UHPLC-Q Exactive高分辨质谱联用法分析淫羊藿中的化学成分

目的 采用UHPLC-Q Exactive四极杆-静电场轨道阱高分辨质谱联用技术鉴定分析淫羊藿药材成分。方法 采用Thermo Hypersil GLOD C₁₈(100 mm×2.1 mm, 1.9μm)色谱柱，体积分数0.1%甲酸水溶液-乙腈溶液为流动相进行梯度洗脱，流速为0.3 mL·min^-1,柱温为40℃,电喷雾正、负离子模式扫描。结果 共分析和鉴定出79个化合物，包括黄酮类、生物碱类、有机酸类和木脂素类化合物，其中16个化合物经与对照品比对鉴定。鉴定的黄酮类包括46个异戊烯基黄酮醇类化合物，异戊烯基黄酮醇类负离子模式二级特征碎片为m/z 367、351、513、353和323。初步鉴定化合物41、54、76、65和69为新的异戊烯基黄酮。结论 淫羊藿中异戊烯基黄酮醇类多具有淫羊藿素或8-异戊烯基山奈酚苷元母核结构，该结果为淫羊藿成分的系统分析提供了数据支持。     

箭叶淫羊藿中一个新的黄酮苷类成分

利用现代柱色谱技术,如大孔吸附树脂、硅胶、ODS、Sephadex LH-20和HW-40C柱色谱及半制备液相等对箭叶淫羊藿正丁醇部位化学成分进行分离纯化,得到14个黄酮类化合物。根据化合物的理化性质和波谱特征,确定其结构分别为3′-羟基宝藿苷-Ⅱ(1)、huazhongilexone-7-O-β-D-glucopyranoside (2)、kaempferol-3-O-α-L-rhamnoside (3)、宝藿苷-Ⅱ(4)、淫羊藿次苷-Ⅱ(5)、kaempferol 3,7-di-O-α-L-rhamnopyranoside (6)、(+)-aromadendrin (7)、kaempferol 3-O-(2-O-β-D-apiofuranosyl)-α-L-rhamnopyranoside (8)、箭藿苷A (9)、2″-O-rhamnosyl icariside-II (10)、apigenin-7-O-β-D-glucoside(11)、quercetin3-O-β-D-apiofuranoyl-(1→2)-α-L-rhamnopyranoside(12)、山柰酚(13)、淫羊...     

朝鲜淫羊藿的化学成分

目的研究朝鲜淫羊藿(Epimedium koreanumNakai)中的化学成分。方法朝鲜淫羊藿干燥地上部分用水回流提取后,经大孔吸附树脂柱色谱分离,用水及不同体积分数的乙醇洗脱;50%乙醇洗脱部分的浸膏用硅胶柱色谱分离,用氯仿甲醇梯度洗脱,得到的第5、8、10流份用羟丙基葡聚糖凝胶柱色谱分离、十八烷基键合硅胶柱色谱分离、制备型HPLC纯化后得到化合物1~8;根据化合物的理化性质和核磁共振氢谱、碳谱数据对所得化合物进行结构鉴定,分析确定化学结构。结果从朝鲜淫羊藿水提取物中分离得到8个化合物:淫羊藿苷(icariin,1)、宝藿苷Ⅰ(baohuosideⅠ,2)、箭藿苷B(sagittatoside B,3)、宝藿苷Ⅱ(baohuosideⅡ,4)a、stragalin(5)、3,5,7三羟基4′甲氧基8异戊烯基黄酮3OαL鼠李吡喃糖基(1→2)α鼠李吡喃糖苷(6)、1,2,3,4 tetrahy-dro 3,7 dihydroxy 1(4 hydroxy 3 methoxyphenyl)6 methoxy 2,3 naphthalenedimethanol(7)、朝藿定C(epimedin C,8)。结论化合物7为首次从该属植物中分离得到,化合物8为首次从该种植物中分离得到。     

