酸碱度对人参皂苷提取率及其稳定性影响分析

目的考察酸碱度对人参皂苷浸渍提取及其稳定性的影响。方法采用高效液相色谱法测定人参皂苷总含量,以人参皂苷Rg1、人参皂苷Re的含量为考察指标,选用不同酸碱度的乙醇浸渍提取法提取人参皂苷,在加速与常温条件下考察人参皂苷稳定性。结果乙醇pH值为8.7时,人参皂苷提取率最高;人参制剂最稳定的pH值为5.8。结论调节提取溶剂及制剂的酸碱度,可有效提高人参皂苷的稳定性。     

不同参类中人参皂苷Rg_1、Re、Rb_1的质量测定

目的:建立超高效液相色谱法(UPLC)同时测定人参、红参、西洋参中人参皂苷Rg1、Re、Rb1质量方法。方法:采用AcquityUPLCBEHC18色谱柱(2.1mm×100mm,1.7μm),以乙腈-水为流动相进行梯度洗脱,流速为0.5mL/min,检测波长为203nm,柱温为40℃。结果:人参、红参、西洋参中人参皂苷Rg1、Re、Rb1在14min内得到完全分离并进行了质量测定。结论:应用UPLC法能够同时测定人参、红参、西洋参中人参皂苷Rg1、Re、Rb1质量,大大提高了人参皂苷类成分的分析速度,减少了溶剂消耗,为人参皂苷类成分质量测定提供了参考。     

HPLC-MS/MS法同时测定三七须根总皂苷中人参皂苷Rg_1和Re的含量

目的:建立以液-质联用法同时测定三七须根总皂苷中人参皂苷Rg1和Re含量的方法。方法:色谱柱为BDS HYPERSUL C18(150mm×2.1mm,5μm),流动相为乙腈-水(梯度洗脱),柱温为30℃;采用负离子多反应监测方法(MRM)测试。用于定量分析的对照品离子对分别为人参皂苷Rg1:m/z799.6→475.5;人参皂苷Re:m/z945.8→783.7;内标物紫杉醇m/z:852.5→525.3。结果:人参皂苷Rg1、人参皂苷Re标准曲线的线性范围分别为0.173～17.3μg·mL-1和0.156～39.1μg·mL-1,精密度和准确度等均符合样品分析的要求。结论:本方法准确、灵敏、特异性强,适用于三七须根总皂苷及其制剂中人参皂苷Rg1、Re含量的同时测定。     

高效液相色谱法测定律复康胶囊中人参皂苷Re和Rg1的含量

目的建立律复康胶囊中红参人参皂苷Rg1与Re的含量测定方法。方法采用HPLC法本品红参中人参皂苷Rg1、人参皂苷Re。Diamonsil C18柱色谱柱,流动相为乙腈-0.05%磷酸水（21.5：79.5）,检测波长203nm。结果人参皂苷Rg1、Re在范围内呈良好线性,分别为2.33-20.95μg（r=0.9994n=5）,0.972-8.748μg（r=0.9992,n=5）,平均回收率分别为100.2%（RSD=1.9%）、97.1%（RSD=0.8%）。结论HPLC方法简便、准确,精密度高、重现性好,适用于含人参皂苷Rg1、Re成分产品含量的测定。     

高效液相色谱法测定参附注射液中人参皂苷3组分的含量

目的建立测定参附注射液中人参皂苷Rg1、Re和Rb1含量的高效液相色谱法。方法采用反相高效液相色谱法进行梯度洗脱,固定相:L ichrospher C18色谱柱;流动相A:水;流动相B:乙腈;流速:1.2 mL.m in-1;检测波长:203 nm;柱温:35℃。结果人参皂苷Rg1、Re和Rb1分别在4~400,4~400和8~800μg.mL-1范围内峰面积与浓度呈良好的线性关系。平均加样回收率分别为101.92%~106.79%(人参皂苷Rg1),94.78%~100.65%(人参皂苷Re),和103.04%~106.62%(人参皂苷Rb1)。结论该方法简便、快速、准确,可同时测定参附注射液中人参皂苷Rg1、Re、Rb1的含量。     

