正交试验优化黑果枸杞果实中花青素的提取工艺

黑果枸杞(Lycium ruthenicum)是西北干旱区特有的小灌木,具有重要的生态和经济价值,而果实中花青素含量多寡是评判其经济价值的重要指标。为找到简单易行的花青素提取技术,本试验采用pH=30的盐酸-乙醇提取法,分别对不同提取溶剂浓度、物料比、提取时间、提取温度以及是否脱脂条件下的花青素提取率进行了研究,结果表明:在黑果枸杞不脱脂的情况下,用乙醇提取花青素的最优条件为pH=30的80%盐酸-乙醇、物料比为1:15、提取时间为5 h、提取温度为40℃,花青素的提取率达到0978%。该试验结果为实验室及产业化提取花黑果枸杞青素提供了较为准确的参数。     

响应面法对黑果枸杞中花青素测定条件的优化

以黑果枸杞为原料,采用pH示差法,通过响应面法优化花青素含量的测定条件。通过单因素试验和Box-Behnken Design试验研究乙醇体积分数、pH 1.0缓冲液平衡时间、pH 4.5缓冲液平衡时间等3个变量对黑果枸杞中花青素响应值的影响程度。根据响应面的最优化分析,得出最佳条件是：用80%酸性乙醇溶液,超声萃取30min,取花青素提取液在pH 1.0和pH 4.5的缓冲液中各平衡80min,在最适波长处测定。通过试验验证,采用pH示差法,通过响应面法优化得到的黑果枸杞中花青素含量的测定条件,可靠准确、具有实用价值。     

柴达木野生黑果枸杞营养成分分析

对柴达木野生黑果枸杞果实中的主要营养成分进行了分析,结果表明,黑果枸杞鲜果含水质量分数85.03%,其干果蛋白质量分数10.61%,脂肪6.66%,多糖4.28%,总黄酮质量分数4.29%,原花青素3.42%.果实含有γ-VE及δ-VE,每百克分别为0.0075和0.016 mg,从黑果枸杞籽中提取的构祀籽油维生素E含...     

黑果枸杞提取物活性的研究进展

黑果枸杞是一种具有良好保健功效的药食两用植物,其富含丰富的营养物质,具有多种生物活性。本文综述了黑果枸杞中花青素、多糖和甜菜碱等活性成分的性质及其保健功效的研究进展,为黑果枸杞的深入研究及进一步的开发应用提供科学依据。     

乳钙泡腾片固体饮料的研制

[目的]开发一种乳钙的便携式健康饮料。[方法]以乳钙、酸奶粉为主要原料,研发乳钙泡腾片固体饮料,从成型性、泡腾性、溶解性和感官等综合评价确定最优配方。[结果]乳钙泡腾片固体饮料的最佳配方为:乳钙粉40%、酸奶粉15%、酒石酸7%、无水碳酸氢钠5%、PVP2%、甘露醇27%、阿斯巴甜0.7%、酸奶香精0.3%、PEG6000 3%,各项评价指标较好。[结论]该配方制得的乳钙泡腾片固体饮料质量稳定,符合固体饮料卫生标准。     

黑果枸杞花青素结构差异对其稳定性及细胞抗氧化活性的影响

本文研究了黑果枸杞花青素化合物结构对其稳定性和细胞抗氧化活性的影响。通过半制备型HPLC分离纯化得到了4种黑果枸杞花青素化合物并鉴定结构,利用比色法评价了化合物的稳定性、细胞毒实验以及H₂O₂诱导的SH-SY5Y细胞损伤模型评价了细胞抗氧化活性,并讨论了化学结构对其影响。结果表明反式结构比顺式结构在中性和弱碱性条件下有更好的稳定性,反式结构、糖基的增加以及母核B环上的甲氧基取代均可以通过缩短母核与糖链的分子距离从而提高稳定性,糖基的增加能显著地降低损伤细胞中的乳酸脱氢酶水平,且反式结构较顺式结构更有活性优势。综上所述,黑果枸杞花青素的结构影响稳定性和细胞抗氧化活性,但糖基的增加和酰基的反式异构对二者有更积极的意义。     

