槲皮素拮抗草鱼呼肠孤病毒感染的药物学研究

前期研究发现槲皮素(Quercetin, Qct)在体外对 I、II、III 型草鱼呼肠孤病毒(grass carp reovirus, GCRV)均有抑制效果。为进一步阐明 Qct 在草鱼(Ctenopharyngodon idella)中拮抗 GCRV 的临床应用潜力, 本研究应用草鱼细胞系测定 Qct 对 I 型 GCRV 的半数有效抑制浓度(EC₅₀), 并利用高效液相色谱法研究 Qct 在草鱼中的药代动力学特征, 在稀有鮈鲫(Gobiocypris rarus)模型中评估 Qct 药效。结果显示, Qct 对 GCRV 的 EC₅₀ 为 4.796 μg/mL; 草鱼单次口灌 20、40、60 mg/kg Qct 粗提物, 48 h 后血液中最大峰值浓度(C_max)分别为 0.129 μg/mL、0.583 μg/mL、0.666 μg/mL; 肝胰脏中 C_max 分别为 3.822 μg/g、5.386 μg/g、6.252 μg/g; 肾脏中 C_max 分别为 2.437 μg/g、3.140 μg/g、3.447 μg/g。 血液中 Qct 的 C_max 远低于 EC₅₀, 40 mg/kg 或 60 mg/kg 槲皮素处理组肝胰腺中的 C_max 高于 EC₅₀。II 型 GCRV 稀有鮈鲫感染模型中, qRT-PCR 显示 Qct 可以抑制病鱼各组织内病毒的复制; 组织病理切片显示 Qct 能减少炎症反应。 综上, Qct 不但抑制 GCRV 复制, 也可能通过降低炎症反应发挥抗病毒作用, 从而降低死亡率, 40 mg/kg 的剂量可以保障 Qct 在草鱼体内的抗病毒活性。     

二种海胆性腺的脂质组成及其抗氧化活性

为分析马粪海胆和光棘球海胆性腺的脂质组成和抗氧化活性，采用核磁共振和气相色谱—质谱技术对2种海胆性腺油脂的脂质成分和脂肪酸组成进行分析，并通过DPPH自由基清除法、羟基自由基清除法和超氧阴离子自由基清除法对其脂质的抗氧化活性进行研究。结果显示，马粪海胆和光棘球海胆性腺脂质均以甘油三酯和磷脂为主，胆固醇、胆固醇酯和游离脂肪酸含量较低。马粪海胆和光棘球海胆性腺总脂富含C20:4n-6和C20:5n-3，且二者总量分别占脂肪酸含量的35.88%和34.98%；同时2种海胆性腺的中性脂和极性脂的脂肪酸组成存在较大差异，中性脂以C14:0和C16:0等饱和脂肪酸为主，而极性脂以C20:4n-6和C20:5n-3等多不饱和脂肪酸为主。马粪海胆和光棘球海胆性腺脂质对DPPH自由基、羟基自由基和超氧阴离子自由基均具有较好的清除能力，DPPH自由基IC₅₀分别为2.75和1.98 mg/mL，羟基自由基IC₅₀分别为0.33和0.29 mg/mL，超氧阴离子自由基IC₅₀分别为0.33和0.31 mg/mL。研究表明，马粪海胆和光棘球海胆性腺脂质具有较高的营养价值和抗氧化活性，可作为C20:4n-6、C20:5n-3和磷脂等功能性脂质因子的重要膳食来源。     

