Improvement of encapsulation efficiency of water-in-oil-in-water emulsion with hypertonic inner aqueous phase.

Water-in-oil-in-water (w/o/w) emulsions encapsulating tryptophan or theophylline were prepared where these compounds are regarded as model drugs. The effects of sodium chloride on the drug entrapment into the w/o/w emulsions and on the separation of aqueous phases were studied. The degree of encapsulation of tryptophan in the w/o/w emulsion increased with the concentration of sodium chloride added in the inner aqueous phase, while it decreased with that in the outer aqueous phase. As for theophylline, although the degree increased with a concentration of sodium chloride in the inner phase, the effect was smaller than that on tryptophan. The difference in the effects between on tryptophan and on theophylline was attributed to their partition coefficients. Theophylline was easily leaked out from the inner phase to the outer aqueous phase after its dissolution and diffusion in the oil phase due to a higher partition coefficient. More than 55% of the aqueous phase was separated from the w/o/w emulsion within 24 h, when sodium chloride was not added in the inner aqueous phase. However, the separation was not observed when more than 0.2M sodium chloride was added. To the contrary, sodium chloride added in the outer aqueous phase accelerated the separation. It was, therefore, concluded that sodium chloride in the inner aqueous phase plays an important role in suppression of the separation and in encapsulation of the drug which does not penetrate into the oil membrane.

Drug release from w/o/w emulsions prepared with different chitosan salts and concomitant creaming up.

Water-in-oil-in-water (w/o/w) emulsions containing chitosan and tryptophan in the inner aqueous phase were prepared. The acids used to dissolve chitosan were formic, acetic, butyric and lactic acids. The effects of these organic acids and pH of the inner aqueous phase on the release of tryptophan and on the separation of the aqueous phase due mainly to creaming up during storage were studied. When the inner aqueous phase was an acidic chitosan solution, the release rate was almost the same irrespective of the acids mentioned above. When acetic and butyric acids were neutralized with sodium hydroxide, the release of tryptophan was prolonged. However, it was enhanced again by an excess amount of sodium hydroxide. On the other hand, the release was enhanced after neutralization of formic and lactic acids. Separation of the aqueous phase from the w/o/w emulsion during storage was remarkably delayed after neutralizing the chitosan solution containing acetic or butyric acids, while not so much in the cases of formic and lactic acids. It is concluded that these w/o/w emulsions were stabilized in the presence of neutralized chitosan gel suspensions containing acetate or butyrate ions in the inner aqueous phase.

Basic study for stabilization of w/o/w emulsion and its application to transcatheter arterial embolization therapy.

Stabilization of w/o/w emulsion and its application to transcatheter arterial embolization (TAE) therapy are reviewed. W/o/w emulsion was stabilized by making inner aqueous phase hypertonic, addition of chitosan in inner phase, and techniques of phase-inversion with porous membrane. Lipiodol w/o/w emulsion for TAE therapy was prepared by using a two-step pumping emulsification procedure. The procedure is so easy that the emulsion could be prepared even during the surgical operation. The deposition after hepatic arterial administration of the emulsion was detected by an X-ray CT scanner. The concentration of epirubicin hydrochloride (EPI) in liver was increased and its residence was prolonged by encapsulating it in the w/o/w emulsion. The toxic effects of EPI and lipiodol on the normal hepatic cells were reduced. The w/o/w emulsion prepared by us is a suitable formulation for the TAE therapy.

Development of a new type nozzle and spray-drier for industrial production of fine powders.

Sophisticated nozzle and spray-drier were newly developed. The nozzle type was that of four-phase spraying, where two liquid streams and two air streams were blown off. The spray pattern from the nozzle was of a hollow-cone type. Mean diameter of droplets in the mist was 13.2 microm in weight average in the condition of blowing at 776 g/min in air flow and 500 ml/min in liquid flow. That is, the weight-based flow ratio of air to liquid was as small as 1. 55. The geometric standard deviation of the droplet size was less than 1.65. This nozzle was still available for a concentrated suspension up to 27% solid without formation of the sludge on the orifice. Thus, fine powder with 1.99 microm in mean diameter was obtained by means of the nozzle and the spray-drier newly developed by us. These are promising for industrial production of the fine powder with low energy and high recovery.

