Solid–Liquid Equilibrium of Maltitol Aqueous Solutions—Implications on the Crystallization Behavior and Process

The solubility of maltitol in pure water and industrial syrup was measured in a temperature range from 10 to 90 °C. Maltitol is highly soluble in water, and this yields high viscosity values for the saturated aqueous solutions at different temperatures. In addition, solubility of maltitol in ethanol/water mixtures was followed at 30, 35, 45, and 55 °C. Results show that maltitol solubility is highly dependent on water content in the solvent mixture. Moreover, it increases monotonically with temperature. The logarithm of viscosity changes linearly against the mole fraction of maltitol in the aqueous solutions up to saturation. The saturated solutions showed a Newtonian behavior in a temperature range from 20 to 90 °C. Maltitol is also characterized in supersaturated solutions by a narrow metastable zone, which slightly increases as temperature is raised. The density of aqueous solutions of maltitol was measured as a function of molality up to saturation at 20 °C, and results show that density can be correlated with concentration according to a linear relation. The obtained results were used to explain maltitol crystallization, which exhibits a high nucleation rate and a slow growth leading to small size crystals.     

Preparation and characterization of physical gels and beads from chitin solutions

A detailed account of physical bulk gel and bead formation from various chitin solutions and nonsolvents is given. Instant gel formation occurs upon contact of chitin solutions in dimethylacetamide (DMAc)/lithium chloride (LiCl) or N-methyl-pyrrolidinone (NMP)/LiCl solvents and nonsolvents such as water, ethanol, or acetone. Ethanol was found to be the optimal nonsolvent for homogeneous spherical bead formation from chitin solutions. Similarly, DMAc-based chitin solutions proved to yield higher quality beads compared to NMP-based solutions. The differences in bead morphology, crystallinity, and thermal degradation are explained in light of the attainment of a balance between attractive hydrogen bonding in the chitin gel network and segment–nonsolvent interactions. The dependence of swelling of chitin gels on pH indicated a maximum of swelling ratio value of 4.3 at pH 11 in aqueous solutions while the equilibrium swelling ratio value of chitin beads formed with ethanol reached a maximum of 2.4. Bulk gels formed under favorable conditions were demonstrated to be recyclable after solvent separation and drying.     

Enhanced glucose to fructose conversion in acetone with xylose isomerase stabilized by crystallization and cross-linking

The effects of acetone and ethanol on glucose to fructose conversion catalyzed by soluble and cross-linked crystalline (CLXIC) xylose isomerase were studied. Relative to pure buffer solvent, the fructose production rate was more than doubled in 50% acetone. The same kind of increase in the isomerization rate was not seen with ethanol. Increase both in acetone and in ethanol concentration in the reaction solvent enhanced the production of fructose. At 50 degrees C in pure buffer solvent the reaction mixture contained 49% fructose in equilibrium and in 90% acetone the fructose equilibrium content was 64%. Furthermore, CLXIC was relatively stable in the presence of high concentration of acetone: 70-80% of activity was left after incubation for 24 h at 50 degrees C in buffer solutions (pH 7.2) containing 10-90% acetone. In buffer containing 50% ethanol only 2% of the initial activity of CLXIC was retained after 24 h at 50 degrees C. Soluble xylose isomerase was considerably less stable than CLXIC in both acetone- and ethanol-containing solutions. These results show that the addition of acetone enhances the production of fructose from glucose by enhancing the reaction rate and shifting the equilibrium toward fructose. However, xylose isomerase must be in the form of cross-linked crystals for maximal activity and stability.     

Infrared spectroscopic studies of solvent-induced conformational changes in globular proteins.