超高效液相质谱法测定增强免疫类中药保健食品中多种功能成分含量

目的：采用超高效液相色谱-串联质谱法(Ultra Performance Liquid Chromatography-tandem mass spectrometry,UPLC-MS/MS)建立了同时测定增强免疫类中药保健食品中葛根素、红景天苷、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、金丝桃苷等功效成分的含量测定方法。方法:样品通过甲醇超声提取,色谱柱分离,水-0.1%甲酸乙腈梯度洗脱,采用负离子多反应监测(multiple reaction monitoring, MRM)模式进行定量定性分析。结果:6种目标化合物在13 min内完成分离且1～100 ng/mL范围内均呈现良好的线性关系（r>0.9979）,0.55、0.60和0.65μg/kg添加水平的平均回收率为83.0%～107.0%,实验室内变异系数为0.74%～3.71%(n=7),方法检出限为0.06～0.44μg/kg。结论:该方法快速准确,灵敏度高,专属性强,适用于同时测定中药保健食品中葛根素、红景天苷、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、金丝桃苷等具有增强免疫力功效成分的含量,可为具有增强免疫...     

反相高效液相色谱法测定含红参组方注射液中人参皂苷3组分含量

目的 测定不同含红参组方注射剂中人参皂苷Rg1、Re和Rb1 的含量。方法 采用反相高效液相色谱法进行梯度洗脱，色谱条件：汉邦 Lichrospher C18色谱柱；流动相A为水，流动相B为乙腈；流速为1.2 mL·min-1；检测波长为203 nm；柱温35 ℃。结果 人参皂苷Rg1、Re和Rb1在4～400，4～400和8～800 μg·Ml^-1 范围内峰面积与其浓度具有良好的线性关系。平均加样回收率分别为97.10 %～108.91% （人参皂苷Rg1），94.78 %～103.00%（人参皂苷Re），和 102.71%～106.83%（人参皂苷Rb1）。结论 该方法简便、快速、准确，可同时测定含红参组方注射剂中人参皂苷Rg1、Re和Rb1的含量。     

正交试验法优选复方参术健脾口服液中药材的提取工艺研究

目的:探讨复方参术健脾口服液中药材的最佳提取工艺.方法:采用正交试验法,以提取液中人参皂苷Rg1、人参皂苷Rb1、人参皂苷Re的总含量以及总固体得率为指标,综合评分优选药材的提取工艺.结果:药材较优的提取工艺为加水8倍,提取3次,每次1h.结论:优选的提取工艺各活性部位提取率高,操作简便,适用于生产.     

四君子汤和理中丸方中人参皂苷的含量测定

目的:测定四君子汤和理中丸中人参皂苷Rg1、Re、Rb1含量。方法:采用高效液相色谱法测定,ZobaxSB-C18(4.6mm×150mm,5μm)色谱柱;流动相为A-乙腈,B-水,梯度洗脱(0min:16%A;30min:19.3%A;33min:30%A;53min:35%A);流速为1.0mL.min-1;检测波长为203nm。结果:四君子汤和理中丸中人参皂苷Rg1含量分别为0.953,0.957mg.g-1,人参皂苷Re分别为1.10,1.09mg.g-1,人参皂苷Rb1分别为0.992,0.975mg.g-1。结论:两复方中人参皂苷Rg1、Re、Rb1含量无明显差异。     

人参皂苷Rg1的提取纯化及其含量测定

目的:提取和纯化人参皂苷Rg1,并测定其含量。方法:采用超声法提取人参总皂苷,使用硅胶柱层析法分离、纯化人参皂苷Rg1,采用高效液相色谱法测定人参皂苷Rg1含量。结果:采用硅胶柱层析分离50g人参,可得到人参皂苷Rg19.91g,其纯度为89.63%。结论:此提取方法简单、准确,成本较低,所获产品纯度较高,可作为获得人参皂苷Rg1的有效方法。     