氨、亚硝酸盐和硝酸盐对凡纳滨对虾幼虾的毒性作用

研究了 NH₃-N、NO₂ ^- -N与 NO₃ ^- -N对凡纳滨对虾幼虾的毒性作用。获得了 NH₃-N_t(NH₃-N_m) 与 NO₂ ^- -N对体长2．4cm幼虾的 24h、48h、72h、96h之 LC₅₀值，两者对幼虾的安全质量分数分别为1．30 (0．101)mg/L和3．80mg/L。当 NH₃-N_t(NH₃-N_m)质量分数在1．3(0．101)~4．3(0．333)mg/L时，存活率为71．4% ~92．9%，体长增长率为36．3% ~57．1%，体重增长率为188．5% ~322．3%。当 NO₂ ^- -N质量分数在3．00~21．00mg/L时，成活率为75．0% ~91．7%，体长增长率为21．2% ~59．2%，体重增长率为72．0% ~311．9%。NO₃ ^- -N对体长7．37cm幼虾的亚急性毒性效应：NO₃ ^- -N的质量分数在 30~195mg/L时，成活率为 35% ~100%，体长增长率为8．5% ~20．5%，体重增长率为29．6% ~56．8% 。三态氮在一定质量分数范围内均对幼虾的存活率和生长率产生影响。     

环境因子对鼠尾藻幼苗叶绿素荧光参数的影响

为了揭示鼠尾藻幼苗的生态适应性,研究了温度(5~34 ℃)、盐度(10~50)和营养盐等环境因子对鼠尾藻幼苗叶绿素荧光参数的影响。结果表明,(1) 氮浓度高于8 mg/L或磷浓度高于1.2 mg/L,或温度高于28 ℃,对鼠尾藻幼苗的光合作用均有显著影响(P<0.05);(2) 短时间的5~15 ℃的低温胁迫或10~50盐度胁迫6 h对鼠尾藻幼苗的F_v/F_m值影响不明显;(3) 氮、磷浓度分别为2~4 mg/L和0.2~0.8 mg/L,且NH⁺₄-N∶NO^-₃-N的比值为1~3时,较利于鼠尾藻幼苗光合作用的进行。     

超声降解法制备可溶性鱿鱼墨黑色素及其抗氧化性

实验研究了一种在碱性条件下利用超声降解制备可溶性鱿鱼墨黑色素的新方法。将鱿鱼墨黑色素溶于NaOH后用超声细胞破碎仪处理1 h,超滤后分别获得不同分子量可溶性黑色素组分。结果显示,超声处理后黑色素平均粒度从17.34 μm下降至1.467 μm,紫外、红外光谱和核磁共振谱图从结构上说明超声作用主要破坏黑色素的高度聚合状态,而黑色素的主要化学结构并没有被破坏,只有少部分基团,尤其是较低分子量组分中的DHI和DHICA结构被氧化;体外抗氧化实验显示经过0.5 mol/L和1 mol/L的NaOH处理,分子量大于10 ku的组分具有很强的抗氧化性,这些组分清除超氧自由基能力(IC₅₀=19～80 μg/mL)远优于作为商品抗氧化剂的肌肽(IC₅₀=355 μg/mL);清除羟基自由基活性(IC₅₀=115～180 μg/mL)与肌肽(IC₅₀=110 μg/mL)相当。可溶性鱿鱼墨黑色素作为一种天然色素和新型自由基清除剂,其良好的可溶性大大提高了机体的吸收利用率。     

北极茴鱼形态性状对体重影响效果分析

为探明北极茴鱼(Thymallus arcticus)形态性状与体重的关系, 以 1⁺ ~3⁺ 龄北极茴鱼为研究对象, 测定了体重(Y) 和体厚(X₁)、眼间距(X₂)、体长(X₃)、体高(X₄)、头长(X₅)、眼径(X₆)、吻长(X₇)、尾柄长(X₈)、尾柄高(X₉)等 9 个形态性状, 利用相关分析、通径分析和多元回归分析等方法, 筛选出影响北极茴鱼体重的主要形态性状, 并建立回归方程。结果显示: (1) 不同年龄阶段, 与北极茴鱼体重显著相关(P＜0.05)的形态性状种类不同, 且数量也随着年龄的增加而降低。(2) 通径分析在 1⁺ ~3⁺ 龄北极茴鱼中分别保留了 4、4 和 2 个形态性状, 1⁺ 龄北极茴鱼中体长(X3)的直接作用最大(0.345), 尾柄高(X9)的间接作用最大(0.745); 2⁺ 龄北极茴鱼中体高(X4)的直接作用最大(0.473), 体厚(X1)的间接作用最大(0.378); 3⁺ 龄北极茴鱼中体厚(X₁)的直接作用最大(0.635), 尾柄高(X₉)的间接作用最大(0.344)。(3) 通径分析保留的形态性状对 1⁺ ~3⁺ 龄北极茴鱼体重的总决定系数较高, 分别为 0.943、0.778 和 0.997。(4) 多元回归分析拟合出 1+ 龄北极茴鱼形态性状(Xi)与体重(Y)回归方程为 Y=?90.510+15.345X₁+3.638X₃+10.473X₄+16.884X₉, 2+ 龄北极茴鱼形态性状(Xi)与体重(Y)回归方程为 Y=?142.449+29.023X₁+81.082X₂+27.126X₄?47.376X₇, 3⁺ 龄北极茴鱼形态性状(Xi)与体重(Y)回归方程为 Y=?228.922+75.063X₁+107.864X₉。本研究丰富了北极茴鱼基础生物学数据, 同时为将来人工养殖利用过程中北极茴鱼的选择育种提供待选形态性状。     

