斑点叉尾鮰鱼皮明胶水解方法的研究

研究了利用斑点叉尾鮰鱼皮明胶水解制取抗氧化活性肽的方法,比较了不同的水解方法,不同底物,不同的蛋白酶对水解产物抗氧化活性的影响。结果表明酶解法适合生产胶原蛋白活性肽,酶解产物具有不连续的分子量分布及较好的抗氧化活性。不同底物酶解制取的产物的抗氧化活力不同,其中以酸法明胶利用碱性蛋白酶酶解的产物活力最高,平均分子量小于4000Da。     

罗非鱼皮明胶酶解物及其模拟消化产物的抗氧化活性

以罗非鱼皮为原料提取罗非鱼皮明胶,选用风味蛋白酶和胰蛋白酶制备罗非鱼皮明胶酶解物,采用ABTS自由基、DPPH自由基、羟基自由基及亚油酸过氧化体系,初步评价罗非鱼皮明胶酶解物的抗氧化活性,再通过模拟体外胃肠道消化实验,结合分子质量分布测定,进一步考察罗非鱼皮明胶酶解物的抗氧化活性。结果显示,在酶解过程中,风味蛋白酶及胰蛋白酶酶解的水解度逐渐升高,在3 h时达到最高,分别达到5.8%和25.36%。在酶解60 min时其TCA可溶性肽得率最高,风味蛋白酶、胰蛋白酶酶解物分别达56.82%和54.44%。通过比较半抑制浓度(IC_50),确定了酶解60 min时风味蛋白酶酶解物的清除DPPH自由基及抑制亚油酸过氧化能力较胰蛋白酶酶解物强。模拟体外胃肠道消化后,酶解物羟基自由基清除活性均显著提高(p0.05),亚油酸脂质过氧化活性明显降低,消化前后样品分子量分布范围均主要集中于3000~5000 Da,消化后风味蛋白酶及胰蛋白酶酶解物3000~5000 Da组分的含量分别提高了45%及13%。以上研究结果表明,罗非鱼皮明胶酶解后制备的明胶水解物具有一定的抗氧化能力,具有潜在的开发价值。     

蜂王浆蛋白肽的制备及其降血糖和抗氧化活性研究

目的:研究蜂王浆蛋白的最佳酶解工艺条件,并分析所制备酶解肽的氨基酸组成、分子量分布、降血糖及抗氧化活性。方法:以蜂王浆为原料,采用碱提酸沉法提取蜂王浆粗蛋白,以α-葡萄糖苷酶抑制率和ABTS自由基清除率为评价指标,筛选出酶解蜂王浆蛋白制备降血糖及抗氧化活性肽的最佳蛋白酶,并研究蜂王浆酶解肽的体外降血糖和抗氧化活性。结果:蜂王浆降血糖及抗氧化活性肽的最佳制备工艺为:以酸性蛋白酶为酶制剂,酶添加量为6000 U/g,酶解温度为43℃,酶解pH为4.0,酶解时间为4 h,料液比为1:10。在上述条件下,制备的蜂王浆蛋白肽的氨基酸总量为82.19%,相对分子质量集中在1000 Da以下。蜂王浆蛋白肽对α-葡萄糖苷酶的半抑制浓度(IC₅₀)为6.94 mg/mL,清除ABTS自由基、羟自由基、1,1-二苯基-2-三硝基苯肼(DPPH)自由基及超氧阴离子自由基的半抑制浓度(IC₅₀)分别为14.18、0.45、11.02和18.38 mg/mL。试验结果表明,蜂王浆蛋白肽具有一定的降血糖和抗氧化活性,可为蜂王浆高值化利用及活性肽产品开发提供理论依据。     

