepared with different monomer/ligand ratios. Upon protonation of the poly(4-vinylpyridine) chains by the wedge-shaped sulfonic acid molecules a hypsochromic and hyperchromic effect was observed with the pi-pi* transition of the azo-chromophor, allowing us to monitor the neutralization process by means of UV-vis spectroscopy in solution. The changes of the absorption characteristics implied a conformational change of the polymer backbone. In the bulk the interaction between pyridine and sulfonic acid moieties was proved by FT-IR spectroscopy. Polarizing optical microscopy, differential scanning calorimetry, and X-ray diffraction measurements were used to study the bulk structure of the complexes. The complexes formed a liquid crystalline lamellar phase at low degrees of substitution, while a hexagonal columnar mesophase was observed at degrees of neutralization of 80% and higher.     

Synthesis and characterization of mesoporous ceria/alumina nanocomposite materials via mixing of the corresponding ceria and alumina gel precursors

Mesoporous ceria/alumina, CeO(2)/Al(2)O(3), composites containing 10, 20 and 30% (w/w) ceria were prepared by a novel gel mixing method. In the method, ceria gel (formed via hydrolysis of ammonium cerium(IV) nitrate by aqueous ammonium carbonate solution) and alumina gel (formed via controlled hydrolysis of aluminum tri-isopropoxide) were mixed together. The mixed gel was subjected to subsequent drying and calcination for 3 h at 400, 600, 800 and 1000 degrees C. The uncalcined (dried at 110 degrees C) and the calcined composites were investigated by different techniques including TGA, DSC, FTIR, XRD, SEM and nitrogen adsorption/desorption isotherms. Results indicated that composites calcined for 3 h at 800 degrees C mainly kept amorphous alumina structure and gamma-alumina formed only upon calcinations at 1000 degrees C. On the other hand, CeO(2) was found to crystallize in the common ceria, cerinite, phase and it kept this structure over the entire calcination range (400-1000 degrees C). Therefore, high surface areas, stable surface textures, and non-aggregated nano-sized ceria dispersions were obtained. A systematic texture change based on ceria ratio was observed, however in all cases mesoporous composite materials exposing thermally stable texture and structure were obtained. The presented method produces composite ceria/alumina materials that suit different applications in the field of catalysis and membranes technology, and throw some light on physicochemical factors that determine textural morphology and thermal stability of such important composite.     

Effects of the number of fatty acid residues on the phase behaviors of decaglycerol fatty acid esters

The effects of the number of fatty acid residues (n) in decaglycerol fatty acid esters, i.e., decaglycerol laurates (abbreviated to (C11)nG10), on the phase behaviors of three laurate esters, (C11)1.9G10, (C11)2.7G10, and (C11)3.4G10, were investigated. The unreacted decaglycerol remaining in each ester was removed by liquid extraction before use. (C11)1.9G10 formed hexagonal liquid crystals in aqueous solutions, while (C11)2.7G10 and (C11)3.4G10, which are more hydrophobic than (C11)1.9G10, formed lamellar liquid crystals. The cloud point in aqueous solution was measured for mixtures of these three esters. The cloud phenomenon was observed when the weight ratio of hydrophilic groups to the total surfactant (WH/WS) was around 0.6. The cloud point shifted to a markedly higher temperature, even with a slight increase in the WH/WS ratio. The solubilization abilities of (C11)nG10 for the oils m-xylene and (R)-(+)-limonene were also examined. When the WH/WS ratio was between 0.60 and 0.64, (C11)nG10 formed microemulsions and lyotropic liquid crystals in the presence of water and the oils. These self-organized structures were stable, even above 90 degrees C. It is concluded that the phase behavior of (C11)nG10 are insensitive to temperature, but strongly dependent on both the WH/WS ratio and the number of fatty acid residues (n).     

In-situ atomic force microscopy observation of enzymatic degradation in poly(hydroxyalkanoic acid) thin films: normal and constrained conditions

