Oil and squalene in amaranthus grain and leaf

Amaranthus grain of 104 genotypes from 30 species was investigated for oil and squalene contents and fatty acid profiles. The overall average oil content in Amaranthus grain was 5.0%, ranging from 1.9 to 8.7%. Squalene concentration in extracted oils ranged from trace to 7.3%, with an average concentration of 4.2%. The average contents of three major fatty acids in Amaranthus grain were 22.2, 29.1, and 44.6% for palmitic, oleic, and linoleic, respectively. The average fat content in dried mature leaves of 45 Amaranthus genotypes was 1.63%, ranging from 1.08 to 2.18%. The squalene concentration in leaf lipid extracts averaged 0.26%, ranging from trace to 0.77%, which is much lower than that from seeds. The major fatty acids of leaf extracts were linolenic, linoleic, and palmitic. Linolenic ranged from 56.5 to 62.0% of total fatty acids; linoleic, from 15.5 to 24.7%; and palmitic acid, from 13.5 to 15.5%. As for the fatty acid compositions at different growth stages, fatty acid content in leaf lipid was lower in mature leaves than in young leaves. The saturated/unsaturated ratio decreased when the leaf grew to maturity. Principal component analysis (PCA) was carried out on compositional characteristics of grain. The first two components accounted for 70% of the total variance (38.3 and 21.7%, respectively). There was a positive correlation between oil content and squalene yield, and a negative correlations were found between linoleic and either of the other two major fatty acids, palmitic and oleic. The taxonomic relationship among the species was also elucidated by PCA.     

Supercritical carbon dioxide extraction of oil and squalene from amaranthus grain

Supercritical carbon dioxide (SC CO(2)) was used for the extraction of oil and squalene from Amaranthus grain. Very small amounts of oil could be extracted by SC CO(2) from undisrupted grains, although SC CO(2) possesses higher diffusivity. Grinding increased the extraction rate and oil yield, and smaller particle size gave higher extraction rate. The oil yield and initial extraction rate increased linearly with the increasing SC CO(2) flow rate from 1 to 2 L/min. Increasing the flow rate of SC CO(2) above 2 L/min resulted in only a slight increase of oil yield and extraction rate. In the pressure range of 150-250 bar, extraction decreased with increasing temperature at a constant pressure, whereas at a pressure of 300 bar, the extraction yield increased with increasing temperature. Possible reasons for this are discussed. Effects of temperature and pressure on squalene yield were different from those on oil yield. A good oil yield (4.77 g of oil/100 g of grain) was obtained at 40 degrees C and 250 bar. The highest squalene yield (0.31 g of squalene/100 g of grain) and concentration (15.3% in extract) were obtained at 50 degrees C and 200 bar, although the oil yield under this condition was low (2.07 g of oil/100 g of grain). The moisture content within 0-10% had little influence on yields of oil and squalene at 40 degrees C and 250 bar. Finally, the oil yield and the squalene concentration in the extracts by SC CO(2) were compared to those by solvent extraction.     

牛骨蛋白与多糖的逐级提取及分离方法研究

为确定牛骨中蛋白质和多糖的提取及分离方法，该文在分析骨胶原蛋白和多糖存在状态的基础上，分别利用不同溶剂探讨了牛骨中蛋白质与多糖的逐级提取方法。首先碱性蛋白、酸性蛋白分别被0.6 mol/L KCl和0.5 mol/L K-P(K₂HPO₄/KH₂PO₄)溶液提取出来；0.1 mol/L HCl脱钙时溶解出酸性、中性及碱性蛋白，再经过NaOH中和后，酸性蛋白随羟基磷灰石沉淀、中性及碱性蛋白存留在上清液中；其他在胶原纤维上的有机物被0.5 mol/L NaCl和1% Na₂CO₃分别提取出来。各级提取有机物用SDS-PAGE电泳以及Stains All染色法、阿里新兰染色和CBB染色进行检测。利用DEAE-葡聚糖凝胶离子交换树脂有效地将碱性蛋白、酸性蛋白和多糖分离精制。     

