Holistic quality evaluation of Panax notoginseng extract and injection using an UPLC-QTOF-MS^E based metabolomics approach

In the present study, we established an UPLC-QTOF-MSE based metabolomic approach in order to evaluate the holistic qualities and compare the quality difference by finding characteristic components of Panax notoginseng extracts （PNE） and Xuesaitong （XST） injection samples from different manufacturers. The data were processed through unsupervised principal component analysis （PCA） and supervised orthogonal partial least squared discrimination analysis （OPLS-DA） to compare the quality differences. Two-dimensional PCA score plots showed a tendency to separate the XST injections and extracts, and most XST injection samples were clearly clustered into two groups. Especially, the injections from He and YB companies were distinguished into two groups. In addition, only injection samples of Hu company were near the cluster of PNE. To explore the potential chemical components contributing most to the differences between XST injection samples from different manufacturers and PNE, an S-plot was constructed following the OPLS-DA. Ginsenoside Rd, ginsenoside Rgl, ginsenoside Re, ginsenoside Rbl, 20（S）-ginsenoside Rhl, gypenoside VII, ginsenoside Rg2, ginsenoside Rh4, ginsenoside Rkl or Rgs, notoginsenoside Fc, 20（R）-ginsenoside Rg3, ginsenoside F2 and protopanaxadiol were recognized as characteristic chemical markers that contributed most to reflect the difference between XST injections and PNE. Ginsenoside Rd, ginsenoside Rgl, ginsenoside Re, ginsenoside Rbl and gypenoside VII were revealed as index components contributing most to the differences of PNE and XST injections, and quantitative analysis of these components could ensure the consistent quality of XST injections. Based on the fact that the injections should be standardized with the characteristic components as quality control chemical markers, it is most important to keep the quality of extracts of raw materials stable and reliable.

Advances in Researches of Extraction, Separation, and Purification Technologies for Total Saponins of Panax notoginseng

Total saponins of Panax notoginseng have the functions of promoting blood circulation and removing phlegm, thus they have high medicinal value. There are many different extraction methods in the extraction and separation of total saponins of P. notoginseng . The extraction methods of total saponins of P. notoginseng are mainly divided into traditional extraction methods, modern extraction methods and compound extraction methods.

Advances in Researches of Extraction Processes of Total Saponins of Pnanx Notoginseng

Traditional extraction methods of total saponins of Panax notoginseng include cold soaking method,water decoction method,alcohol reflux method,percolation method,macroporous resin adsorption method,and accelerated solvent extraction( ASE) method. Modern extraction methods include ultrasonic extraction,microwave assisted extraction,supercritical CO_2 extraction,microbial fermentation assisted extraction,neural network model optimized extraction method,and multi-stage countercurrent extraction method. This paper discussed principles of these methods and compared their advantages and disadvantages.

Design of Traditional Chinese Medicine Extraction Workshop Process and Automation System

Objective: This paper takes the example of a Panax notoginseng extraction workshop and designs an automated production workshop with advanced domestic capabilities. Methods: 1) Based on the small-scale Panax notoginseng extraction process, the feasibility of the workshop production process is demonstrated. 2) The workshop process design for Panax Notoginseng saponin extraction is completed, including production organization plans and the selection of key equipment. 3) For the Panax notoginseng extraction workshop process, an automated production control system is designed. Conclusion: Through optimized design of the production process and automation system, continuous and automated production of traditional Chinese medicine extraction is achieved, leading to improvements in drug quality and production efficiency.

三七茎叶提取物对断奶仔猪生长性能、血清生化指标和肠道健康的影响

试验旨在研究三七茎叶提取物对断奶仔猪生长性能、血清生化指标和肠道健康的影响。选取26日龄断奶仔猪24头随机分为3组,每组8个重复,每个重复1头仔猪。对照组饲喂基础饲粮,低剂量组、高剂量组分别在基础饲粮中添加0.2%和0.4%的三七茎叶提取物。试验期21 d。结果显示,与对照组相比,高剂量组断奶仔猪平均日增重显著升高(P<0.05),低剂量组和高剂量组血清乳酸脱氢酶活性、谷草转氨酶活性和谷草/谷丙的比值显著降低(P<0.05);高剂量组断奶仔猪十二指肠、空肠、回肠的绒毛高度显著增加(P<0.05),十二指肠和回肠的绒隐比显著升高(P<0.05);低剂量组和高剂量组结肠白细胞介素-12 p35(IL-12 p35)、白细胞介素-12 p40(IL-12 p40)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和干扰素-γ(IFN-γ)的mRNA表达量显著降低(P<0.05)。研究表明,在饲粮中添加三七茎叶提取物可以改善断奶仔猪生长性能、肠道健康,添加0.4%三七茎叶提取物较为适宜。

