一种水包油包胶型乳液的制备及其在乳化肠中的应用

以结冷胶和无水氯化钙为内水相凝固剂,酪蛋白酸钠为外水相乳化剂,制备一种水包油包胶（S/O/W）型 乳液。以多重乳液粒径和分布为指标,研究酪蛋白酸钠添加量对S/O/W型多重乳液加工适应性的影响。结果表明： 正交试验得到S/O型单重乳液最佳制备条件为：内水相中结冷胶添加量0.2%、无水氯化钙添加量0.5%;内水相乳化 剂聚甘油蓖麻醇酯添加量2.5%;油相为精炼猪油,油水体积比3∶2;剪切速率17 500 r/min,剪切时间1.5 min。将制 得的S/O型单重乳液与不同添加量酪蛋白酸钠混合制得S/O/W型多重乳液。当酪蛋白酸钠添加量0.1%时,S/O/W型 多重乳液粒径符合加工要求,且贮藏、热处理、剪切稳定性较好。以多重乳液替代猪脂肪制备的低脂乳化肠与高脂 （精炼猪油含量20%）乳化肠外观不存在明显差异;微观结构观察结果表明,多重乳液在乳化肠中包裹良好、分布 均匀。     

Synergistic effects of polyglycerol ester of polyricinoleic acid and sodium caseinate on the stabilisation of water–oil–water emulsions

The inherent thermodynamic instability of water–oil–water (W/O/W) emulsions has restrictions for their application in food systems. The objective of this study was to develop a food grade W/O/W emulsions with high yield and stability using minimal concentrations of surfactants. Emulsions were prepared using soybean oil, polyglycerol ester of polyricinoleic acid (PGPR) alone or in combination with sodium caseinate (NaCN) as emulsifier(s) for primary water-in-oil (W/O) emulsions and NaCN as the sole emulsifier for secondary W/O/W emulsions. Increasing the concentration of PGPR (0.5–8%w/v) had no effect on the droplet sizes of the resulting W/O/W emulsions. However, significant increases in droplet sizes of W/O/W emulsions were observed when the concentration of NaCN in external phase was reduced from 0.5 to 0.03% (w/v) (p<0.05). Percentage yields of emulsions (using a water-soluble dye) improved when PGPR concentration in the inner phase was increased from 0.5 to 8% (w/v). A stable W/O/W emulsion with a yield >90% could be prepared with 4% (w/v) PGPR alone as primary hydrophobic emulsifier and 0.5% (w/v) NaCN as external hydrophilic emulsifier. The concentration of PGPR in the inner phase could be reduced to 2% (w/v) without affecting the yield and stability of the W/O/W emulsion by partially replacing PGPR with 0.5% (w/v) NaCN, which was added to the aqueous phase of the primary W/O emulsion. The results indicate that a possible synergistic effect may exist between PGPR and NaCN, thus allowing formulation of double emulsions with reduced surfactant concentration.     

Release rate profiles of magnesium from multiple W/O/W emulsions

Water-in-oil-in-water (W/O/W) emulsions were formulated based on rapeseed oil, olive oil, olein and miglyol. Polyglycerol polyricinoleate and sodium caseinate were used as lipophilic and hydrophilic emulsifiers, respectively. Magnesium was encapsulated in the inner aqueous droplets. Emulsion stability was assayed through particle sizing and magnesium release at two storage temperatures (4 and 25 °C) over 1 month. Irrespective of the oil nature, both the primary W/O and W/O/W emulsions were quite stable regarding the size parameters, with 10-μm fat globules and 1-μm internal water droplets. Magnesium leakage from W/O/W emulsions was influenced by the oil type used in the formulation: the higher leakage values were obtained for the oils characterized by the lower viscosity and the higher proportion of saturated fatty acids. Magnesium release was not due to droplet–globule coalescence but rather to diffusion and/or permeation mechanisms with a characteristic rate that varied over time. In addition, W/O/W emulsions were resistant to various thermal treatments that mimicked that used in pasteurization processes. Finally, when W/O/W emulsions were placed in the presence of pancreatic lipase, the emulsion triglycerides were hydrolysed by the enzyme. These results indicated a possible use of W/O/W emulsions loaded with magnesium ions in food applications.     

