Study on Optimization of Ethanol Reflux Extraction of Phenolic Acids from Salvia miltiorrhiza

The extraction technology of phenolic acid compounds from Salvia miltiorrhiza by ethanol reflux was studied. In this experiment, salvianolic acid B standard was used to make the standard curve. Single factor experiment and orthogonal experiment were used to study the extraction of different ethanol concentrations, reflux times and material-to-liquid ratios. The OD value of salvianolic acid compounds was measured with a spectrophotometer. The extraction rate of phenolic acid compounds under different extraction conditions was calculated through a regression equation, so as to obtain the optimal conditions for the ethanol reflux extraction process of Salvia miltiorrhiza. The experimental data can provide a reference for the ethanol reflux extraction process of salvianolic acids in the industry. According to the experiment, the extraction rate of phenolic acids in Salvia miltiorrhiza was the highest when the ethanol concentration was 60%, the reflux time was 1.5 hours, and the ratio of material-to-liquid was 1:10.

Effect of salvia miltiorrhiza and ligustrazine hydrochloride injection combined with hydroxyethyl starch injection on serum BNP, Hcy, MMP-2, S100B protein and hemorheology in patients with acute cerebral watershed infarction

Objective:To study the effect of salvia miltiorrhiza and ligustrazine hydrochloride injection combined with hydroxyethyl starch injection on serum BNP, Hcy, MMP-2, S100B protein and hemorheology in patients with acute cerebral watershed infarction.Methods:A total of 90 patientswith acute cerebral watershed infarction in our hospital from August 2014 to December 2016 were enrolled in this study. The subjects were divided into the control group (n=45) and the treatment group (n=45) randomly. The control group was treated with hydroxyethyl starch injection, the treatment group was treated withsalvia miltiorrhiza and ligustrazine hydrochloride injection combined with hydroxyethyl starch injection, and both the two groups were treated for 2 weeks. The serum BNP, Hcy, MMP-2, S100B protein and hemorheology of the two groups before and after treatments were compared.Results:There were no significantly differences of the serum BNP, Hcy, MMP-2, S100B protein and hemorheology of the two groups before treatment. The serum BNP, Hcy, MMP-2, S100B proteinlevels of the two groups after treatment were significantly lower than before treatment, and that of the treatment group after treatment were significantly lower than the control group. The PV, Lr, Mr, Hr and RE of the two groups after treatment were significantly lower than before treatment, and that of the treatment group after treatment were significantly lower than the control group.Conclusion:Salvia miltiorrhiza and ligustrazine hydrochloride injection combined with hydroxyethyl starch injectioncan significantlyimprovetheneurological function and hemorheology, reduce inflammation of the patients with acute cerebral watershed infarction, and it was worthy clinical application.

Determination of Chemical Composition of Salvia miltiorrhiza Cultivated in Jizhou

[Objectives] This study aimed to evaluate the quality of Salvia miltiorrhiza cultivated in Jizhou. [Methods] The contents of chemical ingredients( tanshinone ⅡA,tanshinone Ⅰ and salvianolic acid B) in S. miltiorrhiza were determined by high-performance liquid chromatography( HPLC),and the chemical composition of S. miltiorrhiza planted in Jizhou and purchased from market was compared.[Results]The contents of tanshinone ⅡA,tanshinone I and salvianolic acid B in S. miltiorrhiza planted in Jizhou were 0. 014%,0. 421 2%and 3. 018%,respectively,higher than the standards of the Chinese Pharmacopoeia of the 2015 edition. [Conclusions]The chemical composition of S. miltiorrhiza planted in Jizhou meets the standards of the Chinese Pharmacopoeia of the 2015 edition.

HPLC法同时测定乙肝扶正胶囊中丹酚酸B、隐丹参酮和丹参酮Ⅱ_(A)的含量

目的:建立同时测定乙肝扶正胶囊中丹酚酸B、隐丹参酮和丹参酮Ⅱ_(A)含量的高效液相色谱法。方法:采用Agilent ZORBAX-SB C 18色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%磷酸为流动相,梯度洗脱,检测波长为270 nm。结果:丹酚酸B、隐丹参酮和丹参酮Ⅱ_(A)分别在7.82~313.01μg·mL^(-1)(r 2=0.9998)、0.087~17.324μg·mL^(-1)(r^(2)=1.0000)、0.067~13.468μg·mL^(-1)(r^(2)=1.0000)范围内线性关系良好,平均回收率分别为103.99%、97.76%、100.88%,RSD均≤1.00%。测定样品83批次,丹酚酸B、隐丹参酮和丹参酮IIA的含量分别为0.16~4.06、0.00~103.53、0.00~126.49μg·粒^(-1)。结论:本方法为提升乙肝扶正胶囊的质量控制手段提供了理论依据和有效的技术支撑。

