基于正交实验法优化从栀子黄色素中制备藏红花酸的工艺

基于正交实验法,优化从栀子黄色素中提取制备藏红花酸的碱水解工艺,以期可以简单高效地获得高纯度藏红花酸。建立高效液相色谱法(HPLC)测定藏红花酸含量,以藏红花酸的含量和得率为考察指标,采用正交实验法考察碱水解工艺中的料液比、NaOH浓度、水解温度和水解时间对产品中藏红花酸含量和得率的影响。确定栀子黄色素碱水解的最佳条件:料液比1∶6 g/mL、NaOH浓度3 mol/L、水解温度55℃、水解时间60 min。在该条件下制备获得的藏红花酸得率可达15.33%±1.25%;含量可达到97.24%±0.78%。优化后方法步骤简单易行,绿色无污染,一步制得高纯度藏红花酸,适用于工业化生产。     

藏红花酸及藏红花酸二甲酯的制备与抗氧化性能研究

本文首先从栀子果中提取分离出栀子黄色素,栀子黄色素分别经过水解、酯交换反应得到藏红花酸和藏红花酸二甲酯。将产物分别经过重结晶处理,得到纯度为98.2%的藏红花酸和98.8%的藏红花酸二甲酯,其IR、UV、1HNMR均与文献报道一致。采用羟基自由基体系和超氧阴离子自由基体系对藏红花酸及藏红花酸二甲酯的体外抗氧化活性进行研究,并与BHT进行比较。结果表明,在试验质量浓度范围内,藏红花酸以及藏红花酸二甲酯的清除羟基自由基与超氧自由基的能力都明显强于BHT,为优秀的天然抗氧化剂。     

黄栀子花中的色素

黄栀子(Gardenia sootepensis Hutch)系茜草科栀子属常绿灌木,其果实含黄色色素。主产于湖南、江西、浙江等地的黄栀子的果实常用于食品染色。生长于云南省西双版纳的黄栀子,其花呈亮黄色,民间常将其花水煮后于各种食品上着色。据报道,栀子黄色素主要含藏红花素和藏红花酸,两种物质对碱、热稳定,无毒,着色力强。此次我们从黄栀子的花中分到主要有色成分——藏红花酸(Ⅰ,产率0.02％)和     

响应面法优化混合酶-超声波联合提取栀子黄色素工艺研究

以栀子为原料提取栀子黄色素,将混合酶-超声波技术相结合,对栀子黄色素进行了提取的研究。为了获得提取栀子黄色素最佳工艺条件,采用响应面法组合设计试验方法,建立了酶的用量、提取温度、提取时间之间关系。结果表明,最佳工艺条件为酶的用量4.33%,提取温度61.84℃,提取时间65.06 min,在此最佳提取条件下,栀子黄色素的吸光度为0.914。     

栀子黄色素的醇提制备及其稳定性研究

通过单因素实验,考察栀子黄色素乙醇提取工艺。乙醇提取最优工艺条件为：乙醇浓度30％、液固比8：1、提取温度60℃、提取次数2次、每次1h。在此条件下,栀子黄色素、栀子苷和总三萜酸的提取率分别为1．32、110．76和94．91mg／g。     

稳心草化学成份的研究

从内蒙古野生植物稳心草(Hypericum attenuatum Chois.)全草中分离出四个结晶成份。据光谱分析、酸水解、衍生物制备及理化常数的测定,结晶Ⅰ为金丝桃甙(hyperin 或 hyperoside),结晶Ⅱ为槲皮素(quercetin),结晶Ⅲ经纸层析鉴定,初步证明为含有少量杂质的氯原酸(chlorogenic acid),结晶Ⅳ为尚待鉴定的黄酮类化合物。经测定全草中总黄酮含量平均为1.67%,其花、果、茎、叶不同器官含量差异悬殊,以叶为最高,花果次之,茎最低。稳心草资源丰富,黄酮类化合物含量较高,药理作用广泛,是一个值得进一步研究的药用植物。     

猪毛中胱氨酸的分离和复合氨基酸的制备

本文研究了以猪毛为原料,经过水解、赶酸、中和、结晶、精制提取出胱氨酸纯品;并从分离胱氨酸后的母液中,经过脱色、离子交换、浓缩、结晶、精制,制备出复合氨基酸.在本工艺条件下,胱氨酸产品的收率为4.8%,纯度在99%以上;复合氨基酸产品的收率为41%,纯度在83%以上.本文为扩大试验打下了基础.     