Double emulsions (W/O/W): physical characteristics and perceived intensity of salty taste

This study addressed the correlation between physical characteristics of double emulsions and sensory perception, as the microstructure of these systems may provide the mechanism to understanding the initial cause of the altered salty taste. Thus, double emulsions (W/O/W) were prepared using different volumes of the internal aqueous phase while maintaining the same fat and sodium contents in the evaluated systems. Polyglycerol polyricinoleate (PGPR) and tween 80 were used as hydrophobic and hydrophilic emulsifiers, respectively. After preparation, the samples were stored at 25 °C for 4 days and submitted to analysis of optical microscopy, distribution and polydispersity of the oil droplets size, electrical conductivity, rheological behaviour and sensorial analysis. It was found that the use of different emulsifier concentrations (PGPR) did not influence the physical characteristics of the emulsions with the same formulation, but emulsions with different internal aqueous phase concentrations presented different results. These distinct characteristics may have influenced sensorial perception, as the emulsion with higher internal phase concentration was considered saltier. Thus, it can be concluded that structural differences of the double emulsions can be used to decrease the sodium contents without perceivable changes in salty taste.     

Formulation and rheological characterization of reduced-calorie food emulsions

Water-in-oil-in-water (W/O/W) double emulsions are systems where a water-in-oil emulsion (W/O) is dispersed in a second aqueous phase. The W/O emulsion exists in the suspending aqueous medium as oil globules containing smaller water droplets.
In this work, a selection of both materials and procedures has been made in order to obtain an optimal formulation of a W/O/W food emulsion for both yield and rheologica] properties.
The rheological properties of W/O/W emulsions have been studied by means of both steady-shear and oscillatory measurements, and appeared to be similar to those of a simple O/W emulsion having the same volume fraction of dispersed phase, but lower oil content.
This is of great interest to the food industry, since producing double emulsions with the same texture as simple ones, but a lower oil content, helps to formulate reduced-calorie foods.     

Preparation of Monodisperse Food-Grade Oleuropein-Loaded W/O/W Emulsions Using Microchannel Emulsification and Evaluation of Their Storage Stability

W/O/W emulsion is an emerging system in developing new functional and low-calorie food products. The aim of this study is to produce food-grade monodisperse water-in-oil-in-water (W/O/W) emulsions loaded with a hydrophilic bioactive oleuropein. W/O/W emulsions were prepared via high-pressure homogenization and subsequent microchannel (MC) emulsification. The internal aqueous phase was a 5-mM sodium phosphate buffer containing d(+)-glucose (5 wt.%) and oleuropein (0.1–0.7 wt.%). The oil phase consisted of soybean oil and tetraglycerin monolaurate condensed ricinoleic acid esters (TGCR; 3–8 wt.%). The external aqueous phase was a 5-mM sodium phosphate buffer containing d(+)-glucose (5 wt.%) and decaglycerol monolaurate (1 wt.%). Oleuropein-loaded submicron W/O emulsions with average droplet diameters as small as 0.15 μm and monomodal droplet size distributions were prepared by high-pressure homogenization when applying high TGCR concentrations of 5–8 wt.% and low oleuropein concentrations of 0.1–0.3 wt.%. Monodisperse oleuropein-loaded W/O/W emulsions with average W/O droplet diameters of around 27 μm and coefficients of variation of below 5 % were successfully prepared when using a silicon MC array plate with wide channels of 5-μm depth and 18-μm width. The monodisperse W/O/W emulsions prepared at high TGCR concentrations and low oleuropein concentrations were the most stable during 40 days of storage. The adsorption behavior of oleuropein at the internal aqueous–oil interface was relevant to W/O/W emulsions microstructure and stability. The results are believed to provide useful information for successfully preparing stable monodisperse W/O/W emulsions loaded with hydrophilic functional compounds. The surface activity of the loaded material seems to be a key parameter in optimizing the formulation of W/O/W food emulsion.     

