人参柔顺洗发水中人参皂苷的鉴别和含量测定

建立了人参柔顺洗发水中人参皂苷Rg1,Re和Rb1的鉴别和含量测定方法.采用薄层色谱(TLC)法,以氯仿-甲醇-水(体积比2∶1∶0.1)为展开溶剂,对人参皂苷进行鉴别;通过高效液相色谱(HPLC)法,以乙腈-水为流动相梯度洗脱,测定洗发水中人参皂苷的含量.结果显示TLC法能同时鉴别人参皂苷Rg1,Re和Rb1;HPLC法中人参皂苷Rg1,Re和Rb1的线性回归方程分别为Rg1∶y=246 683x -7 070.6,r=0.999 9;Re∶y=388 224x - 11 320,r=0.999 9;Rb1∶y=216 383x+3 644.9,r=0.999 9;平均回收率分别为99.69％,97.39％和98.05％,RSD值分别为1.62％,1.75％和0.68％.     

HPLC测定不同年份人参药材中3种人参皂苷的含量

本文涉利用高效液相色谱对15年、18年和22年不同年份石柱山参中人参皂苷Rg1、人参皂苷Re和人参皂苷Rb1的含量进行了系统比较。实验结果表明三种人参皂苷人参皂苷Rg1、人参皂苷Re和人参皂苷Rb1在100min内基线分离,线性关系良好。随着年份增加,人参皂苷Rg1、人参皂苷Re和人参皂苷Rb1的含量都有不同程度的提高。     

正交试验优化葶苈生脉方中醇提工艺研究

优选葶苈生脉方中红参、五味子、川芎、葶苈子、泽泻的最佳提取工艺,通过正交试验设计考察乙醇浓度、料液比、提取时间对工艺的影响,采用高效液相色谱法(HPLC)进行梯度洗脱,以人参皂苷Rg1和人参皂苷Rb1为指标进行综合评分,确定最佳提取工艺:6倍量65%乙醇,回流提取3次,第一次3 h,第二次2 h,第三次1 h。通过验证实验表明该工艺科学、合理、稳定、可行,为葶苈生脉方开发为现代中药制剂提供科学依据。     

参苓白术散中甘草黄酮和人参皂苷测定及抗氧化实验

采用高效液相法测定参苓白术散中人参皂苷和甘草黄酮的含量,为下一步探究参苓白术散的体外抗氧化活性研究做准备。方法:采用HPLC法(反相高效液相法),目的:为测定参苓白术散中的甘草黄酮、人参皂苷含量,进行线性范围的考察和实验方法学研究,同时测定参苓白术散中人参皂苷对羟自由基的抑制情况。结果:人参皂苷Rg1、槲皮素、人参皂苷Re、芦丁分别在0.625～5.000μg、0.250～1.000μg、0.375～3.000μg、0.250～1.000μg内对羟自由基的抑制作用显著;芦丁的平均质量分数为0.295 5 mg·g~(-1);人参皂苷Rg1的平均质量分数为0.193 mg·g~(-1);槲皮素的平均质量分数为0.243 9 mg·g~(-1);人参皂苷Re的平均质量分数为0.074 7 mg·g~(-1)。参苓白术散的抗氧化能力经验证也较为明显,人参皂苷对羟自由基的抑制率为79%。     

HPLC法同步测定三七粉中人参皂苷Rg1、Rb1及三七皂苷R1的含量

目的:采用HPLC法同步测定三七粉中人参皂苷Rg1、Rb1及三七皂苷R1含量。方法:采用SGE protecol C18(5μm,4.6×250mm)色谱柱,流动相为乙腈—水梯度洗脱(0~12min,19∶81;12~60min,19~36∶81~64),检测波长为203nm,流速1.0mL/min,进样量20μL。结果:人参皂苷Rg1、人参皂苷Rb1、三七皂苷R1分别在25.625~430mg/L、26.875~410mg/L、10.625~170mg/L范围呈线性,相关系数r分别为0.9999、0.9998、0.9996;该方法重复性及回收率均符合要求。结论:本法用于同步测定三七粉中人参皂苷Rg1、Rb1及三七皂苷R1的含量,具有简便、准确、高效等特点。     

高效液相色谱法测定生脉饮中6种活性成分的含量

采用高效液相色谱法,同时测定生脉饮中人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd、五味子醇甲、五味子乙素6种活性成分的含量.结果表明,各组分线性关系良好(R≥0.9992),精密度、重现性及加样回收率的相对标准误差均小于5.0%.该法操作简便、灵敏度高、重现性好,可用于全面控制和评价生脉饮的质量.     

