UHPLC–Q‐TOF‐MS‐based metabolomics approach to compare the saponin compositions of Xueshuantong injection and Xuesaitong injection

Various traditional Chinese medicine preparations developed from Notoginseng total saponins, including Xueshuantong injection and Xuesaitong injection, are extensively used in China to treat cardiocerebrovascular diseases. However, the difference of their saponin compositions remains unknown. An ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry based metabolomics approach was developed to probe the saponin discrimination between Xueshuantong and Xuesaitong and the related factors by large sample analysis. A highly efficient chromatographic separation was achieved on an HSS T3 column within 20 min with the holistic metabolites information recorded in the negative MS^E mode. A six‐step data pretreatment procedure mainly based on Progenesis QI and mass defect filtering was established. Pattern recognition chemometrics was used to discover the potential saponin markers. The saponin composition of Wuzhou Xueshuantong showed distinct discrimination from the other products. Wuzhou Xueshuantong contains more abundant protopanaxatriol‐type noto‐R₁, Rg₁, Re, and protopanaxadiol‐type Rb₁, but less Rd and other low‐polarity protopanaxadiol‐type ginsenosides. These differences could not directly correlate to the use of different parts of Panax notoginseng , but possibly to the different preparation techniques employed by different manufacturers. These results are beneficial to the establishment of pharmacopoeia standards and the assessment of the efficacy and adverse drug reactions for these homologous products.     

Identification of ginsenoside metabolites in plasma related to different bioactivities of Panax notoginseng and Panax ginseng

Although the chemical components of Panax notoginseng (PN) and Panax ginseng (PG) are similar, their bioactivities are different. In this study, the differential bioactivities of PN and PG were used as the research object. First, the different metabolites in the plasma after oral administration of PN and PG were analyzed using a UPLC-Q/TOF-MS-based metabolomics approach. Afterward, the metabolite-target- pathway network of PN and PG was constructed, and thus the pathways related to different bioactivities were analyzed. As a result, 7 different metabolites were identified in PN group, and 10 different metabolites were identified in the PG group. In the PN group, the metabolite N1 was related to hemostasis, N1 and N3 were related to inhibiting the nerve center, antihypertensive, and abirritation. The metabolites N1, N3, N4, N5, and N6 were related to liver protection. The results showed that the metabolites G1, G2, G3, G5, and G6 in PG group were related to heart protection, and G1, G2, G6, and G9 were related to increased blood pressure. There were 13 signaling pathways related to different biological activities of PN (8 pathways) and PG (5 pathways). These pathways further clarified the mechanism of action that caused the different bioactivities between PN and PG. In summary, metabolomics combined with network pharmacology could be helpful to clarify the material basis of different bioactivities between PN and PG, promoting the research on PN and PG.     

野三七的红外光谱分析

为了证实野三七现有的疗效并推测其未知的潜在的价值,采用傅里叶变换红外光谱仪测试了云南白药、文山三七、川芎及野三七等药材的红外光谱.云南白药与文山三七,川芎与野三七两组红外光谱的峰形、峰位极相似.从特征峰的峰强、峰形、峰位比之,虽有偏差,但很小.此外,四种药物在光谱上也有相当的吻合性.由此可知,野三七和其它三种药物一样,...     

三七炮制前后的红外光谱分析研究

采用红外光谱并结合二维相关红外光谱法对生三七和熟三七进行了研究,分析比较三七炮制前后化学成分的变化。在一维红外谱图上,生三七和熟三七在1 200~400cm-1波段内的相似度较高,不同的是熟三七在2 925,2 855,1 746,1 460,1 376,1 158cm-1出现了花生油的特征峰,这是由于采用煎炸法炮制三七所残留的花生油所引起的。两者药效组分的特征差异在二维相关红外谱图上显示得更为明显和直观,在1 400~1 700cm-1区域,生三七仅在1 650cm-1附近有一个强自动峰,而熟三七在1 469和1 640cm-1附近均有自动峰。在1 120~1 250cm-1区域,生三七和熟三七共有1 139(1 137),1 194(1 196),1 219(1 221)cm-1三个自动峰,不同的是各自动峰的相对强度发生了变化,生三七经炮制后1 139cm-1峰的增强,而1 194cm-1峰减弱。二维相关红外光谱的变化规律说明了三七药材在炮制的过程中发生的主要变化是黄酮、糖类、皂苷等成分的分解。该方法揭示了三七在炮制过程中所发生的物理化学变化过程,从红外光谱的角度解释了三七"生消熟补"的药理。     

Simultaneous determination of saponins in flower buds of Panax notoginseng using high performance liquid chromatography

The flower buds of Panax notoginseng have been commonly used for the treatment of hypertension, vertigo, tinnitus and acute faucitis in China. The amount of total saponins in the flower buds is higher than in any other parts of P. notoginseng. However, the compositions of flower buds have not been quantified clearly until now. A sensitive and efficient high-performance liquid chromatography-ultraviolet (HPLC-UV) method was developed for the first time to simultaneously quantify eight active saponins in the flower buds of P. notoginseng, including notoginsenoside R(1) and ginsenosides Rg(1), Re, Rb(1), Rb(2), Rb(3), Rd and F(2). The analysis was performed on a reversed-phase C(18) column with gradient elution of acetonitrile and 0.01% aqueous formic acid. The proposed method provided good linearity, reproducibility and sensitivity for the simultaneous quantification of the investigated saponins with overall intra- and inter-day precision and accuracy of better than 4.1% (RSD) and higher than 95% (accuracy), respectively. The recoveries for all the saponins determined were in the range 94.7-104.8% with RSD better than 3.1%. Using the optimized method, we were able to analyze samples from different villages of Wenshan Prefecture, China, which is helpful for quality control of flower buds of P. notoginseng.     