中国西北地区次生盐碱水无机氮转化与环境因子的相关关系

中国西北地区次生盐碱水中NO₂^--N长期处于较高浓度，严重制约了盐碱水养殖产业的可持续发展。根据对甘肃省景泰县草窝滩渔农综合示范区（104°7''40″E，37°19''6″N）的定期定点监测，运用配对样本t检验、Duncan''s多重比较和Pearson相关性分析，研究了不同类型次生盐碱水体无机氮转化及其与环境因子的相关关系。结果表明：（1）次生盐碱水无机三态氮（NO₂^--N、NH₄⁺-N、NO₃^--N）及总氮（TN）本底值高，全年均值分别是NO₂^--N（0.3±0.2）mg/L、NH₄⁺-N（1.93±1.25）mg/L、NO₃^--N（2.92±1.5）mg/L、TN（13.91±5.85）mg/L；无机氮占TN比例不超过50%，说明有机氮在次生盐碱水体中所占比例更高；（2）环境因子pH与NO₂^--N正相关，与NO₃^--N负相关，盐度与NO₃^--N负相关，pH和盐度加剧了次生盐碱水NO₂^--N的积累；（3）水产养殖显著降低次生盐碱水体中NO₃^--N浓度和碳酸盐碱度，显示了盐碱水养殖对次生盐碱水的生态改良功能。本研究旨在为中国次生盐碱水的渔业开发利用提供科学依据。     

肌注和口服氟苯尼考在中华鳖体内残留分析及药代动力学

研究不同给药条件下，氟苯尼考在中华鳖体内的残留及药代动力学特征。健康中华鳖160只，随机分为2组，按30 mg·kg^-1剂量分别单次肌注或口服氟苯尼考，高效液相色谱法测定中华鳖血浆和肌肉药物残留浓度，利用3P87药代动力学软件分析数据。肌注和口服时均符合一室开放模型；肌注给药的动力学方程C=16.72(e^-0.15t-e^0.52t) ，主要药代动力学参数：AUC=76.45 μg·mL^-1·h^-1，吸收半衰期（T_1/2Ka）=1.31 h，半衰期(T_1/2Ke)=4.48 h，最高血药浓度C_max=7.09 μg·L^-1；口服给药的动力学 方程为C=39.99(e^-0.19t-e^0.4t)，主要药代动力学参数：AUC=109.42 μg·mL^-1·h^-1，吸收半衰期（T_1/2Ka）=1.73 h，半衰期(T_1/2Ke)=3.63 h，最高血药浓度C_max=10.64 〗μg·L^-1。实验结果表明，氟苯尼考口服情况下，在中华鳖体内吸收快，血药浓度高，维持时间长，生物利用度高；药物在肌肉中消除缓慢。     