体外消化对三文鱼皮胶原低聚肽抗氧化活性的影响

本研究以三文鱼皮为原料通过酶解法制备三文鱼皮胶原低聚肽,并进行体外模拟消化实验,通过分子量分布分析、DPPH自由基清除能力、羟自由基清除能力、总抗氧化能力（ABTS法）以及活性氧ROS实验,探究三文鱼皮胶原低聚肽经模拟胃、肠道消化后抗氧化活性的变化。结果表明:经模拟消化实验后,三文鱼皮胶原低聚肽的重均分子量轻微减少,变化不超过4%;胃蛋白酶消化前后,DPPH自由基清除能力IC₅₀值分别为11.1、12.3 mg/mL,胰蛋白酶消化前后,IC₅₀值分别为12.5、14.6 mg/mL;胃蛋白酶消化前后,羟自由基清除能力IC₅₀值分别为12.2、13.2 mg/mL,胰蛋白酶消化前后,IC₅₀值分别为10.7、11.5 mg/mL;胃蛋白酶消化后,总抗氧化能力下降不超过13%,胰蛋白酶消化后,总抗氧化能力下降不超过19%;胃蛋白酶消化后,ROS清除能力提高不超过3%,胰蛋白酶消化后,ROS清除能力提高不超过5%。这表明三文鱼皮胶原低聚肽具有良好的消化稳定性和抗氧化活性,为其在抗氧化功能性食品的开发提供了理论基础。     

复合酶制备鱼蒸煮液水解肽及其抗氧化活性研究

目的:研究鳀鱼蒸煮液的最佳酶解工艺条件,并分析所制备水解肽的分子量分布和抗氧化特性。方法:以三氯乙酸氮溶解指数(Trichloroacetic acid-nitrogen soluble index,TCA-NSI)为酶解效率的评价指标,在单因素实验基础上,运用中心组合设计法优化水解肽的制备工艺,并对酶解产物的分子量分布及抗氧化活性进行分析。结果:鳀鱼蒸煮液的最佳酶解条件为酶解时间45 min、酶解温度54.5 ℃、酶解pH8.2、碱性蛋白酶与中性蛋白酶比例1:1,在此条件下TCA-NSI可达76.51%。所制备的水解肽中分子量低于1500 Da的多肽含量可达81.941%。水解肽对DPPH自由基、羟自由基和超氧阴离子自由基清除率的IC₅₀分别为2.45、1.37和3.45 mg/mL,表明其具有较强的抗氧化活性。结论:采用复合酶法可高效酶解鳀鱼蒸煮液,并获得具有较强抗氧化活性的水解肽,可为鳀鱼蒸煮液高值化利用及活性肽产品开发提供理论依据。     

复合酶制备鳀鱼蒸煮液水解肽及其抗氧化活性分析

目的:采用复合酶进行鳀鱼蒸煮液制备,分析其水解肽分子量分布及抗氧化活性。方法:利用三氯乙酸氨溶液指数(TCA-NSI)对复合酶制备鳀鱼蒸煮液效率进行评价,以单因素试验为基础,采用中心组合设计法进一步优化鳀鱼蒸煮液中水解肽制备工艺,分析酶解产物的分子量分布及抗氧化活性。结果:鳀鱼蒸煮液制备时的酶解时间最佳为45分钟、酶解温度为55℃水解肽中的分子量在1500Da内的多肽含量为81.941%。同时,结果显示水解肽对自由基清除率IC₅₀为2.45mg/mL,对羟自由基为1.37mg/mL,对超氧阴离子为3.45mg/mL,意味着水解肽具有较强的抗氧化活性。结论:在鳀鱼蒸煮液制备时,应用复合酶能够获取抗氧化活性极高的水解肽,为开发源于海洋生物的天然抗氧化剂提供强有力的依据。     