超声辅助提取野生与人工栽培黑果枸杞花色苷含量研究

黑果枸杞含有的花色苷类成分对人类健康有着重要意义。为探究野生与人工种植黑果枸杞花色苷含量差异,本实验利用超声辅助回流提取,通过单因素实验和Box-Benhnken实验设计优化提取条件,得到最佳提取条件为:提取时间25.62 min、提取温度48.67℃、乙醇浓度84.35%、料液比1∶20 g·m L-1。在此条件下测得野生、人工种植黑果枸杞花色苷平均含量分别为6.141 mg/g、16.014 mg/g。本实验结果为黑果枸杞种植及利用提供了重要依据。     

柴达木地区野生黑果枸杞种群遗传多样性的AFLP分析

采用扩增片段长度多态性(AFLP)分子标记技术对青海省柴达木地区5个野生黑果枸杞(Lycium ruthenicum)种群的120份样品的遗传多样性进行分析。结果表明: 柴达木地区野生黑果枸杞具有很高的遗传多样性, 9对选扩引物共得到1691条清晰条带, 其中多态性条带1678条, 多态性变异率为99.23%, 种群间的有效等位基因数为1.4712, Nei’s基因多样性为0.3245, Shannon信息指数为0.4367。分子方差分析(AMOVA)结果表明: 柴达木地区5个黑果枸杞种群的遗传变异主要存在于种群内部(92%), 种群间的遗传分化较小(8%, 遗传分化系数0.08)。黑果枸杞种群间的遗传相似系数介于0.9709-0.9922之间, 平均值为0.9835。种群间的聚类及Mantel检验(γ = 0.3368, p = 0.8064)均表明柴达木地区黑果枸杞种群地理距离与遗传距离之间的相关性不明显; 黑果枸杞个体间的聚类表明同一种群的个体不能完全聚在一起。对同一种源的遗传多样性分析发现, 诺木洪奥斯勒草场的种源内部的遗传变异更为丰富, 这或许可以推断诺木洪可能为柴达木地区野生黑果枸杞种质资源的中心产区。     

黑果枸杞LrTTG1基因的克隆及表达分析

以黑果枸杞为材料,利用RT PCR和RACE技术克隆了花青素合成相关基因LrTTG1（GenBank登录号为MH633481）。序列分析表明,LrTTG1基因cDNA全长1 453 bp,包含1 029 bp开放阅读框,编码342个氨基酸,含有5个WD40重复基序。同源比对结果表明,LrTTG1与茄子SmTTG1的氨基酸序列相似性较高,达到83.73%。qRT PCR分析显示,LrTTG1基因在茎、叶、花、青果、紫果和黑果中均有表达,且在青果中的表达水平（最高）约为黑果（最低）的4倍;紫外胁迫下LrTTG1基因的表达随胁迫时间的延长呈先降低后升高的变化趋势。花青素含量分析表明,黑果的花青素含量最高（11.3 mg/g）,分别约为紫果（ 1.2 mg/g）和青果（0.53 mg/g）含量的9.4倍和21.3倍。研究表明,随着黑果枸杞果实的发育,LrTTG1基因的表达量呈现下降趋势,而花青素的含量则呈上升趋势,两者呈负相关关系;推测LrTTG1基因在黑果枸杞花青素合成中可能具有重要的调节作用。     

青藏高原黑果枸杞花青素对人肝癌细胞增殖和自噬的影响

探究黑果枸杞花青素在体外对人肝癌HepG2细胞增殖和自噬的影响。利用CCK-8法测定细胞活力,EdU和细胞划痕试验检测细胞增殖和迁移效果,RT-PCR和Western blot检测增殖和自噬相关基因的mRNA和蛋白表达。结果显示,黑果枸杞花青素可有效抑制人肝癌HepG2细胞的增殖和迁移;上调增殖因子(LATS1、LATS2和MOB1)和自噬因子(Beclin-1、LC3-Ⅱ和AMPK),并下调增殖因子YAP的mRNA水平;下调自噬因子p-mTOR和细胞周期因子CDK4,并上调自噬因子p-AMPK和LC3-Ⅱ的蛋白表达。由此推测黑果枸杞花青素可在体外抑制人肝癌HepG2细胞增殖和迁移,并促进人肝癌HepG2细胞发生自噬。     

