淫羊藿总黄酮的碱提工艺研究

以淫羊藿苷和总黄酮的含量为指标,考察了提取溶剂及用量、提取时间、次数以及浓缩比例和酸用量对淫羊藿黄酮类成分提取的影响,确定了淫羊藿总黄酮碱提酸沉的最佳提取条件为:用10倍量的0.1%NaOH溶液加热煮沸2次,每次1 h,滤液浓缩至料液比1∶4后用盐酸调节pH为2,沉淀放置36 h后,真空过滤或离心分离干燥即得高含量提取物。该提取工艺提取率高,稳定性好,操作简便,适合工业化生产。     

油茶茶枯中茶皂素的提取工艺研究

油茶茶枯是南方地区一种常见的加工副产物,含有较高的茶皂素。茶皂素是工业生产中应用广泛的一种表面活性剂,性能优良,但目前茶皂素的提取工艺并不成熟。为了优化油茶茶枯中茶皂素提取的最佳工艺,提高油茶茶枯的利用率,该文以脱脂油茶茶枯粉末为原料,乙醇溶液作为浸提剂,以茶皂素的得率和含量作为提取效果的评价指标,通过单因子实验和正交试验探求油茶茶枯中茶皂素的最佳提取条件。结果表明:以提取时间、3%NaOH添加量、乙醇浓度、料液比、温度和提取次数6个因素作为单因子,对油茶茶枯中茶皂素提取得率和含量进行研究,在单因子实验的基础上进行正交试验得到茶皂素提取的最佳工艺条件为料液比1∶17,乙醇浓度75%,提取温度81℃,提取时间88min,3%NaOH添加量2.5mL(即每10g茶枯粉末中添加0.075g NaOH),提取次数2次。通过论证该结果表明,在最佳的提取条件下,提取得到茶皂素的最高含量为0.54mg·mL-1,得率为21.35%。该实验方法添加了NaOH后,在最佳条件下茶皂素的得率明显提高,对今后优化油茶茶枯中茶皂素的提取工艺具有很好的借鉴意义。     

白桦茸凝集素提取工艺的优化

对白桦茸凝集素最佳提取工艺进行了研究.以2%兔血细胞凝血效价为指标,确定了最佳提取缓冲液.通过正交试验对料液比、提取时间、提取液pH值、NaCl浓度等因素进行了优化分析并确定了提取工艺的最佳参数组合.结果表明,以TBS和PBS为提取缓冲液所得的白桦茸凝集素凝集效价分别为64、16;最佳提取工艺:液料比为50:1,提取时间为20h,NaCl浓度为0mol/L,缓冲液pH值为8.0,按该最佳工艺提取白桦茸凝集素凝血效价为256.所优化的提取工艺稳定、可行,为该凝集素进一步在免疫调节方面的开发应用提供一定基础.     

加热回流法提取淫羊藿苷的试验研究

目的:考察加热回流法提取次数、提取时间、料液比以及乙醇体积分数等实验条件可能对淫羊藿苷的提取率所产生的影响。方法:提取溶剂选择为乙醇,采用加热回流法从淫羊藿中提取淫羊藿苷,采用单因素试验考察淫羊藿苷的提取时间、提取次数、淫羊藿的料液比以及溶剂中乙醇体积分数对淫羊藿苷的提取率所产生的影响,在此基础上进行正交试验的设计,优化提取工艺。结果:结果表明优化的加热回流提取法为料液比1:10(m:V,g/ml),提取2次、提取时间为40min,乙醇体积分数75%。     

野生火棘果红色素提取工艺优化

目的:通过正交实验,确定火棘果红色素的最佳提取工艺。方法:以贵州野生火棘果为原料,首先,以柠檬酸的乙醇溶液为提取剂,在400～800nm范围内对提取液的吸收光谱进行扫描,确定其最大吸收波长;然后,以最大吸收波长处的吸光度为衡量指标,确定最佳提取剂;再次,探讨了提取次数、提取温度、提取时间、料液比等单因素对火棘果红色素提取效果的影响;最后,在单因素实验结果的基础上,通过L9(34)正交实验,确定红色素的最佳提取工艺。结果:1.在400～800nm范围内,火棘果红色素的最大吸收波长为530nm;2.最佳提取剂为9%柠檬酸乙醇溶液;3.火棘果红色素的最佳提取工艺为,料液比1∶10,提取时间60min,提取次数2次,提取温度40℃。     

