同步荧光光谱法研究丹参素-牛血清白蛋白体系的光增强效应

采用紫外光谱法和同步荧光光谱法研究了丹参素与牛血清白蛋白的结合特性.体系的紫外光谱显示,丹参素与牛血清白蛋白(BSA)之间发生了相互作用并生成新的复合物,同步荧光光谱则表明,丹参素对BSA存在荧光增强效应.随着丹参素浓度的增加,Δλ=15 nm时体系荧光强度明显增强,最大发射波长为285 nm,且不随浓度增加而发生变化...     

采用柱切换色谱-离子阱质谱联用法研究复方丹参滴丸中丹参素及其代谢产物

采用柱切换高效液相色谱技术对人血清中的丹参素及其代谢产物进行富集和分离,利用对电喷雾离子阱多级质谱负离子模式进行结构分析。一级流动相为甲醇-水(30：70,V/V),流速0．8mL／min;二级流动相为甲醇-水-甲酸铵(15：85：0．5,V/V/V),流速0．2mL／min;在人血清中检测到丹参素及其代谢产物,并探讨了主要代谢产物乙酰化的丹参素(m/z 240．2)。建立一种复方丹参滴丸中丹参素及其代谢产物在人体内代谢过程的高效分析方法。该方法可用于复方丹参滴丸中丹参素及与其结构类似的其它药物的体内代谢研究。     

丹参水溶性成分的电喷雾质谱行为及其特征图谱的初步研究

应用电喷雾-质谱(ESI-MS)技术研究了原儿茶醛和丹参素的ESI-MS规律,结合ESI-MS技术,对9种丹参水溶性成分的质谱行为进行了研究,并对其中5种成分进行了定性分析。同时利用选择性离子检测质谱技术,初步建立了丹参水溶性成分SIM-MS特征图谱,该图谱简单、易于解析,重现性与特征性较好,可以用于丹参药材及复方丹参的快速指纹鉴别。     

丹参提取液的树脂吸附及醇沉工艺对比研究

以丹参水提取液中丹参素及原儿茶醛含量为指标，HPLC检测不同工艺前后有效成分的含量变化，考察了不同浓度的醇沉除杂工艺，筛选出了大孔吸附树脂富集有效成分的最佳工艺。研究结果显示：单纯采用水醇法提取工艺时，以50％乙醇沉降为最佳工艺，D301大孔吸附树脂能够选择性的吸附丹参素和原儿茶醛，大幅度降低出膏率，提高丹参粗提物中有效成分含量，效果优于醇沉工艺。     

丹参素自氧化与促氧化机理

采用过氧化氢电极与电子自旋共振(ESR)技术对不同pH值条件下丹参素的自氧化及Cu2+对丹参素的促氧化反应机理进行了研究.实验结果发现:在碱性条件下(pH=8.5),丹参素自氧化作用异常显著,氧化首先生成O2-·,随后可歧化为H2O2,并且Cu2+对H2O2信号增强也起着显著的促进作用;在弱酸性条件下(pH=6.5),丹参素自氧化过程受到抑制,主要表现为少许Cu2+的促氧化作用;而在生理条件下(pH=7.4),丹参素的自氧化过程缓慢,有少量H2O2生成,并且Cu2+的促氧化作用非常有限.因此,当丹参素作为一种高效抗氧化剂药用时,其自身的自氧化与金属离子促氧化作用都应有所考虑,尤其在偏碱性体液中,更易产生大量的超氧阴离子自由基,并对生物机体造成某种损害.     

靶细胞提取和高效液相飞行时间质谱联用分析预测丹参中的活性成分

建立靶细胞提取和与高效液相飞行时间质谱联用(HPLC-ESI/TOFMS)分析相结合进行丹参活性成分研究的方法,对丹参中可能的活性成分进行了推测。将靶细胞和丹参样品一起孵育培养,根据活性成分可以与靶细胞膜有特异性的结合或者进入靶细胞内的原理,用HPLC-MS方法对培养后细胞破碎液中的成分进行分析。从加药Raw264.7细胞和Ecv304细胞破碎液中检测到丹参素、原儿茶醛、咖啡酸、丹酚酸D、紫草酸、迷迭香酸及丹酚酸B7种丹参中成分,在加药的HL7702细胞破碎液中检测出丹参素、原儿茶醛、咖啡酸及丹酚酸B4种丹参中的水溶性成分。结果显示:细胞破碎液中检测的成分为丹参在靶细胞中有吸收的成分,本方法可用于预测中药中的活性成分。     