不同光强和温度对长石莼（缘管浒苔）光合作用和叶绿素荧光参数的影响

利用脉冲振幅调制叶绿素荧光仪对不同光照和温度条件下长石莼（缘管浒苔）光合作用和培养条件进行了优化。结果表明，长石莼（缘管浒苔）在25 ℃和72 μmol/(m2·s)条件下，其F_v/F_m、F_m、F_v和α值最高，分别高达0.74、4 567、3 406和0.305，低于该点为光不饱和，高于该点为光抑制，偏离越大，下降越显著（P<0.01），其中在5 ℃和35 ℃时最小，分别是25 ℃最高值的32.24%~64.88%和22.99%~53.44%；光强为18 μmol/(m²·s)和216 μmol/(m²·s)时最小，其F_v/F_m、F_m、F_v和α值分别是25 ℃和72 μmol/(m²·s)条件下最高值的44.94%~82.62%和51.82%~76.72%。F₀变化不太明显，5~30 ℃为先上升后下降或再上升变化趋势，35 ℃为先下降后上升变化趋势。拟合参数α显示，长石莼（缘管浒苔）在达到光饱和点前通过增加光能吸收来增强光合作用，在光抑制后则迅速减少。rETR_maxDuncan检验表明，高温/低温和高光强对长石莼（缘管浒苔）光合作用影响显著（P<0.01）。总体上看，长石莼（缘管浒苔）光合作用和生长适宜条件为15~25 ℃和54~72 μmol/(m²·s)，过高或过低均不适宜。且温度排序为25>20>15>30>10>5>35 ℃，光照强度排序为72>54/108>36/162>18/216 μmol/(m²·s)。     

软骨藻酸对海湾扇贝(Argopecten irradians)血淋巴免疫力和抗氧化力的影响研究

软骨藻酸是一种神经性贝类毒素，大多数研究只报道了对陆生动物神经系统的毒性和在贝类组织内累计的软骨藻酸的浓度，而对软骨藻酸对贝类自身的免疫及抗氧化系统的毒性作用相关的研究不足。扇贝作为无脊椎动物，缺乏适应性免疫，主要依靠先天免疫系统进行防御。因此，本研究以海湾扇贝（Argopecten irradian）为研究对象，在0、10 ng/mL、50 ng/mL和100 ng/mL软骨藻酸浸泡6 h、12 h和24 h后通过测定扇贝血淋巴中超氧化物歧化酶（SOD）、溶菌酶（LZM）活性和谷胱甘肽（GSH）含量以及免疫、抗氧化相关基因（Cu/ZnSOD、MnSOD、GST和ACP）相对表达量来研究其对扇贝免疫力及抗氧化系统的影响。结果发现，LZM活性在10 ng/mL和50 ng/mL软骨藻酸处理后显著提高；ACP基因表达量在6 h和12 h内表达量受到上调，而在24 h表达量显著下调；10 ng/mL、50 ng/mL和100 ng/mL软骨藻酸浸泡扇贝6~24 h后SOD活性受到抑制，并且Cu/ZnSOD和MnSOD基因表达受到调控；GSH含量显著提高，并且GST基因表达量显著上调。以上结果表明软骨藻酸处理虽然会抑制抗氧化酶SOD活性，并且高浓度长时间处理会造成免疫疲劳现象，但机体可通过提高血淋巴GSH水平和GST基因的表达量来抵抗软骨藻酸的毒性。因此，本研究初步揭示了软骨藻酸对扇贝等双壳类免疫力、抗氧化力和解毒力的影响。     

