Ethanol-free Solution Research Articles

Preformulation and Long-Term Stability Studies of an Optimized Palatable Praziquantel Ethanol-Free Solution for Pediatric Delivery.

To date, the treatment for cysticercosis and neurocysticercosis consists of a single oral intake of praziquantel (5-10 mg/kg), which since it is only available as tablets, hinders its administration to pediatric patients. Praziquantel is a poorly water-soluble drug which represents a challenge for its formulation in solution, particularly for the pediatric population. Thus, this study aimed to develop a palatable solution for praziquantel using pharmaceutical-accepted co-solvent systems. A design of experiments approach was applied to identify the optimal conditions for achieving a suitable amount of praziquantel in solution using co-solvent mixtures. Thus, praziquantel solubility increased from 0.38 up to 43.50 mg/mL in the optimized system. A taste masking assay in healthy human volunteers confirmed a successful reduction of drug bitterness after the addition of selected flavors and a sweetener. Stability studies were also conducted at different temperatures (4, 25, and 40 °C) for 12 months Even though the presence of the three known impurities of praziquantel was observed, their amounts never exceeded the acceptance criteria of the USP. Thus, this novel approach should be considered a valuable alternative for further preclinical studies considering the high prevalence of this infection worldwide.

Effects of chronic cocaine and ethanol self-administration on brain dopamine receptors in a rhesus monkey model of polysubstance abuse.

Most individuals with cocaine use disorder also use alcohol; however, little is known about the behavioural and pharmacological mechanisms that promote co‐abuse. For example, although studies in humans and animals have documented that chronic use of either alcohol or cocaine alone decreases D2‐like receptor (D2R) availability, effects of co‐abuse of these substances on dopamine receptor function have not been characterized. These studies examined the effects of long‐term cocaine self‐administration in 12 male rhesus monkeys who also consumed either ethanol or an ethanol‐free solution each day (n = 6 per group). Specifically, all monkeys self‐administered cocaine (0.1 mg/kg per injection) 5 days per week in the morning. In the afternoon, six monkeys consumed 2.0 g/kg ethanol over 1 h to model binge drinking and six monkeys drank an ethanol‐free solution. Assessment of D2R availability using positron emission tomography (PET) and [11C]raclopride occurred when monkeys were drug‐naïve and again when monkeys had self‐administered approximately 400‐mg/kg cocaine. D3R function was assessed at the same time points by determining the potency of the D3R‐preferring agonist quinpirole to elicit yawns. Chronic cocaine self‐administration decreased D2R availability in subregions of the basal ganglia in control monkeys, but not those that also drank ethanol. In contrast, D3R sensitivity increased significantly after chronic cocaine self‐administration in ethanol‐drinking monkeys but not controls. These results suggest that co‐use of ethanol substantially changes the effects of chronic cocaine self‐administration on dopamine receptors, specifically implicating D3R as a target for medications in these individuals.

Hospital preparations of ethanol-free furosemide oral solutions: Formulation and stability study.

Furosemide is a diuretic frequently used in the therapeutic management of edema associated with cardiac, renal, and hepatic failure and hypertension. However, there are a very low number of pharmaceutical dosage forms containing furosemide that are suitable for children under 6- years old. Therefore, there is a real need to develop hospital preparations, especially in the hospital. Four oral pediatric solutions of furosemide (2 mg/Ml) were formulated. Two of those solutions did not contain ethanol. For each formulation, 12 batches of 1600.0 Ml were prepared and packaged in 250.0 Ml brown glass bottles with polypropylene screw caps. The physicochemical properties (visual appearance, pH, osmolarity, drug content) and microbiological quality of the finished product were determined on the freshly prepared solutions and after 90 days of storage at 30°C/65% RH. The physicochemical and microbiological characteristics of the freshly prepared solutions were within the prescribed specifications. After 90 days of storage at 30°C/65% RH, the solutions containing sucrose and those without ethanol showed a slight decrease in pH and furosemide content of about 2.5%–4.5% (w/w). Despite this slight decrease, the characteristics remained within the prescribed specifications. Based on the stability profile of the ethanol-free solution containing sorbitol, it could be implemented in hospitals for the care of pediatric patients.