正交实验法优化碱水解栀子提取物制取京尼平苷酸的工艺

以栀子提取物为原料,利用正交实验设计方法优化其碱水解制取京尼平苷酸的工艺。以京尼平苷酸产率为评价指标,考察了水解温度、NaOH加入量、料液比和水解时间4个单因素,采用四因素三水平正交实验设计优选得出最佳水解工艺:水解温度为70℃、NaOH加入量为14.0 m L、料液比1∶30 g/m L、水解时间为10 min,京尼平苷酸产率为11.53%。水解后溶液经过一次活性炭静态吸附和解吸,京尼平苷酸的含量可达45.33%。     

大孔树脂——溶剂萃取法精制高色价栀子黄色素的集成技术研究

以栀子为原料提取栀子黄色素,采用大孔吸附树脂--有机溶剂萃取相结合的集成技术,从栀子中分离纯化得到高色价的栀子黄色素.先采用大孔吸附树脂对栀子黄色素进行初步精制,以306型大孔吸附树脂为研究对象,探讨了大孔树脂对栀子黄色素的静态吸附率、吸附流速和洗脱剂浓度对吸附的影响,从而得到较为合适的工艺:吸附流速2.O mT/mi...     

栀子黄色素与金属离子效应的研究

本文报导了从栀子(Ga(?)denia Jasminoides Ellis)果实中提取的黄色素,与铁、铜、锡和铝等金属离子的效应。研究表明,栀子黄色素与铁、铜离子反应后,色素的共轭双键的发色基团破坏,栀子黄色素退色;与锡、铝离子反应后,生成黄色络合物,增加了吸光度,保持鲜艳黄色。因此,在提制和使用栀子黄色素时,应注意它们的不同效应。本试验是通过 FeCl_3、SnCl_2、AlCl_3和 CuSO_4四种试剂与栀子黄色的反应,探讨生产上经常遇到的铁、锡、铝和铜离子与栀子黄的效应,为提制栀子黄色素的设备选材、包装材料的选择;为栀子黄色素食品的加工、包装金属材料的选择提供科学依据。     

Crocetin treatment inhibits proliferation of colon cancer cells through down-regulation of genes involved in the inflammation

Background

The current study was designed to investigate the effect of crocetin on the proliferation inhibition of colon cancer cells and the underlying mechanism.

A highly specific glucosyltransferase is involved in the synthesis of crocetin glucosylesters in Crocus sativus cultured cells

Saffron (Crocus sativus) stigmata contain rare water-soluble carotenoids and the major one is crocin, the crocetin digentiobiosyl-ester. Previous studies indicated that two glucosyltransferases might be involved in the formation of crocetin glucosyl- and gentiobiosyl-esters (Dufresne et al. 1997). A UDP-Glc: crocetin 8,8′-glucosyltransferase involved in the biosynthesis of crocetin monoglucosyl- and diglucosyl-esters was extracted from saffron cell cultures and purified 300-fold by gel filtration chromatography and preparative IEF electrophoresis, with a recovery of 13 percnt;. The purified enzyme preparation was highly specific for crocetin and formed ester bonds between the glucose moiety of UDP-Glc and the free carboxyl functions of crocetin. The enzyme did not add other glucose units to the glucosyl-esters to form crocetin gentiobiosyl-esters. A crude desalted extract of the same material was less specific and formed glucosyl-esters with several other compounds, including abscisic and retinoic acids. The purified preparation was active between pH 4.4 and 4.6. SDS-PAGE revealed a major band at 26 kDa while the native molecular mass determined by gel filtration was in the range of 49 to 55 kDa. The study provides concrete evidence for the hypothesis that more than one glucosyltransferase is involved in the biosynthesis of crocetin glycosyl-esters in saffron.     

Direct process integration of extraction and expanded bed adsorption in the recovery of crocetin derivatives from Fructus Gardenia

A process that integrated an extraction tank (EXT) and an expanded bed adsorption (EBA) into a new system EXT-EBA for direct purifying crocetin derivatives from Fructus Gardenia was described. Conditions were set to allow the extraction and purification in a single step. A comparison between the integrated process and the conventional process to purify crocetin derivatives was presented. The integrated process resulted in 52.79% recovery of crocin compared to 24.12% in the conventional process. The process time and solvent used were decreased in the integrated process. The result suggests that the EXT-EBA integrates extraction, clarification, and purification in a single step, greatly simplifying the process flow and reducing the cost and time of extraction and purification of crocetin derivatives from Fructus Gardenia.     

