超声波强化乙醇提取罗汉果甜苷

目的优选超声强化乙醇罗汉果甜苷的提取工艺。方法采用正交试验法考察乙醇溶液提取罗汉果甜苷的主要影响因素,以甜苷提取率为指标,评定优选工艺条件,在此基础上,进行扩展试验。结果在40 kHz、100 W超声波处理下,提取3次,最佳工艺条件为:第1次提取,以体积分数为40%的乙醇溶液,用量为25倍(溶剂体积与罗汉果质量之比),提取温度60℃,提取时间40 min;第2次提取,以体积分数为30%的乙醇溶液,用量为20倍,提取温度65℃,提取时间30 min;第3次提取,以体积分数为20%的乙醇溶液,用量为20倍,提取温度75℃,提取时间20 min。结论采用超声波可以有效地提高罗汉果的提取率。     

正交试验优选药膳罗汉膏的提取工艺研究

目的优选罗汉膏的提取工艺。方法以罗汉果的有效成分罗汉果皂苷V的含量和浸膏得率为评价指标,以提取时间、提取次数、加水量为考察因素,采用正交试验对提取方法的工艺条件进行优化。结果最佳工艺为加8倍量水,提取3次,每次0.5 h。结论优选出的提取工艺合理、稳定可行。     

超高效液相色谱法测定罗汉果中7种成分含量

目的 建立超高效液相色谱法(UPLC)测定罗汉果中11-O-罗汉果苷Ⅴ、罗汉果皂苷Ⅴ、赛门苷Ⅰ、罗汉果苷Ⅳ、罗汉果苷Ⅲ、罗汉果苷Ⅲe、罗汉果苷Ⅱe的含量。方法 采用Waters BEH C₁₈色谱柱(50 mm×2.1 mm, 1.7μm),流动相为水-乙腈溶液,梯度洗脱;柱温35℃;流速为0.3 mL·min^-1;进样量为1μL;采用紫外检测器进行定量测定,检测波长203 nm;采用质谱检测器进行定性鉴别,检测模式为选择离子监测,监测离子的质荷比为1307.8,1309.8,1147.8,1147.8,985.7,985.7和823.7。结果 11-O-罗汉果苷Ⅴ、罗汉果皂苷Ⅴ、赛门苷Ⅰ、罗汉果苷Ⅳ、罗汉果苷Ⅲ、罗汉果苷Ⅲe、罗汉果苷Ⅱe分别在0.50～20.10,5.03～201.20,0.51～20.24,0.52～20.63,0.51～20.22,0.51～20.56,0.50～19.92μg·mL^-1范围内线性关系良好(r≥0.999);平均回收率分别为100.7%,98.4%,100.4%,99.0%,100...     

罗汉果甜苷干预棕榈酸致胰岛B细胞氧化应激相关损伤的机制研究

目的:研究罗汉果甜苷干预棕榈酸致胰岛B细胞氧化应激相关损伤的机制。方法:以小鼠胰岛B细胞系NIT-1细胞作为细胞模型。实验分为空白对照、罗汉果甜苷对照、模型、罗汉果甜苷干预4组,使用流式细胞术测定细胞内活性氧自由基(ROS)含量和细胞凋亡率,半定量逆转录PCR测定葡萄糖转运受体(GLUT)-2和丙酮酸激酶编码基因表达水平。结果:与模型组比较,罗汉果甜苷组NIT-1细胞内ROS含量显著降低,GLUT-2和丙酮酸激酶编码基因的表达水平显著增强(P<0.05),细胞凋亡无显著改变(P>0.05)。结论:罗汉果甜苷可显著降低NIT-1细胞内ROS水平,显著上调受抑制的GLUT-2和丙酮酸激酶编码基因表达,其机制可能是通过降低细胞内ROS含量,逆转由氧化应激激活的转录因子叉头框蛋白-1(FOXO1)导致的葡萄糖代谢障碍和其他效应,产生对胰岛B细胞的保护作用。     

罗汉果甜苷V特异性抗血清的制备与应用

目的制备罗汉果甜苷V(MogV)特异性抗血清,建立MogV含量竞争性抑制酶联免疫检测法。方法用罗汉果甜苷V-丁二酸酯-牛血清蛋白(MogV-HS-BSA)结合比约为44∶1为免疫抗原制备抗血清,用罗汉果甜苷V-丁二酸酯-人血清蛋白(MogV-HS-HSA)结合比约为64∶1为包被抗原,建立MogV含量竞争性抑制酶联免疫检测法。结果包被抗原工作浓度为1μg.mL 1,血清最佳稀释倍数为1∶8 000,在此工作浓度下,MogV浓度在1×10-4~1μg.mL 1内回归方程Y=0.158X+0.298,r=0.991 0。结论成功制备了MogV抗血清,MogV浓度在1×10-4~1μg.mL 1内对抗血清的OD(405 nm)值抑制作用呈良好的线性关系。     