二至密黑洗发乳的研制

采用正交试验设计法,以特女贞苷、蟛蜞菊内酯含量为指标,优选最佳提取工艺;根据洗发水配制原则,筛选基本配方,考察二至组方提取液添加量对洗发乳调理性能和配伍性能的影响。结果表明,最佳提取工艺为:用8倍量90%乙醇回流提取两次,每次2 h,再以8倍量水提取2 h;提取液添加量在4%~6%时,与配方中其他组分相容性良好,理化测试各项指标达到国家标准。证明二至组方可以用于草本洗发乳的开发。     

乙醇体系中金属离子催化转化人参皂苷Rb1的研究

人参皂苷Rb1是人参中的主要皂苷之一,但人体吸收量极少,而其衍生物Rg3、Rk1、Rg5却易于人体吸收,但其制备和分离成本较高。针对以上问题将人参皂苷Rb1在50%乙醇水溶液体系和无水乙醇体系中进行催化条件的优化,并分析其产物。实验发现,通过对金属离子催化效果的筛选,选取NbCl₅为反应催化剂;在50%乙醇水溶液体系中最优条件下反应,Rg3得率为18.29%;在无水乙醇体系中最优条件下反应,Rg3得率为49.45%。根据两组反应产物的对比,推测在50%乙醇水溶液体系反应时,水可能供给电子基团,并生成更多水合化合物即20(S)-25-OH-Rg3和20(R)-25-OH-Rg3;在无水乙醇体系反应时,电子基团可能无法供给,形成双键,生成其他化合物即Rk1和Rg5。该发现为金属离子催化人参皂苷以制备稀有人参皂苷的方法奠定了理论基础。     

高效液相色谱法同时测定三七提取物中三七皂苷R1与人参皂苷Rg1、Rb1含量

建立了高效液相色谱法同时测定三七提取物中三七皂苷R1与人参皂苷Rg1、Rb1含量的方法。采用高效液相色谱仪,Agilent ODS-C18(5μm,4.6mm×250mm)液相色谱柱,流动相为乙腈-水梯度洗脱,流速为1.0 mL/mim,检测波长为203 nm,柱温为35℃。结果表明,三七皂苷R1在度15.29~244.60 mg/L的范围内(r=0.9998),人参皂苷Rg1在14.30~228.90 mg/L范围内(r=1.0000),人参皂苷Rb1在13.13~210.11 mg/L范围内(r=1.0000),线性关系良好。三七皂苷R1和人参皂苷Rg1、人参皂苷Rb1平均加标回收率分别为99.54%、101.61%、102.99%,峰面积相对标准偏差RSD(n=6)分别为1.67%、0.93%、0.92%。该方法简便、准确,重现性好,可以对三七提取物中三七皂苷R1与人参皂苷Rg1、Rb1进行定量分析。     

大孔吸附树脂对两种人参皂苷的竞争性吸附研究

人参皂苷Re和人参皂苷Rg1是天然产物人参提取物中重要的药用成分,具有多种药理作用。目前关于两种皂苷的分离纯化普遍采用大孔吸附树脂分离的方法。但由于两种皂苷结构相似,结晶分离纯化难度较大,尤其是在树脂吸附过程中存在竞争吸附问题。本文利用利用D101大孔吸附树脂考察了树脂吸附人参皂苷Re和Rg1混合物的动力学性能和竞争吸附作用,同时分析了吸附作用机理,为系统利用吸附树脂纯化天然产物提供技术参考。     

常温超高压提取人参总皂苷

在常温条件下使用超高压提取了人参皂苷;采用均匀设计法对提取工艺条件作了优化;常温超高压提取人参皂苷最优工艺条件为：溶剂50%乙醇、压力200 MPa、固（g）液（ml）比1∶100、提取时间1 min;提取收率为7.32%.与煎煮法、回流提取法、超声提取法、超临界CO2提取等方法做了比较,结果表明常温超高压提取具有提取温度低（常温）、提取时间短、提取得率高、能耗低、绿色环保等优点,为人参皂苷的提取提供了一种新技术、新工艺.     