Thermostable ethanol tolerant xylanase from a cold-adapted marine species Acinetobacter johnsonii

Abstract A xylanase-producing bacterium, isolated from deep sea sediments, was identified as the cold-adapted marine species Acinetobacter Johnsonii. A cold-adapted marine species Acinetobacter Johnsonii could grow at 4 °C. The optimum temperature and pH of xylanase from a cold-adapted marine species Acinetobacter Johnsonii were 55 °C and pH 6.0. Xylanase from a cold-adapted marine species Acinetobacter Johnsonii remained at 80% activity after incubation for 1 h at 65 °C. The xylanase activity was 1.2-fold higher in 4% ethanol solution than in ethanol free solution. Gibbs free energy of denaturation, ΔG, was higher in 4% ethanol solution than in ethanol free solution. Thermostable ethanol tolerant xylanase was valuable for bioethanol production by simultaneous saccharification and fermentation process with xylan as a carbon source.

Ethanol tolerant endoglucanase from Aspergillus niger isolated from wine fermentation cellar

Abstract The endoglucanase from Aspergillus niger isolated from a wine fermentation cellar was ethanol tolerant. Optimum pH and temperature of the endoglucanase activity was 5.0 and 60 °C. The endoglcuanase activity in 8% ethanol solution was higher than that in ethanol-free solution. Melting point temperature of the endoglucanase in 8% ethanol solution was 1.9 °C higher (40.0 °C) than that in ethanol-free solution (37.1 °C). At 45–60 °C, Gbbs free energe, ΔG, in 8% ethanol solution was higher than that in ethanol-free solution. The α-helix content increased in 8% ethanol solution. The increasing of α-helix content led to higher activity and better thermostability in high concentration ethanol solution. The acidic, thermostable and ethanol tolerant endoglucanase was valuable for bioethanol production by simultaneous saccharification and fermentation process.

A cold adapt and ethanol tolerant endoglucanase from a marine Bacillus subtilis

Abstract The catalytic properties and thermodynamic kinetics of the endoglucanase from a marine Bacillus subtilis were analyzed. Optimum pH and temperature of the endoglucanase activity were 5.0 and 35 °C. The endoglucanase activity, melt point temperature was 1.13 folds (247.02 U·ml− 1), 2.1 °C higher (39.2 °C) in 6% ethanol solution than that (218.60 U·ml− 1), (37.1 °C) in free ethanol. At 40 °C–55 °C, Gibbs free energy, ΔG, and the content of α-helix was higher in 6% ethanol solution than that in ethanol free solution. The increasing of α-helix content led to higher activity and better thermostability in ethanol solution. The cold adapt ethanol tolerant endoglucanase was valuable for bioethanol product by simultaneous saccharification and fermentation process.

Efficient automated synthesis of 2-(5-[18F]fluoropentyl)-2-methylmalonic acid ([18F]ML-10) on a commercial available [18F]FDG synthesis module

[18F]ML-10 (2-(5-[18F]fluoro-pentyl)-2-methylmalonic acid) is a small molecule positron emission tomography (PET) probe for apoptosis imaging. Automated synthesis of [18F]ML-10 was developed by using two different purification methods through a direct saponification procedure on a modified commercial [18F]Fluoro-2-Deoxyglucose ([18F]FDG) synthesizer. C18 purification method 1: The final [18F]ML-10 solution containing ethanol was obtained with radiochemical yields of 60±5% (n=5) at the end of bombardment (EOB) and radiochemical purity of 98% in 35min. Al2O3 and SCX purification method 2: To avoid possible side effects of a conventional ethanol-containing formulation, an new ethanol-free solution of [18F]ML-10 was also developed, the radiochemical yields was 50±5% (n=5, EOB) within 45min and the radiochemical purity was 98%.

