黄金分割法结合动态设计优选盐黄柏提取工艺

采用黄金分割法结合正交设计以及动态设计对盐黄柏提取工艺参数进行优选。以HLPC法测定盐酸小檗碱、木兰花碱、盐酸小檗红碱、黄柏碱、巴马汀、药根碱得率,黄金分割法初选正交设计时间范围,单因素设计优选溶剂比,基于黄金分割法及单因素设计采用正交设计优选提取溶剂比、提取次数、提取时间。以正交设计优选方案为基础,用动态设计进一步优化盐黄柏的提取工艺参数。结果表明,盐黄柏的优选提取工艺为:提取次数:2次;容积比:1∶25;超声时间:第一次超声60 min,第二次超声20 min;盐酸小檗碱、木兰花碱、盐酸小檗红碱、黄柏碱、巴马汀、药根碱提取率分别为6,51、1,48、0,67、0,62、5,01、73,22 mg/g。因此,黄金分割法结合正交设计、动态设计对盐黄柏提取工艺参数进行优选基本可行,可作为中药提取工艺的筛选方法之一。     

一测多评法测定不同产地黄柏炭中4种生物碱含量

为建立同时测定不同产地黄柏炭中4种生物碱成分(黄柏碱、木兰花碱、小檗红碱、小檗碱)的一测多评法。实验以小檗碱为内参物,基于HPLC法建立小檗碱与其它成分的相对校正因子,并用校正因子计算4种成分的含量;用外标法和一测多评法同时测定30批黄柏炭样品的含量。结果表明30批黄柏炭样品的外标法与一测多评法结果之间无明显差异,说明了一测多评法的准确性和可行性。因此在缺乏对照品的情况下,以小檗碱为内参物同时测定黄柏碱、木兰花碱、小檗红碱的含量是可行的。该方法准确性和重复性良好,可用于黄柏炭的质量控制。     

一种提取中草药化学成分的方法——超声提取法

本文阐述利用超声技术提取中草药材中化学成分的新方法。该方法能提高提出率 ,且不改变化学成分的结构 ,也大大地加速了提取速度。用提取川黄柏中的小檗碱做实验 ,观察了川黄柏皮在超声作用下细胞结构的变化 ,还用现代仪器检测了小檗碱的结构 ,为超声提取的机理研究提供了有力的依据。     

HPLC法同时测定黄柏中盐酸药根碱、盐酸巴马汀及盐酸小檗碱的含量

分离并测定黄柏中盐酸药根碱、盐酸巴马汀、盐酸小檗碱的含量.方法:Shim-Pack型C18柱,0.5% w/v SDS-Britton-Robinson缓冲液 pH2.5 -乙腈-三乙胺 0.5∶56∶34∶0.5 为流动相,检测波长:345nm;柱温:40℃;流速1mL·min-1.盐酸药根碱回归方程为Y=38315164.0X-1897.7,r=0.9991 n=6 ,线性范围0.05～0.30μg,平均回收率为95.90%,RSD=3.10%;盐酸巴马汀回归方程为Y=1982872.0X-2471.1,r=0.9998 n=6 ,线性范围0.09～0.54μg,平均回收率100.2%,RSD=2.01%;盐酸小檗碱回归方程为Y=9450164.2X-3456.9,r=0.9999 n=6 ,线性范围0.50～3.00μg,平均回收率103.1%,RSD=1.12%;结果表明:在以上条件下,盐酸药根碱、盐酸巴马汀、盐酸小檗碱成分可以完全分离,并测定了黄柏药材含量.本法可用于黄柏药材、饮片、含黄柏中成药的质量控制.     

酸酯碱成分结合指纹图谱优选盐黄柏炮制工艺

通过测定三大类成分含量,结合盐黄柏指纹图谱优选盐黄柏炮制工艺。采用HLPC-DAD法测定盐黄柏中6种生物碱以及2种酸类、1种内酯,共9种,3大类成分含量。以AHP法确定各指标成分权重,综合评分结合指纹图谱,通过单因素及正交设计实验,定性定量的分析炮制温度、炮制时间、炒药量对盐黄柏炮制的影响,优选盐黄柏炮制工艺。优选炒制工艺为:180℃,20 min,300 g药材(占容器体积1/4)。三大类成分结合指纹图谱优选的盐黄柏炮制工艺稳定、可行;此外,该方法可用于类似工艺的筛选。     

高乌头活性成分提取工艺的优化北大核心CSCD

为开发并优化高乌头(Aconitum sinomontanum)中有效成分高乌甲素及冉乌头碱的提取工艺,采用高效液相色谱法测定高乌甲素及冉乌头碱含量,以二者综合提取率为考察指标,采用正交实验设计优选最佳提取工艺。结果表明,优选工艺为药材粉末加10倍量的80%甲醇及2%药材量的浓盐酸,在75°C下回流提取6小时。经验证,优选所得的工艺稳定可行,可作为高乌头活性成分提取的新工艺。     