化学-酶催化制备天然丹参素

丹参素是中药丹参的水溶性化学成分之一,具有很多潜在的药理活性。 以3,4-二羟基苯甲醛和乙酰甘氨酸为原料,经Erlenmeyer反应并开环得到α-乙酰氨基-β-(3,4-二乙酰氧基苯基)丙烯酸、盐酸水解制得β-(3,4-二羟基苯基)丙酮酸,再经乳酸片球菌1.2696静息细胞生物酶催化还原合成了天然右旋丹参素,总收率69.4%,丹参素对映体过量值97.5%。     

丹参素分子印迹聚合物的制备及吸附性能研究

在硅胶表面接枝乙烯基三乙氧基硅烷得到接枝硅胶,将其与模板分子丹参素、功能单体4-乙烯基吡啶、交联剂乙二醇二甲基丙烯酸酯等,在偶氮二异丁腈引发作用下进行聚合反应,制备丹参素分子印迹聚合物。采用红外光谱、元素分析对分子印迹聚合物进行表征,静态平衡吸附法和Scatchard分析法研究印迹聚合物的吸附特征。结果表明:该印迹化合物存在两种结合位点,它们的离解常数分别为714.29 mg·L-1和467.74 mg·L-1,饱和吸附量分别为9.93 mg·g-1和61.80 mg·g-1,印迹因子分别达1.93和2.19,对丹参素具有较好的选择吸附性能,可用于丹参素的分离和富集。     

大孔树脂分离纯化丹酚酸的研究

比较了D301R、D392、D380大孔阴离子交换树脂和X-5.AB-8、NKA-9、SP825大孔吸附树脂对丹参水溶性成分的吸附和解吸能力,筛选出效果较好的SP825进行分离纯化丹酚酸的研究.实验表明,大孔吸附树脂SP825能分离出纯度为95.32%的丹参素,在梯度洗脱条件下可得到以丹参素(水洗脱)和丹酚酸B(乙醇洗脱)为主的产品.在最佳吸附与解吸工艺参数下,丹参素、紫草酸、迷迭香酸、丹酚酸A和丹酚酸B的收率分别为:36.92%、80.39%、82.45%、43.07%和41.03%.     

反相高效液相色谱法测定冠心宁注射液中的丹参素和原儿茶醛

冠心宁注射液是新中药制剂,处方中含有丹参和川芎,丹参为该制剂的君药。该制剂的活性成分主要为丹参的酚酸类成分,包括丹参素（3,4-二羟基苯基乳酸）、原儿茶醛和迷迭香酸等。目前,关于该药品的质量控制标准尚未见任何报道。本文建立了测定冠心宁注射液中丹参素和原儿茶醛的反相高效液相色谱法,用以控制该药品的质量。     

Separation of salvianic acid A from the fermentation broth of engineered Escherichia coli using macroporous resins

Salvianic acid A (also known as danshensu) is a plant‐derived polyphenolic acid, and has a variety of physiological and pharmacological activities. Our laboratory previously constructed an unprecedented artificial biosynthetic pathway in Escherichia coli and established the fermentation process to produce salvianic acid A. Here, we developed an efficient method for separating salvianic acid A from the fermentation broth of engineered Escherichia coli by macroporous resins. Among ten tested macroporous resins, the static and dynamic adsorption/desorption experiments demonstrated that X5 resin was the best to separate salvianic acid A from fermentation broth. Other parameters during static and dynamic procedures were also investigated. Under the optimum separation conditions, the average adsorption capacity of SAA were 10.66±0.54 mg/g dry resin and the desorption ratio was 85.6±4.1%. The purity and recovery yield of salvianic acid A in the final dry product were 90.2±1.5 and 81.5±2.3%, respectively. The results show that adsorption separation with macroporous resin X5 was an efficient method to prepare salvianic acid A from fermentation broth. This work will benefit the development and application of plant‐derived salvianic acid A and its derivatives.     