UPLC法测定人参总皂苷提取物中7种人参皂苷的含量

目的建立一种同时测定人参总皂苷提取物中7种人参皂苷的超高效液相色谱分析方法,该方法对人参提取物的质量评价更准确、更快捷。方法采用Waters Acquity UPLC BEH C18色谱柱(100mm×2.1mm,1.7μm);乙腈-水为流动相;检测波长:203nm;流速:0.4mL·min~(-1);测定人参皂苷Rg1、人参皂苷Re、人参皂苷Rf、人参皂苷Rb1、人参皂苷Rb2、人参皂苷Rc和人参皂苷Rd的含量。结果测定的各色谱峰均能达到基线分离、分离度大于1.5,7种人参皂苷在各自的范围内有良好的线性关系,且RSD值符合要求。结论 UPLC能代替HPLC测定人参皂苷的含量,该方法灵敏、简单、准确、重复性好。     

Ginsenoside Rh2 reduces ischemic brain injury in rats

Ginseng was incubated under mildly acidic conditions and its inhibitory effect on a rat ischemia-reperfusion model was investigated. When ginseng was treated with 0.1% hydrochloric acid at 60 degrees C, its protopanaxadiol saponins were transformed to diasteromeric ginsenoside Rg3 and Delta20-ginsenoside Rg3. When the transformed ginseng extract, of which the main component was ginsenosides Rg3, was treated with human intestinal microflora, the main metabolite was ginsenoside Rh2. Orally administered acid-treated ginseng (AG) extract and ginsenoside Rh2 potently protect ischemia-reperfusion brain injury. The ginsenoside Rh2 also inhibited prostaglandin-E2 synthesis in lipopolysaccharide-stimulated RAW264.7 cells, but showed no in vitro antioxidant activity. These results suggest that AG and ginsenoside Rh2 can improve ischemic brain injury.     

Comparison of the pharmacological effects of Panax ginseng and Panax quinquefolium

Medical application of Panax ginseng was first found in "Shen-Nong Herbal Classic"around 200 AD Panax quinquefolium was first introduced in "Essential of Materia Medica" in 1694 in China. The most important bioactive components contained in P ginseng and P quinquefolium are ginseng saponins (GS). The contents of ginsenoside Rb1, Re, and Rd in P quinquefolium are higher than they are in P ginseng. In P ginseng, the contents of Rg1,Rb2, and Rc are higher than they are in P quinquefolium. P ginseng had a higher ratio of Rg1: Rb1, and which was lower in P quinquefolium. After steaming for several hours, the total GS will decrease. However, some ginsenosides (Rg2, 20R-Rg2, Rg3, Rh1 and Rh2) increase, while others (Rb1, Rb2, Rb3, Rc, Rd, Re, and Rg1) decrease. However, variation, especially in P quinquefolium, is high. P ginseng and P quinquefolium are general tonics and adaptogens. Rg1 and Rb1 enhance central nervous system (CNS) activities, but the effect of the latter is weaker. Thus, for the higher contents of Rg1, P ginseng is a stimulant, whereas the Rb1 contents of P quinquefolium are mainly calming to the CNS. Re, Rg1, panaxan A and B from P ginseng are good for diabetes. Re and Rg1 enhance angiogenesis, whereas Rb1, Rg3 and Rh2 inhibit it. Rh2, an antitumor agent, can be obtained from Rb1 by steaming. The content of Re in P quinquefolium are higher than in P ginseng by 3-4 times. The vasorelax, antioxidant, antihyperlipidemic, and angiogenic effects of Re are reported. Thus, for the CNS "hot," wound healing and hypoglycemic effects, P ginseng is better than P quinquefolium. For anticancer effects, P quinquefolium is better.     