黑果枸杞化学成分研究进展

黑果枸杞是一种独特的药食两用植物,具有抗氧化、抗动脉粥样硬化、增强免疫力、抗衰老、降血脂等药理作用。综述近年来 黑果枸杞的化学成分、生物活性、提取方法和质量分析等方面的研究进展,为黑果枸杞药效成分的进一步开发和利用提供参考。     

Development and evaluation of acid-buffering bioadhesive vaginal tablet for mixed vaginal infections

An acid-buffering bioadhesive vaginal tablet was developed for the treatment of genitourinary tract infections. From the bioadhesion experiment and release studies it was found that polycarbophil and sodium carboxymethylcellulose is a good combination for an acid-buffering bioadhesive vaginal tablet. Sodium monocitrate was used as a buffering agent to provide acidic pH (4.4), which is an attribute of a healthy vagina. The effervescent mixture (citric acid and sodium bicarbonate) along with a superdisintegrant (Ac-Di-sol) was used to enhance the swellability of the bioadhesive tablet. The drugs clotrimazole (antifungal) and metronidazole (antiprotozoal as well as an antibacterial) were used in the formulation along with Lactobacillus acidophilus spores to treat mixed vaginal infections. From the ex vivo retention study it was found that the bioadhesive polymers hold the tablet for more than 24 hours inside the vaginal tube. The hardness of the acid-buffering bioadhesive vaginal tablet was optimized, at 4 to 5 kg hardness the swelling was found to be good and the cumulative release profile of the developed tablet was matched with a marketed conventional tablet (Infa-V). The in vitro spreadability of the swelled tablet was comparable to the marketed gel. In the in vitro antimicrobial study it was found that the acid-buffering bioadhesive tablet produces better antimicrobial action than marketed intravaginal drug delivery systems (Infa-V, Candid-V and Canesten 1).     

红色诺卡菌固体培养基配方的优化

应用响应面优化设计法优化固体培养基配方,增大红色诺卡菌的固体培养细胞生物量。首先用Plackett-Burman法从现有培养基组分中找到影响红色诺卡菌细胞生物量的关键因素,再通过最陡爬坡法确定细胞生物量最大的配方,用作中心组合设计(Central Composite Design, CCD)实验的基础起始值,拟合数学模型方程,最后找到最优组分的组合。优化的配方转移至企业实施放大实验,对结果进行验证和比较。试验结果表明,培养基各组分中影响红色诺卡菌细胞生物量的关键因素为蛋白胨、NaCl、牛肉膏;最优固体培养基配方:蛋白胨42 g/L、牛肉膏8 g/L、NaCl 1.2 g/L、甘油10 mL/L、Na_2HPO_4·12H_2O 0.3 g/L、琼脂20 g/L。在细胞生物量方面最优固体培养基配方比原配方高104%。响应面优化设计可用于提高红色诺卡菌细胞生物量固体培养基的优化,也为红色诺卡菌培养条件、液体发酵的优化研究提供参考。     

Compressional Behavior of a Mixture of Granules Containing High Load of Phyllanthus niruri Spray-Dried Extract and Granules of Adjuvants: Comparison between Eccentric and Rotary Tablet Machines

The purpose of this paper was to evaluate the compressional behavior of granules containing high load of a Phyllanthus niruri spray-dried extract in eccentric (ETM) and rotary (RTM) tablet presses. Tablets were constituted by spray-dried extract granules (SDEG, 92%), excipient granules (EXCG, 7.92%), and magnesium stearate (0.08%). SDEG was obtained by dry granulation and EXCG, composed of microcrystalline cellulose (62.9%) and sodium starch glycolate (37.1%), by wet granulation. Particle size distribution was fixed between 0.250 and 0.850 mm. Tablets did not evidence any mechanical failures, such as lamination or capping, or anomalous weight variation in either tablet machine types. Upper and lower tablet surface photomicrographs from ETM and RTM tablets showed differences in porosity and texture. Different RTM speeds suggested the visco-plastic behavior of the formulation, since, by slowing down rotation speeds, the tensile strength of the tablets increased significantly, but the porosity and disintegration time were not affected. Tablets produced in RTM showed lower friability and porosity than ETM tablets, which did not reflect on higher tensile strength. The EXCG distribution at upper and lower surfaces from ETM and RTM tablets was quantified by image analysis and evaluated through statistical methods. Spray-dried extract release was not influenced by the type of equipment or operational conditions to which the compacts were submitted. Construction and operation differences between both tablet presses influenced the final product, since tablets with similar tensile strength, made by distinct tablet machines, exhibited different quality parameters.     