罗非鱼皮/鳞胶原蛋白肽加工性能及抗氧化性质比较研究

为探究企业提供的5种不同标称分子量的罗非鱼鱼皮和鱼鳞胶原蛋白肽的加工性能及其抗氧化活性,对几种胶原蛋白肽产品的吸湿性、吸油性、乳化性、起泡性、还原力、超氧阴离子自由基(O2-·)以及羟自由基(·OH)清除效果进行了系统的研究比较。结果表明,采用凝胶渗透色谱(gel permeation chromatography,GPC)测得标称分子量为1、3、5 k Da罗非鱼鱼皮胶原蛋白肽的重均相对分子质量分别为823、1 196、2 753 Da,标称分子量为3、5 k Da罗非鱼鱼鳞胶原蛋白肽重均相对分子质量分别为1 189、1 926 Da;标称为5 k Da的鱼鳞肽吸湿性最强,为0.26 g/g;标称为1 k Da的鱼皮肽吸油性最强,为2.57 m L/g;标称为5 k Da的鱼鳞肽乳化性最强,达到51.8%,5 h内乳化稳定性最优;5种产品的起泡性很接近,在165%~170%之间;60 min内,5 k Da的鱼皮肽和鱼鳞肽起泡稳定性均较好;以铁氰化钾还原法、超氧阴离子自由基(O2-·)和羟自由基(·OH)法对5种肽产品的抗氧化性进行分析显示,肽产品3种抗氧化能力均随样品浓度增大而增强;肽产品的还原力和O2-·清除效果略差于维生素C,5种产品中标称为1 k Da鱼皮肽的还原力最佳,标称5 k Da罗非鱼鱼鳞肽O2-·清除能力最佳;而标称3 k Da和5 k Da的罗非鱼鱼鳞胶原蛋白肽对羟自由基(·OH)的清除效果则比维生素C显优,其中5 k Da的鱼鳞肽对羟自由基(·OH)的清除效果最高达到84%;肽产品的抗氧化性与肽的分子量和来源有密切关系。研究表明,5种肽产品的标称分子量与实际分子量均有一定的差异,但具有良好的加工性和较好的抗氧化性,研究结果可为这5种肽的进一步开发利用提供支持。     

刺参低聚肽的质量评价和抗疲劳作用研究

研究p H渐变条件下采用碱性蛋白酶和菠萝蛋白酶分步酶解猪皮得到的不同分子量胶原蛋白肽的体内外抗氧化活性。猪皮预处理后,采用碱性蛋白酶和菠萝蛋白酶分步进行酶解,酶解液经超滤得到M1 k Da、1 kDaM3 kDa、M3 kDa 3个不同分子量肽段,以福林酚比色法测定各组分肽含量,通过测定各组分肽段对DPPH自由基、ABTS+自由基、O2-自由基及OH自由基的清除实验评价各组分肽的体外抗氧化活性,再以不同分子量肽段酶解液对小鼠进行灌胃,以试剂盒测定小鼠灌胃后血清的总超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶的活性、总抗氧化能力和微量丙二醛含量评价各组分肽的体内抗氧化活性。综合体内外抗氧化实验数据,各组分肽的抗氧化能力强弱顺序为:小于1 kDa组分1 k Da～3 kDa组分大于3 kDa组分。pH渐变条件下采用碱性蛋白酶和菠萝蛋白酶分步酶解的各组分猪皮胶原蛋白肽在体内外均有不同程度的抗氧化能力,且分子量越小,抗氧化活性越强,可对机体起到保护作用。     

黄豆酱中低聚肽的分离及抗氧化活性研究

为研究黄豆酱中低聚肽的抗氧化性质,以黄豆酱为原料,通过超滤、离子色谱、凝胶色谱等技术手段分离出黄豆酱中高抗氧化活性组分,并确定其抗氧化活性与分子量分布。获得最佳分离条件:DEAE离子交换树脂上样最佳pH为8.6,葡聚糖凝胶G-15分离最佳流速为2mL/min,上样浓度为1.2mg/mL,并确定高抗氧化活性的两组黄豆酱低聚肽分子量范围为925~1 124Da和580~925Da。     