Transformation of ginseng saponins to ginsenoside rh<Subscript>2</Subscript> by acids and human intestinal bacteria and biological activities of their transformants

When ginseng water extract was incubated at 60 degrees C in acidic conditions, its protopanaxadiol ginsenosides were transformed to ginsenoside Rg3 and delta20-ginsenoside Rg3. However, protopanaxadiol glycoside ginsenosides Rb1, Rb2 and Rc isolated from ginseng were mostly not transformed to ginsenoside Rg3 by the incubation in neutral condition. The transformation of these ginsenosides to ginsenoside Rg3 and delta20-ginsenoside Rg3 was increased by increasing incubation temperature and time in acidic condition: the optimal incubation time and temperature for this transformation was 5 h and 60 degrees C resepectively. The transformed ginsenoside Rg3 and delta20-ginsenoside Rg3 were metabolized to ginsenoside Rh2 and delta20-ginsenoside Rh2, respectively, by human fecal microflora. Among the bacteria isolated from human fecal microflora, Bacteroides sp., Bifidobacterium sp. and Fusobacterium sp. potently transformed ginsenoside Rg3 to ginsenoside Rh2. Acid-treated ginseng (AG) extract, fermented AG extract, ginsenoside Rh2 and protopanaxadiol showed potent cytotoxicity against tumor cell lines. AG extract, fermented AG extract and protopanaxadiol potently inhibited the growth of Helicobacter pylori.     

人参叶中三萜皂苷类成分的研究

目的 研究人参叶中的微量皂苷类化合物.方法 利用正相硅胶柱层析和反相硅胶制备色谱技术,分离人参叶中的微量成分,并运用质谱和核磁共振技术鉴定其结构.结果 从人参叶中分离得到5个微量成分,分别鉴定为:人参皂苷Rk1(Ⅰ)、Rk3(Ⅱ)、Rh4(Ⅲ)、RgS(Ⅳ)和Rg6(Ⅴ).结论 化合物Ⅰ～Ⅳ为首次从人参叶中分离得到.     

人参与附子配伍后不同制备条件下人参皂苷含量变化的研究

目的:研究人参与附子配伍后不同制备条件对人参皂苷Rg1、Re、Rb1含量的影响。方法:采用高效液相色谱法测定人参配伍附子后水煎液、浓缩膏及不同温度下的干燥细粉中人参皂苷的含量。结果:人参与附子配伍后,浓缩、干燥是人参皂苷类成分损失最多的制备工序。结论:人参与附子配伍浓缩、干燥温度不宜高于80℃。     

Pharmacokinetic study of ginsenoside Re with pure ginsenoside Re and ginseng berry extracts in mouse using ultra performance liquid chromatography/mass spectrometric method

Ginsenoside Re is the major ginsenoside in ginseng berry(GB) extract and its pharmacokinetics were studied following the intravenous and oral administration of pure Re or ginseng berry extract in mouse with doses of 10 and 50 mg/kg using ultra performance liquid chromatography mass spectrometric (UPLC/MS) method which can simultaneously determine ginsenoside Re, Rg1 and Rh1 in mouse serum. The serum samples were pretreated by protein precipitation and chromatographic separation was performed on AQUITY UPLC BEH C₁₈ column using gradient elution with the mobile phase of 5 mM ammonium formate and acetonitrile. Analytes and digoxin (I.S.) were analyzed and identified using an electrospray negative ionization mass spectrometry in the selected ion monitoring mode with the linear concentration range of 5.0–5000 ng/mL and lower limits of detection (LLOD) under 2.5 ng/mL. Ginsenoside Re was rapidly cleared from the body with a short half-life (0.2 ± 0.03 h for male and 0.5 ± 0.08 h for female mice after i.v.) and oral absorption was generally poor (F% 0.19–0.28). Notably, GB extract showed a superior oral absorption of ginsenoside Re (F% 0.33–0.75) at equivalent ginsenoside Re dose to pure ginsenoside Re, indicating that GB extract might be a good form for ginsenoside Re intake.     