竹柏叶总黄酮不同提取方法比较

本研究采用正交试验设计分别优选回流提取和超声波提取竹柏叶总黄酮的最佳工艺条件。在单因素试验的基础上,以总黄酮得率为参考指标,选取液料比、乙醇体积分数、温度和时间4个影响因子,进行正交试验,分别确立回流提取和超声波提取最佳提取工艺条件。结果表明,回流提取优化工艺条件为:液料比30∶1(m L∶g),乙醇体积分数为70%,回流时间1.0 h,回流温度80℃;超声波提取优化工艺为:液料比25∶1(m L∶g),乙醇体积分数为70%,超声时间30 min,超声水浴温度60℃。两种提取方法的最终提取效果比较,回流提取的总黄酮得率为15.522%,超声波提取总黄酮得率为13.637%,回流提取法总黄酮得率高于超声波提取法。本实验结果为后续竹柏叶总黄酮的开发利用提供了参考。     

锁阳多糖提取工艺的研究

以浸膏得率和浸膏中多糖质量分数综合评价,利用L9(34)正交实验从水提和醇沉两方面优选锁阳中水溶性多糖最佳提取工艺条件,为锁阳活性多糖的开发提供科学依据和实验基础。最佳水提工艺条件为:m(锁阳):v(水)=1:12,回流提取0.5 h,提取2次;最佳醇沉工艺条件为:在m(生药):v(水提液)=1 g:2 mL溶液中加入乙醇,使φ(乙醇)=70%,静置6 h。     

双指标优选肠安颗粒中有机酸提取工艺条件(I)

以总有机酸和绿原酸的提取率双指标作为提取条件考核对象,来探讨肠安颗粒中所含有机酸成分的中药材的最佳提取工艺。采用L9(34)正交设计,考察提取溶媒所需加水量、浸泡时间、煎煮时间、煎煮次数四个因素,选择进行3个水平试验。得出结论:以加10倍量的水(第一次煎煮多加1.5倍量),浸泡1 h,煎煮2次,每次1h为最佳提取工艺条件,亦即以A2B2C2D2组合为合理。     

水蒸气蒸馏法提取大蒜挥发油的工艺研究

以挥发油提取率为指标,通过单因素和L9(34)正交试验设计确定了水蒸气蒸馏法提取大蒜挥发油的最佳提取工艺及条件。结果表明,各因素作用主次顺序为蒸馏时间(D)>料液比(C)>发酵温度(A)>浸泡时间(B),各因子的最优组合为A2B2C2D3,即以1∶3.5的料液比(m∶V),在35℃时浸泡3h后蒸馏2h为大蒜挥发油的最佳提取工艺,此时挥发油的提取率达到0.32%,证明此方法是大蒜挥发油提取的可靠、易行的科学方法。     

微波辅助提取石榴皮多糖工艺优化及其免疫调节作用研究

优化石榴皮多糖的微波辅助提取工艺并初步探究石榴皮多糖的免疫调节作用。在单因素试验的基础上,以液料比、微波提取时间、微波功率为自变量,石榴皮多糖得率为响应值,利用响应曲面分析方法,确定石榴皮多糖提取工艺的最佳条件为:液料比44∶1 m L/g,微波提取时间10 min,微波功率450 W,在该条件下石榴皮多糖的得率为10.48%。通过MTT法、中性红法、NO释放量测定和黏附试验初步表明石榴皮多糖对RAW264.7巨噬细胞具有一定的免疫调节作用。     

An improved method for the extraction of corticosterone from cell homogenates and subcellular fractions of the rat adrenal cortex