Infrared absorption spectroscopy has been used to study the effect of organic solvents on the conformation of myoglobin, apomyoglobin, hemoglobin, lysozyme and ribonuclease. Beta structure can easily be induced by specific solvent effects. Films prepared from a 50% (v/v) mixture of alcohol, acetone, pyridine, tetrahydrofuran or dimethylsulfoxide/water mixtures show a high proportion of beta structure. The degree of induction of beta structure depends on the hydrocarbon content of the alcohol in the order methanol greater than ethanol greater than butanol. No beta structure was observed in films prepared from aqueous octanol solutions. Lyophilization tends to decrease secondary structure. The conformation of the proteins depends on the particular solvent system and the solvent composition. Solution studies of myoglobin in pure dimethylsulfoxide show that the conformation is a mixture of random and beta forms while in dimethylsulfoxide/2H2O mixtures the conformation is a mixture of alpha-helical and beta forms.     

Swelling behavior and controlled release of theophylline and sulfamethoxazole drugs in beta-lactoglobulin protein gels obtained by phase separation in water/ethanol mixture

Physically cross-linked beta-lactoglobulin (BLG) protein gels containing theophylline and sulfamethoxazole low molecular weight drugs were prepared in 50% ethanol solution at pH 8 and two protein concentrations (6 and 7% (w/v)). Swelling behavior of cylindrical gels showed that, irrespective of the hydrated or dehydrated state of the gel, the rate of swelling was the highest in water. When the gels were exposed to water, they first showed a swelling phase in which their weight increased 3 and 30 times for hydrated and dehydrated gels, respectively, due to absorption of water, followed by a dissolution phase. The absorption of solvent was however considerably reduced when the gels were exposed to aqueous buffer solutions. The release behavior of both theophylline and sulfamethoxazole drugs from BLG gels was achieved in a time window ranging from 6 to 24 h. The drug release depended mainly on the solubility of the drugs and the physical state of the gel (hydrated or dry form). Analysis of drug release profiles using the model of Peppas showed that diffusion through hydrated gels was governed by a Fickian process whereas diffusion through dehydrated gels was governed partly by the swelling capacities of the gel but also by the structural rearrangements inside the network occurring during dehydration step. By a judicious selection of protein concentration, hydrated or dehydrated gel state, drug release may be modulated to be engineered suitable for pharmaceutical as well as cosmetics and food applications.     

Fibrillar beta-lactoglobulin gels: Part 3. Dynamic mechanical characterization of solvent-induced systems

Oscillatory shear rheometry has been used to study the gelation of beta-lactoglobulin at ambient in 50% v/v trifluoroethanol (TFE)/pH 7 aqueous buffer and in 50% v/v ethanol (EtOH)/water at pH 2. In contrast to what was found on heating aqueous solutions at pH 2 (Part 2 of this series), a more expected "chemical gelation"-like profile was found with modulus components G' and G' ' crossing over as the gels formed and then with G' ' passing through a maximum. In addition, for the EtOH system, there was a significant modulus increase at long time, suggestive of a more complex two-step aggregation scheme. Modulus-concentration relationships were obtained for both systems by extrapolating cure data to infinite time. For the TFE gels, this data was accurately described by classical branching theory, although it could also be approximated by a constant power--law relationship. Only the latter described the modulus--concentration data for the gels in ethanol, but there were problems here of greater frequency dependence of the modulus values and much less certain extrapolation. Gel times for the TFE systems showed higher power laws in the concentration than could be explained by the branching theory in its simplest form being similar, in this respect, to the heat-set systems at pH 2. Such power laws were harder to establish for the EtOH gels as for these there was evidence of gel time divergence close to a critical concentration. Reduced G'/G'inf versus t/tgel data were difficult to interpret for the gels in ethanol, but for the TFE system they were consistent with previous results for the heat-set gels and approximated master curve superposition. The frequency and temperature dependences of the final gel moduli were also studied. In general, the networks induced by alcohols appeared more flexible than those obtained by heating.     