Mechanism and Cellular Kinetic Studies of the Enhancement of Antioxidant Activity by Using Surface‐Functionalized Gold Nanoparticles

The enhanced antioxidant activity of surface‐functionalized gold nanoparticles (AuNPs) synthesized by self‐assembly has attracted great attention, but little is known about the mechanism behind the enhanced activity. To address this challenge, the antioxidant activity of Au@PEG3SA (i.e., surface‐functionalization of spherical AuNPs with the antioxidant salvianic acid A) was used as an example to illustrate the mechanism of the enhanced activity. Evaluation of the antioxidant activity was performed in a radical‐scavenging reaction between Au@PEG3SA and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical. As expected, the rate constant for the reaction of Au@PEG3SA with DPPH was about nine times greater than that for the salvianic acid A monomer. A comparative analysis of the spectral characteristics of Au@PEG3SA and the salvianic acid A monomer further imply that the enhancement of the antioxidative reaction kinetics may be ascribed to the variation in the transition state for the DPPH‐radical scavenging reaction through π–π stacking interactions between and among adjacent groups on the surface of Au@PEG3SA. On the other hand, the kinetic enhancement of Au@PEG3SA on reactive‐oxygen‐species (ROS) scavenging can be observed in living cells and in vivo, which possibly provides new insight for the bioapplication of self‐assembly of surface‐functionalized AuNPs.     

药物抗氧化作用对DNA发光动力学行为的影响

本文应用CuSO₄-Phen-VC-H₂O₂-DNA化学发光体系观察药物对发光动力学行为的影响,研究了九种药物对DNA损伤的保护作用,并对其药物结构对抗氧化能力的影响进行了分析。发现药物在该系统中的抗氧化作用有三种类型,提示它们可能在体内不同的反应过程中起不同的作用。在该系统中具有抑制DNA损伤发光能力的药物,按其作用强弱依次为阿魏酸、丹参素、芦丁、茶多酚、SOD、丹参酮Ⅱ-A、丹皮酚、五味子乙素,具有延迟DNA损伤发光能力的药物,按其作用强弱依次为阿魏酸、芦丁、茶多酚、Trolox、丹参素、丹皮酚。分子结构中具有共轭大π键、羟基、羧基的药物具有较强抑制DNA损伤的能力,这可能与药物分子的氧化还原电位或自由基生成热等多种因素有关。     

Qualitative and quantitative determination of YiXinShu Tablet using ultra high performance liquid chromatography with Q Exactive hybrid quadrupole orbitrap high‐resolution accurate mass spectrometry

To clarify and quantify the chemical profile of YiXinShu Tablet rapidly, a feasible and accurate strategy was developed by applying ultra high performance liquid chromatography with Q Exactive hybrid quadrupole orbitrap high‐resolution accurate mass spectrometry. A total of 105 components were identified, including 25 phenanthraquinones, 11 lactones, 19 lignans, 24 acids, and 26 other compounds. Among them, 26 major compounds were unambiguously detected by comparing with reference standards. And 19 of these compounds in three batches of YiXinShu Tablet were selected for quantitative determination. (Z )‐Ligustilide, salvianic acid A, salvianolic acid A, salvianolic acid B, and rosmarinic acid were abundant in these three batches with contents over 1 mg/g. The established analysis methods were examined to be accurate and feasible. The results show that the ultra high performance liquid chromatography with Q Exactive hybrid quadrupole orbitrap high‐resolution accurate mass spectrometry method has a powerful qualitative ability and promising quantitative application.     

Evaluation of affinity interaction between small molecules and platelets by open tubular affinity capillary electrochromatography

In this paper, an open tubular affinity capillary electrochromatography (OT‐ACEC) was developed by physical adsorption of rabbit platelets on the inner surface of capillary. The interactions between small molecules include adenosine diphosphate (ADP) (positive control), protocatechuic acid (negative control) and seven natural products (salvianolic acid B, salvianic acid A sodium, hydroxysafflor yellow A, ferulic acid, chlorogenic acid, sinapic acid, caffeic acid) and platelets were evaluated by their retention factors and binding constants obtained based on peak‐shift assay. Then, the activities of anti‐platelet aggregation induced by thrombin (THR), ADP and arachidonic acid (AA) for those small molecules (except ADP) were evaluated by turbidimetric method. The results indicate that: (i) ADP, a platelet aggregation inducer, had strong interaction with platelet, while protocatechuic acid that had no inhibition on platelet aggregation behaved no specific interaction; (ii) there was a positive correlation between the anti‐platelet aggregation activities of small molecules and their interactions with platelet, generally those compounds with higher binding constants with platelet exhibited higher activities. Therefore, the OT‐ACEC method developed in the present study can be a potential method to evaluate affinity interactions between small molecules and platelets, so as to predict the biological activities such as anti‐platelet aggregation for the small molecules.     

