人参中人参皂苷的直接高压微波辅助降解

采用高效液相色谱-电喷雾质谱联用法测定了人参提取液中的人参皂苷. 考察了天然人参皂苷发生降解的条件, 同时研究了单体人参皂苷Rg1, Re, Rb1, Rc, Rb2和Rd的降解, 并对降解产物进行了分析. 结果表明, 随着提取压力的升高, 提取液中天然人参皂苷的含量逐渐减少, 同时产生多种次级人参皂苷. 当微波提取压力达到600 kPa, 提取时间为10 min时, 提取液中的主要天然人参皂苷达到完全降解, 次级人参皂苷Rg3含量达到最高. 在单体人参皂苷Rb1, Rc, Rb2和Rd的降解产物中均得到人参皂苷Rg3.     

基于多壁碳纳米管的人参皂苷快速分离

将多壁碳纳米管(MWCNTs)作为选择性吸附材料,用于快速分离人参提取物中的人参皂苷.人参经甲醇溶液超声提取后,提取物中主要为人参皂苷和糖类.人参中的糖类与人参皂苷的极性相近,是提取分离人参皂苷时的主要干扰物. MWCNTs可以快速吸附和脱附人参皂苷,但是对糖类无吸附作用.利用其选择性吸附性能,建立了MWCNTs快速分离人参提取物中人参皂苷和糖类的方法.在优化的分离条件下,MWCNTs对人参提取物中糖类的分离度高于90%,对8种主要人参皂苷[Rb1,Rb2,Rc,Rd,Re,Rg1,20(S)-Rf和Ro]的吸附容量为15. 0~24. 0μg/mg,回收率高于90%.进一步研究表明,人参皂苷在3 min内即可达到吸附和脱附平衡,并且人参皂苷的回收率受脱附溶剂极性的影响.相比于常规材料大孔树脂,MWCNTs可以更快速、简便地分离人参皂苷.     

人参皂苷Rb1在大鼠体内的药物代谢研究

人参皂苷Rb1是人参中的达玛烷型三萜皂苷类化合物, 具有多种生物活性. 对人参皂苷Rb1代谢产物的分析已有报道, 在大鼠尿液、粪便、胃和大肠中共检出了5种代谢产物. 本文采用高效液相色谱-飞行时间串联质谱进行人参皂苷Rb1的体内代谢研究, 通过口服和静脉给予药物, 在大鼠尿液中共检出了人参皂苷Rb1的14种代谢产物, 并系统分析和推断了这些代谢物的转化规律和可能结构.     

田七花提取物中22种皂苷类成分的液相色谱-质谱测定

在甲酸体系中以高效液相色谱负离子模式电喷雾电离质谱以及碰撞诱导裂解技术研究了12种人参皂苷(Re、Rg1、Rg2、Rg3、Rf、Rb1、Rb2、Rb3、Rc、Rd、Rh1 和Rh2).结果表明,应用皂苷化合物(包括人参皂苷、田七皂苷和绞股蓝皂苷)的质谱及裂解规律可在缺少相应对照品的情况下对其进行可靠的鉴定.在此基础上,对田七花样品以加压溶剂萃取法提取,然后以LC-MS/MS分析,从中鉴定出22种皂苷,其中六糖皂苷Ⅰ和Ⅱ、乙酰基Rb1为首次报道,并且定量测定了其中10种皂苷的含量.     

在线二维柱切换高效液相色谱法同时测定牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1

使用双梯度液相色谱系统紫外检测器,建立了二维液相色谱法全自动快速同时测定牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1的含量。样品经超声提取后,以Syncronis C18为一维分析柱,ODS C18为二维分析柱,利用一维色谱柱完成三七皂苷R1和人参皂苷Rb1分离测定以及人参皂苷Rg1和人参皂苷Re的净化;利用二维色谱柱完成人参皂苷Rg1和人参皂苷Re的分析。一维分析和二维分析均以乙腈-水体系作为流动相,梯度洗脱,检测波长为203 nm,整个分析过程仅需30 min。三七皂苷 R1、人参皂苷 Rg1、Re 和 Rb1在0.5~200 mg/L范围内线性良好,相关系数R2分别为0.9994,0.9996,0.9995和0.9994,平均回收率均在86.4％~95.1％之间。本方法简便快速,测定结果准确可靠,可用于牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1含量的测定。     