Investigation and optimisation of the drying of reduced glutathione by steered molecular dynamics simulation

The drying of reduced glutathione from a series of aqueous–ethanol binary solutions at 300 K (below human body temperature) and 330 K (above human body temperature) was investigated in detail by steered molecular simulation and an umbrella sampling method with the Gromacs software package and Gromos96(53a6) united atomic force field. The results show that electrostatic interactions between glutathione and solvent represent the main resistance to drying. When the aqueous solution was gradually changed to pure ethanol, the energy of electrostatic interaction between glutathione and solvent molecules increased by 445.088 kJ/mol, and the drying potential of mean force (PMF) free energy also fell by 253.040 kJ/mol. However, an increase in temperature from 300 to 330 K in the aqueous solution only results in an increase of 23.013 kJ/mol in electrostatic interaction energy and a decrease of 34.956 kJ/mol in drying PMF free energy. Furthermore, we show that hydrogen bonding is the major form of electrostatic interaction involved, and directly affects the drying of glutathione. Therefore, choosing water-miscible solvents that minimise hydrogen-bond formation with glutathione will enhance its drying rate, and this is likely to be more efficient than increasing the temperature of the process. Thus, a power-saving technology can be used to produce the high bioactivity medicines.     

Reentrant condensation of lysozyme: Implications for studying dynamics of lysozyme in aqueous solutions of lithium chloride

Recent studies have outlined the use of eutectic solutions of lithium chloride in water to study microscopic dynamics of lysozyme in an aqueous solvent that is remarkably similar to pure water in many respects, yet allows experiments over a wide temperature range without solvent crystallization. The eutectic point in a (H₂O)_R(LiCl) system corresponds to R ≈ 7.3, and it is of interest to investigate whether less‐concentrated aqueous solutions of LiCl could be used in low‐temperature studies of a solvated protein. We have investigated a range of concentrations of lysozyme and LiCl in aqueous solutions to identify systems that do not show phase separation and avoid solvent crystallization on cooling down. Compared to the lysozyme concentration in solution, the concentration of LiCl in the aqueous solvent plays the major role in determining systems suitable for low‐temperature studies. We have observed interesting and rich phase behavior reminiscent of reentrant condensation of proteins. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 624–629, 2014.     

Effects of ethanol on the secondary structure specific hydration properties of Chymotrypsin Inhibitor 2 in its folded and unfolded forms

Molecular dynamics simulations were performed for the protein Chymotrypsin Inhibitor 2 in water–ethanol binary mixtures at several ethanol concentrations at two different temperatures to explore the differential role of ethanol on the hydration properties of the secondary structural segments of the protein. We find that, compared to the water structure around the secondary structure segments of the unfolded protein in pure water, water molecules preferred to be structured more in presence of ethanol. The structuration of ethanol is noticed to be higher around helix and sheet, as compared to that around the loop. The differential slow structural relaxations of the hydrogen bonds involving the secondary structural segments of protein in water–ethanol mixed solutions, as compared to that in pure water infer that ethanol alters the water properties in a remarkable manner. The slow relaxations of the hydrogen bonds formed between ethanol and the secondary structural segments of the protein, in general, suggests that ethanol interacts directly with the protein. However, the relatively faster relaxation of the ethanol-segment hydrogen bonds involving unfolded protein as compared to that involving the folded protein suggests that the unfolded protein surface can remain dynamically free to interact with water more.     

Spectrophotometric characteristics and assay of pure pyronin Y

The spectrophotometric characteristics of analytically pure pyronin Y have been investigated. Addition of metal ions (Fe3+, Zn2+, and Mg2+) and of dextrin were shown not to influence the absorption characteristics. The composition of the solvent strongly influenced the value of the extinction coefficient. Aqueous ethanolic solutions with a content of about 50% ethanol gave higher epsilon-values than those found for more concentrated ethanol solutions. The difference can be explained by the existence of a solvent-solute complex in the less concentrated ethanol solutions. A new spectrophotometric assay is proposed using the epsilon-value 11.7 X 10(4) lmol-1 cm-1 found in aqueous ethanol (52%) as standard.     