Separation of ovotransferrin and ovomucoid from chicken egg white

Ovotransferrin and ovomucoid were separated using 2 methods after extracting the ovotransferrin- and ovomucoid-containing fraction from egg white. Diluted egg white (2×) was added to Fe3+ and treated with 43% ethanol (final concentration). After centrifugation, the supernatant was collected and treated with either a high-level ethanol (61% final concentration) or an acidic salt combination (2.5% ammonium sulfate and 2.5% citric acid) to separate ovotransferrin and ovomucoid. For the high-level of ethanol method, ovotransferrin was precipitated using 61% ethanol. After centrifugation, the precipitant was dissolved in 9 vol. of distilled water and the residual ethanol in the solution was removed using ultrafiltration. The supernatant, mainly containing ovomucoid, was diluted with 4 vol. of water, had ethanol removed, and was then concentrated and used as the ovomucoid fraction. For the acidic salt precipitation method, the ethanol in the supernatant was removed first. The ethanol-free solution was then concentrated and treated with a 2.5% ammonium sulfate and 2.5% citric acid combination. After centrifugation, the precipitant was used as the ovotransferrin and the supernatant as the ovomucoid fraction. The ovomucoid fraction from both of the protocols was further purified by heating at 65°C for 20 min and the impurities were removed by centrifugation. The yields of ovomucoid and ovotransferrin were >96 and >92%, respectively. The purity of ovomucoid was >89% and that of the ovotransferrin was >88%. The ELISA results confirmed that the activity of the separated ovotransferrin was >95%. Both of the protocols separated ovotransferrin and ovomucoid effectively and the methods were simple, fast, and easy to scale up.

Preparation and stability of ethanol-free solution of [18F]florbetapir ([18F]AV-45) for positron emission tomography amyloid imaging

We have developed an ethanol-free formulation method of [(18) F]florbetapir ([(1) (8) F]AV-45) using a commercially available automated JFE multi-purpose synthesizer. We have also evaluated the radiochemical stability in an ethanol-free solution of [(18) F]AV-45 under visible light irradiation and dark conditions by comparison with a conventional 10% ethanol solution of [(18) F]AV-45. [(18) F]AV-45 was obtained with a radiochemical yield of 55.1 ± 2.2% (decay-corrected to end of bombardment), specific activity of 591.6 ± 90.3 GBq/µmol and radiochemical purity of >99% within a total synthesis time of about 73 min. The radiochemical purity of [(18) F]AV-45 formulated by dissolving the ethanol-free solution was found to decrease as a function of the period of exposure to visible light. In contrast, the visible light photolysis could be suppressed by adding 10% ethanol to the formulation or by avoiding exposure to visible light. In the radiosynthesis of [(18) F]AV-45 formulated by dissolving the ethanol-free solution, [(18) F]AV-45 could be obtained with high radiochemical purity and high stability by avoiding exposure to visible light.

Simple preparation and purification of ethanol-free solutions of 3′-deoxy-3′-[18F]fluorothymidine by means of disposable solid-phase extraction cartridges