Effect of a new emulsifier containing sodium stearoyl-2-lactylate and carrageenan on the functionality of meat emulsion systems

The emulsion capacity and stability of a new emulsifier containing sodium stearoyl lactylate plus iota carrageenan (SSL/iC) in comparison to caseinate and soy isolate was analysed. The emulsion capacity and stability of SSL/iC in oil/water (O/W) model system emulsions was higher than shown by caseinate and soy isolate. However, the O/W emulsion stability was negatively affected by sodium chloride addition, but positively affected by an increase in temperature. Meat batters were made with caseinate, soy isolate, and SSL/iC at the minimum concentration that showed a good performance (>75% stability) in the O/W emulsions. The emulsifier SSL/iC produced high cook yields and good stability when used in meat batters. However, the cooked meat batters containing SSL/iC showed texture characteristics highly detrimental to the sensory analysis. On the other hand, the addition of 2% potato starch reduced the differences in texture parameters among the samples made with the different emulsifiers.     

A liquid crystal application in skin care cosmetics

Intercellular lipids of the stratum corneum contribute threefold to the maintenance of a healthy skin: by hydration, cell adhesion, and reduction of transepidermal water loss. All of these functions can be attributed to the self-organizing property of the amphiphilic molecules of the stratum corneum lipids. A new type of skin care product called Lamellar Gel was developed, which contains a (synthesized) pseudo-ceramide. Its structure is similar to that of ceramide found among the stratum corneum lipids, which allows it to control intramolecular interactions. Compared to regular emulsions the Lamellar Gel demonstrated better skin care characteristics regarding permeability, skin hydration, and skin occlusion. This was attributed to the fact that it formed the same self-organizing structure as natural stratum corneum lipids, hence showing a high affinity to the skin. A high moisturizing effect was observed as the Lamellar Gel combines the benefits of both O/W and W/O emulsions: it provides the same initial hydration as an O/W emulsion and at the same time the same occlusivity as a W/O emulsion. Transepidermal water loss increases under dry environmental conditions. This especially affects the skin around the eyes, where the skin is very thin, and wrinkles are very easily formed. Treatment with the Lamellar Gel recovered these wrinkles promptly and hydrated the stratum corneum for a long time.     

Occlusive power evaluation of O/W/O multiple emulsions on gelatin support cells

The water content of the stratum corneum plays an important role in providing skin suppleness and smoothness. The diffusion of water through the skin is limited primarily by the stratum corneum and the noncornified part of the epidermis has negligible water barrier properties. Multiple emulsions are vesicular systems utilized for the prolonged liberation of active ingredients. The O/W/O multiple emulsion type is employed in cosmetics because its high oil content is able to maintain an occlusive film (barrier) on the skin surface. The objective of this study was to determine the occlusive power of O/W/O multiple emulsions on gelatin support cells. The results showed that occlusive products form a uniform layer on the surface of gelatin after the test, whereas nonocclusive products form two layers: an aqueous phase on the gelatin, and an oil phase above the aqueous phase. Thus, the different occlusive powers are due to the homogeneity of this layer and to its ability to prevent water evaporation.     

Review on the Stability Mechanism and Application of Water‐in‐Oil Emulsions Encapsulating Various Additives

Water‐in‐oil (W/O) emulsions can be used to encapsulate and control the release of bioactive compounds for nutrition fortification in fat‐based food products. However, long‐term stabilization of W/O emulsions remains a challenging task in food science and thereby limits their potential application in the food industry. To develop high‐quality emulsion‐based food products, it is essential to better understand the factors that affect the emulsions’ stability. In real food system, the stability situation of W/O emulsions is more complicated by the fact that various additives are contained in the products, such as NaCl, sugar, and other large molecular additives. The potential stability issues of W/O emulsions caused by these encapsulated additives are a current concern, and special attention should be given to the relevant theoretical knowledge. This article presents several commonly used methods for the preparation of W/O emulsions, and the roles of different additives (water‐ and oil‐soluble types) in stabilizing W/O emulsions are mainly discussed and illustrated to gain new insights into the stability mechanism of emulsion systems. In addition, the review provides a comprehensive and state‐of‐art overview of the potential applications of W/O emulsions in food systems, for example, as fat replacers, controlled‐release platforms of nutrients, and delivery carrier systems of water‐soluble bioactive compounds. The information may be useful for optimizing the formulation of W/O emulsions for utilization in commercial functional food products.     