高乌头活性成分提取工艺的优化

为开发并优化高乌头(Aconitum sinomontanum)中有效成分高乌甲素及冉乌头碱的提取工艺,采用高效液相色谱法测定高乌甲素及冉乌头碱含量,以二者综合提取率为考察指标,采用正交实验设计优选最佳提取工艺。结果表明,优选工艺为药材粉末加10倍量的80%甲醇及2%药材量的浓盐酸,在75°C下回流提取6小时。经验证,优选所得的工艺稳定可行,可作为高乌头活性成分提取的新工艺。     

正交实验法优化黄柏中黄连素的酶碱法提取工艺

为了提高黄柏中黄连素的提取得率与效率,采用传统方法碱提法与酶解辅助提取相结合来优化黄柏中黄连素的提取工艺。以黄连素浸提率为评价指标,考察纤维素酶浓度、酶解时间、料液比及浸提时间对提取效率的影响。在单因素实验的基础上,通过正交实验分析各因素的作用大小以及它们之间的交互作用。优选出黄柏中黄连素的酶碱法提取最佳工艺条件为:纤维素酶浓度15‰、酶解时间1 h、料液比1∶7、浸提时间7 h,在该条件下黄连素的浸提率可达8.99%。该提取工艺浸提率高,成本低,稳定性好,可用于生产上黄柏中黄连素的提取。     

黄柏再生皮小蘖碱含量研究

黄柏(Phellodendron amurense)树皮为常用清热解毒中药材。黄柏剥皮再生新皮试验的成功,为扩大黄柏药源提供了新途径。为了大面积推广,作者对黄柏再生皮(二年生、三年生)主要药用成分进行了定性定量检查,检测结果表明,再生皮和原生皮与商品黄柏的成分和含量基本一致,作药材是可行的。     

王不留行刺桐碱提取工艺的比较研究

本实验采用高效液相色谱(HPLC)法检测王不留行刺桐碱含量为评价指标,分别采用单因素试验法和正交试验法对水浴回流提取和超声提取对王不留行刺桐碱的提取工艺进行比较。结果表明:超声提取王不留行刺桐碱的最佳工艺参数为液固比10∶1 mL/g,60%乙醇在350 W功率、80℃下提取40 min,在该工艺条件下王不留行刺桐碱含量为0.395 mg/g;水浴回流提取王不留行刺桐碱的最佳工艺参数为乙醇浓度75%,提取温度95℃,提取时间2 h,液固比12∶1 mL/g时,王不留行刺桐碱含量为0.345 mg/g。超声提取王不留行刺桐碱的提取效率、稳定性和重复性都优于水浴回流提取法。     

Bioreactor operating strategy inThalictrum rugosum plant cell culture for the production of berberine

Optimal substrate feeding strategy in bioreactor operation was investigated to increase the production of secondary metabolite in a high density culture of plant cell. It was accomplished by the previously proposed structured kinetic model that describes the cell growth and synthesis of the secondary metabolite, berberine, in a batch suspension culture ofThalictrum rugosum. Four types of operation strategies for sugar feeding intoT. rugosum culture were proposed based on the model, which were the periodic fedbatch operations to maintain the cell activity, the cell viability, and the specific production rate, and the perfusion operation to maintain the specific production rate. From the simulation results of these strategies, it could be found that the periodic fed-batch operation and the perfusion operation could achieve the higher volumetric production of berberine (mg berberine/L) and specific production yield (mg berberine/g dry cell weight) than those of batch cultures. Although the highest productivity (mg berberine/day) of berberine could be achieved by the periodic fed-batch operation to maintain the cell activity compared with the other strategies in the periodic fed-batch operations, the specific production yield was low due to the higher maximum dry cell weight than other cases. The periodic fed-batch operation to maintain cell viability resulted in the highest volumetric production of berberine and specific production yield compared with the other strategies. In the cases of maintaining the specific production rate, the per-formance of the periodic fed-batch operation was better than that of the perfusion operation in the respect of the volumetric production and productivity of berberine. In order to increase the volumetric production of berberine and to get the highest specific production yield, the periodic fed-batch operation to maintain cell viability could be chosen as the optimal operating strategy in high density, culture ofT. rugosum plant cell.     

双频超声强化提取黄柏中小檗碱的研究

设计25kHz和40kHz的双频超声强化装置,以黄柏为对象提取小檗碱,结果表明：25kHz与40kHz双频超声强化5min,提取率为64．1％,25kHz及40kHz超声30min,提取率分别为52．9％和45．2％,双频超声强化的提取率显著高于单频超声强化的提取率。研究还表明,双频超声强化可以降低提取温度,缩短提取时间,为热敏性药物的提取提供了新的强化方法。     

Berberine inhibits TPA-induced MMP-9 and IL-6 expression in normal human keratinocytes

Berberine is a plant ingredient that has anti-inflammatory and anti-oxidative effects. Matrix metalloproteinase-9 (MMP-9) and interleukin-6 (IL-6) are known to be highly induced by ultraviolet (UV) light and may play important roles in UV-induced skin inflammation and the skin aging process. In this study, we investigated the effects of berberine on MMP-9 and IL-6 expression in normal human keratinocytes (NHK). Our results demonstrated that berberine dose-dependently inhibited basal and TPA-induced expression and activity of MMP-9, and also suppressed TPA-induced IL-6 expression. Berberine prevented TPA-induced ERK activation and AP-1 DNA binding activity. Therefore, berberine may be used as an effective ingredient for anti-skin aging products, which can prevent skin inflammation and the degradation of extracellular matrix proteins, including collagen, by MMPs.     