奥斯曼生眉液超滤工艺的研究

目的：研究奥斯曼生眉液的超滤工艺。方法采用截留相对分子量分别为6000和30000的聚丙烯中空纤维膜,以超滤前后药液中吲哚苷和鞣质含量为指标,采用单因素考察和正交实验方法优选超滤的最佳工艺参数。结果截留相对分子质量为30000的超滤膜有较好的分离提纯效果,最佳工艺参数为：料液加水稀释5倍,调pH至6.0,常温、操作压力0.10Mpa的条件下膜分离3遍。结论超滤工艺能有效提高产品有效物的含量和解决产品稳定性问题。     

罗汉果中的葫芦烷型糖苷及其对EB病毒活性的抑制作用

罗汉果Siraitia grosvenorii(Swingle)Lu & Zhang中的葫芦烷型糖苷有化学防癌、抑制肠麦芽糖酶和血糖升高等作用。从该植物果实的甲醇提取物中分离出6个新的葫芦烷型糖苷:罗汉果苷(mogrosdie)ⅡB(2)、11-脱氧罗汉果苷Ⅲ(4)、7-氧代罗汉果苷ⅡE(5)、7-氧代罗汉果苷Ⅴ(6)、11-氧代罗汉果苷ⅡA_1(7)和11-氧代罗汉果苷ⅣA(8),并首次发现2个天然来源的、已知的葫芦烷型糖苷:罗汉果苷ⅡA_1(1)和罗汉果苷ⅢA_2(3)。     

罗汉果的质量标准

目的：建立测定罗汉果中多糖、总糖及罗汉果苷V含量的方法,并测定其不同部位的含量。方法：采用苯酚-硫酸比色法测定多糖及总糖含量,测定波长为487nm;用HPLC测定罗汉果苷V含量,Phenomenex C18柱,乙腈-水（22：78）为流动相,流速1．0ml·min^-1,检测波长208nm。结果：无水葡萄糖在32．0～192．2μg,r=0．9998,罗汉果苷V在0．81—12．1μg围内具有良好的线性关系（r=0．9999）,平均回收率分别为100．7％和102．0％,RSD为2．33％和1．03％（n=6）;其含量均以果肉最高而种子较低。结论：方法灵敏、准确,重复性好,适用于该药材的质量评价。     

乙醇提取番泻叶中总番泻苷的工艺优选

目的：优选乙醇提取番泻叶中总番泻苷的最佳工艺。方法：采用正交设计法，以乙醇为溶媒对番泻叶中总番泻苷的提取工艺进行优选。结果与结论：最佳提取工艺条件为：70%乙醇提取3次，每次提取1h。     

罗汉果果实的化学成分与应用研究

罗汉果 [Siraitia grosvenorii (Swingle) C.Jeffrey],又名光果木鳖、拉江果、假苦果 ,原产我国 ,是我国特有的果树。清光绪 11年 (1885年 )广西永宁州志 (即广西永福县 )就有罗汉果作药的记载。清光绪 31年 (190 5年 )广西临桂县县志记述 :“罗汉果 ,大如柿 ,椭圆中空 ,味甜性凉 ,治痨嗽”。可见罗汉果在医药上的应用至少有一百年以上的历史。李典鹏和张厚瑞 [1 ] 指出 ,罗汉果在广西民间的药用历史已有 30 0多年。近年来许多学者对罗汉果果实化学成分、药理作用、应用及产品开发进行了研究 ,为了促进我国对罗汉果果实的开发利用 ,本文对…     

罗汉果皂苷Ⅳa碳氢NMR信号的归属

目的　研究传统中药罗汉果的皂苷类化学成分罗汉果皂苷IVa的结构 ,总结其结构的NMR特点。方法　采用常法提取、分离、纯化罗汉果皂苷IVa ,应用NMR方法 ( 1 HNMR、1 3CNMR、DEPT、1 H 1 HCOSY、HSQC、HMBC和NOESY)和计算机分子模拟研究其结构。结果　对罗汉果皂苷IVa的1 H和1 3C信号进行了归属 ,并为该类型化合物的结构确定提供了波谱学依据。结论　包括1 H 1 HCOSY、HSQC、HMBC、NOESY的一维和二维核磁共振波谱技术是对结构进行无创伤性分析的有力工具。NMR方法确定的结构与计算机分子模拟的最优构象一致。     