Influence of Emulsification Technique and Wall Composition on Physicochemical Properties and Oxidative Stability of Fish Oil Microcapsules Produced by Spray Drying

Encapsulation of fish oil is an effective way to protect it against oxidation and masking its fishy odor. One of the possible ways to produce fish oil microcapsules is to produce an oil-in-water (O/W) emulsion followed by spray drying. This study compares the production of the O/W emulsion by mechanical homogenization (rotor–stator) with membrane emulsification and examines the effect of the type and amount of wall material added before drying. The membrane emulsification process selected for the emulsion production is premix membrane emulsification (ME), which consists of the production of a coarse emulsion by mechanical means followed by droplet breakup when the coarse emulsion is forced through a membrane. The emulsions produced had an oil load of 10 and 20 % and were stabilized using whey protein (isolate and hydrolyzate at 1 or 10 %) and sodium caseinate with concentrations of 2 and 10 %. Regarding the material used to build the microcapsule wall, whey protein, maltodextrin, or combinations of them were used at three different oil/wall ratios (1:1, 1:2, 1:3). The results clearly show that premix ME is a suitable technology for producing O/W emulsions stabilized with proteins, which have a smaller droplet size and are more monodisperse than those produced by rotor–stator emulsification. However, protein concentrations of 10 % are required to reduce the droplet size down to 2–3 μm. Small and monodisperse emulsions have been found to produce microcapsules with lower surface oil content, which increases oil encapsulation efficiency and presents lower levels of oxidation during storage at 30 °C. Of all the possible combinations studied, the one with the highest oil encapsulation efficiency is the production of a 20 % O/W emulsion stabilized with 10 % sodium caseinate followed by the addition of 50 % maltodextrin and drying.     

Dispersion Stability of O/W Emulsions with Different Oil Contents Under Various Freezing and Thawing Conditions

Freezing and thawing of oil‐in‐water (O/W) emulsion‐type foods bring about oil–water separation and deterioration; hence, the effects of freezing and thawing conditions on the destabilization of O/W emulsions were examined. The freezing rate and thawing temperature hardly affected the stability of the O/W emulsion. O/W emulsions having different oil fractions were stored at temperatures ranging from –30 to –20 °C and then thawed. The stability after thawing depended on the storage temperature, irrespective of the oil fraction of the emulsion. A good correlation was found between the time at which the stability began to decrease and the time taken for the oil to crystalize. These results indicated that the dominant cause for the destabilization of the O/W emulsion during freezing and thawing is the crystallization of the oil phase and that the effects of the freezing and thawing rates on the stability are insignificant.     

Impact of iron encapsulation within the interior aqueous phase of water-in-oil-in-water emulsions on lipid oxidation

Iron (Fe³⁺) was encapsulated within the internal aqueous phase of water-in-oil-in-water (W/O/W) emulsions, and then the impact of this iron on the oxidative stability of fish oil droplets was examined. There was no significant change in lipid droplet diameter in the W/O/W emulsions during 7 days storage, suggesting that the emulsions were stable to lipid droplet flocculation and coalescence, and internal water diffusion/expulsion. The initial iron encapsulation (4 mg/100 g emulsion) within the internal aqueous phase of the water-in-oil (W/O) emulsions was high (>99.75%), although, a small amount leaked out over 7 days storage (≈10 μg/100 g emulsion). When W/O/W emulsions were mixed with fish oil droplets the thiobarbituric acid-reactive substances (TBARS) formed decreased (compared to fish oil droplets alone) by an amount that depended on iron concentration and location, i.e., no added iron < iron in external aqueous phase < iron in internal aqueous phase. These differences were attributed to the impact of W/O droplets on the concentration and location of iron and lipid oxidation reaction products within the system.     

米糠蛋白O/W及W/O/W乳液制备及界面稳定性

为对比不同米糠蛋白质量浓度下O/W及W/O/W乳液的稳定性,以米糠蛋白作为基料,采用双乳化法制备O/W及W/O/W乳液,考察不同米糠蛋白质量浓度下乳液的微观形态和稳定性并探究其界面稳定机理。结果表明：W/O/W乳液的贮存稳定性显著优于O/W乳液;与相同蛋白含量的O/W乳液相比,W/O/W乳液的黏度显著提高;当米糠蛋白质量浓度为0.4 g/100 mL时,W/O/W乳液的稳定性较O/W乳液提高了1 倍以上;乳液内部包裹更多的W/O液滴,W/O/W乳液的粒径较大;而此时静电斥力也较大,起到稳定乳液的目的。同时,米糠蛋白质量浓度不小于0.4 g/100 mL时,O/W及W/O/W乳液中蛋白质的吸附率较高,达到78%以上。本研究为天然米糠蛋白质在食品级乳液中的开发提供参考,为粮食副产物的综合利用提供了新思路。     