An improved technique is described for the extraction and analysis of corticosterone (11β,21-dihydroxy-4-pregnene-3,20-dione) from homogenates and subcellular fractions of the rat adrenal cortex. Factors influencing complete extraction of corticosterone were the nature of the organic solvent system and the concentration of the tissue being extracted. The continued activity of steroidogenic enzymes during subcellular fractionation was prevented by 0.1 mM 1-benzylimidazole. For optimum extraction, homogenates were diluted 1:12 (v/v) in 0.25 M sucrose, containing 0.1 M potassium hydroxide. Dilute homogenate was mixed with absolute ethanol (1:10, v/v) and extracted three times with diethyl ether (1:5, v/v). Following extraction, corticosterone in each sample was isolated by thin-layer chromatography (TLC), quantitated by radioimmunoassay (RIA), and corrected by measuring the recovery of added ³H corticosterone. With these procedures, 90–100% of corticosterone found in extracts of adrenal homogenates was recovered in extracts of subcellular fractions of the homogenates.     

Optimization of extraction process by response surface methodology and preliminary structural analysis of polysaccharides from defatted peanut (Arachis hypogaea) cakes

In this work, response surface methodology was used to determine optimum conditions for extraction of polysaccharides from defatted peanut cake. A central composite design including independent variables, such as extraction temperature (x₁), extraction time (x₂), and ethanol concentration (x₃) was used. Selected response which evaluates the extraction process was polysaccharide yield, and the second-order model obtained for polysaccharide yield revealed coefficient of determination of 97.81%. The independent variable with the largest effect on response was ethanol concentration (x₃). The optimum extraction conditions were found to be extraction temperature 48.7 °C, extraction time 1.52 h, and ethanol concentration of 61.9% (v/v), respectively. Under these conditions, the extraction efficiency of polysaccharide can increase to 25.89%. The results of structural analysis showed that the main composition of defatted peanut cake polysaccharide was α-galactose.     

Cryopreservation of Plasmodium chabaudi. II. Cooling and warming rates

Various cooling and warming rates were investigated to determine the optimum conditions for cryopreserving the intraerythrocytic stages of Plasmodium chabaudi. Infected blood, equilibrated in 10% v/v glycerol at 37 degrees C or in 15% v/v Me2SO at 0 degree C for 10 min, was cryopreserved using cooling rates between 1 and 5100 degrees C min-1. After overnight storage in liquid nitrogen the samples were warmed at 12,000 degrees C min-1. Warming rates between 1 and 12,000 degrees C min-1 were investigated using samples previously cooled at 3600 degrees C min-1. After thawing, the glycerol and Me2SO were removed by dilution in 15% v/v glucose-supplemented phosphate-buffered saline. Survival was assayed by inoculation of groups of five mice each with 10(6) infected cells and the time taken to reach a level of 2% parasitemia estimated. The optimum cooling rate was 3600 degrees C min-1 for parasites frozen using either 10% glycerol or 15% Me2SO; the pre-2% patent periods were 0.90 and 1.01 days above control values (representing survival levels of 21 and 17.5%, respectively). The optimum warming rate was 12,000 degrees C min-1; the pre-2% patent periods were 1.01 and 1.32 days above control values, respectively (18 and 10% survival), for glycerol and Me2SO. With ethanediol (5% v/v) and sucrose (15% w/v) as cryoprotectants the optimum warming rates were also 12,000 degrees C min-1 while the optimum cooling rates were 330 and 3600 degrees C min-1, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Composting of goat manure and wheat straw using pine cones as a bulking agent

This study aimed to determine the optimum mixture ratio of pine cones, goat manure and wheat straw and obtained optimal Free Air Space (FAS) values for composting. For this aim, pine cones were added at different ratios into goat manure and wheat straw mixtures. So, the FAS value of mixtures was fixed at four different levels. According to the results, the highest organic matter degradation and temperature value were obtained at the mixture ratio of 10% pine cones, 45% goat manure and 45% wheat straw. FAS value of this mixture was 32.8.     

