Multiple chromatographic fingerprinting and its application to the quality control of herbal medicines

Recently, chromatographic fingerprinting has become one of the most powerful approaches to quality control of herbal medicines. However, the performance of reported chromatographic fingerprinting constructed by single chromatogram sometimes turns out to be inadequate for complex herbal medicines, such as multi-herb botanical drug products. In this study, multiple chromatographic fingerprinting, which consists of more than one chromatographic fingerprint and represents the whole characteristics of chemical constitutions of the complex medicine, is proposed as a potential strategy in this complicated case. As a typical example, a binary chromatographic fingerprinting of “Danshen Dropping Pill” (DSDP), the best-sold traditional Chinese medicine in China, was developed. First, two HPLC fingerprints that, respectively, represent chemical characteristics of depsides and saponins of DSDP were developed, which were used to construct binary chromatographic fingerprints of DSDP. Moreover, the authentication and validation of the binary fingerprints were performed. Then, a data-level information fusion method was employed to capture the chemical information encoded in two chromatographic fingerprints. Based on the fusion results, the lot-to-lot consistency and frauds can be determined either using similarity measure or by chemometrics approach. The application of binary chromatographic fingerprinting to consistency assessment and frauds detection of DSDP clearly demonstrated that the proposed method was a powerful approach to quality control of complex herbal medicines.     

蛋白质组学技术鉴定与分析三七粉诱导海兔神经连索表达的差异蛋白质

RP-HPLC分离三七粉提取液,并鉴定含有Rb1、Rg1、Re、R1等皂甙成分。以蓝斑背肛海兔(Notarcusleachii cirrosus Stimpson,NLCS)为分析模型,三七粉提取液为诱导剂,选用蛋白质组技术研究NLCS神经连索诱导前后所表达的差异蛋白质。通过优化双向凝胶电泳分离NLCS神经连索全蛋白质组技术,获得496个蛋白质斑点。采用肽指纹图谱技术和数据库检索比对法,初步鉴定了NLCS受三七粉提取液诱导前后,其神经连索表达13个差异蛋白质,其中较高的匹配率蛋白质为肌动蛋白、3-羟酯酰辅酶A脱氢酶、ATP结合转运子和甲基转移酶12。选用LOC tree软件对13个差异蛋白质进行亚细胞定位,认为它们在保护神经系统中发挥重要的调节作用。     

The Antiviral Effect of Panax Notoginseng Polysaccharides by Inhibiting PRV Adsorption and Replication In Vitro

Porcine pseudorabies (PR) is an important infectious disease caused by pseudorabies virus (PRV), which poses a major threat to food safety and security. Vaccine immunization has become the main means to prevent and control the disease. However, since 2011, a new PRV variant has caused huge economic losses to the Chinese pig industry. Panax notoginseng polysaccharides have immunomodulatory activity and other functions, but the antiviral effect has not been reported. We studied the anti-PRV activity of Panax notoginseng polysaccharides in vitro. A less cytopathic effect was observed by increasing the concentration of Panax notoginseng polysaccharides. Western blot, TCID₅₀, plaque assay, and IFA revealed that Panax notoginseng polysaccharides could significantly inhibit the infectivity of PRV XJ5 on PK15 cells. In addition, we also found that Panax notoginseng polysaccharides blocked the adsorption and replication of PRV to PK15 cells in a dose-dependent manner. These results show that Panax notoginseng polysaccharides play an antiviral effect mainly by inhibiting virus adsorption and replication in vitro. Therefore, Panax notoginseng polysaccharides may be a potential anti-PRV agent.     

三七不同部位成分的谱学性质研究

采用不同溶剂提取了三七的不同部位,即提取三七皮和芯的化学成分,通过研究提取液的紫外-可见光谱(UV-Vis),傅里叶变换红外光谱(FTIR)、荧光光谱(FS)、电喷雾离子质谱(ESI-MS)及高效液相色谱(HPLC),寻找谱学特征与其化学成分的相关性,从化学成分的角度解释了三七皮与芯的药理效应差异的可能原因。     

离子液体-基质固相分散-超声雾化-固相萃取结合高效液相色谱法检测人参中三嗪类除草剂

采用固载1-甲基-3己基咪唑六氟磷酸([C6MIM][PF6])的硅胶作为基质固相分散剂,与超声雾化提取及C18固相萃取相结合,对人参提取物进行提取和净化;采用高效液相色谱法,建立了人参中5种三嗪类除草剂的检测方法.样品前处理的最优条件为:300 mg人参粉末与150 mg硅胶([C6MIM][PF6]含量2.0 mmol/g)研磨5 min,以20.0 mL去离子水(pH 7,含1.0％ NaCl)超声雾化提取10 min,再以5.0 mL乙腈洗脱萃取小柱(300 mg C18).实验结果表明,目标物的检出限为0.020～0.035 g/g,线性相关系数r2≥0.9992,实际样品加标回收率为78.2％～95.4％,相对标准偏差为3.5％ ～6.0％.本方法快速高效、净化效果好、提取率高,为中药中农药残留的检测提供了新的样品前处理方法.     