O/W emulsion and W/O/W multiple emulsion: physical characterization and skin pharmacokinetic comparison in the delivery process of caffeine

The aim of this study was to compare the characteristics of an O/W emulsion and a W/O/W emulsion, formulated with similar ingredients. The physical properties of these two types of vehicles were characterized first. Using three types of in vitro models, human skin biopsies, nitrate acetate cellulose membranes and human reconstituted epidermis, the delivery potential of each vehicle for one hydrophilic drug, caffeine, was compared. The assessment of physical parameters, such as particle size, conductivity and rheological behavior enabled the nature of the test emulsions to be clearly identified. Clear differences were observed in the ability of each type of emulsion to deliver caffeine. Whatever the nature of the membrane used for the pharmacokinetic study, the absorption of caffeine was roughly two-fold lower (2.6 for human skin) when the W/O/W multiple emulsion was used as the vehicle. The concomitant determination of physical and kinetic properties of these two test emulsions allowed the W/O/W multiple emulsion and a simple O/W emulsion to be clearly differentiated.     

In vitro skin permeation of sunscreen agents from O/W emulsions

The effects of different emulsifiers on the in vitro permeation through human skin of two sunscreen agents [octylmethoxycinnamate (OMC) and butylmethoxydibenzoylmethane (BMBM)] were investigated from O/W emulsions. The test formulations were prepared using the same oil and aqueous phase ingredients and the following emulsifier and coemulsifier systems: Emulgade SE((R)) (ceteareth-12 and ceteareth-20 and cetearyl alcohol and cetyl palmitate) and glycerylmonostearate (emulsion 1); Brij 72((R)) (steareth-2), Brij 721((R)) (steareth-21) and cetearyl alcohol (emulsion 2); Phytocream((R)) (potassium palmitoyl-hydrolysed wheat protein and glyceryl stearate and cetearyl alcohol) and glycerylmonostearate (emulsion 3); Montanov 68((R)) (cetearyl glucoside and cetearyl alcohol) (emulsion 4); Xalifin-15((R)) (C(15-20) acid PEG-8 ester) and cetearyl alcohol (emulsion 5). The cumulative amount of OMC that permeated in vitro through human skin after 22 h from the formulations being tested decreased in the order 3 > 1 congruent with 4 > 5 > 2 and was about nine-fold higher from emulsion 3 compared with that from emulsion 2. As regards BMBM, no significant difference was observed as regards its skin permeation from emulsions 1, 3, 4 and 5, whereas formulation 2 allowed significantly lower amounts of BMBM to permeate the skin. In vitro release experiments of OMC and BMBM from emulsions 1-6 through cellulose acetate membranes showed that only emulsions 4 and 5 provided pseudo-first-order release rates only for OMC. The results of this study suggest that the type of emulsifying systems used to prepare an O/W emulsion may strongly affect sunscreen skin permeation from these formulations. Therefore, the vehicle effects should be carefully considered in the formulation of sunscreen products.     

Stability of Bisphenol A (BPA) in Oil-In Water Emulsions under Riboflavin Photosensitization

Effects of riboflavin photosensitization on the degradation of bisphenol A (BPA) were determined in oil-in-water (O/W) emulsions containing ethylenediaminetetraacetic acid (EDTA) or sodium azide, which are a metal chelator or a singlet oxygen quencher, respectively. Also, the distribution of BPA between the continuous and dispersed phases in O/W emulsions was analyzed by high-performance liquid chromatography (HPLC). The concentration of BPA in O/W emulsions significantly decreased by 38.6% after 2 h under visible light irradiation and in the presence of riboflavin (P < 0.05). Addition of EDTA and sodium azide protected the decomposition of BPA significantly in a concentration dependent manner (P < 0.05), which implies both transition metals and singlet oxygen accelerate the photodegradation of BPA in O/W emulsions. Approximately 21.7% of the BPA was distributed in the 2.5% (w/v) dispersed lipid particles and 78.3% was in the continuous aqueous phase of the emulsions. The amount of BPA in aqueous phase decreased faster than the amount of BPA in the lipid phase during riboflavin photosensitization (P < 0.05). Thus, the BPA in the aqueous phase was the major target of riboflavin photodegradation in O/W emulsions. Practical Application: Concentration of BPA, an endocrine disrupting chemical, was decreased significantly in oil-in-water emulsions under riboflavin and visible light irradiation. BPA in continuous aqueous phase was major target of riboflavin photosensitization. However, BPA was distributed more densely in lipid phase and more protected from riboflavin photosensitized O/W emulsions. This study can help to decrease the level of BPA in foods made of O/W emulsions containing riboflavin, which could be displayed under visible light irradiation.     