Biotransformation of mogrosides from Siraitia grosvenorii by Ganoderma lucidum mycelium and the purification of mogroside III E by macroporous resins

Mogrosides are the major triterpenoidal saponins found in swingle, the fruit of Siraitia grosvenorii, which have recently been widely used throughout the world as natural food sweeteners. Among this class of compounds, mogroside III E (MG III E) exhibits the most intense sweetness, and it was also found to effectively regulate blood glucose levels. However, the relative abundance of naturally occurring MG III E is low compared to other mogrosides. Therefore, the purpose of this study was to enrich MG III E through biotransformation of fruit extracts and to develop a reliable method for its purification. We used HPLC coupled with mass spectrometry and nuclear magnetic resonance spectroscopy for metabolite analysis and identified MG III E as a major metabolite of Ganoderma lucidum mycelium. This organism converts the most abundant mogroside, mogroside V, to MG III E via a deglycosylation reaction; high levels of β-glucosidase activities were also detected. In addition, we established an efficient purification method for MG III E using HP-20 macroporous resin. Optimization of the method was accomplished by kinetic model fitting, dynamic adsorption studies, and desorption experiments. The purity of MG III E was increased from 11.71% to 54.19%, with a 70%–76% recovery rate, and the scaled-up purification process allowed us to harvest 17.38 g of MG III E with 55.14% purity and a 74.71% of recovery rate. Therefore, our low cost, time-saving, easy to scale-up procedure for isolating MG III E could be applicable in industrial processes.     

正交实验法优选苦瓜根总皂苷提取工艺

目的 优选苦瓜根中苦瓜总皂苷的最佳提取工艺. 方法 以提取液中苦瓜总皂苷含量为考察指标,分光光度法测定其含量,采用L9(34)正交实验法优选苦瓜根总皂苷的提取工艺. 结果苦瓜根中苦瓜总皂苷较优的提取工艺为提取2次,每次2 h,乙醇浓度60%,乙醇用量8倍,提取温度80 ℃. 结论 优选得到的提取工艺稳定、可行、重复性好,可为苦瓜根总皂苷的提取提供参考.     

中药罗汉果抗血脂紊乱的实验研究

目的观察中药罗汉果对实验性血脂紊乱SD大鼠血清总胆固醇(TC)、低密度脂蛋白(LDL-c)、三酰甘油(TG)及高密度脂蛋白(HDL-c)水平的影响。方法 50只雄性SD大鼠(体质量180g-220g)适应性饲养7d后随机分为空白对照组和造模组,造模组上午8时至下午18时给予普通饲料喂养,18时至次日上午8时高脂饲料喂养,共4周。4周后禁食12h断尾采血测两组空腹血清TC、LDL-c、TG、及HDL-c水平,造模组血脂各指标显著升高具有统计学意义者,确定为模型建立成功。将造模成功的模型组大鼠再随即分为药物实验组(20只,给予含生药4g/(kg d)罗汉果水溶液灌胃)和模型对照组(20只),后者继续高脂饲料喂养。至第八周后测血清TC、LDL-c、TG、及HDL-c水平。结果实验组血清TC)和LDL-c)的水平显著低于对照组(P<0.05);TG水平较对照组有下降趋势,HDL-c水平较对照组有上升趋势。结论中药罗汉果可显著降低实验大鼠血清TC和LDL-c,可通过调节血脂代谢紊乱发挥抗动脉粥样硬化作用。     

人参黄芪有效成份提取的最佳工艺

目的：优选人参黄芪有效成分水提的最佳工艺条件。方法：采用正交试验进行优选。结果：人参黄芪水提的最佳工艺条件为加水量10倍，煎煮时间2小时、煎煮次数2次。结论：通过正交试验优选出的人参黄芪水提的最佳工艺可节约能源，降低成本，减少工时，并且有效成分提取率最高。     