Compatible stability study of panax notoginseng saponin injection (xueshuantong®) in combination with 47 different injectables

Panax Notoginseng Saponin (PNS) injection (Xueshuantong®, XST) plays an important role in acute and chronic treatment of cardiovascular disease in China. XST, a freeze‐dried powder injection of PNS, contains ginsenosides, Rg1, Rb1, Re, Rd. and notoginsenoside, R1. Study of the stability of herbal medicine injections is limited. The degradation products may contribute to adverse drug reactions (ADRs). The stability kinetics and degradation mechanisms of the five ingredients of PNS in XST injection were systematically explored in aqueous solution. The compatible stability of XST injection in combination with 47 injectables was evaluated using biophysical analysis. A principal components analysis model provided a good clustering result after compatibility. The degradation of the five ingredients in aqueous solution was found to be acid, temperature, oxidation and trivalent ion‐dependent. All ingredients were much more stable in XST injection than in aqueous solutions. Nine degradation products were identified by adopting LC‐Q‐TOF/MS. Standards exceeding the osmotic pressure and/or particle size were found to be the probable causes of the ADR, which were related to drug combination. To minimize the risk of ADRs, drug combinations should be avoided until the complicated chemical matrices of the Chinese medicine injections are more clearly understood. Copyright © 2016 John Wiley & Sons, Ltd.     

秦巴山区中草药钮子七中微量元素的测定

用ICP-AES测定了秦巴山区独有的中草药钮子七中的2种常量营养性元素(钙、镁)和5种微量元素(铜、铬、铁、锌、锰)含量。钙4790.66μg.g-1、镁55.32μg.g-1、铜18.44μg.g-1、铬2.1μg.g-1、铁264.50μg.g-1、锌56.71μg.g-1、锰17.83μg.g-1,回收率均在96.2%—100.5%之间,结果表明,此方法简便、可靠,具有良好的精确度和准确性。并初步讨论了中草药钮子七中有益元素与治疗疾病的关系,并与中药材汉三七中微量元素含量相比较,二者微量元素含量略有差异。     

液相色谱-串联质谱法同时测定三七中508种农药残留

采用液相色谱-串联质谱(LC-MS/MS)建立了可在20 min内快速测定三七中508种农药残留的方法。样品经改良的QuEChERS方法提取,分散固相萃取法(d-SPE)净化后,在多反应监测(MRM)模式下进行LC-MS/MS分析,内标法定量。在优化实验条件下,508种农药均在0.005～0.4 mg/L范围内线性关系良好,方法检出限(LOD)为0.005～0.1 mg/kg;在100μg/kg加标水平下,除8种农药外,其余500种农药的回收率均在50.2%～117%之间,且相对标准偏差(RSD)均不大于15%;其中有468种农药的回收率为70%～120%。该方法样品前处理简单快速,灵敏度、准确度和精密度均符合农药多残留检测技术的要求,适用于三七中508种农药残留的快速筛查。     

Determination of Saponins in Leaf of Panax Ginseng C. A. Mey. by High Performance Liquid Chromatography

A high performance liquid chromatography(HPLC) with UV detection was established for simultaneous determination of saponins in the leaf of Panax ginseng C. A. Mey. Nine ginsenosides(Rb1,Rb2,Rb3,Rc,Rd,F1,F2,F3,F5) and notoginsenoside Fe(NFe) were studied. Among the saponins,the ginsenosides F1,F2,F3,F5 and NFe were determined by HPLC-UV method for the first time. The determination of the ginsenosides via the HPLC-UV method was performed on a reversed-phase C18 column with gradient elution in 40 min. The line...     

Identification of Ginsenosides in Panax quinquefolium by LC-MS

An HPLC-APCI-MS method for the identification of ginsenosides in Panax quinquefolium has been developed. HPLC-APCI-MS could effectively identify ocotillol, protopanaxadiol, protopanaxatriol and oleanane-type ginsenosides in a single MS experiment since [M-H]⁻ ions and characteristic thermal degradation ions of ginsenosides could be simultaneously observed under negative and positive ionization conditions. Nine ocotillol-type ginsenosides including 24(R)-pseudoginsenoside F₁₁ were firstly identified and a total of 30 ginsenosides were identified in Panax quinquefolium. The ginsenoside profile differences between Chinese and American P. quinquefolium were investigated by HPLC-APCI-MS.