HPLC-MS结合多元统计分析区分人参产地及筛选皂苷类标志物

利用高效液相色谱-质谱联用(HPLC-MS)技术结合多元统计分析方法, 区分中国人参主产区5个不同产地的45个人参样本, 筛选出差异性皂苷类标志物. 根据人参总皂苷在反相C₁₈色谱柱中的洗脱顺序, 结合串联质谱分析和标准品比对, 在提取的人参总皂苷中鉴定出15种原人参三醇型、 24种原人参二醇型和2种齐墩果酸型共41种皂苷. 对人参总皂苷的HPLC-MS全扫描数据进行了多元统计分析. 正交偏最小二乘-判别分析(OPLS-DA)结果表明, 所建立的分析模型具有良好的数据描述能力和预测能力. 所有人参样本能够根据产地被区分, 并筛选得到同时区分5个产地的差异性皂苷类组分18种; 能够区分任意2个产地人参样本的差异性组分主要为在人参中含量较高的人参皂苷Rb1, Rg1, Re, Rc, Rd, Ro和m-Rb1等. 分层聚类分析(HCA)结果显示, 黑龙江和吉林两省的样本能够独自聚类, 但是绥化市的样本更接近于吉林省. 初步推断原因为绥化市地理位置较接近吉林省, 两地人参生长环境相似并可能存在种质资源交换.     

高效液相色谱法测定竹节参中多种人参皂苷含量

建立了高效液相色谱法(HPLC)测定竹节参中人参皂苷Rg1、Re、Rb1、Rb2、Rg2、Rd含量的方法.运用二极管阵列检测器(DAD)峰纯度和光谱检索功能,结合保留时间定性,外标峰面积法定量.采用C18反相柱,以乙腈-水梯度洗脱测定了同一批竹节参总皂苷中人参皂苷Rg1、Re、Rd的含量分别为0.81%、0.15%、2.99%,回收率为93.46%~94.02%,含量及回收率的RSD均小于5%,该方法简便、灵敏,精密度及准确度在允许范围内,可作为竹节参皂苷提取物中多种人参皂苷的同时测定方法.     

密闭式微波降解法促进常见人参皂苷向稀有人参皂苷转化的规律

采用密闭微波技术对7种常见人参皂苷单体(Rb1,Rb2,Rb3,Rc,Rd,Re和Rg1)进行降解,通过高效液相色谱(HPLC)分析并与相同条件下非微波降解物对比,研究了密闭微波降解人参皂苷的产物在化学结构及组成上的变化规律,以期快速、高效地制备生物活性高的稀有人参皂苷.结果表明,密闭式微波降解法能够使常见人参皂苷基本降解完全,而相同条件下非微波降解法则基本不发生降解.原人参二醇型人参皂苷易水解掉C20位糖,并发生C20位构型变化,生成20(R)-Rg3和20(S)-Rg3,其中20-(R)为优势构型,C20位羟基进一步脱水产生稀有人参皂苷Rk1和Rg5.同时,20(S/R)-Rg3失去C3位的1分子葡萄糖转化为20(S/R)-Rh2,C20位羟基再进一步脱水生成了Rk2和Rh3.此外,人参皂苷C20位所连的糖种类与构型影响了降解产物中各稀有皂苷的组成与比例,但7种原人参二醇型人参皂苷密闭式微波降解产物中Rg5含量均为最高.密闭式微波降解对原三醇型人参皂苷的转化作用与原二醇型人参皂苷具有相似的规律,人参皂苷Re和Rg1的密闭式微波降解产物中Rh4含量均为最高.本文结果进一步说明在相同的降解条件下,密闭式微波降解法的降解效率远高于高温高压非微波降解法,密闭式微波降解可明显促进常见人参皂苷向稀有人参皂苷转化,因此采用密闭微波技术对常见人参皂苷进行降解可以大量获得稀有人参皂苷.     

泡沫浮选法分离富集三七中的4种人参皂苷

采用泡沫浮选法对三七提取液中的人参皂苷Rg1、Re、Rb1和Rd进行了分离富集,并用高效液相色谱法分别测定了含量.考察了浮选液浓度、浮选时间、浮选液pH值、氮气流速和电解质NaCl浓度对浮选效率的影响.结果表明:泡沫浮选法对4种皂苷均有较好的分离富集效果,尤其是对人参二醇型皂苷(Rb1,Rd)效果更为明显.当浮选液浓度为2.0 mg/mL,pH值为2～3,氮气流速为20 mL/min,浮选时间10 min,电解质氯化钠浓度0.20 mol/L,泡沫浮选效果最佳.     

电喷雾质谱法研究人参皂苷Rb1和Rd与细胞色素c的非共价复合物

用质谱法研究了人参皂苷Rb1和Rd与细胞色素c的非共价相互作用, 证明主-客体之间形成了多种化学计量比的复合物, 用直接计算的方法得出人参皂苷Rb1和Rd与细胞色素c形成的非共价复合物的各级解离常数KD, 用竞争体系验证了计算结果, 二者结果相互吻合.     