3'-Deoxy-3'-[(18)F]fluorothymidine ([(18)F]FLT) shows great potential as a tracer for proliferative studies with PET. However, its regular application is often limited by low radiochemical yields and the use of a troublesome HPLC separation. Moreover, a high content of ethanol, at least one-fold higher than the European Pharmacopoeia and US Pharmacopoeia's established limit, is always present in the final product. The present study reports an optimization of the reaction conditions and a simple and straightforward purification step which affords a solution of [(18)F]FLT suitable for human use. Several conditions and materials were tested for both the nucleophilic substitution and purification step. The latter was achieved by means of a series of commercial solid-phase extraction cartridges. Very conveniently, the whole one-pot synthesis was carried out on commercial automated modules using basically the same setup employed for the synthesis of [(18)F]FDG. Under routine conditions, radiochemical yields of 37% [decay-corrected to start of synthesis (SOS)] were achieved in ca. 39 min from SOS, with radiochemical purities >98% (usually >99%). The negligible radiolysis observed could be easily suppressed by adding 0.5% of EtOH. Typical unlabelled chemical impurities detected were thymidine (0.15 ppm), thymine (0.28 ppm) and stavudine (0.05 ppm). A reliable, simple and efficient preparation of [(18)F]FLT has been developed, able to afford an ethanol-free solution of the tracer with no need for any HPLC purification. Because of its similarity to the [(18)F]FDG synthesis, the method can be readily implemented on basically all the commercial modules developed for this common radiotracer.

SFG Study of the Ethanol in an Acidic Medium−Pt(110) Interface: Effects of the Alcohol Concentration

Ethanol in an acidic solution-Pt(110) interface was studied by SFG spectroscopy (between 1820 and 2325 cm(-1)) to explore primarily the effects of the alcohol concentration. Stretching bands of H-Pt (ca. 1970 or 2050 cm(-1)) and CO (ca. 1980 and 2040 cm(-1)) species, produced by the ethanol oxidation, were detected during the adsorption and oxidation of 0-1 mol L(-1) ethanol in a 0.1 mol L(-1) HClO(4) solution on the electrode surface. Hydrogen and CO coadsorb stably on Pt(110) between 0.05 and 0.15 V in ethanol-containing solutions. In this potential range, the blue shift of the hydrogen resonance (ca. 80 cm(-1)) reveals a weakening of the hydrogen bonding between adsorbed hydrogen and water molecules in the double layer. After the hydrogen desorption (0.15 V), the formation of compact CO islands, depending on the ethanol concentration, lifts the Pt(110) surface reconstruction. In ethanol-free solution, the surface remains reconstructed. The lower-frequency CO band is assigned to the CO species adsorbed on (1 x 2) reconstructed Pt(110) domains, having smaller local coverages, while the higher-frequency CO band is attributed to the close-packed CO species adsorbed on (1 x 1) patches. The reaction pathway forming CO(2) is less favored with increasing ethanol concentration.

Chronic ethanol ingestion modifies the renin-aldosterone axis independent of alterations in the regulation of atrial natriuretic peptide.

Using an animal model, we have investigated the effects of chronic ethanol ingestion on the regulation of atrial natriuretic peptide (ANP) synthesis and release. Male Sprague-Dawley rats were maintained for 6 weeks on a liquid diet of ethanol (up to 20% v/v) as part of a 2% solution of calf milk replacer. Weight-matched controls received an equal volume of ethanol-free solution, and normal animals drank ad libitum. All animals received rat chow throughout the experiment. This model produced physiologically relevant levels of blood ethanol, as concentrations at the time of sacrifice were 171.98 +/- 39.26 mg/dl. Plasma renin activity was significantly elevated in response to ethanol treatment, whereas circulating aldosterone concentration was reduced. No alterations in the plasma or atrial tissue levels of ANP were evident, although we did observe a significant increase in the ventricular tissue levels of ANP from 45.1 to 71.8 ng/g as a consequence of ethanol treatment. Levels of both atrial and ventricular ANP mRNA were not different between alcohol-treated and liquid-restricted control animals, although both groups showed significant increases in the amount of transcript in comparison with rats drinking ad libitum. No significant increases in either arterial blood pressure or heart/body weight ratio were observed for ethanol-treated rats. These results suggest that modifications in the renin-aldosterone axis can occur independently of alterations in the regulation of ANP under the influence of chronic ethanol ingestion.

Chronic ethanol treatment increases the circulating plasma levels of B-type natriuretic peptide (BNP-45) in the rat.