Crocetin protects against cardiac hypertrophy by blocking MEK-ERK1/2 signalling pathway

Oxidative stress plays a critical role in the progression of pathological cardiac hypertrophy and heart failure. Because crocetin represses oxidative stress in vitro and in vivo , we have suggested that crocetin would repress cardiac hypertrophy by targeting oxidative stress-dependent signalling. We tested this hypothesis using primary cultured cardiac myocytes and fibroblasts and one well-established animal model of cardiac hypertrophy. The results showed that crocetin (1–10 μM) dose-dependently blocked cardiac hypertrophy induced by angiogensin II (Ang II; 1 μM) in vitro . Our data further revealed that crocetin (50 mg/kg/day) both prevented and reversed cardiac hypertrophy induced by aortic banding (AB), as assessed by heart weight/body weight and lung weight/body weight ratios, echocardio-graphic parameters and gene expression of hypertrophic markers. The inhibitory effect of crocetin on cardiac hypertrophy is mediated by blocking the reactive oxygen species (ROS)-dependent mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase-1/2 (MEK/ERK1/2) pathway and GATA binding protein 4 (GATA-4) activation. Further investigation demonstrated that crocetin inhibited inflammation by blocking nuclear factor kappa B (NF-κB) signalling and attenuated fibrosis and collagen synthesis by abrogating MEK-ERK1/2 signalling. Overall, our results indicate that crocetin, which is a potentially safe and inexpensive therapy for clinical use, has protective potential in targeting cardiac hypertrophy and fibrosis by suppression of ROS-dependent signalling pathways.     

转基因烟草表达西红花CSzCD基因产生西红花酸

为研究转基因烟草中产生西红花酸的可行性,在本研究中,西红花玉米黄素裂解酶（CSzCD）基因插入到pBI121载体的花椰菜花病毒（CaMV）35S启动子下游,通过农杆菌介导整合到烟草基因组中。通过Southern blotting 分析得到21株转基因烟草植株系; 转基因烟草叶片提取物Western blotting 和HPLC分析显示有西红花酸的产生,然而阴性对照中并没有发现西红花酸的存在。     

The pharmacokinetic profile of crocetin in healthy adult human volunteers after a single oral administration

Crocetin, a unique carotenoid with a short carbon chain length, is an active compound of saffron and Gardenia jasminoides Ellis used as traditional herbal medicine. The present study was undertaken to investigate the pharmacokinetic profiles of crocetin in healthy adult subjects. The study was conducted as an open-label, single dose escalation with 10 Filipino volunteers (5 men and 5 women). The subjects received a single dose of crocetin at three doses (7.5, 15 and 22.5 mg) in one week interval. Blood samples were collected from the brachial vein before and at 1, 2, 4, 6, 8, 10 and 24 h after administration. Plasma concentrations of crocetin were determined by high-performance liquid chromatography (HPLC). Crocetin was rapidly absorbed and detected within an hour of administration with a mean time to reach maximum concentration (T_max) of crocetin ranging from 4.0 to 4.8 h. The mean values of C_max and AUC_0-24 h ranged from 100.9 to 279.7 ng/ml and 556.5 to 1720.8 ng^.h/ml respectively. C_max and AUC values increased with dose proportional manner. Crocetin was eliminated from human plasma with a mean elimination half life (T_1/2) of 6.1 to 7.5 h.In summary, there were no serious adverse events up to 22.5 mg dose of crocetin while crocetin was found to be absorbed more quickly than the other carotenoids such as β-carotene, lutein and lycopene.     

Crocetin inhibits beta-amyloid fibrillization and stabilizes beta-amyloid oligomers

Aggregation of a peptide, beta-amyloid (Aβ), is a hallmark molecular process found in Alzheimer’s disease (AD). During Aβ aggregation, oligomeric and fibrillar Aβ are formed, and these molecular self-assembly steps are implicated in generation of toxic effects in AD. Crocetin is a natural carotenoid dicarboxyl acid displaying various pharmaceutical effects and may be co-localized with Aβ mediated by human serum albumin. In the study presented here, we examined the effects of crocetin on Aβ aggregation in three different molecular pathways. Our results demonstrate that crocetin inhibited Aβ fibril formation and destabilized pre-formed Aβ fibrils. Moreover, crocetin caused stabilization of Aβ oligomers and prevented their conversion into Aβ fibrils. Our study reveals potential pathological and pharmaceutical implication of crocetin in AD and suggests possible application of crocetin for currently limited structural studies on unstable Aβ oligomers.