固相萃取-超高效液相色谱-串联质谱法检测保健食品中9种人参皂苷

建立了以固相萃取结合超高效液相色谱-串联质谱(UPLC-MS/MS)同时检测保健食品中9种原人参二醇型和原人参三醇型人参皂苷的方法.保健食品中人参皂苷经过提取后,通过Alumina-N/XAD-2 SPE柱净化,在Hypersil Gold C18色谱柱(100 mm×2.1 mm,1.9μm)上分离,利用乙酸铵溶液(...     

Studies on Intestinal Transport of Ginsenoside Compatibility with Veratrum nigrum via Caco-2 Cell Monolayer Model Coupled with UPLC-ESI-MS Method

The Caco-2 cells have been recognized as effective tools to be applied to imitating the drug absorption in human intestine for the transport of drug. Herein, Caco-2 cell monolayer model was used to study the transport of the ginsenoside compatibility with Veratrum nigrum in different proportions. A specific high performance liquid chromatography-electrospray ionization-mass spectrometry(HPLC-ESI-MS) method was developed for the semiquantitative determination of ginsenoside in intestinal transport with Dioscin as an internal standard. For the Caco-2 model constructed, two influencing factors were investigated, including time and concentration. The results suggest that the absorption of ginsenoside Re, Rg1, Rb1, Rc, Rb2 and Rd are time- and concentration-dependent and the excretions of Rb1, Rc, Rb2 and Rd have a relatronship with some transport proteins. The bioavailability of the ginsenosides has reduced compared to the single Panax ginseng extract when compatibility with a certain amount of Veratrum nigrum.     

Determination of Seven Major Ginsenosides in Different Parts of Panax quinquefolius L.(American Ginseng) with Different Ages

Ginsenosides Rg1,Re,Rb1,Rc,Rb2,Rb3,and Rd in different parts of the American ginseng plant were investigated.The extraction process was a pressurized microwave-assisted extraction(PMAE).The seven ginsenosides were separated and determined by high-performance liquid chromatography(HPLC) with a ultraviolet(UV) detector,at 203 nm.The experiment results showed significant variations in the individual ginsenoside contents of the American ginseng in different parts and ages of the plant.The results demonstrated that the leaves,root hairs,and rhizomes of Panax quinquefolius L.contained higher ginsenoside contents,followed by the main roots and stems.The leaves contained dramatically higher levels of ginsenoside Rg1,Rb3,and Rd than the other four parts.Higher contents of Rb1 and Re were present in the main roots,root hairs,and rhizomes.The amount of ginsenoside content in the stems was the lowest.The total content of the seven ginsenosides in main roots,root hairs and rhizomes increased with the age of the plant.In contrast,the ginsenoside contents in the leaves and stems decreased with a year of growth.     

动态泡沫浮选法分离富集人参提取液中的二醇型人参皂苷

采用动态泡沫浮选法分离富集人参提取液中的二醇型人参皂苷, 用高效液相色谱法测定6种人参皂苷Rg₁, Re, Rb₁, Rc, Rb₂和Rd的含量. 考察了浮选液pH值、电解质NaCl浓度、载气流量、料液浓度及料液流速对人参皂苷浮选率的影响, 确定了动态泡沫浮选的最佳条件, 并与溶剂提取法、溶剂浮选法以及静态泡沫浮选法进行了比较. 结果表明, 动态泡沫浮选法对二醇型人参皂苷Rb₁, Rc, Rb₂和Rd具有高富集效率, 回收率分别为93.3%, 98.6%, 96.9%和98.3%, 而对三醇型人参皂苷Rg1和Re的富集效率却很低, 回收率分别为4.8%和4.2%. 该法是分离纯化二醇型人参皂苷的一种简便有效的方法.     

Anti-allergic Effects of Ginsenosides Extracted by High Temperature and High Pressure Method

Ginseng(Panax ginseng C. A. Mey.) is a traditional medicinal herb in Asia. Studies have shown that ginsenosides significantly affect immunoregulation and rare ginsenosides have anti-allergic effects. In this research, a high temperature and high pressure method was utilized to increase the contents of rare ginsenosides in the ginseng extract(GE). The anti-allergic effects of this extract were investigated in vivo. Water was used as the extraction solvent in extracting the rare ginsenosides via the high temperature and high pressure method. Extraction time and temperature were investigated in order to increase the contents of rare ginsenosides. Rare ginsenosides were qualitatively analyzed by HPLC-ESI-MS and quantitatively analyzed by HPLC-UV. Anti-allergic effects of the extracts were assessed using the ovalbumin(OVA)-induced allergic asthma model in vivo. An extraction temperature of 145℃ and extraction time of 2.0 h were chosen as the optimal conditions. Compared with traditional method, the contents of total rare ginsenosides extracted were considerably higher using the new method, that is, 14.74 times that extracted by the traditional method. In our in vivo experiments, treatment with high concentration GE may have anti-allergic effects in decreasing the total amount of IgE in serum and IL-4 in bronchoalveolar lavage fluid(BALF), and in improving the ratio of CD4⁺ to CD8⁺ T cells. The high temperature and high pressure method was an effective method to obtain GE containing more rare ginsenosides, which maybe become anti-allergic agents.     