Chronic ethanol ingestion is associated with a number of cardiovascular disorders, including stroke, heart failure, and hypertension. Given that the regulation of A-type natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) is known to be altered in both congestive heart failure and essential hypertension, we have investigated the regulation of BNP under the influence of ethanol ingestion. Sprague-Dawley rats were given ethanol in drinking fluid for a 6-week period, while a weight-matched liquid-restricted group received an equivalent volume of ethanol-free solution. Plasma BNP levels were increased in ethanol-treated animals relative to both liquid-restricted and normal control groups. No changes in cardiac BNP gene expression were observed, but an increased trend in atrial tissue BNP levels was evident. No changes in either the mRNA, tissue, or plasma levels of ANP were evident. These results suggest a differential regulation of natriuretic peptides under the influence of ethanol, and implicate chronic ethanol ingestion as a further clinical condition under which the plasma levels of a natriuretic peptide may be elevated.

Contractile, metabolic and electrophysiologic effects of ethanol in the isolated rat heart

The metabolic, functional and electrical effects of ethanol were studied in the isolated isovolumic rat heart retrogradely perfused at constant flow using phosphorus-31 nuclear magnetic resonance spectroscopy and surface electrogram recordings. Ethanol (0.75 to 6.0 vol%; 128 to 1024 m m) caused a concentration-dependent decline in developed pressure without a change in adenosine triphosphate, phosphocreatine, inorganic phosphate or pH. Ethanol (6%) caused abolition of electrical activity. The functional decline could be rapidly and completely reversed by perfusing with ethanol-free solution and, significantly although not completely, reversed by increasing perfusate calcium to 4 m m. Furthermore, ethanol shifted the perfusate calcium-tetanic pressure relationship in the presence of ryanodine (1 μ m) downwards and to the right. The results suggest ethanol's acute effects in this model are not mediated by changes in energy metabolism or cellular pH, but rather by sarcolemmal effects and by a decrease in both myofilament calcium sensitivity and maximal force generating ability.

16,16-Dimethyl prostaglandin E2 alleviates jejunal microvascular effects of ethanol but not the ethanol-induced inhibition of water, sodium, and glucose absorption

To examine the relation between ethanol-induced microvascular and absorptive changes, we have investigated the effect of 16,16-dimethyl prostaglandin E2 on the jejunal intraluminal plasma albumin loss (which was taken as a measure of microvascular changes) and the inhibition of water, sodium, and glucose transport caused by intraluminal ethanol. A group of 8 dogs received intravenously 16,16-dimethyl prostaglandin E2 at a dose of 0.1 μg/kg as a bolus followed by 0.05 μg/kg · hour for 2 h (prostaglandin-treated group). A second group of 8 dogs received no 16,16-dimethyl prostaglandin E2 (untreated group). In each dog of both groups, one jejunal segment was perfused with an ethanol-free solution (control segment) and an adjacent segment was perfused with the same solution containing 6% (wt/vol) ethanol (ethanol-perfused segment). The albumin loss (mg/g dry gut wt · 90 min, mean ± SE) by the control and the ethanol-perfused segments was 0.76 ± 0.23 and 8.29 ± 1.27, respectively, in the untreated group, and 0.66 ± 0.23 and 4.81 ± 0.67, respectively, in the prostaglandin-treated group. The ethanol-induced increase in albumin loss was significant in both groups, but was significantly lower (p < 0.05) in the prostaglandin-treated group than in the untreated group. Intraluminal ethanol depressed net water, sodium, and glucose transport by 74%, 52%, and 22%, respectively, in the untreated group, and by 92%, 65%, and 38%, respectively, in the prostaglandin-treated group. The magnitude of this depression did not differ significantly between the two groups. As 16,16-dimethyl prostaglandin E2 attenuated the ethanol-induced plasma albumin loss, but not the inhibition of water, sodium, or glucose transport, we conclude that the microvascular and the absorptive changes produced by ethanol are not mediated by the same mechanism.