铜藻岩藻黄素提取及纯化工艺研究

为了对岩藻黄素的提取、纯化进行系统研究,进而为高纯度岩藻黄素的工业化生产提供研究基础,筛选了适用于提取铜藻（Sargassum horneri）鲜藻中岩藻黄素的有机溶剂,并通过单因素实验和正交实验确定了最佳的提取溶剂浓度、提取温度、提取时间、料液比等工艺参数。随后采用硅胶柱层析法进行纯化,并通过单因素实验确定了最佳的硅胶柱床高度、上样量和洗脱流速。最后采用制备液相法对经层析纯化的岩藻黄素进一步纯化。结果表明,有机溶剂萃取的最佳工艺条件为：甲醇浓度90%,提取温度50 ℃,提取时间1 h,料液比1∶10,此条件下岩藻黄素提取率达到(0.258 9±0.003 6) mg·g-1鲜重（FW）［(1.078 8±0.015 0) mg·g-1干重（DW）］。硅胶柱层析的最佳工艺条件为：硅胶柱床高度10 cm,上样量6 g,洗脱流速10 mL·min-1,此条件下岩藻黄素得率为0.176 5 mg·g-1FW（0.735 3 mg·g-1 DW）,纯度为87.01%±0.88%。经制备液相进一步纯化后,岩藻黄素得率为0.127 1 mg·g-1 FW（0.529 4 mg·g-1 DW）,纯度为99.27%±0.22%。研究所用工艺简单,岩藻黄素得率高,为高纯度岩藻黄素的制备提供了实验基础。     

Effect of macromolecule supplementation during in vitro maturation of goat oocytes on developmental potential

In vitro maturation (IVM) of goat oocytes with serum-supplemented media results in oocytes with reduced developmental potential. The objective of this study was to develop a defined medium for IVM of goat oocytes that better supports subsequent embryonic development. Cumulus oocyte complexes (COC) were matured for 18-20 hr in: Experiment (1), tissue culture medium 199 (TCM199) with 10% (v/v) goat serum or modified synthetic oviduct fluid maturation medium (mSOFmat) with 2.5, 8.0, or 20.0 mg/ml bovine serum albumin (BSA); Experiment (2), mSOFmat with 4.0, 8.0, 12.0, or 16.0 mg/ml BSA; or Experiment (3), 1.0 mg/ml polyvinyl alcohol (PVA; control), 4.0 mg/ml BSA, 0.5 mg/ml hyaluronate plus 0.5 mM citrate, or hyaluronate, citrate, and BSA. Mature COC were coincubated for 20-22 hr with 12-15 x 10(6) sperm/ml in modified Brackett and Oliphant (mBO) medium. Embryos were cultured for a total of 7 days in G1/2, and evaluated for cleavage, and blastocyst development, hatching, and total cell numbers. In the first experiment, more (P < 0.05) blastocysts developed per cleaved embryo following maturation in mSOFmat with 2.5 or 8.0 mg/ml BSA than with 20.0 mg/ml BSA or TCM199 with 10% goat serum. The various concentrations of BSA used in the second experiment did not affect (P > 0.05) any of the developmental endpoints examined. In the third experiment, developmental potential of oocytes matured with PVA or hyaluronate with citrate was not different (P > 0.05) from oocytes matured in the presence of BSA. These results demonstrate that developmentally competent goat oocytes can be matured under defined conditions.     

Microwave-assisted extraction of glycyrrhizic acid from licorice root

In the present study, a microwave-assisted extraction (MAE) technique has been developed for the extraction of glycyrrhizic acid (GA) from licorice root. Various experimental conditions, such as extraction time, different ethanol and ammonia concentration, liquid/solid ratios, pre-leaching time before MAE and material size for the MAE procedure were investigated to optimize the efficiency of the extraction. Under appropriate MAE conditions, such as extraction times of 4-5min, ethanol concentrations of 50-60% (v/v), ammonia concentrations of 1-2% (v/v) and liquid/solid ratios of 10:1(ml/g), the recovery of GA from licorice root with MAE was equivalent with conventional extraction methods. Those methods include extraction at room temperature (ERT), the traditional Soxhlet extraction, heat reflux extraction and ultrasonic extraction. Due to the considerable savings in time and solvent, MAE was more effective than the conventional methods. This novel method is suitable for fast extraction of GA from licorice root.     