金枪鱼鱼骨胶原肽的制备及抗氧化活性研究

为制备金枪鱼鱼骨胶原肽,并对其抗氧化活性进行研究,利用酶解、超滤、凝胶色谱和反相高效液相色谱制备抗氧化胶原肽,采用氨基酸序列分析仪测定其氨基酸序列,利用质谱(ESIMS)确定其分子量,采用羟自由基、DPPH自由基、ABTS自由基和超氧阴离子自由基清除实验和脂质过氧化抑制实验对胶原肽抗氧化能力进行评价。结果显示,金枪鱼鱼骨胶原蛋白经胃蛋白酶和胰蛋白酶2步酶解和分离纯化得到1个十肽(TFCH-P2),经氨基酸序列分析和质谱(ESIMS)确定其氨基酸序列为Gly-Pro-Ala-Gly-Pro-Ala-Gly-Glu-Gln-Gly(GPAGPAGQEG),分子量为839.87 u([M+H]+840.68 u)。体外抗氧化实验结果表明,GPAGPAGQEG对羟自由基(EC500.18 mg/mL)、DPPH自由基(EC500.97 mg/mL)、ABTS自由基(EC500.52 mg/mL)和超氧阴离子自由基(EC500.38 mg/mL)具有良好的清除作用;GPAGPAGQEG亦显示出良好的脂质过氧化抑制作用。研究表明,胶原肽GPAGPAGQEG抗氧化活性良好,可以用于抗氧化相关的功能食品、药物或者食品添加剂。     

Influence of Different Hydrolysis Processes by Trypsin on the Physicochemical,Antioxidant, and Functional Properties of Collagen Hydrolysates from Sphyrna lewini,Dasyatis akjei,and Raja porosa

ABSTRACT

Acid soluble collagen hydrolysates from the cartilages of Sphyrna lewini, Dasyatis akjei, and Raja porosa on different hydrolysis conditions by trypsin were prepared and named as S, D, and R, respectively. The hydrolysates (S1, D1, and R1) obtained from hydrolysis in pH 2.5, 3 h, 37°C presented wonderful foaming and emulsifying capacities, caused by their high average molecular weights (AMWs), high degree of Pro and Lys hydroxylation, and imino acid contents (17.1, 15.3, and 14.1%). At the concentration of 0.1% (w/v), the foaming capacities of S1, D1, and R1 were 104.75 ± 2.57, 63.87 ± 2.73, and 76.87 ± 2.02%; and the emulsifying activity indexes of them were 116.07 ± 1.89, 91.04 ± 1.79, and 123.85 ± 2.14 m²/g, respectively. Conversely, hydrolysates (S2, D2, and R2) obtained from hydrolysis in pH 7.8, 3 min, 37°C exhibited strong scavenging activity on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (EC₅₀ 0.568, 0.680, and 1.634 mg/mL), hydroxyl radical (EC₅₀ 0.253, 0.376, and 0.438 mg/mL), and moderate reducing power (0.0465–0.4702 at the concentration of 5 mg/mL), due to their low AMWs. The results indicated that S1, D1, and R1 could be used as emulsifiers and foaming agents, and S2, D2, and R2 could be used as natural antioxidants in food systems.     

Antioxidant Activity of Brown Seaweeds

Antioxidant potential of three brown seaweeds, Anthophycus longifolius, Sargassum plagiophyllum, and Sargassum myriocystum, obtained from the Gulf of Mannar region of India were evaluated utilizing different in vitro systems. Ethyl acetate (EtOAc) fraction of Anthophycus longifolius registered significantly greater hydroxyl radical scavenging ability (IC₅₀ 0.19 mg/mL) and was effective in stabilizing the 2,2′-azino-bis-3-ethylbenzothiozoline-6-sulfonic acid (IC₅₀ 1.23 mg/mL) and 1,1-diphenyl-2-picryl-hydrazil (DPPH) radicals (IC₅₀ 0.48 mg/mL) (p < 0.05). No significant differences in hydrogen peroxide (H₂O₂) scavenging and ferrous ion chelating properties of the EtOAc extracts of the seaweeds were apparent. The utilities of the reverse-phase high-performance liquid chromatography (RP-HPLC) method hyphenated to diode-array detection for analyzing the fingerprints of phenolic constituents in the solvent extracts and fractions of the seaweeds were denoted. High-performance liquid chromatographic analysis indicated the presence of phenolic acids in the solvent extracts of seaweeds. This study demonstrated the potential use of A. longifolius as candidate species to be used as a nutritional food supplement/functional foods to increase the shelf life of food items for human consumption.     