木鳖子中总皂苷的提取分离工艺及其急性毒性的研究

目的优选木鳖子中总皂苷的最佳提取工艺,并观察木鳖子总皂苷对小鼠的急性毒性作用,评价其安全性。方法采用正交实验,以木鳖子总皂苷含量为指标,考察溶剂用量、提取次数、提取时间对木鳖子中总皂苷提取率的影响,并采用D101大孔吸附树脂结合正丁醇萃取分离提取液中木鳖子总皂苷。采用小鼠半数致死量(LD50)测定方法,观察小鼠灌胃给予木鳖子总皂苷的急性毒性反应。结果最佳提取工艺条件为:8倍量水提取三次,每次煮沸提取1 h。大孔树脂分离纯化的总皂苷纯度可达72.5%。小鼠单次灌胃给药的LD50为1.490 g.kg-1。结论优选的提取工艺结合大孔吸附树脂和正丁醇分离纯化木鳖子中总皂苷的方法,成本低且操作简便可行,适用于工业化生产木鳖子总皂苷。木鳖子总皂苷有一定的毒性。     

Impact of sample preparation in peptide/protein profiling in human serum by MALDI-TOF mass spectrometry

The low molecular weight (LMW) serum proteome (<15 kDa) is the most generally informative from a medical point of view. Different sample pre-treatment approaches and devices for serum depletion in high-abundant proteins were tested in order to analyze, by MALDI-TOF-MS (both in "linear" and "reflectron" acquisition mode), the serum low molecular weight proteins/peptides. The best results in terms of detected ions number and abundance were obtained by using ultrafiltration of serum on 30 kDa molecular weight cut off membranes followed by miniaturized reverse-phase solid-phase extraction (mu-SPE) as sample pre-treatment; this procedure yielded also satisfactory within-sample and sample-to-sample repeatability (on both m/z values and peak intensity of the main observable ions). The procedure was finally applied to serum samples of breast cancer patients, and the relevant results compared to "normal" samples seem to be promising for the individuation of different profiles ("linear" and "reflectron" mode) and/or peptides capable of differentiating for malignancies ("reflectron" mode).     

Systematic optimisation of ethyl glucuronide extraction conditions from scalp hair by design of experiments and its potential effect on cut‐off values appraisal

The quantitative determination of ethyl glucuronide (EtG) in hair samples is consistently used throughout the world to assess chronic excessive alcohol consumption. For administrative and legal purposes, the analytical results are compared with cut‐off values recognised by regulatory authorities and scientific societies. However, it has been recently recognised that the analytical results depend on the hair sample pretreatment procedures, including the crumbling and extraction conditions. A systematic evaluation of the EtG extraction conditions from pulverised scalp hair was conducted by Design of Experiments (DoE) considering the extraction time, temperature, pH, and solvent composition as potential influencing factors. It was concluded that an overnight extraction at 60°C with pure water at neutral pH represents the most effective conditions to achieve high extraction yields. The absence of differential degradation of the internal standard (isotopically‐labelled EtG) under such conditions was confirmed and the overall analytical method was validated according to SGWTOX and ISO17025 criteria. Twenty real hair samples with different EtG content were analysed with 3 commonly accepted procedures: (a) hair manually cut in snippets and extracted at room temperature; (b) pulverised hair extracted at room temperature; (c) hair treated with the optimised method. Average increments of EtG concentration around 69% (from a to c) and 29% (from b to c) were recorded. In light of these results, the authors urge the scientific community to undertake an inter‐laboratory study with the aim of defining more in detail the optimal hair EtG detection method and verifying the corresponding cut‐off level for legal enforcements.     

Liver stiffness measurement as a predictive tool of clinically significant portal hypertension in patients with compensated hepatitis C virus or alcohol-related cirrhosis

Background Hepatic venous pressure gradient (HVPG) is the gold standard for assessing the presence and the severity of portal hypertension (PHT). Liver stiffness measurement (LSM) is a non‐invasive method for liver fibrosis assessment. Aims To assess the relationship between LSM and HVPG in patients with compensated cirrhosis related to hepatitis C virus (HCV) or alcohol and to define the performance and the best cut‐off of LSM for the diagnosis of PHT in these patients. Methods Between January 2004 and September 2006, we studied all the consecutive patients with compensated HCV or alcohol‐related‐cirrhosis referred for transjugular liver biopsy with HVPG measurement and LSM performed the same day. Results Ninety‐two patients were eligible, 44 had HCV related‐cirrhosis and 48 alcoholic cirrhosis. LSM was positively correlated to HVPG in both groups. The area under the receiver operating characteristic curve for the diagnosis of significant PHT was 0.76 ± 0.07 in HCV patients (best cut‐off at 20.5 kPa) and 0.94 ± 0.03 (best cut‐off at 34.9 kPa) in alcoholic patients. Conclusions Liver stiffness measurement and HVPG were significantly correlated in patients with compensated cirrhosis because of HCV infection or alcohol. LSM could predict significant PHT in both these groups of patients with a higher cut‐off and a better performance in alcoholic patients.