南极磷虾多肽的组成及其抗氧化与A C E抑制活性

以冷冻南极磷虾为原料,经碱溶酸沉法与酶解法联用制备南极磷虾多肽,采用紫外-可见光谱、红外光谱、凝胶色谱和氨基酸分析解析了多肽的组成和结构,并重点评价了多肽的抗氧化活性和血管紧张素I转化酶（angiotensin-I converting enzyme,ACE）抑制活性。结果表明：采用碱溶酸沉法与碱性蛋白酶酶解法联用制备南极磷虾多肽,水解度和肽得率可达到（17.83±3.73）%和（57.20±0.24）%。南极磷虾多肽的紫外最大特征吸收波长在273.5 nm;多肽在3 390.84、2 917.66、1 648.36、1 547.59 cm-1和 1 402.21 cm-1处具有红外特征吸收峰;多肽的分子量主要在189 Da～6 500 Da范围内（98.93%）,其中,451 Da～1 450 Da占比最高（41.00±0.05）%;多肽的氨基酸组成理想,符合联合国粮农组织/世界卫生组织规定的标准。南极磷虾多肽具有显著的还原能力和良好的清除DPPH·、ABTS+自由基、·OH的能力,自由基清除IC50值分别为5.65、0.17、4.46 mg/mL;此外,多肽还具有良好的ACE抑制活性,当多肽浓度为0.5 mg/mL时,ACE抑制率为（55.91±2.63）%。     

羧甲基壳聚糖在秀丽线虫体内的抗衰老作用

目的:基于模式动物秀丽线虫探究羧甲基壳聚糖的抗衰老能力。方法:检测羧甲基壳聚糖处理前后,秀丽虫寿命、运动能力、吞咽频率以及排泄周期的变化,同时测定过氧化氢酶和谷胱甘肽过氧化物酶的活力,判断羧甲基壳聚糖线虫体内抗氧化的能力。结果:500和1000 mg/L羧甲基壳聚糖分别能显著(P<0.05)和极显著(P<0.01)延长秀丽线虫的寿命,而500 mg/L能够极显著(P<0.01)提高秀丽线虫运动能力活跃度,加速咽泵运动,缩短排泄周期。同时,100、500和1000 mg/L羧甲基壳聚糖均能极显著(P<0.01)提高秀丽线虫过氧化氢酶和谷胱甘肽过氧化物酶的活力。结论:500 mg/L的羧甲基壳聚糖抗衰老作用较好强,具有抗氧化、抗衰老的应用前景。     

羧甲基籼米淀粉的理化性质及其在冰淇淋中的应用

以实验室制备得到的羧甲基籼米淀粉(DS=0.49)为原料,对其主要的性质作了详细的研究包括溶解度、膨胀度、冻融稳定性、动态流变性、静态流变性及质构特性。同时,以膨胀率,融化度和感官评价为指标,研究了其作为增稠稳定剂在冰淇淋的应用效果。结果表明,羧甲基化后的籼米淀粉的溶解度、膨胀度均有明显提高,冻融稳定性也明显改善。由静态流变曲线可知,羧甲基淀粉具有更高的表观黏度及凝胶稳定性;动态流变曲线表明,羧甲基淀粉的糊化温度降低,抗老化能力提高。质构数据说明羧甲基淀粉的凝胶强度较低。在冰淇淋制备中添加羧甲基籼米淀粉作为增稠稳定剂,能显著改善冰淇淋的品质特性,添加量在0.3%时得到的冰淇淋膨胀率大于90%,具有较好的抗融化能力,外观和口感较好。     

Kinetics of moisturizing and firming effects of cosmetic formulations

The assessment of cosmetic efficacy is rarely performed in studies comparing different concentrations of active compounds. The aim of the present study was to determine the skin hydrating and the skin firming dose-response effects of cosmetic formulations enriched in compounds derived from algae and fish collagen. A series of factors were studied including the type of formulation (cream or serum), the concentration in active ingredients, the effect of repetitive applications, as well as any residual effect of the formulations after stopping their applications. The serum enriched in marine compounds showed a better moisturizing effect in short term. The cream appeared more active later, particularly following repeat applications. A sustained tensor (firming) effect was observed during treatment with both the lotion and the cream. However, no remnant firming effect was perceived after stopping treatment.     