三七药渣中多糖提取工艺优化及其动力学分析

以三七药渣为原料,探讨了超声辅助提取三七药渣中三七多糖的优化工艺,并对提取过程进行动力学分析。在单因素实验的基础上,利用BBD(Box-Behnken Design)响应面法优化工艺条件,得到最优提取条件为:超声温度60℃、超声功率136.8 W、超声时间53 min、液固比43 mL/g,此时多糖得率为6.79%。通过测定不同超声功率、不同超声时间下提取液中的多糖浓度,得到超声辅助提取三七药渣中三七多糖的提取速率常数、相对萃余率,验证了所推导的动力学模型,拟合得到了半衰期以及有效扩散系数回归方程。

不同提取方式对三七改善小鼠睡眠功能的影响

目的:观察不同提取方式对三七改善小鼠睡眠功能的影响。方法:通过直接睡眠试验、延长戊巴比妥钠睡眠时间试验、戊巴比妥钠阈下剂量催眠试验、巴比妥钠睡眠潜伏期试验,评价三七水提物、三七30%乙醇提取物、三七70%乙醇提取物对小鼠睡眠的影响。结果:与对照组比较,三七70%乙醇提取物各剂量组、三七30%乙醇提取物高剂量组、三七水提物高剂量组均可延长戊巴比妥钠诱导的小鼠睡眠时间(P<0.05或P<0.01),增加戊巴比妥钠阈下催眠剂量入睡小鼠只数(P<0.05或P<0.001),缩短小鼠的睡眠潜伏期(P<0.05或P<0.001)。以小鼠睡眠时间、入睡只数、睡眠潜伏期时间作为评估不同提取物对改善小鼠睡眠作用的主要指标。三七30%乙醇提取物低剂量组、三七水提物低剂量组与对照组比较改善小鼠睡眠作用不显著,与三七水提取物高剂量组比较三七70%乙醇提取物高剂量组对小鼠睡眠改善作用差异显著(P<0.05),三七30%乙醇提取物高剂量组以及三七70%乙醇提取物低剂量组改善小鼠睡眠作用无显著性差异(P>0.05)。三七70%乙醇提取物高剂量组、三七30%乙醇提取物高剂量组、三七水提物高剂量组小鼠下丘脑及血清中的5-羟色胺(5-HT)及γ-氨基丁酸(GABA)含量均显著高于对照组(P<0.05或P<0.01),多巴胺(DA)含量均显著低于对照组(P<0.05或P<0.01)。结论:三七水提物、三七30%乙醇提取物、三七70%乙醇提取物均可在一定程度上改善小鼠的睡眠,其中三七70%乙醇提取物效果最佳。

Smashing Tissue Extraction and HPLC Determination of Active Saponins from Different Parts of Panax notoginseng

Objective To optimize the extraction technology used for extracting active saponins from the roots,fibrous roots,basal part of stems,root verrucae,fruits,flowers,stems,and leaves of Panax notoginseng based on the contents of ginsengsides Rg1,Rb1,and notoginsengside R1 as evaluation indexes.Methods Different parts of P.notoginseng were extracted by smashing tissue extraction(STE),ultrasound extraction,and reflux extraction.The contents of ginsengsides Rg1,Rb1,and notoginsengside R1 in 24 kinds of extracts were determined by HPLC-UV.Hypersil C18 column(200 mm × 4.6 mm,5 μm) and acetonitrile-warter(20:80 for 30 min→45:55 for 18 min→70:30 for 2 min→80:20 for 10 min→100:0) were used;UV detector was set at 203 nm;The flow rate was set at 1.0 mL/min.Results STE was the most efficient technology with the highest yield of active saponins among the three tested extraction technologies.Conclusion STE is a fast,effective,and economical method to extract the active saponins from different parts of P.notoginseng.It could significantly shorten the extraction time and simplify the determination of the pre-processing work on identifying P.notoginseng.Such quick and effective extraction provides a powerful tool for analyzing P.notoginseng in the future.

三七根提取物的表面活性及其复配体系的性能研究

对三七根提取物的表面活性和复配增效性能进行研究。结果表明,三七根提取物在25℃的cmc为0.1 g/L,对应的γ_(cmc)为43.67 mN/m。在盐度质量分数范围为0%~1%,其表面活性随盐度的增加而增加。在pH范围为4.0~8.0时,三七根提取物的表面张力随pH的降低而降低。根据Rosen理论推算出三七根提取物与聚氧乙烯失水山梨醇月桂酸酯(Tween 20)的最佳复配质量比为4∶1,二元复配体系形成胶束的能力比理论结果增效41.18%。乳化力测试结果显示,三七根提取物与Tween 20复配后的乳化能力和乳化稳定性均与单一使用Tween 20接近,表明三七根提取物与Tween 20的复配具有较好的协同增效能力。而三七根提取物对Tween 60无协同增效作用。