Chinese Prescription Kangen-karyu as Potential Anti-Alzheimer’s Disease Therapeutic:Analyses of BACE1 and GSK-3βInhibitory Activities

Inhibition ofβ-site amyloid precursor protein-cleaving enzyme 1(BACE1)or glycogen synthase kinase-3β(GSK-3β)is estimated to be the central therapeutic approach for Alzheimer’s disease(AD).In this study,water extract of Kangenkaryu,its crude drug and chemical composition used in oriental medicine were evaluated regarding their BACE1 and GSK-3βinhibitory activities.Fluorescence resonance energy transfer was used to characterize the BACE1 inhibitory effect of Kangen-karyu,its crude drug and chemical composition.GSK-3βactivity was determined using the Kinase-Glo Luminescent Kinase Assay Platform.The water extract of Kangen-karyu inhibited BACE1 and GSK-3βin concentration-dependent manners when compared with reference drugs,quercetin and luteolin.Among six components of Kangen-karyu,the water extracts of Salviae Miltiorrhizae Radix or Cyperi Rhizoma exhibited significant inhibitory effects on BACE1 and GSK-3β.Among the constituents of Salviae Miltiorrhizae Radix extract,salvianolic acid C,salvianolic acid A,rosmarinic acid,and magnesium lithospermate B significantly inhibited BACE1.In addition,they inhibited GSK-3βwith an IC50 value range of 6.97 to 135.35μM.From these results,one of the effectiveness and its mechanisms of action of Kangen-karyu against AD may be the inhibition of BACE1 and GSK-3β,and one of the active ingredients of Kangen-karyu is Salviae Miltiorrhizae Radix and its constituents.

高效液相色谱法测定丹参中丹酚酸B

目的 :建立丹参中丹酚酸B高效液相色谱测定方法。方法 :75 %甲醇回流提取 ,色谱柱YWGC18,流动相甲醇 5 %乙酸 (35∶6 5 ) ,检测波长 2 81nm。结果 :丹酚酸B 0 .17～ 1.70 μg线性关系良好 (r =0 .9997) ,回收率98.9% ,RSD1.82 %。结论

UV-B辐射和干旱对丹参生长和叶片中酚酸类成分的影响

在3种UV-B辐射强度以及3种干旱的单因子以及复合作用下,采用盆栽法对丹参植株生物量、叶面积、株高等形态指标和光合参数以及叶片中丹酚酸B、迷迭香酸、咖啡酸和丹参素含量的变化进行研究。旨在探索西北干旱、半干旱地区强光和干旱对丹参生物量和叶片中酚酸类含量的影响,为丹参的深入开发利用提供依据。结果表明:干旱和UV-B辐射单因子作用均对丹参生长指标有抑制作用,而对酚酸积累有促进作用;相比于干旱,UV-B辐射的影响更加显著。UV-B辐射显著引起植株矮化、叶面积减小、生物量降低、净光合速率降低以及叶片中酚酸类物质含量增加,两种因子复合作用对酚酸类物质积累的效果表现为复杂的加强或交互抗性作用。两种因子单独和复合作用处理下,适宜水分和重度UV-B辐射复合处理(75%T2)最有利于丹参叶片中酚酸类物质积累。

高速逆流色谱法分离制备丹酚酸B

采用高速逆流色谱法分离纯化丹参水溶性成分丹酚酸类物质,制备丹酚酸B化学对照品。分离采用的溶剂系统为正己烷-乙酸乙酯-水-甲醇(1.5:5:5:1.5),上相做固定相,下相做流动相,流速为1.7 mL/min,仪器转速850 rpm,进样量80 mg,纯度用HPLC方法测定。结果表明:一次分离可制备63.4 mg丹酚酸B,其纯度为98.6%。该方法操作简单,可作为高纯度丹酚酸B化学对照品的制备分离方法。

丹参水提液中丹酚酸B湿热降解动力学研究

目的:研究丹参水提液中丹酚酸B的湿热降解动力学。方法:以HPLC测定不同温度不同加热时间丹参水提液中丹酚酸B湿热降解动力学相关参数。结果:丹参水提液中丹酚酸B湿热降解符合一级动力学。结论:缩短丹参水提、浓缩、干燥时间,尽快使丹参水提液固体化可减少丹酚酸B损失。

丹酚酸B对缺氧损伤心脏微血管内皮细胞细胞间粘附分子表达的影响

目的 :探讨丹参有效成分丹酚酸B在心脏微血管内皮细胞 (CMEC)缺氧 /复氧损伤中的保护作用。方法 :通过心脏微血管内皮细胞体外培养技术 ,建立缺氧预适应 (HPC)模型并给予丹酚酸B ,应用酶联免疫方法检测细胞间粘附分子 1(ICAM 1)表达。结果 :在CMEC缺氧 3h ,复氧 6h后ICAM 1表达升高 ;缺氧预适应和应用丹酚酸B对CMEC进行预处理则能有效降低CMECICAM 1的表达。结论 :丹酚酸B在CMEC缺氧 /复氧损伤中可产生与缺氧预适应类似的保护效果。