黑果枸杞汁对大鼠酒精性肝损伤的保护作用*

目的: 研究黑果枸杞汁对大鼠酒精性肝损伤的保护作用,探讨其中Toll样受体4(TLR4)/p38 丝裂原活化蛋白激酶(p38 MAPK)信号通路的调节机制。方法: 60只雄性SD大鼠随机分为空白对照组(C)、模型组(M)、低剂量黑果枸杞汁组(LLM)、中剂量黑果枸杞汁组(MLM)、高剂量黑果枸杞汁组(HLM),每组12只。M、LLM、MLM和HLM组每天以20 ml/kg(8 g/(kg·d))剂量分2次灌胃400 g/L酒精,C组于相同时间点灌胃等体积蒸馏水。每日首次酒精灌胃前4 h,黑果枸杞汁各剂量组分别以2.4、4.8、9.6 ml/(kg·d)进行灌胃,其他组于相同时间点灌胃等体积蒸馏水。实验周期4周,末次实验结束后24 h处死大鼠,取血液和肝脏,计算肝脏指数,HE染色观察肝脏组织形态,比色法检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性,免疫组化法检测肝脏TLR4、p38 MAPK及磷酸化p38 MAPK(p-p38 MAPK)蛋白质表达,酶联免疫吸附试验检测肝脏肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)、白细胞介素-18(IL-18)水平。结果: 与C组比较,M组成功建立酒精性肝损伤模型。与M组比较,黑果枸杞汁各剂量组的相关指标出现改善,其中HLM组肝脏组织形态改善最为显著,其肝脏系数、血清ALT和AST、肝脏TLR4蛋白质表达、p-p38 MAPK/p38 MAPK比值、TNF-α、IL-1β和IL-18水平均显著降低(P＜0.05或P＜0.01),肝脏IL-10水平显著升高(P＜0.01)。黑果枸杞汁各剂量组组间比较,肝脏系数、血清AST、肝脏TLR4蛋白质表达、p-p38 MAPK/p38 MAPK比值、TNF-α和IL-18水平,HLM组较LLM组均显著降低(P＜0.05或P＜0.01);肝脏IL-10水平,HLM组较LLM和MLM组均显著升高(P＜0.05或P＜0.01);其余主要指标组间比较均无统计学差异(P＞0.05)。结论: 黑果枸杞汁可以通过调控TLR4/p38 MAPK信号通路,改善炎症应激,缓解大鼠酒精性肝损伤,高剂量组效果优于其他剂量组。     

Formulation and Optimization of Orodispersible Tablets of Diazepam

Diazepam is one of the most prescribed benzodiazepines. The purpose of the present research was to optimize the formulation of orodispersible tablets of diazepam. Orodispersible tablets of diazepam were prepared using different types of superdisintegrants (Ac-Di-Sol, sodium starch glycolate, and crospovidone (CP)) and different types of subliming agents (camphor and ammonium bicarbonate (AB)) at different concentrations and two methods of tablets preparations (wet granulation and direct compression methods). The formulations were evaluated for flow properties, wetting time, hardness, friability, content uniformity, in vivo disintegration time (DT), release profiles, and buccal absorption tests. All formulations showed satisfactory mechanical strength except formula F5 which contains camphor and formula F9 which is prepared by direct compression method. The results revealed that the tablets containing CP as a superdisintegrant have good dissolution profile with shortest DT. The optimized formula F7 is prepared using 10% CP as a superdisintegrant and 20% AB as a subliming agent by wet granulation method which shows the shortest DT and good dissolution profile with acceptable stability. This study helps in revealing the effect of formulation processing variables on tablet properties. It can be concluded that the orodispersible tablets of diazepam with better biopharmaceutical properties than conventional tablets could be obtained using formula F7.     