湘西慈竹叶中黄酮的提取与测定

设计正交试验,以溶剂浸提-回流法提取,以分光光度法测定,研究了湘西慈竹叶中总黄酮提取和测定的方法。结果表明:溶剂、溶剂浓度、水浴温度、回流时间4种因素对提取效果影响大小依次为,溶剂>溶剂浓度>回流时间>水浴温度,其中溶剂及其浓度是主要因素,回流时间和水浴温度是次要因素。不同溶剂对慈竹叶总黄酮提取效果影响大小顺序为丙酮>乙醇>乙醚。浸提-回流法提取湘西慈竹叶中黄酮类化合物的优化提取条件(固定料液比为1∶20)是:以75%的丙酮为浸提剂,于70℃下回流加热4 h。在优化提取条件下,测得湘西慈竹叶总黄酮含量为13.0 mg.g-1(春)~12.4 mg.g-1(秋),春季新叶中黄酮含量略高于秋季老叶。湘西慈竹叶总黄酮含量略低于四川南充慈竹叶。     

In vivo measurement of flavour release from mixed phase gels

Flavour release was investigated from pure gelatin, pure agarose and mixed gelatin-agarose gels, all containing 25% sucrose and flavoured with p-cymene, ethyl butyrate, pyrazine and ethanol. Gels were characterised by optical microscopy, and rheological techniques to determine phase separation, elastic modulus and melting temperature. Volatile release was measured by monitoring the four volatiles in the expired air from one individual eating the gels, using Atmospheric Pressure Chemical Ionisation-Mass Spectrometry. The release pattern of p-cymene was not affected by gel type. The release of ethanol, ethyl butyrate and pyrazine was affected to different extents by the matrix suggesting that both the properties of the volatile and the matrix determine volatile release in vivo.     

Regenerated cellulose-silk fibroin blends fibers

Fibers made of cellulose and silk fibroin at different composition were wet spun from solutions by using N-methylmorpholine N-oxide hydrates (NMMO/H(2)O) as solvent and ethanol as coagulant. Different spinning conditions were used. The fibers were characterized by different techniques: FTIR-Raman, scanning electron microscopy, wide-angle x-ray diffraction, DSC analysis. The results evidence a phase separation in the whole blends compositions. The tensile characterization, however, illustrates that the properties of the blends fibers are higher respect to a linear behaviour between the pure polymers, confirming a good compatibility between cellulose and silk fibroin. The fibers containing 75% of cellulose show better mechanical properties than pure cellulose fibers: modulus of about 23 GPa and strength to break of 307 MPa.     

Low Temperature Absorption Spectra of Chlorophyll a in Polar and Nonpolar Solvents

Absorption spectra of chlorophyll a were measured in polar and non-polar solvents, as a function of temperature from 298 degrees to 77 degrees K. Both dilute and concentrated solutions were examined. In both types of solvents at room temperature, the absorption spectra of concentrated solutions differ from dilute ones in that the half width of the main red absorption band is greater, and all bands are shifted to longer wavelengths. These differences are largely due to the presence of dimers when the pigment concentration is high. In dilute ethanol solutions, where the chlorophyll is unassociated, cooling causes a red shift in all bands which is due to the increased polarity of the solvent at low temperature. On cooling at high concentrations in ethanol and EPA, a new band appears near 700 nm. This band is attributed to dimers present prior to cooling, but absorbing at shorter wavelengths at room temperature. In nonpolar solvents, a band near 700 nm appears at the solvent freezing point. In these solvents, the "700" nm absorption is attributed to dimers, and/or small polymers, partly formed by cooling. A change in aggregate geometry when the solvent becomes viscous or frozen can account for the appearance of this "700" nm absorption band at low temperature, in polar and nonpolar media.     