Ginsenoside Rg3 Reduces Lipid Accumulation with AMP-Activated Protein Kinase (AMPK) Activation in HepG2 Cells

Cardiovascular disease (CVD) is one of the main causes of mortality worldwide, and dyslipidemia is a major risk factor for CVD. Ginseng has been widely used in the clinic to treat CVD. Ginsenoside Rg3, one of the major active components of ginseng, has been reported to exhibit antiobesity, antidiabetic, and cardioprotective effects. However, the effect of ginsenoside Rg3 on hepatic lipid metabolism remains unclear. Therefore, we investigated whether ginsenoside Rg3 would regulate hepatic lipid metabolism with AMP-activated protein kinase (AMPK) activation in HepG2 cells. Ginsenoside Rg3 significantly reduced hepatic cholesterol and triglyceride levels. Furthermore, ginsenoside Rg3 inhibited expression of sterol regulatory element binding protein-2 (SREBP-2) and 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGCR). Ginsenoside Rg3 increased activity of AMPK, a major regulator of energy metabolism. These results suggest that ginsenoside Rg3 reduces hepatic lipid accumulation with inhibition of SREBP-2 and HMGCR expression and stimulation of AMPK activity in HepG2 cells. Therefore, ginsenoside Rg3 may be beneficial as a food ingredient to lower the risk of CVD by regulating dyslipidemia.     

生姜6-姜辣素的提取及姜汁去屑洗发水的研制

优选生姜6-姜辣素的提取方法,研究生姜提取液在去屑洗发水中的应用。采用HPLC法,以6-姜辣素为考察指标,分别对榨汁法、加热回流提取法、微波提取法、超声提取法和超声-微波协同萃取法的生姜提取液进行含量测定;通过抑菌实验,考察不同添加量的生姜提取液对洗发香波性能影响,确定生姜提取液的最佳添加量,最小抑菌浓度。结果显示,榨汁法为生姜的最佳提取方法,姜汁的最佳添加量是15%,最小抑菌浓度是:糠秕马拉色菌MIC为18.75 mg/mL,金黄色葡萄球菌、大肠埃希菌和白色念珠菌MIC均为9.375 mg/mL。     

Oral Rg1 supplementation strengthens antioxidant defense system against exercise-induced oxidative stress in rat skeletal muscles

ABSTRACT: BACKGROUND: Previous studies reported divergent results on nutraceutical actions and free radical scavenging capability of ginseng extracts. Variations in ginsenoside profile of ginseng due to different soil and cultivating season may contribute to the inconsistency. To circumvent this drawback, we assessed the effect of major ginsenoside-Rg1 (Rg1) on skeletal muscle antioxidant defense system against exhaustive exercise-induced oxidative stress. METHODS: Forty weight-matched rats were evenly divided into control (N = 20) and Rg1 (N = 20) groups. Rg1 was orally administered at the dose of 0.1 mg/kg bodyweight per day for 10- week. After this long-term Rg1 administration, ten rats from each group performed an exhaustive swimming, and remaining rats considered as non-exercise control. Tibialis anterior (TA) muscles were surgically collected immediately after exercise along with nonexercise rats. RESULTS: Exhaustive exercise significantly (p <0.05) increased the lipid peroxidation of control group, evidenced by elevated malondialdehyde (MDA) levels. The increased oxidative stress after exercise was also confirmed by decreased reduced glutathione to oxidized glutathione ratio (GSH/GSSG ratio) in control rats. However, these changes were completely eliminated in Rg1 group. Catalase (CAT) and glutathione peroxidase (GPx) activities were significantly (p <0.05) increased by Rg1 in non-exercise rats, while no significant change after exercise. Nevertheless, glutathione reductase (GR) and glutathione S-transferase (GST) activities were significantly increased after exercise in Rg1 group. CONCLUSIONS: This study provide compelling evidences that Rg1 supplementation can strengthen antioxidant defense system in skeletal muscle and completely attenuate the membrane lipid peroxidation induced by exhaustive exercise. Our findings suggest that Rg1 can use as a nutraceutical supplement to buffer the exhaustive exercise-induced oxidative stress.     

用于渗漉提取过程分析的中药有效组分近红外光谱快速测定法

研究提出一种快速测定中药渗漉提取液中有效组分的近红外（NIR）光谱分析方法,可用于中药渗漉提取过程在线分析.以中药三七为对象,采用人参皂苷Rg1、Rb1、Rd的HPLC测定值及三七总皂苷（PNS）的比色法测定值作对照值,建立了NIR光谱与对照值之间的校正模型.比较研究了径向基函数神经网络（RBFNN）和偏最小二乘回归（PLSR）两种建模方法,其中RBFNN校正模型对Rg1、Rb1、Rd和PNS 4种组分的交叉验证均方差（RMSECV）分别为1.120、1.230、0.267、4.749,预测均方差（RMSEP）分别为0.677、0.969、0.155、8.065.研究结果表明,本文方法方便、准确、无损,可推广用于中药提取过程分析及质量控制.     