Influence of the permeation enhancers 1-alkyl-2-pyrrolidones on permeant partitioning into the stratum corneum.

In a previous study, the enhancing effects of a series of 1-alkyl-2-pyrrolidones (APs; 1-ethyl, 1-butyl, 1-hexyl, and 1-octyl-2-pyrrolidone) on the transport of steroidal permeants across hairless mouse skin were investigated via a parallel pathway skin model. Isoenhancement concentration conditions were deduced under which different APs induce essentially the same transport enhancement for the lipoidal pathway of the stratum corneum (s.c.). As a continuing effort to understand the mechanism of action of permeation enhancers, the influence of the APs on permeant partitioning into hairless mouse s.c. was investigated under the isoenhancement concentration conditions using beta-estradiol (E2 beta) as the model permeant. The amount of E2 beta uptake into s.c. was found to be essentially the same for all the APs under these isoenhancement conditions. This result suggests that inducing a higher partitioning tendency for E2 beta into the lipoidal pathway of hairless mouse s.c. is a principal mechanism of action of the APs in enhancing transdermal transport. The uptake of the APs into s.c. lipoidal domains was also determined, and the results show only a modest (approximately 2-fold) increase in the uptake of the APs in going from 1-ethyl-to 1-octyl-2-pyrrolidone under isoenhancement conditions. This indicates the potency of the APs as permeation enhancers is only very modestly dependent upon the alkyl chain length in this chain length region when compared at concentrations in the microenvironment where the action occurs in the lipid domains.

[Effect of formation of surface complex between hydroxypropylcellulose and sodium dodecylsulfate on dispersion/flocculation of O/W-type emulsion of soybean oil].

An O/W-type emulsion of soybean oil (theta v = 0.05) was prepared in the presence of various concentrations of hydroxypropylcellulose (HPC) and sodium dodecylsulfate (SDS). The mean diameter (d) of secondary particles of the oil droplets decreased with concentrations of SDS and HPC after attaining a maximum at [SDS] = 5 mmol/dm3 and [HPC] = 10(-3) g/dl, while the relative viscosity (eta rel) of the emulsion increased with an SDS concentration after attaining a minimum around 5 mmol/dl3 when the HPC concentration was kept constant. These facts were explained in terms of the formulation of a surface complex by hydrophobic interaction between SDS and HPC adsorbed on the surface of the droplets. When the concentration of SDS is low, it bridges between the HPC segments intra- and intermolecularly on and between the particles, resulting in shrinking of the HPC surface layers as well as in bridging among the particles. Therefore, the total volume of the secondary particles effective on eta rel decreases while the d-value increases with the SDS concentration. When the concentration becomes high, binding ratio of SDS to HPC also increases and the segments are repulsive each other owing to the negative charges given by the bound SDS. As a result, the adsorption layer swells and the secondary particles are redispersed. This fact causes an increase in eta rel and a decrease in d-value with a concentration of added SDS.

Hemin binding to DNA with bis-dentate acridine intercalator.

Hemin was covalently connected to two 9-aminoacridines (9AA) through its two propionates, and the binding properties of this bis-dentate compound (hemin(9AA)2) to DNA were examined by visible absorption spectroscopy. The binding affinity of the hemin(9AA)2 was found to be higher than that of the hemin, and this should be attributed to the two linked acridine moieties. The binding constant (K) and the number of binding sites per nucleotide (n) were estimated by Scatchard plot analyses. Though the order of the K value of hemin(9AA)2 was similar to that of 9AA, the hemin(9AA)2 was analyzed to have a smaller n value, the order of which was of about 10(-4). The small n value may reflect the sequence specificity of the bis-dentate hemin(9AA)2 on binding to the DNA.

[Effects of temperature and pH on hydrolysis of monoacetoacetin in an aqueous phase].