Cation effects on swelling of kappa-carrageenan: a photon transmission study

Swelling behavior of kappa-carrageenan gels in water and KCl solutions was investigated by photon transmission experiments following the preparation of gels in the presence and absence of externally added K+ ion as a gel promoting agent. Transmitted photon intensity, Itr, increased continuously during swelling depending on the carrageenan and ion content in the gel. This increase in Itr was modeled using the Li-Tanaka equation. Both the experimental work and the model showed that the swelling of low carrageenan and ion content gels took less time than that of high ion content gels. It is confirmed that double helices in a swollen gel move much faster in pure water than in KCl solution during swelling processes. Swelling time constants, tau1, and collective diffusion coefficients, Do, were measured for the gels swollen in water and KCl solutions.     

Sequence analysis of end-labeled DNA fragments by solvolysis in hot aqueous piperidine solutions

One-lane DNA sequencing by solvolysis in hot aqueous piperidine solutions, originally described for 5'-32P-labeled DNA (B. Ambrose and R. Pless (1985) Biochemistry 24, 6194-6200), is extended to 3'-labeled fragments. A salt-free sample for electrophoresis can be obtained by using 1 M LiCl in the solvolysis mixture and removing this salt from the dried hydrolysate by washing with ethanol. Rate and distribution of DNA cleavage in hot aqueous piperidine, containing 0.3 M NaCl, are studied in dependence of temperature, solvent, amine concentration, and reaction time. An increase in temperature strongly accelerates overall DNA degradation, but leaves the distribution of cleavage essentially unchanged. When 50% aqueous ethanol is substituted for water as the reaction solvent, the overall cleavage is slower, and scission at G-sites is enhanced relative to cleavage at the other bases. A rise in the piperidine concentration strongly accelerates the reaction, except at very high amine concentration. Cleavage at A-, G-, and C-sites increases steadily with reaction time, while the T-cleavage observed takes place primarily at the very beginning of the solvolysis.     

水/乙醇溶剂中阿奇霉素的热力学及多晶型研究

在278.2~308.2 K温度范围内,测定阿奇霉素在水/乙醇混合溶剂中的溶解度,根据固液平衡理论建立了该体系的溶解度修正模型。采用X线粉末衍射法和差示扫描量热法,对阿奇霉素在不同温度、不同体积比的水/乙醇混合溶剂中得到的晶体进行鉴别。同时利用溶解度数据估算了阿奇霉素在水/乙醇体系中的溶解热(-25.26~-16.11 k J/mol)、混合热(-9.94~-3.25 k J/mol)。通过溶液化学理论推导了阿奇霉素溶剂化平衡常数K与活度系数γ2的方程:γ2=1/(1+K),建立了溶剂化焓与温度、水/乙醇两者体积比(φ)之间的关系式,为ΔH=RTln(17.86exp(3.4φ)-1)。采用溶析结晶方法得到的6种阿奇霉素晶体,均属单斜晶系,但具有不同的晶胞参数且其密度和熔点也不同。同时发现温度越高,水/乙醇体积比越大,得到的晶体稳定性越差(晶体的熔点和密度降低)。在水/乙醇混合溶剂的溶析结晶体系中,产生阿齐霉素多晶型的现象与溶剂化作用的强弱有关。     

Dielectric studies on water in solutions of purified lecithin vesicles

The complex permittivity of sonicated aqueous solutions of purified dimyristoylphosphatidylcholine has been measured as a function of frequency between 3 kHz and 40 GHz. The dielectric spectrum of the samples shows two dispersion/absorption regions, one centered at about 80 MHz the other at about 20.GHz (30°C). Otherwise than in previous studies no additional dispersion/absorption process has been found at frequencies below 10 MHz.The complex dielectric spectrum of the samples is discussed with respect to the dynamical state of solvent water in solutions of single-bilayer vesicles. The main relaxation time of the solvent water, τ₁ ((2πτ₁)^?1 ≈ 20 GHz), is smaller than that of pure water, τ_W, at the same temperature. This effect results from the action of internal depolarizing fields which obviously overcompensate and enhancement of τ₁ due to specific solute/solvent interactions (hydration) as had been previously found with micellar solutions of lysolecithins.It cannot be excluded, that some solvent water shows unusual dynamical behaviour. If there exists a substantial amount of such motionally perturbed water, however, it must be characterized by a relaxation time close to that of the phosphorylcholine zwitterions, τ₂ ((2πτ₂)^?1 ≈ 80 MHz).     