Optimization of extraction and purification of glucoamylase produced by Aspergillus awamori in solid-state fermentation

Various parameters such as solvent selection, concentration, soaking time, and temperature were tested in a single bioreactor in order to determine optimum extraction conditions of glucoamylase, when produced simultaneously with protease by Aspergillus awamari nakazawa MTCC 6652. Optimum conditions were achieved in a 10% glycerol solution soaked for 2 h at 40°C, followed by concentration of extracted glucoamylase (9,157 U/gds) by acetone precipitation (1:2, v/v), which yielded 51.9% recovery. Ion exchange chromatography and gel filtration showed specific activities of 270.5 and 337.5 U/mg, respectively, while SDS-PAGE and zymogram analysis of glucoamylase indicated the presence of three starch-hydrolyzing isoforms with molecular weights of approximately 109.6, 87.1, and 59.4 kDa, respectively     

The effects of buck teasing on synchronization of estrus in goats after intravulvo-submucosal administration of cloprostenol

A study was conducted on 35 East African shorthorned female goats to determine if a combination of buck teasing and low doses of a prostaglandin (PGF(2) alpha) analogue, cloprostenol, given intravulvo-submucosally (i.v.s.m.) would be suitable for synchronization of estrus. Goats were allotted, with the onset of estrus, to seven groups (n = 5 goats per group). Five of the seven groups received varying doses of cloprostenol: Group 1 (125 mug cloprostenol i.m. per goat); Group 2 (62.5 mug cloprostenol i.v.s.m. per goat); Group 3 (62.5 mug cloprostenol i.v.s.m. per goat plus buck teasing); Group 4 (31.25 mug cloprostenol i.v.s.m. per goat); Group 5 (31.25 mug cloprostenol i.v.s.m. per goat plus buck teasing); Group 6 (buck teasing); Group 7, (2 ml physiological saline i.v.s.m. per goat, control group). Plasma progesterone concentration was measured on day of treatment and for 6 d thereafter. All goats in groups 1, 2, 3 and 5 exhibited estrus within 68 h. Thus, the number of goats receiving low doses of PG-cloprostenol intravulvo-submucosally observed in estrus increased (P < 0.05) with exposure to bucks. Exhibition of behavioral signs of estrus was maximal between 2 and 20 h after onset of signs of estrus. The exposure of females to males prior to intrauterine penetration was an advantage because copious mucus eased penetration.     

Hybridoma cell lines secreting monoclonal antibodies against foot-and-mouth disease virus (FMDV). II. Cloning conditions

Three methods of cloning hybridoma cells--picking colonies from the masterplate, limit dilution cloning, and cloning in semi-solid medium over macrophage (m phi) feeder layers--were compared. Cloning in semi-solid medium was found to be the most efficient and reliable, especially with our relatively slow growing anti-foot-and-mouth disease virus (FMDV) antibody secreting hybridoma cells. The optimum culture dish for this cloning was the 6-well (6W) dish (well diameter 1.5 cm), while the optimum cloning procedure was 10 to 10(2) hybridoma cells in 1% (w/v) methylcellulose in Dulbecco's minimal essential medium (DMEM) supplemented with 50% (v/v) conditioned medium, 10% (v/v) foetal or donor calf serum and 2 mM glutamine, over a 24-48 h old culture of syngeneic m phi in each well of the 6W plate. This could, however, produce problems in that colony formation was sometimes 'loose', and confident picking of individual colonies was impaired. Such a problem could be avoided by using a solid agar interface of 1% (w/v) agar Noble in DMEM or RPMI 1640 medium between the feeder cells and the hybridoma cells in semi-solid medium.