淫羊藿苷的提取、分离和抗肿瘤作用机制的研究进展

综述了近年来淫羊藿苷的提取分离方法和抗肿瘤活性及其机制.目前报道的提取方法有醇提法、超声法、微波法、高压法,分离方法有柱色谱法(聚酰胺法和大孔吸附树脂法)、聚酰胺色谱联用醋酸乙酯萃取法等.淫羊藿苷及其衍生物对肺癌、胃癌、肝癌、乳腺癌、白血病细胞均有一定的抑制活性,主要作用机制为淫羊藿苷通过影响肿瘤细胞周期,下调与肿瘤细...     

阿可拉定的有关物质研究

目的分离纯化阿可拉定(淫羊藿苷,IC-162)的主要有关物质,进行结构确证。方法利用反相硅胶和凝胶柱色谱分离阿可拉定中的2个主要杂质,经化合物谱图数据(LC-DAD、LC-MS、1H-NMR和13C-NMR)鉴定其结构。结果阿可拉定中2个主要杂质分别为8-异戊烯基山柰酚和朝藿素B。结论阿可拉定有关物质的定量和定性研究可以为制定合理的杂质限度提供依据。     

8—异戊烯基黄酮类化合物的波谱学特征

综合了从朝鲜淫羊藿（ＥｐｉｍｅｄｉｕｍｋｏｒｅａｎｕｍＮａｋａｉ）的地上部分分离获得的８个８－异戊烯基黄酮类化合物的波谱学特征。     

淫羊藿苷转化产物的制备及活性研究

目的旨在分析淫羊藿苷及转化产物抑制α-葡萄糖苷酶活性研究。方法对淫羊藿苷进行酶水解、酸水解、酶水解与酸水解相结合的方式进行结构转化,采用薄层色谱法、理化方法、核磁共振波谱法对转化产物进行结构鉴定,分别得到淫羊藿次苷Ⅰ、淫羊藿次苷Ⅱ、水合淫羊藿素、环淫羊藿素及脱水淫羊藿素。采用pNPG法建立α-葡萄糖苷酶筛选模型,对淫羊藿苷及其产物进行α-葡萄糖苷酶抑制活性筛选。结果体外活性试验果表明,脱水淫羊藿素、淫羊藿次苷Ⅰ及淫羊藿次苷Ⅱ体外活性较淫羊藿苷好,其中脱水淫羊藿素(IC_(50)=0.102 mg/mL)的活性最好,与拜糖平(IC_(50)=0.850 mg/mL)相比抑制活性更强。结论淫羊藿苷转化产物可作为继续进行结构修饰的先导化合物,具有深入研究和开发的前景。     

RP-HPLC测定淫羊藿不同品种和部位中柔藿苷和淫羊藿苷

目的:为了对中药淫羊藿中柔藿苷和淫羊藿苷含量进行系统研究,测定了淫羊藿不同品种和药用部位的柔藿苷和淫羊藿苷含量。方法:采用RP-HPLC法对3个品种多个样品进行测定,以BECKMAN COULTER_(TM)-C_(18)柱(带预柱)为色谱柱,流动相为乙腈-水(25∶75),流速1.0mL·min~(-1),检测波长为270nm。结果:柔藿苷在0.19642-1.9642μg范围内呈良好的线性关系,淫羊藿苷在0.19335-1.9335μg范围内呈良好的线性关系。巫山淫羊藿同一植株体的不同部位中柔藿苷和淫羊藿苷的分布均为叶>根>茎。巫山淫羊藿同一植株的任何部位都是柔藿苷含量高于淫羊藿苷,而淫羊藿和箭叶淫羊藿叶中都是淫羊藿苷含量高于柔藿苷。结论:巫山淫羊藿中以柔藿苷为主要成分,淫羊藿和箭叶淫羊藿中以淫羊藿苷为主要成分。     