The enzymatic degradation of lamellar crystals in poly(hydroxyalkanoic acid) thin films has been visualized by using in-situ dynamic force mode (tapping mode) atomic force microscopy (AFM) in buffer solution. It was found that poly(hydroxybutyric acid) (PHB) depolymerase from Ralstonia pickettii T1 degraded the thin surface layers formed at room temperature first, and that lamellar crystals formed at the crystallization temperature (110 degrees C) were eroded from the crystallographic a-axis to show splintered morphologies at the tips of the crystals. In some cases, lamellar crystals were hydrolyzed from the crystallographic b-axis, resulting in the formation of small crevices. These results suggest that disordered molecular chain-packing regions exist in the crystal along the crystallographic a- and b-axes, and that enzymatic degradation predominantly occurs from these defective regions. In addition, cantilever-tip-induced enzymatic degradation was carried out in the presence of PHB depolymerase. A concave area was artificially formed on the stacked lamellar crystals by the AFM tip. In-situ AFM observation has revealed that enzymatic degradation proceeds along both the longitudinal and lateral directions of the lamellae. At the same time, the PHB depolymerase preferentially eroded the concave area along the crystallographic c-axis. These results demonstrated that the PHB depolymerase predominantly degrades the less-ordered molecular chain-packing regions in the crystals.     

Self-assembly of mixed anionic and nonionic surfactants in aqueous solution

We present the phase diagram and the microstructure of the binary surfactant mixture of AOT and C(12)E(4) in D(2)O as characterized by surface tension and small angle neutron scattering. The micellar region is considerably extended in composition and concentration compared to that observed for the pure surfactant systems, and two types of aggregates are formed. Spherical micelles are present for AOT-rich composition, whereas cylindrical micelles with a mean length between 80 and 300 ? are present in the nonionic-rich region. The size of the micelles depends on both concentration and molar ratio of the surfactant mixtures. At higher concentration, a swollen lamellar phase is formed, where electrostatic repulsions dominate over the Helfrich interaction in the mixed bilayers. At intermediate concentrations, a mixed micellar/lamellar phase exists.     

High-performance liquid chromatographic determination of polysorbate 80 in pharmaceutical suspensions

A quick HPLC method is reported for the analysis of polysorbate 80 in pharmaceutical suspensions. A typical pharmaceutical suspension was mixed with dilute potassium hydroxide, and heated at 40 degrees C for 6 h. This procedure resulted in quantitative hydrolysis of polysorbate 80 to release oleic acid. A quick HPLC procedure was used to analyze the hydrolyzed samples without further sample treatment. Polysorbate 80 USP, treated in the same way as the pharmaceutical suspensions, was used as standard. Full validation tests were carried out and the validation studies demonstrated that this method is suitable for accurate and reproducible analysis of polysorbate 80 in pharmaceutical suspensions.     

Synthesis and analytical properties of a novel columnar metallomesogenic polymer

A metallomesogenic side-chain polymer with copper carboxylato discotic units in stacks prepared by covalent bonding of 14-pentadecenoic acid, stearic acid and poly(methylhydrosiloxane) is described. The physico-chemical and thermal properties of both monomeric and polymeric metallomesogens were determined by elemental analysis, IR, polarizing optical microscopy, thermal gravimetric analysis and differential scanning calorimetry. The polymeric states showed a discotic lamellar phase at 50-95 degrees C and an ordered discotic hexagonal phase at 95-200 degrees C. By dynamic coating, the metallomesogenic polymer was crosslinked to the capillary wall via benzoyl peroxide. The wall-coated capillary columns (15 m x 0.25 mm I.D.) were used for the separation of phenols. Factors affecting the retention and the sample selectivity were examined. Van 't Hoff plots as a function of temperature indicated that phase transitions were occurring. Thermodynamic properties of the analytes in this system were also studied. For the determination of a mixture of 3-aminophenol, 2-chlorophenol, 2-nitrophenol, 4-nitrophenol, o-methylphenol, m-methylphenol, p-methylphenol, 2,4-dichlorophenol, 2,4-dimethylphenol, 2,4-dinitrophenol, 2,4,6-trichlorophenol, 2,4,6-trimethylphenol, 4-bromophenol, 3-methyl-4-chlorophenol, pentachlorophenol, and unsubstituted phenol, the calibration graphs for most phenols were linear over the range of 10-1000 microg ml(-1) and the mass detection limits were in the ng range based on three times standard deviation of seven measurements of the lowest peak that could be detected.     