不同产地丹参药材中丹参酮ⅡA和丹酚酸B含量比较

目的:测定和比较不同大规模栽培产地丹参药材中丹参酮ⅡA和丹酚酸B含量。方法:采用高效液相色谱法,选用Synchropak PR100-5色谱柱,检测波长分别为270nm和286nm。结果:丹参酮ⅡA和丹酚酸B含量均较高的是四川中江栽培品。山西芮城栽培品丹参酮ⅡA含量尽管最低,但丹酚酸B含量却最高。结论:丹参中丹参酮ⅡA和丹酚酸B的含量与产地关系较大,此研究结果可作为丹参药材采购及临床选用的参考依据。

丹参氮、磷、钾积累分配特点及其与干物质、丹酚酸B积累的关系

田间条件下研究了丹参的干物质和丹酚酸B积累与植株氮、磷、钾积累的特点及其相互关系。结果表明,丹参干物质积累总量随氮、磷、钾积累总量的增加呈直线增加趋势,相关系数达显著水平。丹参的叶、茎的干物质积累在整个生育期内呈先上升后下降的趋势;根部干物质积累呈上升趋势。在整个生育期内,丹酚酸B的积累趋势与丹参干物质积累趋势相似。不同时期,丹参对氮、磷、钾的吸收量不同。在移栽后30 d内对氮、磷、钾的吸收较少,分别占总吸收量的2.68%,2.23%,2.20%;移栽后60～120 d吸收量迅速增加,氮、磷、钾的吸收量占全部吸收量的58.57%,39.92%,50.18%;移栽120 d后又有所下降。总的来说,植株对氮的吸收量最多,钾次之,磷最少,整个生育期内对氮、磷、钾的吸收比例为1∶0.12∶0.46。

丹参酮ⅡA和丹酚酸B在丹参根组织中的分布特征研究

对生长在粘土、沙土和马骨石土上1～2年生丹参不同根组织的分析表明，丹参酮ⅡA主要分布在木栓层，丹酚酸B分布在木质部和韧皮部．1年生丹参根木栓层的丹参酮ⅡA含量较2年生丹参为高，而1年生丹参根木质部和韧皮部中的丹酚酸B含量与2年生丹参没有显著差异．粘土地栽培丹参根中的丹参酮ⅡA含量较沙土、马骨石土地栽培丹参为高，但不同类型土壤上丹参根中丹酚酸B含量的差异不显著．

丹参酮IIA与丹酚酸B在丹参药材中的分布研究

目的对丹参酮IIA与丹酚酸B在丹参药材中的分布规律进行研究,为丹参药材的GAP栽培与采收加工提供科学依据。方法在丹参药材的采收季节,分别在不同的主产地采集药材样品,按实验需要分成不同部分,晒干后,以HPLC法测定药材中活性成分丹参酮IIA和丹酚酸B的含量。结果研究表明,丹参酮IIA在皮层含量最高,木质部含量甚微,根的下端含量高于上端,须根的含量高;丹酚酸B在皮层的含量高于木质部,根下端含量高于上端,同样以须根的含量高。结论脂溶性的丹参酮IIA与水溶性的丹酚酸B的在丹参根中的分布有一定的规律;这两个成分在丹参根中的横向分布以皮层为高,纵向分布均以根的上端高于下端,须根含量最高。

丹参和丹酚酸B对抗泼尼松致大鼠皮肤衰老作用的观察

目的:探讨丹参(Salvia Miltiorrhiza)和丹酚酸B(Salvianolic acid B,Sal B)对用泼尼松引起的大鼠皮肤衰老作用的影响。方法:32只SD雄性大鼠随机分为4组(n=8):正常对照组,激素模型组(GC)、激素加用丹参组(GC+DS),激素加用丹酚酸B组(GC+Sal B)。正常对照组每天灌胃生理盐水,后3组每天分别灌胃泼尼松3.5mg.kg-1.d-1造模,GC+DS组再给予丹参水提液5g.kg-1.d-1;GC+Sal B组再给予丹酚酸B原料药125mg.kg-1.d-1。喂养90d后处死大鼠取皮肤进行组织病理学检查,并用计算机图像分析系统定量分析表皮厚度、弹力纤维的面积;另外以高效液相色谱法对药品进行丹酚酸B含量测定。结果:GC组可见大鼠皮肤表皮显著变薄,弹力纤维减少(P<0.01),胶原纤维减少,断裂等皮肤衰老样的表现,GC+DS、SalB组可使大鼠表皮增厚、弹力纤维总面积增大(P<0.01),皮肤衰老样改变明显减轻;丹参水提液、丹酚酸B原料药分别检测出相当含量的丹参水溶性成份丹酚酸B。结论:长期灌胃给药泼尼松可使大鼠出现皮肤衰老样改变,丹参和丹酚酸B对其有较好的对抗作用,丹酚酸B可能是丹参对皮肤作用的重要有效成分。