Cream or foam in pedal skin care: towards the ideal vehicle for urea used against dry skin

The aim of this study was to evaluate different urea‐containing cosmetic preparations designed for foot care regarding skin occlusion. The primary aim was therefore to screen the short‐term transepidermal water loss (TEWL) as a parameter for skin barrier function and skin occlusion and to characterize the relative role of the vehicle, i.e. cream or foam in the context of cosmetics containing urea in the 2–10% range addressing the cosmetic products urea 2% cream (GEHWOL FUSSKRAFT blau), petrolatum containing cream (GEHWOL med Schrundensalbe), urea 10% cream (GEHWOL med Lipidro‐Crème), urea 10% foam (Allpresan Fuss Schaum) and vaseline (positive control) compared with an untreated area on the volar forearms of volunteers. Moreover, the short time (24 h) kinetics regarding the moisturizing effect of cream and foam formulations in diabetic patients were compared. The efficacy of a cream on reduction of skin thickness of hyperkeratotic skin in the heel region before and after a period of product application was also evaluated. In some of the trials, healthy individuals and in others, diabetic patients (type I and II) were enrolled. TEWL was determined before product application, as well as at given points of time thereafter. In this study, no excessive occlusion effects comparable with a blockage of the skin’s natural water evaporation could be observed for any of the test products. To the extent to be expected, this was found neither for the cream products nor for the foam product. Slightly lowered TEWL values after application of the 10% urea cream can be interpreted as a beneficial effect in terms of an improved barrier function. Regarding skin moisture, the urea‐containing cream formulation appeared equal or slightly superior to the foam formulation. The thickness of the horny layer was found reduced after application of 10 % urea‐containing cream. At present it looks as if cream vehicles would still be vehicles of choice in general, when it comes to the formulation of skin care preparations for not only dry skin but also in the context of pedal skin care.     

羧甲基壳聚糖的降解及其抗氧化性能的研究

羧甲基壳聚糖是壳聚糖的一种重要衍生物。本研究采用过氧化氢氧化法对羧甲基壳聚糖进行降解,并考察了降解时间及过氧化氢用量对降解的影响。用粘度法和改进的Schales’方法测定了降解产物的分子量。并用流动注射化学发光分析法检测了降解产物的抗氧化能力--对羟基自由基的清除活性。结果表明:随着过氧化氢在降解体系中的浓度的增大,降解产物的分子量将趋于一固定值;当H2O2在总反应体系中浓度大于1.3mol/L,降解时间大于16h,将得到分子量低至1100左右的羧甲基壳聚糖;经降解得到的羧甲基壳聚糖均具有一定的羟基自由基清除活性,且分子量越小,其抗氧化性能越强。     

The effect of rheological properties of experimental moisturizing creams/lotions on their efficacy and perceptual attributes

Although anecdotal evidence suggests that the rheology of creams and lotions might affect their moisturizing efficacy and perceptual qualities, to date no systematic investigation of these effects has been carried out. We prepared 12 prototype moisturizing creams that had identical overall formulae but differed in as far as they contained three different polymers (Carbopol 981, Carbopol 5984 and Permulen TR.-2) each at four concentration levels. Using these creams, we carried out two series of experiments. In the first series, we applied pre-determined quantities of creams to defined areas of skin on the forearms of eight panelists and then measured the subsequent changes in Transepidermal Water Loss (TEWL) and Skin Capacitance (SC) values as a function of time. In the second series, the panelists were asked to spread and rub over their entire two hands as much of the creams as they deemed necessary for achieving satisfactory perceived moisturization. In this latter series, we also weighed the amounts of cream used and measured the TEWL and SC values at several skin sites as a function of time subsequent to the cream application. The panelists also scored 14 skin attributes for each of the 12 creams used. The same procedures were also carried out with a commercial skin moisturizing cream (Keri(R)) that was used as a control. In separate laboratory experiments, the rheological properties (i.e. the elastic and viscous moduli) of the various creams at a number of frequencies were measured. The analysis of the experimental results suggested that the overall rheology of creams had little direct effect on both, the moisturizing efficacy and the perceived perceptual attributes. Neither did the overall viscoelasticity or the esthetic attributes of the preparations seem to affect the amounts of creams/lotions that the panelists judged to be necessary for achieving satisfactory perceived hand moisturization. Although all preparations contained the same occlusive material (oil) at identical concentration, nevertheless some of the preparations appeared to show different efficacies depending on the nature and the levels of the polymeric ingredient present.     