Isolation and determination of ginsenosides in American ginseng leaves and root extracts by LC-MS

Ginseng saponins (ginsenosides) were extracted from the root and leaves of locally cultivated American ginseng (Panax quinquefolium L.). For the isolation of compounds from plant samples three different extraction methods were utilized: accelerated solvent extraction, the ultrasound-assisted solvent extraction and mechanical shaking assisted solvent extraction. The separation of compounds was achieved with a water–acetonitrile gradient system using a C18 reversed-phase column. Target compounds were identified in MS² and MS³ experiments. The relative distribution of these ginsenosides in each root and leaf extract was established. The limit of detection of the method was less than 30 ng/ml. Recovery of ginseng saponins in spiked samples exceeded 80%, while the relative standard deviation ranged from 7.1 to 9.1%. The total concentrations of ginsenosides were 41 and 13 mg/g in root and leaves.     

Comparative study on chemical components and anti‐inflammatory effects of Panax notoginseng flower extracted by water and methanol

Methanol and water are commonly used solvents for chemical analysis and traditional decoction, respectively. In the present study, a high‐performance liquid chromatography with ultraviolet detection method was developed to quantify 11 saponins in Panax notoginseng flower extracted by aqueous solution and methanol, and chemical components and anti‐inflammatory effects of these two extracts were compared. The separation of 11 saponins, including notoginsenoside Fc and ginsenoside Rc, was well achieved on a Zorbax SB C18 column. This developed method provides an adequate linearity (r ² > 0.999), repeatability (RSD < 4.26%), inter‐ and intraday variations (RSD < 3.20%) with recovery (94.7–104.1%) of 11 saponins concerned. Our data indicated that ginsenoside biotransformation in PNF was found, when water was used as the extraction solvent, but not methanol. Specifically, the major components of Panax notoginseng flower, ginsenosides Rb1, Rc, Rb2, Rb3, and Rd, can be near completely transformed to the minor components, gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, notoginsenoside Fd, and ginsenoside F2, respectively. Total protein isolated from Panax notoginseng flower is responsible for this ginsenoside biotransformation. Additionally, methanol extract exerted the stronger anti‐inflammatory effects than water extract in lipopolysaccharide‐induced RAW264.7 cells. This difference in anti‐inflammatory action might be attributed to their chemical difference of saponins.     

反相高效液相色谱法测定人参皂甙Compound-K的含量

人参皂甙compound-K(C-K)在人参中的含量极低,但它是其他含量较高的人参皂甙Rb1和Rb2等在人体肠道内的主要 降解产物和最终吸收形式,具有很高的生物活性。采用反相高效液相色谱法测定了人参总皂甙发酵液中C-K的含量。色谱 条件为:反相C18柱;乙腈-水(体积比为48∶52)溶液为流动相,流速1.0 mL/min;紫外检测波长203 nm;柱温35 ℃;外标法 定量。结果表明:C-K的质量浓度为0.05~0.8 g/L时,其峰面积与质量浓度具有良好的线性关系,相关系数为0.9998。方法 的检测限(S/N=3)为2.5 mg/L,峰面积测定值的相对标准偏差(n=6)为2.20%。测定栽培人参总皂甙及三七茎叶总皂甙微生 物发酵液中C-K的平均加标回收率(n=3)分别为98.6%和99.7%。该方法快速简便,准确可靠,可用于C-K的制备研究及药物 开发。     

Determination of ginsenoside content in Asian and North American ginseng raw materials and finished products by high-performance liquid chromatography: single-laboratory validation

A single-laboratory validation study was conducted for the quantification of Rg1, Re, Rb1, Rc, Rb2, and Rd in Asian ginseng (Panax ginseng C.A. Meyer) and North American ginseng (Panax quinquefolius L.) raw materials and finished products by RP-HPLC. The extraction with aqueous methanol was optimized for whole root, powdered extract, and finished product (raw, tablet, and capsule matrixes) test articles. Root materials were treated with base to hydrolyze acidic malonyl ginsenosides to their neutral counterparts. Calibration curves for each ginsenoside were linear over the following ranges (microg/g): 5-394 for Rg1, 15-1188 for Re, 39-2981 for Rb1, 6-499 for Rc, 5-406 for Rb2, and 7-600 for Rd, all having a coefficient of determination (r2) of > or = 99.5%. The LOD for Rg1, Re, Rb1, Rc, Rb2, and Rd was determined to be 1.06, 1.25, 2.19, 1.24, 1.27, and 1.70 microg/mL, respectively. Quantitative determinations performed with eight test materials by two analysts over 3 days (n = 12) resulted in RSDr values that ranged from 1.11 to 7.61%.