表面活性剂对林可霉素发酵生产的影响

在林可霉素发酵过程中,当向培养基中加入表面活性剂十二烷基磺酸钠(SDS)、吐温80(Tween 80)和曲拉通(Triton X-100)时,林可霉素的产量受到较大影响。本研究应用响应面设计法(Response surface design)对表面活性剂的配比进行了优化,得到的优化配比为:十二烷基磺酸钠为31.13 mg/100 mL,吐温80为51.97 mg/100 mL,曲拉通为16.9 mg/100 mL。将该优化配比应用于林可霉素发酵,产量提高了36.67%。     

Aqueous two-phase assisted by ultrasound for the extraction of anthocyanins from Lycium ruthenicum Murr.

In this study, an efficient ultrasound-assisted aqueous two-phase extraction method was used for the extraction of anthocyanins from Lycium ruthenicum Murr. An ethanol/ammonium sulfate system was chosen for the aqueous two-phase system due to its fine partitioning and recycling behaviors. Single-factor experiments were conducted to determine the optimized composition of the system, and the response surface methodology was used for the further optimization of the ultrasound-assisted aqueous two-phase extraction. The optimal conditions were as follows: a salt concentration of 20%, an ethanol concentration of 25%, an extraction time of 33.7?min, an extraction temperature of 25°C, a liquid/solid ratio of 50:1 w/w, pH value of 3.98, and an ultrasound power of 600?W. Under the above conditions, the yields of anthocyanins reached 4.71?mg/g dry sample. For the further purification, D-101 resin was used, and the purity of anthocyanins reached 25.3%. In conclusion, ultrasound-assisted aqueous two-phase extraction was an efficient, ecofriendly, and economical method, and it may be a promising technique for extracting bioactive components from plants.     

Design and Optimization of Sustained-Release Divalproex Sodium Tablets with Response Surface Methodology

Response surface methodology is defined as a collection of mathematical and statistical methods that are used to develop, improve, or optimize a product or process. In the present study, a statistical design (Mixture Design) was employed for formulation and optimization of a sustained-release hydrophilic divalproex sodium matrix tablet. Different excipients were used to improve the drug’s poor flowability. The hardness of the prepared tablets and also their release pattern were tested. The formulation design was carried out employing mixture design using four excipients in three levels. The Carr’s index of formulations and tensile strength were determined and analyzed using Minitab software. The suitable formulations regarding flowability and tablet tensile strength were selected by this software for subsequent drug release studies. The dissolution tests were carried out in acidic and basic phases which were previously proved to be biomimetic. Samples were analyzed using HPLC, and release data were compared to Depakine® (sustained-release divalproex from Sanofi). Release kinetics was also determined for selected formulations. Selected formulations were subjected to dissolution test and showed similar dissolution profiles with Depakine® based on difference and similarity factor calculations. The software selected an optimized formulation which had a slightly different release pattern in vitro compared to innovator but of nearly zero-order kinetics. It can be concluded that application of Mixture Design is a shortcut method to design suitable formulations of sustained-release divalproex sodium containing hydrophilic matrix tablets by direct compression method.     

Artificial neural networks in the modeling and optimization of aspirin extended release tablets with eudragit L 100 as matrix substance

The purpose of the present study was to model the effects of the concentration of Eudragit L 100 and compression pressure as the most important process and formulation variables on the in vitro release profile of aspirin from matrix tables formulated with Eudragit L 100 as matrix substance and to optimize the formulation by artificial neural network. As model formulations, 10 kinds of aspirin matrix tablets were prepared. The amount of Eudragit L 100 and the compression pressure were selected as causal factors. In vitro dissolution time profiles at 4 different sampling times were chosen as responses. A set of release parameters and causal factors were used as tutorial data for the generalized regression neural, network (GRNN) and analyzed using a computer. Observed results of drug release studies indicate that drug release rates vary widely between investigated formulations, with a range of 5 hours to more than 10 hours to complete dissolution. The GRNN model was optimized. The root mean square value for the trained network was 1.12%, which indicated that the optimal GRNN model was reached. Applying the generalized distance function method, the optimal tablet formulation predicted by GRNN was with 5% of Eudragit L 100 and tablet hardness 60N. Calculated difference (f ₁ 2.465) and similarity (f ₂ 85.61) factors indicate that there is no difference between predicted and experimentally observed drug release profiles for the optimal formulation. This work illustrates the potential for an artificial neural network, GRNN, to assist in development of extended release dosage forms.