裙带菜中褐藻糖胶的提取纯化及其组成的研究

该实验优化了裙带菜褐藻糖胶的提取工艺,对粗多糖进行了分离纯化,并研究了各组分的单糖组成和硫酸根含量。利用响应面优化法,依据二次回归分析确定裙带菜褐藻糖胶的最佳提取工艺为：提取温度76℃,提取时间2.5 h,料液比1︰6.5。在此条件下提取2次,粗多糖的提取率可达到7.53%。提取的粗多糖用DEAE-Sepharose Fast Flow离子交换层析分离纯化,共得到5个峰,干燥后得到5个组分。用硫酸钡比浊法测定5个组分的硫酸根含量分别为8.3%、19.2%、30.1%、38.4%和33.0%。糖腈乙酸酯衍生物气相色谱测得各组分的单糖组成分别为：组分1主要含有甘露糖和半乳糖;组分2主要含有鼠李糖,岩藻糖,木糖,甘露糖,葡萄糖和半乳糖;组分3主要含有鼠李糖,岩藻糖和甘露糖;组分4主要含有岩藻糖和半乳糖;组分5主要含有岩藻糖,甘露糖和半乳糖。     

油梨皮黄酮提取及大孔树脂纯化

该文研究了油梨皮黄酮的提取及大孔树脂纯化条件.结果表明:油梨皮黄酮的最佳提取条件为乙醇浓度70%、提取温度70℃、提取时间1.5 h、料液比(m/V)1:20.乙醇浓度和提取温度对提取得率有显著性(P<0.05)影响.在此条件下,黄酮的提取得率为1.12%;AB-8型树脂对油梨皮黄酮有较好的吸附和洗脱效果,其纯化油梨皮黄酮的条件为柱体积250 mL,上样量2.03 g,水洗,接着用75%的乙醇洗脱(约500 mL),在此条件下AB-8型树脂可重复使用6次.经纯化后油梨皮黄酮相对纯度为82.37%,纯化后总黄酮回收率为71.65%.     

菜籽蛋白的提取和纯化

为了对菜籽饼进行综合利用,以低温压榨制油的双低菜籽饼为原料,探讨了菜籽蛋白的提取和纯化工艺.通过料液pH值的先后调节对比、正交、提取次数和纯化试验,得到较佳工艺条件:采用先调pH值法,水溶液的pH值为12,提取时间为20 min,温度为室温,提取3次,通过pH5和7的两步沉淀,经干燥得到分离蛋白,蛋白纯度达84.8%,得率为38.9%,且有害物质残留较少.先调pH值法与后调法相比,总的提取率可提高20个百分点,达90%以上.     

鸡胸软骨硫酸软骨素的提取及分离纯化

为优化硫酸软骨素的提取工艺和初步分析其结构,采用中心组合设计和响应面分析优化鸡胸软骨硫酸软骨素的酶法提取工艺,三氯乙酸除蛋白质、乙醇沉淀以及柱色谱精细纯化得到高纯度硫酸软骨素,并采用高效凝胶过滤色谱判定其纯度,红外光谱和核磁共振初步分析其结构。结果表明,硫酸软骨素的最佳提取工艺参数为温度55.68℃,pH值为6.40,时间为3.85 h,葡萄糖醛酸和硫酸软骨素提取率分别达到9.82%和28.05%。高效凝胶过滤色谱分析其纯度为99.01%。红外光谱和核磁共振显示硫酸软骨素主要是4硫酸化软骨素。     

Extraction and purification of isoflavones from soybeans and characterization of their estrogenic activities

Soybean isoflavones have multiple beneficial health effects especially on estrogen-deficient diseases such as menopausal symptoms. In this study, isoflavones were produced from soybean flour, and the extraction and purification parameters were optimized to give a high yield of total isoflavones, about 0.62 mg of aglycones/g of soybean flour, which is >2 times the initial yield. HPLC analysis and MTT cell proliferation assay using MCF-7 cells revealed that the product thus obtained not only contained a high content of isoflavone aglycones but also had estrogenic activity. MTT data also revealed that both genistein and daidzein exhibited estrogenic effects at lower concentrations and antiproliferative effects at higher concentrations, and 1 microM genistein and 10 microM daidzein exerted significant estrogenic activities, which were not more than that of the endogenous level of 17beta-estradiol (E2). The production method developed can be used as a guideline for manufacturing soy isoflavones, and the MTT assay was demonstrated to be suitable for quality control on isoflavone products. The results on the estrogenic properties of isoflavones can be used as reference data for their effective and safe usages in estrogenic therapy.     