The kinetics of hydrolysis of monoacetoacetin (MAA; 1,2-dihydroxy-3-acetoacetyloxypropane) in 300 mM phosphate buffer was studied by means of high performance liquid chromatography. MAA was hydrolyzed to glycerin (Glyc) and acetoacetate (AA), the latter of which furthermore decomposed to acetone (Acet) and CO2 at high temperature. The rate constant, k1, for the reaction of MAA-->AA+Glyc increased with increases in temperature and pH. On the other hand, another one, k2, for the reaction of AA-->Acet+CO2 increased with an increase in temperature but showed a minimum around pH 7.5 when pH was changed at a given temperature. The activation energy for k1 was almost constant in the range of studied pHs (5-8) but the pre-exponential factor, k1,0, increased with an increase in pH. It was concluded that the decomposition reaction of MAA is consecutive in a sequence of MAA-->AA-->Acet. Calculated concentrations of MAA, AA, and Acet through the obtained k1 and k2 together with a given initial concentration of MAA (= 40 mM) were compared with those experimentally observed on the time course of the hydrolysis. They were in good agreement with each other.

Effect of phosphorylated organic compound on the adsorption of bovine serum albumin by hydroxyapatite.

The amount of adsorption of bovine serum albumin (BSA) by hydroxyapatite (HAP) increased with a concentration of CaCl2 due to the bridging effect of Ca2+ between adsorbate BSA and adsorbent HAP. On the other hand, it decreased remarkably with a concentration of K2HPO4. This was explained in terms of the effects of ionic strength and competitive adsorption between inorganic phosphate anion (Pi) and BSA, because BSA is in negatively charged over the examined pHs. A similar effect was observed in the presence of phosphorylated compounds such as phosphoserine, phytate, and phosphorylated polyvinylalcohol. The inhibiting effect of these compounds was stronger than that of their mother compounds (serine, inositol, and polyvinylalcohol). This result shows that phosphate groups bound to the mother compounds interfere with the adsorption of BSA by HAP in the same manner that Pi does. Although the adsorption of BSA was almost irreversible with respect to dilution with water, desorption was performed when these organic phosphorylated compounds were added after the accomplishment of the adsorption of BSA. However, the effective concentration of the phosphorylated compounds for the desorption of BSA was fairly higher than that for the competitive inhibition against the BSA adsorption.

Decrease in the solubility product of hydroxyapatite by the adsorption of surface-active ion.

The solubility product (Ksp) of hydroxyapatite (HAP) in ionic surfactant solutions decreased with an increase in the adsorbed amount of the surface-active ions (dodecylammonium and dodecyl sulfate ions). In the presence of nonionic surfactant (polyoxyethylene(10) octylphenyl ether), Ksp was almost constant within the limit of experimental error. It was concluded that the decrease in Ksp is due to the decrease in the chemical potential of HAP through the adsorption of the surface-active ion on HAP, and due to the spontaneous change in the constituent ions of the HAP surface (i.e., surface complex formation) by ion-exchange with the surface-active ion.

Interaction of hydroxyapatite with sodium chondroitin sulfate and calcium chondroitin sulfate in an aqueous phase.

The suspension pH at a given concentration of hydroxyapatite (HAP) decreased with the concentration of calcium chondroitin-6-sulfate (CaChs), whereas it increased with the concentration of sodium chondroitin-6-sulfate (Na2Chs). The former effect is due to the increase in the concentration of H+ by ion exchange between H+ on the surface of HAP and Ca2+ of CaChs, and the latter is due to the protonation of phosphate ion (Pi) released from the surface of HAP. The absorbed amount of chondroitin-6-sulfate anion (Chs) by HAP was higher with CaChs than with Na2Chs over the concentration range examined. The equilibrium concentration of Pi decreased with increasing concentration of added CaChs because the concentration of free Ca2+, which dissociates from CaChs, regulates the free concentration of Pi through the restriction of the solubility product of HAP (Ksp). In contrast, that in the presence of Na2Chs increased with increasing concentration of added Na2Chs owing to the anion exchange between Chs and Pi of the HAP surface. The total concentration of Ca2+, which was released from HAP into the solution phase, increased after passing through a minimum with increasing concentration of added Na2Chs. That is, the concentration of Ca2+ free from Chs decreased with an increase in the concentration of released Pi owing to the restriction of the solubility product, whereas that of Ca2+ bound by Chs increased with increasing concentration of added Na2Chs through the ion exchange of Na+ with Ca2+. It was confirmed by the dialysis method that the value of Ksp was almost constant around 10, although HAP dissolves incongruently in the presence of Na2Chs.