Biological Activities of Freshwater Algae,Spirogyra singularis Nordstedt

Spirogyra is commonly found as accessible algae in freshwater areas all over the world. Some medical uses have been reported from this genus. Biological activities of Spirogyra singularis were investigated employing eight in vitro assays. The extract showed different antioxidant activity. IC₅₀ for DPPH radical-scavenging was 4.71 ± 0.11 μg mL^?1. The extract showed very strong nitric oxide-scavenging activity with IC₅₀ = 77.3 ± 2.07 μg mL^?1, but its Fe²⁺ chelating ability was weak. The extract also exhibited high antioxidant activity in hemoglobin-induced linoleic acid peroxidation tests and scavenging of hydrogen peroxide. The extract also showed good antihemolytic activity. The total amount of phenolic content in the extract was determined as gallic acid equivalents, and total flavonoid content was calculated as quercetin equivalents. Biological activities may be attributed, at least in part, to the presence of phenol and flavonoid contents in the extract.     

大眼金枪鱼头蛋白酶解物1 ku超滤组分的抗氧化活性及其理化性质

研究大眼金枪鱼头蛋白酶解物1 ku超滤组分体外的还原力、自由基清除能力及对衰老小鼠体内抗氧化能力的影响,分析1 ku超滤组分的一般成分、氨基酸组成及分子量分布,为进一步分离纯化金枪鱼头抗氧化肽提供基础。体外结果显示,1 ku超滤组分对羟基自由基、超氧阴离子和DPPH自由基的清除活性随浓度的增加而增强,IC50分别为1.38、0.73与0.93mg/mL,还原力也随浓度的增加而增大,在浓度为12.5 mg/mL时为0.763;体内结果显示,灌胃30 mg/kg的1 ku超滤组分连续42 d,D-半乳糖致衰老小鼠肝组织的超氧化物歧化酶(SOD)活性、肝组织和血清的谷胱甘肽过氧化物酶(GSH-PX)活性显著提高(P<0.05),血清丙二醛(MDA)含量显著降低(P<0.01);理化分析结果显示,1 ku超滤组分(干基计)蛋白质含量为96.40%,脂肪0.11%,灰分4.86%,疏水性氨基酸占氨基酸总量的35.8%,活性组分分子量在1 802~2 519 u和422~922 u。     

Antioxidant Properties of Protein Hydrolysate Obtained from Oyster Saccostrea cucullata (Born, 1778)

The antioxidant activities of enzymatically hydrolyzed (protease from Bacillus cereus SU12) oyster (Saccostrea cucullata) protein were studied. The hydrolysate exhibited a strong antioxidant potential in 1, 1-diphenyl-2-picrylhydrazyl (DPPH, 85.7 ± 0.37%), followed by hydrogen peroxide radical scavenging activity (81.6 ± 0.3%), hydroxyl radical scavenging activity (79.32 ± 0.6%), and reducing power assay (2.63 ± 0.2 OD at 700 nm) at a concentration of 1 mg/mL. Due to the high antioxidant potential, the hydrolysate was purified in Sephadex G-25 gel filtration chromatography. The active peptide fraction was identified by DPPH and reducing power assay. The amino acid content of the purified active peptide fraction was analyzed by high performance liquid chromatography. The active fraction contained a good quantity of both essential and nonessential amino acids. The present study revealed that oyster (S. cucullata) protein hydrolysate is a potential source for natural antioxidants.     

Antioxidant and ACE Inhibitory Activities of Kawakawa (Euthynnus affinis) Protein Hydrolysate Produced by Skipjack Tuna Pepsin

ABSTRACT

Pepsin enzyme from skipjack tuna was extracted for the production of kawakawa (Euthynnus affinis) fish protein hydrolysate. Using ultra-fractionation, Kawakawa protein hydrolysates were separated into four different fractions, including fractioned protein hydrolysate I (FPH I) (< 1 kDa), FPH-II (1–3 kDa), FPH-III (3–10 kDa), and FPH-IV (> 10 kDa). The antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, inhibition of linoleic acid oxidation, reducing power tests, and chelating activity of metal ions. Results indicated that FPH II fraction peptides had higher antioxidant activity in comparison with the other fractions, followed by FPH I. Further, the fractions were evaluated for angiotensin converting enzyme (ACE) inhibition, and IC₅₀ value ranged from 0.45 to 1.86 mg/ml with higher activity in FPH I (IC₅₀ 0.45). Finally, the amino acid profile of different fractions was analyzed. The fractions exhibited significant amounts of hydrophobic amino acids, which could perform as hydrogen donors, frustrate the free radicals, and inhibit the ACE. The recovered pepsin from the viscera was used to produce hydrolysates with good biological activities. Peptides lower than 3 KDa had antioxidant activity as positive controls and significant ACE activity. These are very important findings that could be used to conduct further research in a preclinical study of these peptides.     