Grape skin extracts as a sustainable source of antioxidants in an oil-in-water emulsion: an alternate natural approach to synthetic antioxidants using principal component analysis

Grape (Kyoho) skin, used to retard lipid oxidation in edible oil foods, was investigated to reduce lipid oxidation in an oil-in-water (O/W) emulsion during 20 day of storage. The antioxidant efficacy of Kyoho skin extracts in O/W emulsions was determined by the measurement of secondary oxidation products. Moreover, principal component analysis (PCA) was conducted to determine similarities between emulsions treated with or without Kyoho skin extracts and standards. The data revealed that Kyoho skin extracts exhibited >93% inhibition and reported a similar p-anisidine (4.30–20.71) and TBARS (6.08–11.15 mg MDA L⁻¹) values over the standards during 20 day of storage. PCA (PCs 1 (51.83%) and 2 (18.85%)) demonstrated a similarity in the contribution of Kyoho skin extracts over the synthetic antioxidants in O/W emulsion. Overall, these findings highlighted the possibility of using Kyoho skin extracts as natural antioxidants to decrease oxidative rancidity in foods.     

枯草芽孢杆菌对大豆蛋白-磷脂复合乳液在体外消化中稳定性的影响

为探讨枯草芽孢杆菌在消化过程中对大豆蛋白-磷脂复合乳液稳定性的影响。本试验设计胃肠消化模型,通过对加菌与不加菌的O/W型和W/O型乳液的乳化活性指数（Emulsifying activity index,EAI）、乳化稳定指数（Emulsification stability index,ESI）、浊度、粒径、Zeta电位检测和显微观察,考察乳液是否出现分离、絮凝、聚集上浮等现象。结果表明,在胃消化阶段,O/W型和W/O型乳液EAI、ESI、粒径、电位都明显减小,显微观察显示液滴密度减小,体积增大;加入枯草芽孢杆菌的乳液与不加菌乳液相比,各指标减小的幅度更大。在小肠消化阶段,O/W型和W/O型乳液EAI和ESI整体呈上升趋势,而粒径和电位则明显下降,另外,两者的显微观察显示液滴密度减小,体积也减小。与不加菌的相比,加入枯草芽孢杆菌后乳液EAI、ESI、粒径显著降低,电位显著升高（P<0.05）。综合上述结果,枯草芽孢杆菌促进了大豆蛋白-磷脂复合乳液在消化过程中的破乳现象。     

Stability and release properties of double-emulsions stabilised by caseinate–dextran conjugates

The subject of the present paper was to investigate the possibility of stabilising water-in-oil-in-water emulsions (W/O/W) by using sodium caseinate (SC)–dextran (Dex) conjugates in order to influence the release of vitamin B₁₂ from the inner water phase (W₁) to the outer aqueous phase (W₂).To prepare the conjugate the SC was combined with Dex (M_r 250,000 or 500,000 g/mol) and incubated at 60 °C and a humidity of 79% for 8 h.The double emulsions, with encapsulated vitamin B₁₂, were prepared using a two-step emulsification technique. Whereas different amounts of polyglycerin polyricinoleate (PGPR, E476) were the hydrophobic emulsifier, the conjugate and the SC alone were used as the hydrophilic emulsifiers. The investigations comprised the determination of the particle size distribution of the W/O/W emulsion and measurement of the amount of vitamin B₁₂ migration from W₁- to the W₂-phase during the second stage of emulsion preparation and after heating or pH changing of emulsion.The water-containing oil droplets of the W/O/W emulsions were smaller and distributed more narrowly using SC–Dex conjugate as emulsifier instead of pure protein. Under acidic conditions, the conjugate-containing emulsions were more coalescence stable than the emulsions with SC, and the vitamin B₁₂ release from the inner W₁-phase was significantly decreased.