HPLC指纹图谱法测定不同粉碎度野山花旗参粉末中人参皂苷含量

目的：建立高效液相色谱法（HPLC）指纹图谱法,测定野山花旗参100目、200目、1000目粉三种粉末中人参皂苷Re、Rb1、Rg1的总含量,通过指纹图谱相似度比较,考察粉碎对野山花旗参中人参皂苷的影响。方法：采用梯度洗脱、HPLC测定三种粒径粉末的HPLC指纹图谱,计算主要药效成分人参皂苷Re、Rb1、Rg1的含量,并进行比较,通过“中药色谱指纹图谱相似度评价系统”评价三个样品的相似度。结果：野山花旗参100目、200目、1000目粉的主要有效成分人参皂苷Re、Rb1、Rg1的总含量均大于3.96%,三种粉末的指纹图谱重叠率高,共有峰的相对保留时间吻合,与生成的对照指纹图谱相似度分别为0.946,0.979,0.970。结论：野山花旗参100目、200目与1000目粉的HPLC指纹图谱相似度大于0.946,化学成分的种类没有变化,主要有效成分人参皂苷Re、Rb1、Rg1的总含量基本相等,野山花旗参超微粉碎成1000目粉,化学成分保持不变。     

Preparation of monoclonal antibody against ginsenoside Rf and its enzyme immunoassay

A rapid and sensitive indirect competitive enzyme immunoassay method has been developed for quantitating ginsenoside Rf (Rf) in crude total Panax ginseng saponins and in rat plasma using high titer mouse monoclonal antibody (mAb) raised against a conjugate of Rf and bovine serum albumin (BSA). The isotype of mAb against Rf was IgG3 with a K chain. The presence of Rf inhibited the binding of the mouse anti-Rf mAb to a Rf-BSA solid phase coating antigen. The working range was 0.01-10 ng/assay and detection limits were 20 pg in various ginseng extract fractions or 34 pg in rat plasma per assay. The anti-Rf mAb cross-reacted with ginsenoside Rg2 by 57.5%, but not with other ginsenosides. However, this anti-Rf mAb did not cross-react with BSA or cellubiose, which is a carbohydrate component of Rf. Using this standard curve, we could measure the amount of Rf in ginseng total extract, ginseng total saponins, protopanaxadiol saponins, and propanaxatriol saponins. We could also measure the amount of Rf in rat plasma after the oral administration of Rf and found that Rf reached a maximum level in rat plasma after 16 h. These results indicate that the anti-Rf mAb could be useful for the quantitation of Rf in crude ginseng fractions and in body fluids.     

Ginseng and ginsenoside Rg3, a newly identified active ingredient of ginseng, modulate Ca2+ channel currents in rat sensory neurons

There is increasing evidence that ginseng influences pain modulation. In spite of extensive behavior studies, the detailed mechanism of ginseng actions at the cellular level and the identity of the active substance have not been elucidated yet. Whole-cell patch-clamp recordings were used to examine the modulation of high-voltage-activated Ca2+ channel currents by ginseng total saponins and its various individual ginsenosides in rat dorsal root ganglion neurons. Application of ginseng total saponins suppressed Ca2+ channel currents in a dose-dependent manner. Occlusion experiments using selective blockers revealed that ginseng total saponins could modulate L-, N-, and P-type currents. The co-application of ginseng total saponins and the gamma-opioid receptor agonist, D-Ala(2), N-MePhe(4), Gly(5)-ol-enkephalin (DAMGO), produced non-additive effects in most cells tested and each effect was significantly relieved by a depolarizing prepulse. Overnight treatment of cells with pertussis toxin profoundly reduced the inhibition. Furthermore, we now report that ginsenoside Rg3, among the major fractions of ginseng saponins, is a newly identified active component for the inhibition. These results suggest that the modulation of Ca2+ channels by ginseng total saponins, in particular by ginsenoside Rg3, could be part of the pharmacological basis of ginseng-mediated antinociception.