三七多糖的研究进展

三七是我国传统的名贵中草药,药食两用,补血第一。三七多糖具有抗氧化、抗肿瘤、抗炎症、保护肝脏、活血化淤、消肿镇痛、降糖降脂和免疫调节等活性作用。对三七多糖的提取工艺、测量方法、分子结构、药理活性进行了综述,为进一步研究三七及提高综合利用率提供了理论依据。

Combination of Total Astragalus Extract and Total Panax Notoginseng Saponins Strengthened the Protective Effects on Brain Damage through Improving Energy Metabolism and Inhibiting Apoptosis after Cerebral Ischemia-Reperfusion in Mice

Objective： To explore the effects and molecular mechanisms of the combination between total Astragalus extract （TAE） and total Panax notoginseng saponins （TPNS） against cerebral ischemia- reperfusion injury. Methods： C57BL/6 mice were randomly divided into sham-operated group, model group, TAE （110 mg/kg） group, TPNS （115 mg/kg） group, TAE-TPNS combination group and Edaravone （4 mg/kg） group, treated for 4 days, then, cerebral ischemia-repeffusion injury was established by bilateral common carotid artery （CCA） ligation for 20 min followed by reperfusion for 1 and 24 h. Results： TPNS could increase adenosine triphosphate （ATP） level, TAE and TAE-TPNS combination increased ATP, adenosine diphosphate （ADP） contents and Na＋-K＋-ATPase activity, and the effects of TAE-TPNS combination were stronger than those of TAE or TPNS alone after reperfusion for 1 h. After reperfusion for 24 h, TAE, TPNS and TAE-TPNS combination significantly increased neurocyte survival rate and decreased the apoptosis rate as well as down-regulated the expression of phosphorylated c-June N-terminal kinasel/2 （p-JNK1/2）, cytochrome C （Cyt C）, cysteine aspartic acid-specific protease （Caspase）-9 and Caspase-3. Furthermore, the effects in TAE-TPNS combination were better than those in TAE or TPNS alone. Conclusion： The combination of TAE 110 mg/kg and TPNS 115 mg/kg could strengthen protective effects on cerebral ischemia injury, the mechanism underlying might be related to improving jointly the early energy metabolism, and relieving the delayed apoptosis via inhibiting the mitochondrial apoptosis pathway of JNK signal transduction.

复合酶解法提取三七皂苷的实验研究

以三七提取液中总皂苷的含量和提取物得率为指标,考察了乙醇回流法、渗漉法、纤维素酶解法、果胶酶解法、复合酶解法的优劣,并采用单因素法和四因素(纤维素酶用量、果胶酶用量、酶解温度、乙醇浓度)三水平正交设计法对复合酶解法提取工艺条件进行优选,得到如下较理想的提取工艺条件:纤维素酶用量为15U/g生药)、果胶酶用量为140U/g(生药),酶解pH值为4.5,酶解温度为50℃,乙醇浓度为80%,提取时间为2.5h。所得三七提取液中总皂苷的含量为12.01%,提取物得率为35.82%。

三七提取物吸湿性及其数学模型研究

目的:筛选可准确描述三七提取物吸湿过程的数学模型,科学量化吸湿过程。方法:25、35和45℃下,用重量法测试三七提取物吸湿过程动力学曲线和吸附/解吸等温线,并用1stOpt统计分析软件进行拟合,以均方根误差(RMSE),残差平方和(RSS)和决定系数(R^2)为评价指标,筛选最佳拟合模型,结合25、35和45℃下吸附等温线计算吸附热。结果:25、35和45℃下,三七提取物吸湿动力学双指数模型、吸附/解吸等温线Peleg模型拟合效果较好,吸附/解吸等温线都属于第Ⅱ种类型,解吸等温线因较高的平衡含水量而出现明显的滞后现象,该滞后现象属于H3型。随着温度升高这种滞后现象变弱。吸附热随着平衡吸湿量的增大而增大。结论:双指数模型和Peleg模型分别较好地拟合三七提取物的吸湿动力学曲线及吸附/解吸等温线,且吸附/解吸等温线存在滞后现象,该滞后现象随温度升高而变弱,吸附热与平衡吸湿量密切相关。

人参、三七提取物与牛磺酸对兔心肌缺血再灌注损伤中ET、CGRP影响的实验研究

目的 :探讨人参、三七提取物与牛磺酸配伍防治心肌缺血再灌注损伤 (MIRI)的作用机理。方法 :雄性比利时兔 18只 ,随机分为对照组 (生理盐水组 )与治疗组 (益气活血解毒组 )各 9只 ,均于同等条件下口服给药 7d。结扎冠脉造成MIRI模型 ,术中动态监测心电 ;分别在服药前 (T1)、冠状动脉结扎 10min(T2 )、再灌注 10min(T3)取血 ,抗凝 ,测定血清内皮素 (ET)、降钙素基因相关肽 (CGRP)含量。结果 :治疗组与对照组比较 ,可显著降低ET、ET CGRP水平 ,CGRP水平有升高趋势。结论 :人参提取物、三七提取物、牛磺酸对MIRI有良好的保护作用 。