结合预处理方法的酶法制备美藤果肽工艺的优化

以美藤果粕为原料,制备美藤果肽。通过对比多种预处理方式,筛选出可促进蛋白水解的预处理方法,在此基础上利用响应面实验优化美藤果肽的酶法制肽工艺,并分析此法制得的肽相对分子量。结果表明:碱性蛋白酶最适宜用于美藤果肽的制备,而添加0.15%~0.45%亚硫酸钠、表面活性剂十六烷基三甲基溴化胺(CTAB)或采用不同频率的超声预处理,能使美藤果蛋白酶解效果提升10%~50%,其中40 Hz的超声预处理效果最佳。响应面实验优化得到的最佳工艺为:40 Hz 15 min超声处理美藤果粕,添加7600 U/g碱性蛋白酶,52℃下酶解6 h,美藤果蛋白水解度为18.21%,得到相对分子量小而集中的寡肽(<1000 Da占94.64%),其中分子量在180~500 Da范围的占过半比例。     

香蕉皮多糖的提取及其在护手霜中的应用

采用水提醇沉法提取香蕉皮多糖,选择苯酚—硫酸法测定香蕉皮多糖含量,并评价其保湿性能,进而用于制备香蕉皮多糖护手霜。结果表明,香蕉皮多糖含量为3.99%,24h内保湿性能良好,保湿率均在80%以上,且保湿性能明显强于甘油。此外,所制备的香蕉皮多糖护手霜其理化指标、感官指标及卫生指标均符合中国轻工行业标准(QB/T1857—2004),具有推广价值。     

Changes in surface configuration of the skin caused by ageing and application of cosmetics: three-dimensional analysis according to a new system based on image analysis and Fourier transformation

The surface configuration of the skin, as characterized by surface furrows and plateaux, is known to change with ageing and skin condition. There have been many attempts to analyse the surface configuration in order to evaluate the effect of skin care products. However, the systems proposed up to now are insufficient to obtain three-dimensional data.
A system based on image analysis and Fourier transformation has been developed. By using this system, we can make a quick and quantitative analysis of the three-dimensional surface configuration of the skin. This system was applied to measure changes in the surface configuration with ageing, and to clarify the effects of cosmetic cream.
As a result, it was found that the furrows become less clear, become parallel in one direction and the texture becomes coarse and irregular, as ageing continues. These changes are believed to correspond to deterioration of the metabolic and moisturizing functions of the skin.
It was also found that the furrows become clear and the texture becomes finer after the application of cream than before. This is believed to be the result of the improvement of the skin condition due to the moisturizing effect of the cream.     

罗非鱼皮胶原酶解物对HaCat细胞生长的影响

研究经不同商业酶水解后得到的罗非鱼皮胶原酶解物对人皮肤角质细胞HaCaT生长的影响,并对胶原酶 解物的抗氧化活性与促进细胞生长作用的关系进行探讨。结果表明：罗非鱼皮经中性蛋白酶（neutral enzyme, NE）、碱性蛋白酶、胰蛋白酶水解得到的酶解产物中,NE酶解物（neutral enzymatic hydrolysate,NEH）具有显著的 体外抗氧化活性和促进HaCaT细胞增殖的活性（P＜0.05）;NEH（1～5 ku）在质量浓度为25 mg/mL时对HaCaT细胞 的生长具有显著的促进作用（P＜0.05）,并在24 h内能显著促进细胞划痕处伤口愈合（愈合率（81.10±5.77）%, P＜0.05）。经氨基酸组成分析和凝胶色谱分析发现,NEH（1～5 ku）主要包含2 个肽组分,NEH（1～5 ku）的 抗氧化氨基酸苯丙氨酸、亮氨酸、酪氨酸、赖氨酸、精氨酸和组氨酸的质量分数要显著高于其他分子质量的肽 组分（P＜0.05）。研究认为,经NE水解得到胶原肽分子质量为1～5 ku组分对HaCaT细胞生长具有促进作用,这可 能与肽组分中抗氧化氨基酸质量分数较高有关。