Molecular packing in sphingomyelin bilayers and sphingomyelin/phospholid mixtures

The molecular packing properties of sphingomyelin (SM) from egg yolk were studied. The influence of the spontaneous curvature of SM on the phase behaviour of SM/dodecane/water systems was investigated. A comparison was made to a previous study by Lindblom et al. (Liq. Cryst. 3 (1988) 783), of the phase behaviour of dipalmitoylphosphatidylcholine (DPPC)/dodecane/water systems, where a reversed hexagonal liquid crystalline was shown to form at high water contents (60-80%, w/w). In contrast, SM/dodecane/water systems mainly maintained a lamellar liquid crystalline phase at all compositions and temperatures >35 degrees C. This suggests that the spontaneous curvature of SM is larger than for DPPC. To further examine the packing properties of SM and DPPC, the phase behaviour of SM/dioleoylphosphatidylethanolamine (DOPE)/water and DPPC/DOPE/water systems were investigated. Aqueous dispersions of DOPE normally form a reversed hexagonal liquid crystalline phase, while an isotropic phase was formed at small additions (20 mol.%) of SM or DPPC and a lamellar liquid crystalline phase was maintained at higher fractions (>35 mol.%) of SM or DPPC.     

Cryo-fracture TEM: direct imaging of a random mesh phase

A novel and a simple method that allows direct imaging of viscous samples by cryo-TEM (cryo-transmission electron microscopy) is presented. A fracture on the vitrified sample is created in a controlled way. In the fracture, some edges are thin enough to allow direct imaging in transmission mode. The method was used to directly image a nonionic surfactant lamellar phase where a random mesh structure is formed at lower temperatures (<10 degrees C). A so-called random mesh phase, characterized by the presence of perforated surfactant bilayers, is imaged here for the first time. Images from the mesh structure are compared with images from the classical lamellar structure formed at room temperature.     

Formation of Cubic-Phase Microemulsions with Anionic and Cationic Surfactants at Equal Amounts of Oil and Water

The formation and microstructure of cubic phases were investigated in anionic and cationic surfactant-containing systems at 25 degrees C. In the system sodium dodecyl sulfate(SDS)-dodecyltrimethylammonium bromide(DTAB)-water, mixing of two surfactants shows the phase transition hexagonal phase (H(1))-->surfactant precipitate, accompanied by an obvious decrease in the cross-sectional area per surfactant in the rod micelles of the hexagonal liquid crystal. In the mixed systems brine(A)-dodecane(B)-SDS(C)-DTAB(D)-hexanol(E), the isotropic discontinuous cubic phase is formed from the H(1) phase at a low cationic surfactant weight fraction, Y=D/(C+D), and from the lamellar phase at high Y upon dilution with equal amounts of oil and brine, respectively. The minimum surfactant concentration to form the cubic phase decreases with increases both in cationic surfactant weight fraction Y from 0 to 0.30 and in hexanol weight fraction, W(1)=E/(C+D+E), accordingly. The maximum solubilization for oil of the cubic phase reaches 43 wt% at 14 wt% of mixed surfactants and alcohol. Copyright 2000 Academic Press.     

Physical properties of diglycerol esters in relation to rheology and stability of protein-stabilised emulsions

The surface activity and lyotropic phase behaviour of concentrated diglycerol-esters of fatty acids with chain length of C14, C16, C18 and C18:1 (cis-oleic acid) are investigated. Diglycerol-esters show a much stronger reduction in the interfacial tension at a low concentration (0.01–0.1%) than corresponding monoglycerides. The diglycerol-esters form lamellar mesophases above their Krafft point, and no other types of mesophases are found in the temperature region examined (0–80°C). The lamellar phases show a limited swelling capacity, corresponding to a water layer thickness of ≈24 Å, which is found when the ratio of diglycerol-ester to water is 60:40, or lower. At high water concentrations (>90%) multi-lamellar liposomes are formed. The diglycerol-monooleate form lamellar phases in water in the temperature region from zero to 80°C. This is in strong contrast to the corresponding glycerol-monooleate, which forms cubic and reversed hexagonal mesophases in water. Oil in water emulsions are stabilised by diglycerol-esters by formation of liquid crystalline interfacial films around the oil droplets, which can be seen in polarised light microscopy. In presence of milk proteins in the aqueous phase the emulsion stability is depending on the protein to emulsifier ratio. At 40°C a mixed interfacial film of diglycerol-monooleate (DIGMO) and protein is present at the oil–water interface, but when cooled to 5°C, the proteins are displaced by DIGMO. This behaviour affects the stability and rheological properties of emulsions stored at low temperatures.     