Modification of the Solid-State Nature of Sulfathiazole and Sulfathiazole Sodium by Spray Drying

Solid-state characterisation of a drug following pharmaceutical processing and upon storage is fundamental to successful dosage form development. The aim of the study was to investigate the effects of using different solvents, feed concentrations and spray drier configuration on the solid-state nature of the highly polymorphic model drug, sulfathiazole (ST) and its sodium salt (STNa). The drugs were spray-dried from ethanol, acetone and mixtures of these organic solvents with water. Additionally, STNa was spray-dried from pure water. The physicochemical properties including the physical stability of the spray-dried powders were compared to the unprocessed materials. Spray drying of ST from either acetonic or ethanolic solutions with the spray drier operating in a closed cycle mode yielded crystalline powders. In contrast, the powders obtained from ethanolic solutions with the spray drier operating in an open cycle mode were amorphous. Amorphous ST crystallised to pure form I at ≤35 % relative humidity (RH) or to polymorphic mixtures at higher RH values. The usual crystal habit of form I is needle-like, but spherical particles of this polymorph were generated by spray drying. STNa solutions resulted in an amorphous material upon processing, regardless of the solvent and the spray drier configuration employed. Moisture induced crystallisation of amorphous STNa to a sesquihydrate, whilst crystallisation upon heating gave rise to a new anhydrous polymorph. This study indicated that control of processing and storage parameters can be exploited to produce drugs with a specific/desired solid-state nature.KEY WORDS: amorphous state, dynamic vapour sorption, particle habit, physical stability, polymorphism, sulfathiazole     

In vitro and in vivo release of albumin using a biodegradable MPEG-PCL diblock copolymer as an in situ gel-forming carrier

An MPEG-PCL diblock copolymer was synthesized as an in situ gel carrier, and its phase transition behavior in aqueous solutions was examined. For comparison, aqueous solutions of Pluronic F-127, a widely used injectable gel-forming solution, were also studied. Both MPEG-PCL copolymer and Pluronic aqueous solutions were sols at room temperature. As the temperature was increased above room temperature, the diblock copolymer and Pluronic solutions underwent a sol-to-gel phase transition, which manifested as an increase in viscosity indicative of the formation of a gel. All of the copolymer solutions became gels at body temperature, although the gel viscosity increased with the increasing concentration of the MPEG-PCL diblock copolymer in the solution. In in vitro experiments, in which the gels were exposed to PBS, the MPEG-PCL gels maintained their structural integrity for more than 28 days, whereas the Pluronic gel disappeared within 2 days. The same results were observed when the polymer solutions were subcutaneously injected into rats. The MPEG-PCL gels maintained their structural integrity longer than 30 days, while the Pluronic gel could not be observed after 2 days. The ability of the gels as drug carriers was studied by measuring the release of fluorescein isothiocyanate-labeled bovine serum albumin (BSA-FITC) from MPEG-PCL diblock copolymer gels in vitro as well as in vivo. In vitro, BSA release was sustained above 20 days, with a greater release at lower diblock copolymer concentration; by contrast, Pluronic gels exhibited almost complete release of BSA-FITC within 1 day. When the BSA-FITC-loaded diblock copolymer and Pluronic solutions were subcutaneously injected into rats, they immediately transformed into a gel. In vivo, sustained release of BSA-FITC over 30 days was observed from the MPEG-PCL gel, whereas BSA-FITC release from the Pluronic gel ceased within 3 days. Collectively, the present findings show that MPEG-PCL diblock copolymer solutions are thermo-responsive and maintain their structural integrity under physiological conditions, indicating that they are suitable for use as injectable drug carriers.