Antioxidant Assessment of Schizochytrium Meal Protein Enzymatic Hydrolysate and Its Potential Application

Schizochytrium meal protein (SMP) extracted from Schizochytrium meal was hydrolyzed by flavourzyme. Response surface methodology (RSM) was used to optimize the extraction conditions for the protein extraction yield from Schizochytrium meal. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (DRSA) was used to evaluate the antioxidant activity of hydrolysates. The orthogonal test was used to investigate the effects of three independent variables, namely protease dosage, hydrolysis time, and pH. The optimum conditions obtained were as follows: protease dosage of 7%, hydrolysis time of 1.5 h, pH of 6, under which, DRSA at the concentration of 2 mg/mL was 89.38%. Aspartic and glutamic acid constituted approximately 26.32% of the total amino acids, and glutamic acid was the most abundant amino acid of Schizochytrium meal protein hydrolysate (SMPH) by amino acid composition analysis, which may have contributed to the scavenging activity of SMPH. Moreover, SMPH was made into chewable tablets with suitable formula and high humidity stability. These findings indicate that Schizochytrium meal can be reused as a raw material for preparation of antioxidant peptides.     

Optimized Extraction,Preliminary Characterization and Evaluation of the in Vitro Anticancer Activity of Phlorotannin-Rich Fraction from the Brown Seaweed,Cystoseira sedoides

Brown seaweeds produce useful bioactive substances with high cosmetic and pharmacological values due to the presence of antioxidant derivatives, mainly phlorotannins (PHT), which are of particular interest. A central composite rotatable design (CCRD) with three variables (extraction time, dry material-to-solvent ratio and ethanol concentration) and two responses was performed to optimize the microwave-assisted extraction (MAE) of phlorotannins from Cystoseira sedoides using response surface methodology (RSM). Results showed that the independent variables significantly affected both phlorotannin content and the scavenging capacity. The optimum operating conditions were extraction time, 101.74 sec; dry material-to-solvent ratio, 1:10 g/mL; and ethanol extraction, 50%. Under these conditions, the predicted values of PHT content and radical scavenging activity-IC50 were close to the observed values and were 383.887 µg PGE/g Dm and 18.353 µg/mL, respectively. Characterization of the phlorotannin-rich fraction was conducted by high-performance liquid chromatography and Fourier transform infrared spectroscopy. The evaluation of the anticancer activity against MCF-7 breast cancer cell line showed a potent activity to trigger apoptotic death in more than a half of the MCF-7 cells, with an estimated IC50 value of 78 μg/mL. In addition, this fraction induced a notable growth regression effect on 3D spheroids model in a concentration-dependent manner, with a growth rate of about 1.17, at 200 µg/mL.

Abbreviations: CCRD: Central composite rotatable design; Dm: Dry material; DMBA: 2,4-dimethoxy benzaldehyde; DMSO: Dimethyl sulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EDTA: Ethylene diamine tetraacetic acid; FBS: Fetal bovine serum; FT-IR: Fourier transform infrared; HPLC: High-performance liquid chromatography; IC₅₀: Half-maximal inhibitory concentration; MAE: Microwave-assisted extraction; Min: minutes; PBS: Phosphate buffered saline; PGE: Phloroglucinol equivalent; PHT: Phlorotannins; PHT-SED: phlorotannins derived from Cystoseira sedoides; PI: Propidium iodide; RSA: Radical scavenging activity; RSM: Response surface methodology; Sec: seconds