沙枣果肉原花青素的提取、纯化及抗氧化性能的研究

该文对沙枣果肉中原花青素的提取工艺进行了研究,通过单因素试验,在中心组合试验基础上得出了提取沙枣果肉原花青素的最佳工艺条件：提取温度76℃、提取时间86 min、乙醇浓度72％、提取液pH值5.0、料液比1∶9、提取3次。在此条件下提取率达95.9%。采用固相萃取法对沙枣果肉提取物中原花青素进行纯化,产物纯度为89.91％,原花青素的得率为4.23％;同时对纯化后沙枣果肉原花青素的抗氧化性能进行了测定,结果显示沙枣果肉原花青素有较好的抗脂氧化能力。     

Extraction,purification and properties of soil hydrolases

Invertase, cellulase, phosphatases, protease and β-glucosidase were extracted from permanent pasture soil with 0.2 M phosphate buffer (pH 8) in the presence of 0.2 M EDTA. This extract was further treated with ammonium and salmine sulphates. Attempts were made to fractionate these enzyme activities by gel and anion-exchange chromatography. Specific activities were estimated in all fractions and some characteristics of the purified enzymes (optimum pH, temperature and substrate concentration, and K_m and V_max) were investigated. The results indicated that extracted enzyme activities occurred partly in soil as a carbohydrate-enzyme complex and partly as a humo-carbohydrate complex.     

欧李多酚提取纯化及抗氧化性研究

欧李富含钙素和多酚类物质,具有提高人体免疫力等保健功能,近年来在中国得到了大力发展。欧李多酚提取、纯化及其抗氧化功能,为欧李开发利用提供理论依据。该研究采用单因素及响应面优化试验研究了欧李多酚超声辅助酶解提取工艺,采用大孔树脂纯化多酚粗提物,利用高效液相色谱法测定酚类物质的种类,利用分光光度法测定欧李多酚类物质总还原力和对自由基·OH、DPPH·、ABTS·的清除作用。试验得出欧李多酚提取最优工艺参数为：超声波功率105 W、酶解温度50 ℃、酶解时间80 min、液料比30∶1 mL/g,多酚提取量为42.63 mg/g。经大孔树脂纯化后纯化物中欧李总多酚含量为73.42 mg/g,从欧李果实中检测出6种多酚类物质,其含量顺序为绿原酸>没食子儿茶素没食子酸酯>咖啡因>对羟基苯甲酸>原儿茶酸>没食子酸。欧李多酚的总还原力与维生素C相当,对·OH、DPPH·、ABTS·等自由基清除作用显著。     

碱性土壤基因组DNA的分离纯化和基因文库的构建

直接从广西南宁市凤凰纸业排污沟碱性土壤样品中抽提和分离纯化混合基因组DNA，所获得DNA的产量为每克土壤样品10～30pg。采用限制性内切酶EcoR1酶处理后，构建了以pGEM-3Zf( )为载体的DNA部分文库。文库的容量为23650个转化子。外源片段DNA平均大小为3．2kb。建库效率为每克环境样品获得6000个左右的含1～15kb外源随机插入片段的克隆。通过DNA序列测定和同源性比较，对从文库随机词取的16个转化子序列进行分析，发现13个外源插入片段包含序列尚未确定的DNA片段。     

伊犁绢蒿种子黏液提取工艺及对畜禽粪便污水的净化效果

伊犁绢蒿种子遇水能分泌黏液,黏液物质具有良好的保水性、成膜性、增稠性和分散性,可作为新的天然絮凝剂应用于污水处理中。该文就影响黏液提取率的3个因素提取时间、液料比、提取温度进行探讨,同时用所提取的种子黏液对牛、猪粪便污水进行处理。响应面法结果表明,绢蒿种子黏液在该试验条件下的最高提取率为16.76%,提取条件为：提取时间3 h、液料比60∶1、提取温度64 ℃;畜禽粪便废水处理试验表明,牛粪便废水处理在黏液投加量为50 mg/L时,絮凝率为28%和COD去除率为32.7%;pH值等于6时,絮凝率为46%,COD去除率为44.6%。明矾的联合使用对絮凝有显著的效果,絮凝率和COD去除率最大分别为72.3%和61.3%。猪粪便处理过程中,在黏液投加量为20 mg/L时,絮凝率为30.9%,COD去除率为32.7%,在pH值为7时,絮凝率为29.3%,COD去除率为50%,明矾的联合使用下,絮凝率和COD去除率最大分别为62.6%和68.3%,达到了预想的处理效果。研究可为伊犁绢蒿种质资源在畜禽粪便污水处理方面的开发应用提供参考。     