Study on the interaction between salvianic acid A sodium and bovine serum albumin by spectroscopic methods

The interaction between salvianic acid A sodium (SAS) and bovine serum albumin (BSA) was investigated using fluorescence and ultraviolet spectroscopy at different temperatures under imitated physiological conditions. The experimental results showed that the fluorescence of BSA was quenched by SAS through a static quenching procedure. The binding constants of SAS with BSA were 2.03, 1.17 and 0.71×10(5) L mol(-1) at 291, 298 and 305 K, respectively. Negative values of ΔG, ΔH, and ΔS indicate that the interaction between SAS and BSA is driven by hydrogen bonds and van der Waals forces. According to F?rster non-radiation energy transfer theory, the binding distance between BSA and SAS was calculated to be about 2.92 nm. The effect of SAS on the conformation of BSA was analyzed using synchronous fluorescence spectroscopy. In addition, the effect of some metal ions Cu(2+), Ca(2+), Mg(2+), and Zn(2+) on the binding constant between SAS and BSA was examined.     

Study on the Electrochemical Properties of Salvianic Acid A Sodium and its Analytical Application

The electrochemical behavior of salvianic acid A sodium (SAS), a main active content in Chinese herbal medicine (CHM), was studied for the first time by cyclic voltammetry (CV). A new electroanalytical method of SAS was erected using differential pulse voltammetric (DPV) technique. In pH 3.3 britton‐robinson (B‐R) buffer solution, the medicine showed a pair of redox peaks driven by adsorption. The electrode process involved two electrons and two protons transformation with apparent rate constant (ks) of 2.85 s^?1 and transfer coefficient (α) of 0.81. Based on understanding the electrochemical process of SAS at the glassy carbon electrode (GCE), analysis of SAS can be realized. The oxidation peak currents showed linear with the concentrations of SAS in the range of 5.45 × 10^?8 to 1.09 × 10^?5 M. The limit of detection was 5.45 × 10^?8 M. The proposed method has high sensitivity, wide linear range, and was successfully applied to quantitative determination of the SAS in Rukuaixiao Tablets.     

毛细管区带电泳法考察牛凝血酶、核酸适配体与丹参活性分子间的相互作用

采用毛细管区带电泳法(CZE)研究牛凝血酶(B-Thr)、凝血酶核酸适配体(Apt29)与丹参中3种活性物质(丹参素钠(SAS)、原儿茶酸(PA)、阿魏酸(FA))间的相互作用。电泳条件为:毛细管长度40 cm,紫外检测波长214 nm,分离电压15 kV,压力进样3.45 kPa,进样时间5 s。分别固定3种活性药物分子的浓度,改变B-Thr或Apt29的浓度,采用Scatchard方程和非特异性结合方程计算结合常数(K_b),表征药物分子和B-Thr或Apt29的相互作用大小。结果表明,PA与B-Thr结合能力最强,K_b为3.39×104L/mol,FA与B-Thr的K_b为1.05×104L/mol,SAS与B-Thr未表现出结合能力;在与Apt29的相互作用的研究中显示,SAS基本未与Apt29结合,FA与Apt29结合能力强于PA与Apt29的结合能力,FA、PA与Apt29的K_b分别为1.48×104L/mol和1.32×104L/mol。该研究报道了丹参活性分子与B-Thr、Apt29的相互作用结果,可为核酸适配体(蛋白质)与中药分子相互作用的探索提供新方法,适配体与中药分子相互作用亦可为中药分子的靶点发现、靶向转运提供借鉴。     

有机酸对土壤中镉形态及其生物有效性影响的研究进展

根系分泌的低相对分子质量有机酸与土壤中镉之间相互作用影响着土壤中镉的吸附、解吸，使镉在土壤-水-植物系统中的迁移、转化规律发生重大改变，从而影响到镉在土壤中的形态及其生物有效性。对近二十年来有关有机酸与重金属镉交互作用的研究进展、有机酸对土壤中镉的有效性、毒性的影响以及作用机制进行了综述，并对现在研究存在的问题以及今后的研究趋势做了初步展望。