三七的提取动力学研究

研究三七的提取动力学。以Fick第一扩散定律为基础建立三七中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1三种成分的提取动力学模型,根据方程计算获得提取速率常数、表观活化能、相对萃余率等一系列动力学参数。结果表明,温度升高,三种成分提取速率加快,提取浓度升高,且温度降低各成分浓度达平衡所需时间延长。本研究所建立的三七中三种成分的提取动力学模型均符合一级动力学方程特征,该模型较好的描述了三七药材各成分的动态溶出过程,以期为三七及复方丹参提取工艺的设计和优化提供理论依据。

三七皂苷类成分的加速溶剂提取法研究

目的 :探讨中药质量控制中应用加速溶剂提取法提取三七皂苷类成分的可行性。方法 :以三七总皂苷、人参皂苷Rb1和人参皂苷Rg1含量为指标 ,用单因素考察法对影响加速溶剂提取三七皂苷类成分的因素进行优化。结果 :优选出了三七皂苷类成分加速溶剂提取的最佳条件 :以甲醇为溶剂 ,药材粒径为 0 3～ 0 4 5mm ,提取温度 15 0℃ ,提取压力 6 895MPa ,提取 15min ,提取 1次。结论 :加速溶剂提取技术能够快速高效地提取三七皂苷类成分。

溶剂pH及与三七合煎对丹参药材水提效果影响的研究

目的考察提取溶剂的pH值以及与三七共煎对丹参水溶性成分提取效果的影响。方法使用HPLC测定当溶剂pH值分别为4.0,7.0,8.2和9.2时丹参药材提取液以及丹参、三七合煎液中丹参素、原儿茶醛和丹酚酸B的含量,并比较了其他水溶性成分的化学组成差异。结果酸性条件下丹酚酸B的提取效果较好,而中性和碱性环境对原儿茶醛和丹参素的溶出较为有利。在酸性、中性和弱碱性溶剂中,与三七共煎对丹参中丹参素、原儿茶醛、和丹酚酸B的溶出产生抑制作用;而在pH为9.2时,效果则相反。结论此结果可为丹参提取工艺的优化提供实验依据。

三七总皂苷的减压内部沸腾提取及树脂吸附分离

目的提高三七总皂苷的收率及纯度。方法三七粉末经少量70%乙醇解吸后,以水为溶剂在45℃及-0.084 MPa下进行减压内部沸腾提取,提取两次,4 min/次,提取液经浓缩后,直接采用大孔树脂进行吸附分离。结果得到的浸膏中三七皂苷R1、人参皂苷Rb1及Rg1三者含量之和达到70.0%,得率达到5.4%,大大高于传统方法。结论减压内部沸腾提取与大孔树脂吸附分离相结合的工艺具有提取速度快、无需水沉或醇沉工序、收率高及质量好等特点,具有良好的应用前景。

三七水提液对几种植物种子萌发和幼苗生长的化感作用

通过比较不同部位、不同浓度、不同生长年限的三七水提液化感作用强弱,筛选适宜与三七轮作的植物。以小麦、玉米、油菜为受体植物,采用正交试验法比较三七水提液对这3种供试植物种子萌发和幼苗生长的化感作用。(1)三七各部位水提液对小麦、油菜种子萌发及幼苗生长均具有抑制作用。(2)低浓度的三七水提液可以促进玉米种子萌发和幼苗生长,高浓度时则起到抑制作用。(3)不同生长年限及不同部位的三七水提液对受体植物的化感作用也有显著差异。各供试因素中,以不同浓度三七水提液对受体植物的影响最为显著。玉米是3种受体植物中最适合与三七轮作的植物,这可能与玉米对三七化感作用的耐受性较强有关。

顶空毛细管气相法测定三七提取物中有机溶剂残留量

目的：建立三七提取物中7种有机溶剂残留量的测定方法。方法：采用顶空毛细管气相色谱法，以标准溶液加入法测定三七提取物中有机溶剂残留量。结杲：在考察的浓度范围内7种有机溶剂呈现良好的线性关系（乙醇r=0．9994、正己烷r=0．9982、苯r=0．9999、甲苯r=0．9991、二甲苯r=0．9994、苯乙烯r=0．9997、二乙烯苯r=0．9932），回收率81．74%～111．2%，理论板数均大于15000，相邻峰的分离度均大于2，日内、日间精密度RSD均小于10％。结论：该方法简单、准确、灵敏度高、重现性好，排除了基质的干扰，适用于三七提取物中有机溶剂残留量的测定。