板栗壳色素的提取、纯化及稳定性（简报）

该文以板栗壳为材料,研究板栗壳色素的提取、纯化工艺及纯化色素的稳定性,为板栗壳色素的推广应用提供理论基础和实验数据.结果表明,板栗壳色素易溶于碱性水溶液,不溶或难溶于非极性溶剂.用1%的NaOH提取色素,石油醚和乙酸乙酯分别笨取3次后获得粗提色素,粗提色素经醇沉、酸沉及霞结晶法三种方法纯化后通过薄层层析证明色素纯度较高.紫外-可见光谱分析表明,纯化色素的0.01%水溶液在223 nm和264 nm处有明显吸收峰,0.01%甲醇溶液在218 m和264 nm处有明显吸收峰,推测板栗壳色素含有苯环及酚羟基.纯化色素的稳定性试验表明,色素264.nm处吸光度对pH值较敏感.510 nm处吸光度在pH为2.0至6.0时.呈逐渐增加的趋势,在pH为8.0至14.0时变化不大,pH=10.0条件下测定510 nm处吸光度可以作为色素定量的方法;不同金属离子对色素有不同影响;色素对光、热、氧化剂及还原剂的耐受力较强.     

On the role of squalene in olive oil stability.

The role of squalene in olive oil stability was studied for various concentrations and experimental conditions. No effect was found in induction periods of olive oil at elevated temperatures using the Rancimat apparatus. Samples were then stored at 40 and 62 degrees C in the dark, and the extent of oxidation was followed by periodic measurements of peroxide value and conjugated dienes. A concentration dependent moderate antioxidant activity was evidenced which was stronger in the case of olive oil compared to that found for sunflower oil and lard. In the presence of alpha-tocopherol (100 mg/kg) and caffeic acid (10 mg/kg) the contribution of squalene (7000 mg/kg) was not significant. No radical scavenging activity was observed using DPPH(*) in 2-propanol. The weak antioxidant activity of squalene in olive oil may be explained by competitive oxidation of the different lipids present which leads to a reduction of the oxidation rate. Squalene plays a rather confined role in olive oil stability even at low temperatures.     

Recovery of squalene from wine lees using ultrasound assisted extraction-a feasibility study

The present work is a systematic approach for valorization of wine lees regarding the recovery of squalene, a bioactive lipid. Such a study is presented for the first time in literature. Separate examination of squalene content in "light" and "heavy" lees from different vinification processes by RP-HPLC demonstrated that these waste streams can be used as a source for this lipid, despite variations due to technological or genetic effects. Next, ultrasound assisted extraction of squalene from the "industrial waste" (the mixture of wine lees generated from different wines) using n-hexane was optimized with the aid of response surface methodology (independent variables: sonication duration and duty cycles). Autolysis was monitored through optical microscopy. Squalene yield (0.6 ± 0.08 g SQ/kg dry lees) was comparable to that of recently examined potential sources (0.2-0.35 g SQ/kg dry olive pomace and 0.06 g SQ/kg olive leaves).     

鱼腥草叶总黄酮的提取分离

采用微波辅助提取鱼腥草叶总黄酮,并用正交试验进行工艺参数的优化。同时选择7种大孔树脂,比较其对鱼腥草叶总黄酮的吸附量和解吸率,筛选出较优的大孔树脂并对其动态吸附及解吸性能进行考察。结果表明:优化的工艺条件为乙醇浓度50%,固液比1∶50(w/v),预浸泡30 min,微波作用时间30 s,微波功率540 W;与传统乙醇提取法相比,微波法使每次提取时间由3 h减少为30.5 min,提取率从92.14%增加到95.93%。NKA-9型大孔树脂对鱼腥草叶总黄酮有较好的吸附和解吸效果;较优的吸附分离工艺参数为:样液pH值在3.00～3.50,上样液流速1 mL/min,上样液浓度2.21～3.10 mg/mL,用70%乙醇洗脱时,解吸率达97.8%,3BV洗脱液基本上能将鱼腥草叶总黄酮洗脱下来。     

Characterization and purification of glycosaminoglycans from crude biological samples

Chondroitin sulfate (CS) is a glycosaminoglycan derived from cartilage and commonly used to treat osteoarthritis, psoriasis, and other conditions. The dimethylmethylene blue (DMMB) assay has been used often to measure glycosaminoglycan levels in relatively pure samples. In this study, we verified the accuracy of the DMMB assay in measuring CS levels in unpurified extract from bovine trachea and shark cartilage, despite potential interference from salts, proteins, and DNA. We found that the glycosaminoglycan signal obtained was due to CS and not to other glycosaminoglycan species. This was confirmed using fluorophore-assisted carbohydrate electrophoresis, which also revealed that the majority of the CS was monosulfated at the C4 or C6 position. Finally, we used anion-exchange chromatography to purify the bovine extract and obtained complete recovery of the glycosaminoglycans, with no contaminating protein. The results of this study should be very useful for future purification and analysis of this common supplement.