Determination of binding sites in carboplatin-bound cytochrome c using electrospray ionization mass spectrometry and tandem mass spectrometry

Interaction of carboplatin with cytochrome c (Cyt. c) has been investigated by electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS). ESI-MS studies revealed that the ring-opened adducts of carboplatin with Cyt. c were formed in the stoichiometric ratio of 1:1 and 2:1 at pH 5.0 and 37 degrees C and in the stoichiometric ratio of 1:1 only at pH 7.0 and 37 degrees C. It was also found that Cyt. c could be cleaved by carboplatin at pH 2.5 and 50 degrees C. The cleaved fragments of Cyt. c were determined by ESI-MS and MS/MS analysis to be Glu66 approximately Met80, Ac-Gly01 approximately Met65, Glu66 approximately Glu104, Ac-Gly01 approximately Met80 and Ile81 approximately Glu104. The carboplatin prefers to anchor to Met65 first, then to Met80. To further confirm the binding site of Met, AcMet-Gly was used as the model molecule to investigate its interaction with carboplatin and its hydrolysis reaction. On the basis of species detected during the reaction monitored by ESI-MS, a possible pathway of the cleavage reaction was proposed.     

From decanoate micelles to decanoic acid/dodecylbenzenesulfonate vesicles

Different aspects of mixtures of decanoic acid and sodium decanoate were investigated in aqueous solution up to a total concentration of 300 mM. Depending on the ratio of ionized to nonionized decanoic acid, micelles or vesicles form above the critical concentrations of micelle (cmc) or the critical concentration for vesicle formation (cvc). The micelles and the vesicles are always present together with nonmicellized or nonvesiculized decanoate. The latter was determined for different total concentrations. On the basis of titration curves, by application of the Gibbs phase rule, and on the basis of differential scanning calorimetry measurements and an electron microscopy analysis, the pH region within which vesicles exist was identified (pH 6.8-7.8). At pH 7.0, the concentration of nonvesiculized decanoate is approximately 20 mM. Decanoic acid/decanoate vesicles can be sized down by the extrusion technique to form stable and mainly unilamellar vesicles with a mean diameter of less than 100 nm. By coaddition of an equimolar amount of sodium dodecylbenzenesulfonate (SDBS) to decanoic acid, vesicles also formed below pH 6.8. These mixed vesicles were investigated as potential templates for the peroxidase-catalyzed polymerization of aniline at pH 4.3. Furthermore, decanaote micelles (at pH 11.0) were applied as reaction modifiers for the simultaneous competitive alkaline hydrolysis of p-nitrophenylacetate and fluorescein diacetate. While the rate of hydrolysis of fluorescein diacetate is slowed considerably in the presence of the micelles in comparison with the micelle-free system, the rate of hydrolysis of p-nitrophenylacetate remains almost unaffected.     

Rippled domain formation in phase-separated mixed Langmuir-Blodgett films

The morphology and composition of phase-separated Langmuir and Langmuir-Blodgett films of stearic acid (C17H35COOH) (SA) mixed with perfluorotetradecanoic acid (C13F27COOH) (PA) have been investigated using a combination of atomic force microscopy (AFM) measurements and surface pressure-area isotherms. At elevated surface pressures, the mixed film phase-separated to form a distinct series of lines (ripples), as opposed to the hexagons that have previously been observed with mixed films with longer alkyl chain fatty acids. At low surface pressures, phase separation is still observed, though a range of different domain structures was formed. The chemical composition of the phase-separated domains has been investigated by AFM-based compositional mapping, which has allowed unambiguous identification of the chemical composition of the domains. A simple mechanistic model describing how domain formation takes place in this system is presented.     

A capillary zone electrophoresis method for determining N-acetylneuraminic acid in glycoproteins and blood sera

A simple capillary zone electrophoresis (CZE) method for the determination of the content of the major sialic acid form N-acetylneuraminic acid (Neu5Ac) in glycoproteins was established. The present method utilizes a simplified hydrolysis-purification procedure consisting of mild acid hydrolysis (25 mM trifluoroacetic acid for 2h at 80 degrees C) to release Neu5Ac and ultrafiltration on Centricon-3 membrane to remove the obtained asialoglycoproteins and other macromolecules present in biologic samples. Derivatization with benzoic anhydride at 80 degrees C for 20 min resulted in complete conversion of Neu5Ac to per-O-benzoylated Neu5Ac. CZE analysis was performed using the operating buffer 25mM phosphate, pH 3.5, containing 50% (v/v) acetonitrile as organic modifier at 30 kV, and detection of the per-O-benzoylated Neu5Ac at 231 nm. The method showed excellent repeatability (RDS<1.98%) and a linearity range from 5 microg/mL to 5mg/mL with a detection limit of 2 microM. Application of the method to microanalysis of human alpha(1)-acid glycoprotein and blood serum samples showed excellent agreement with previously published values, suggesting a high precision for the developed CZE method.