Effects of pre-fermentation enzyme treatments of leaves,stems, and roots of ginseng on <Emphasis Type="Italic">Lactobacillus</Emphasis> fermentation characteristics and ginsenoside metabolite formation

The extraction and enzymatic treatment conditions of ginseng leaves, stems and roots for the production of fermented ginseng were optimized in order to enhance the extraction of oligosaccharide, which is a Lactobacillus growth-activating factor. Additionally, the effects of enzymatic hydrolysis on Lactobacillus fermentation characteristics and metabolites of ginsenoside were investigated. The ginseng leaves were found to be more suitable for the raw material of fermented ginseng products because ginseng leaves have higher carbohydrate and crude saponin content than ginseng roots. The optimized conditions were found as particle size of ginseng raw material below 0.15 mm, pH 5.0–5.5, reaction temperature of 55–60 °C, Ceremix concentration of 1%, and reaction time of 2 h. It was shown that the polysaccharides of ginseng were hydrolysed to oligosaccharide by the enzymatic hydrolysis of ginseng leaves, stems and roots. The total oligosaccharide content increased by the enzyme treatment up to 2.2-fold, 5.3-fold and 2.3-fold in ginseng leaves, stems and roots, respectively, compared to control (no treatment). It was found that the enzymatic treatment promoted the Lactobacillus growth, resulting in more significant change in total oligosaccharide consumption and total acidity. The content of several metabolites of ginsenoside, such as Compound K, Rg₁, Rh₁ and Rg₃, was selectively increased by combining the enzymatic treatment and Lactobacillus fermentation. Especially, in the case of enzyme treatment using ginseng leaves, Compound K formation was enhanced up to three-fold compared to control (no treatment). Moreover, in case of combined treatment of enzyme and fermentation, Compound K formation was significantly promoted up to ten-fold.     

人参皂苷Rg1诱导人红白血病K562细胞株衰老与p16-Rb信号通路的相关性

目的探讨人参皂苷Rg1(以下简称Rg1)诱导人红白血病K562细胞株衰老与p16-Rb信号通路的相关性。方法以不同浓度的Rg1(0、5、10、20、40和80μmol/L)作用于K562细胞不同时间(24、48、72 h),MTT法筛选Rg1抑制K562细胞增殖的最佳作用浓度及作用时间,以该浓度干预K562细胞不同时间,流式细胞术检测细胞的细胞周期;以最佳浓度Rg1干预K562细胞最佳时间,集落培养法检测细胞集落形成能力;衰老相关-β-半乳糖苷酶(SA-β-Gal)染色检测阳性细胞百分率;透射电镜观察细胞的超微结构;Southern blot检测细胞的端粒长度;Western blot检测细胞中P16和RB蛋白的表达。结果 Rg1在体外可明显抑制K562细胞增殖,其最佳作用浓度及作用时间分别为20μmol/L和48 h;经20μmol/L Rg1诱导48 h的K562细胞与常规培养对照组相比,细胞出现G2/M期阻滞(P<0.05),集落形成能力明显减弱(P<0.05),SA-β-Gal染色阳性率增加(P<0.01),细胞体积增大,且溶酶体,线粒体体积增大,数目增多,端粒缩短加速(P<0.05),P16和RB蛋白表达上调(P<0.05)。结论 Rg1能诱导人红白血病K562细胞株衰老,可能与Rg1激活了p16-Rb信号通路有关。     

Rapid and Sensitive Determination of Timosaponin AIII in Rat Plasma by LC-MS/MS and Its Pharmacokinetic Application

A rapid sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for determination of timosaponin AIII (TA-III) in rat plasma, using ginsenoside Re as an internal standard (IS). TA-III and the IS were detected in MRM mode with a negative ionization electrospray mass spectrometer. The calibration curves were linear over the concentration ranges from 11.14 to 1114 ng/mL and the lower limit of quantification (LLOQ) was 11.14 ng/mL. Intra-day and inter-day precisions (RSD) were within 10%, and accuracy ranged from 6.4% to 9.1%. The extraction recovery at three concentrations ranged from 92.3% to 95.5%. The validated method was successfully applied to monitor the concentrations of TA-III in rat plasma after intragastric administration. The best fit pharmacokinetic model to estimate the pharmacokinetic parameters was a single compartment model with weight of 1/x² for oral administration groups of rats for TA-III.