Berberine-induced pigment dispersion in Bufo melanostictus melanophores by stimulation of beta-2 adrenergic receptors

Abstract

Reduced production of melanin by decreased or the absence of melanocytes leads to various hypopigmentation disorders, and the development of melanogenetic agents for photoprotection and hypopigmentation disorders is one of the top priority areas of research. Hence, the present study was carried out to elucidate the ability of berberine, a principal active ingredient present in the roots of the herb Berberis vulgaris to stimulate pigment dispersion in the isolated skin melanophores of the toad Bufo melanostictus. In the present study, mean melanophore size index of the isolated skin melanophores of B. melanostictus was assayed after treating with various concentrations of berberine. A marked melanin dispersion response leading to skin darkening was observed in the isolated melanophores of toad in response to berberine, which was found to be mediated through beta-2 adrenergic receptors. The physiologically significant dose-related melanin dispersion effects of berberine per se were found to be completely abolished by propranolol, which is a specific beta-2 adrenergic receptor blocker. These per se melanin dispersal effects were also found to be markedly potentiated by isoprenaline, which is a specific beta-adrenoceptor agonist. The results indicate that berberine causes a tremendous, dose-dependent, physiologically significant pigment dispersing in the isolated skin melanophores of B. melanostictus.     

Effect of oxygen supply on berberine production in cell suspension cultures and immobilized cells ofThalictrum minus

The ample supply of O₂ proved to be of great importance for berberine production in cell suspension culture ofThalictrum minus, as the specific O₂ consumption rate of berberine-producing cells was twice as high as that of non-producing cells. Furthermore, berberine yield increased with increases in the volumetric O₂ transfer coefficient (K_La). Estimation of the optimum conditions of oxygen supply in suspension cultures and immobilized cells according to a known theoretical model assuming O₂ uptake by cells to be a zero-order reaction was in good agreement with the experimental data. The O₂ supply to immobilized cells could be improved by reducing the cell density and radius of the bead.     

产小檗碱诱变菌株培养条件的优化

S-NU-3-2菌株是对源自药用植物黄檗的内生真菌S6进行诱变获得的小檗碱产量比出发菌株有明显提高的突变株,本研究对其培养条件进行了优化,以期进一步提高其产量,为开发利用真菌发酵生产植物活性成分的新途径奠定基础。以菌丝生长和小檗碱产量为指标,筛选出了适宜该菌株发酵的基本培养基、碳源、氮源、光照和培养温度,并经进一步的正交试验优化,得出该高产菌株的最佳培养条件为豆芽汁基本培养基含蔗糖3%,酵母膏0.2%,pH7.0,温度26℃,全光照培养。与初始培养条件相比,在优化后的条件下,该菌株的小檗碱得率提高了47.2%,菌体生物量提高了24%。     

Microbiota transplantation reveals beneficial impact of berberine on hepatotoxicity by improving gut homeostasis

Berberine has been shown to reduce acute liver injury although the underlying mechanism is not fully understood. Because of the anatomic connection, the liver is constantly exposed to gut-derived bacterial products and metabolites. In this study, we showed that berberine has beneficial effects on both hepatotoxicity and intestinal damage in a rat model of chronic or acute liver injury. Microbiota transplantation from the rats with chronic hepatotoxicity could aggravate acute hepatotoxicity in mice treated with diethylnitrosamine (DEN). In rat models with gut homeostasis disruption induced by penicillin or dextran sulfate sodium (DSS), their fecal microbiota could also cause an enhanced hepatotoxicity of recipient mice. When treated with berberine, the DSS-induced enteric dysbacteriosis could be mitigated and their fecal bacteria were able to reduce acute hepatotoxicity in recipient mice. This study indicates that berberine could improve intestinal dysbacteriosis, which reduces the hepatotoxicity caused by pathological or pharmacological intervention. Fecal microbiota transplantation might be a useful method to directly explore homeostatic alteration in gut microbiota.     

Cultivation of Thalictrum rugosum cell suspension in an improved airlift bioreactor: stimulatory effect of carbon dioxide and ethylene on alkaloid production

Airlift bioreactor operations have been studied for the growth-associated production of secondary metabolites from plant cell suspension cultures. The model system used in this work was Thalictrum rugosum producing berberine, an isoquinoline alkaloid. The airlift system was well suited for growth of Thalictrum cell suspension cultures unless the cell density was high. At high cell density, the airlift system with a draught tube was not adequate due to large aggregates clogging the recirculation paths. This was overcome by use of a cell scraper in the reactor. For berberine production, gas-stripping also played a significant role and it was discovered that CO(2) and ethylene were important for product formation. By supplying a mixture of CO(2) and ethylene into the airlift system, the specific berberine content was increased twofold. It is evident that continuous gas sparging was harmful for the production of berberine without supplementation with other gases.