丹酚酸B的体外抗氧化活性

为考察丹酚酸B的体外抗氧化活性,应用比色法测定丹酚酸B对Fenton体系产生的.OH、邻苯三酚自氧化体系产生的O2-.、DPPH.的清除能力及其还原能力、抗脂质过氧化能力,并考查丹酚酸B与抗氧化活性之间的量效关系。结果表明,丹酚酸B有很强的.OH、O2-.、DPPH.清除能力、还原能力和抗脂质过氧化能力,且其抗氧化活性随丹酚酸B浓度的增大而增强,呈明显的量效关系。

丹酚酸B、丹参酮ⅡA对家兔动脉粥样硬化模型内皮细胞功能的影响

[目的]探讨丹参水溶性成分丹酚酸B和脂溶性成分丹参酮ⅡA对家兔动脉粥样硬化模型内皮细胞功能的影响,以寻找其作用靶点。[方法]复制家兔动脉粥样硬化模型,放射性免疫方法检测血浆血栓素(TXB2)、6酮前列腺素F1α(6-keto-PGF1α)、内皮素(ET)浓度,观察丹酚酸B和丹参酮ⅡA对内皮细胞功能的影响。[结果]丹酚酸B能够减少动脉粥样硬化家兔血浆TXB2、ET浓度,增加6-keto-PGF1α浓度,丹参酮ⅡA对动脉粥样硬化家兔血浆TXB2、6-ke-to-PGF1α、ET没有明显影响。[结论]丹参水溶性成分与脂溶性成分在治疗动脉粥样硬化过程中作用靶点及机制不同。

高效液相色谱-质谱联用法研究丹酚酸B的大鼠小肠吸收

目的使用高效液相色谱-质谱联用技术,研究丹酚酸B的大鼠小肠吸收。方法分别在大鼠外翻空肠中加入丹酚酸B溶液和丹参提取液,比较两种给药方式丹酚酸B的吸收,同时观察加入P-糖蛋白抑制剂维拉帕米后对丹酚酸B吸收的影响。结果丹酚酸B在单体形式下吸收较好,维拉帕米对丹酚酸B的吸收未见显著影响。结论丹参提取液中的其他成分抑制了丹酚酸B的吸收,丹酚酸B不是P-糖蛋白的底物。

丹参水溶性有效部位群与丹酚酸B对去卵巢大鼠血脂的调节作用

目的:探讨两种丹参水溶性有效部位群与其主要成分丹酚酸B等对去卵巢大鼠血脂水平的调节作用。方法:40只SD雌性大鼠随机分为5组,每组8只,以去卵巢大鼠为血脂紊乱模型,预防用药90天,观察以上3种丹参制剂分别对大鼠血脂谱各值以及体重、肝脏、子宫重量等的影响。结果:与假手术组比,去卵巢组大鼠体重、总胆固醇(Total cholesterol;TC)明显升高(P<0.01、P<0.05),甘油三酯(Total triglycerides;TG)升高不明显,高密度脂蛋白胆固醇(High density lipoprotein cholesterol;HDL-c)和子宫重量显著下降(P<0.01、P<0.05);与去卵巢组相比,3个丹参药物预防组可显著降低TC和TG水平、升高HDL-c水平(P<0.05或P<0.01),对体重、子宫重量没有显著影响。结论:两种丹参水溶性有效部位群与丹酚酸B能有效降低去卵巢大鼠血脂,其主要成分丹酚酸B在其中有确切的药理学作用。

超临界二氧化碳萃取丹酚酸B的提取工艺研究

目的研究超临界CO2中加入非离子表面活性剂的多元醇混合体系萃取丹酚酸B的新工艺。方法以丹酚酸B的提取率为指标,采用正交实验法进行丹参提取工艺的优选,采用HPLC法测定其含量。结果超临界CO2萃取中加入非离子表面活性剂的多元醇混合体系可大大提高丹酚酸B的提取率。在本实验条件下,丹参的超临界萃取最佳工艺为:夹带剂选择10%乙醇+5%吐温-80,用量与药材相等,萃取温度55℃,压力30MPa,萃取时间1.5h。结论加入非离子表面活性剂的多元醇混合体系比只用乙醇作夹带剂进行超临界CO2提取丹酚酸B的效率高约6倍。