三七总皂甙对牛血清白蛋白溶液构象的影响

应用衰减全反射傅立叶变换红外光谱结合荧光光谱和紫外光谱研究了中药三七 的有效成分三七总皂甙与牛血清白蛋白（BSA）的相互作用，采用对蛋白质红外光 谱酰氨Ⅰ带和酰氨Ⅲ带进行曲线拟合的方法，定量分析了不同浓度三七总皂甙对 BSA二级结构的影响，发现随着三七总皂甙浓度的增加，蛋白分子结构逐渐发生了 由螺旋向折叠的转化。a-螺旋结构减少了3%，β-折叠结构增加了约5%，其它二级 结构没有明显的变化，红外差谱和荧光光谱的结果为药物与蛋白质的作用引起牛血 清白蛋白溶液构象的变化提供了佐证，紫外光谱反映了单体皂甙与蛋白质的结合常 数的差异。     

气相色谱-三重四极杆串联质谱结合QuEChERS测定三七中有机磷农药残留

建立了改良的QuEChERS样品前处理法,结合气相色谱-三重四极杆串联质谱(GC-MS/MS)同时测定三七中18种有机磷农药残留的方法。样品经1%乙酸-乙腈提取,改良QuEChERS技术净化,利用GC-MS/MS采取多反应离子监测模式测定,基质匹配校准曲线外标法定量。18种有机磷农药残留在2.5~120.0μg/kg范围内线性良好,相关系数(R2)大于0.99;检出限为0.4~1.5μg/kg,定量限为0.8~2.5μg/kg。在3个加标水平(2.5,8.0,60.0μg/kg)下平均回收率为70.5%~118.9%,相对标准偏差为1.0%~9.8%。     

Analysis of bioactive components and multi‐component pharmacokinetics of saponins from the leaves of Panax notoginseng in rat plasma after oral administration by LC–MS/MS

In this study, simple ultra‐high performance liquid chromatography coupled with quadrupole time‐of‐flight mass tandem mass spectrometry is used to characterize the absorbed components in rat plasma after the oral administration of saponins from the leaves of Panax notoginseng. Seventeen prototype compounds are structurally characterized. Furthermore, a simple and sensitive liquid chromatography with tandem mass spectrometry method is also used for the simultaneous determination of notoginsenoside Fc, ginsenoside Rb1, ginsenoside Rc, ginsenoside Rb3, ginsenoside Rd, and notoginsenoside Fe in rat plasma within 5 min. After n‐butanol mediated liquid–liquid extraction, all analytes were separated on a C₁₈ column and monitored in negative ion mode. Linearity, sensitivity, intra‐ and inter‐assay precision, accuracy, recovery, matrix effect, and stability were all within acceptable ranges. The validated liquid chromatography with tandem mass spectrometry method is successfully applied to the pharmacokinetic study of saponins from the leaves of Panax notoginseng in rats after oral administration. The results suggest that notoginsenoside Fc and ginsenoside Rb3 showed relatively higher exposure compared with other saponins. All saponins showed a long duration in plasma with a t_1/2 longer than 15 h, except notoginsenoside Fe (t_1/2 = 2.78 h). This study provides important information about the metabolism of saponins from the leaves of Panax notoginseng, which is useful for completely understanding its mechanism of action.     

不同消解对原子吸收光谱法测定三七中重金属铬的影响

以云南文山三七为材料,分析研究了用干法灰化、湿法消解前处理样品对原子吸收光谱法测定三七中重金属C r含量的影响,方法的相关系数为0.9993,干法灰化回收率为95.02%,湿法消解回收率为93.03%,分析结果满意。结果表明,干法灰化的最佳条件为灰化温度500—600℃、灰化时间6—8h。湿法消解的最佳消解体系为HNO3:HC lO4(V/V)=4:1体系。用干法灰化或常压下湿法消解处理样品,原子吸收光谱法可以准确测定三七中重金属C r含量,且方法简单、快捷。云南文山三七粉中重金属C r含量为1.8178μg/g。     

反相高效液相色谱法同时测定三七药材中4种皂苷的含量

建立了以0．02％磷酸-乙腈为流动相，梯度洗脱反相高效液相色谱同时测定中药材三七中三七皂苷R1、人参皂苷Rg1、Rb1和Rd 4种皂苷的新方法。R1、Rg1、Rb1和Rd 4种皂苷的加样回收率分别为89．54％、90．08％、82．82％与84．46％；线性范围分别为0．244-6．110、0．820-20．510、0．396-9．890与0．260-6．500μg。测定了不同规格、部位和来源的三七药材里的4种皂苷R1、Rg1、Rb1和Rd。方法准确可靠，结果稳定，重现性好，可用于三七及其制剂的质控。