脱水淫羊藿素与山柰素对体外培养成骨细胞成熟矿化影响的比较研究

比较研究脱水淫羊藿素与山柰素对体外培养大鼠成骨细胞(rat osteoblasts,ROB)增殖与成熟矿化的影响。取新生SD大鼠颅骨,多次酶消化法获得成骨细胞,培养于含10%FBS的MEM培养液中,3天后首次换液,铺满皿底90%以上后传代培养。分别用1×10 4、1×10 5、1×10 6和1×10 7mol.L 1的脱水淫羊藿素和山柰素进行药物干预,增殖分析采用MTT法,诱导培养9天后检测碱性磷酸酶(alkaline phosphatase,ALP)活性,筛选出最佳浓度。采用最佳浓度的脱水淫羊藿素和山柰素进行药物干预,比较脱水淫羊藿素、山柰素和对照组之间的ALP、钙盐沉积量、骨钙素分泌量、CFU-FALP和矿化结节数量等成骨性指标。提取总RNA,实时荧光定量PCR(real time PCR)检测BMP-2、Osterix(OSX)及Runx-2的mRNA表达情况,同时提取总蛋白,Western blotting法检测Ⅰ型胶原蛋白的分泌量。结果表明,1×10 4mol.L 1脱水淫羊藿素和山柰素对细胞增殖有抑制作用;1×10 5mol.L 1脱水淫羊藿素虽对ROB增殖无明显影响,但能显著提高成骨细胞中ALP活性、钙盐沉积量和骨钙素分泌量,增加CFU-FALP和矿化结节数量,与成骨细胞成熟矿化相关的因子BMP-2、OSX和Runx-2的基因表达量也显著升高,同时I型胶原蛋白的分泌量也明显增多;而山柰素并没有表现出促进体外培养ROB成熟矿化的特性。脱水淫羊藿素促进成骨细胞成熟矿化的作用明显高于山柰素,很可能是由脱水淫羊藿素8位C原子上存在的异戊烯基引起的。     

药典内5种淫羊藿中黄酮类成分的反相高效液相色谱分析

报道了药典规定的5种淫羊藿──淫羊藿、箭叶淫羊藿、巫山淫羊藿、柔毛淫羊藿和朝鲜淫羊藿中9种黄酮──淫羊藿甙(icariin)、宝藿甙-Ⅰ(baohuosideI)、宝藿甙-Ⅱ(baohuosideⅡ)、淫羊藿甙A(epimedosideA)、箭藿甙B(sagittatosideB)、朝藿定B(epimedinB)、朝藿定C(epimedinC)、大花淫羊藿甙C(ikarisosideC)和大花淫羊藿甙F(ikarisosideF)的反相高效液相色谱测定。色谱柱为DISK-C_phenlyl,流动相为乙腈-乙酸液(水:36%乙酸=100:4),梯度洗脱,检测波长为272nm。     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg x kg(-1)) or i.p. (50 mg x kg(-1)) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) 1 x h(-1) x kg(-1) in the male rat and 10.6 (95% CI: 7.5, 15.0) 1 x h(-1) x kg(-1) in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was approximately 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p < 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p < 0.001) in plasma obtained from the male (8.8 +/- 2.0%) compared with the female rat (11.7 +/- 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Hyperkalaemia in patients in hospital

A survey of all laboratory blood specimens with a plasma potassium concentration greater than or equal to 5.5 mmol/L was conducted over a three month period. Of 331 specimens with hyperkalaemia, 71 were excluded because the specimens was haemolysed, old or contaminated. The laboratory served a population of 348,561 and during this time measured the plasma potassium on 25,016 occasions. Sixty-six outpatients and 20 neonates were not evaluated. The survey was undertaken on 86 of 102 inpatients (46 males), 48 of whom were over 66 years of age. Fifty-seven patients were admitted under a medical service and 29 under a surgical service. Fifty-nine had a single episode of hyperkalaemia. Thirty-two underwent a surgical procedure. The commonest contributing factor was impaired renal function which was present in 71 (83%) patients. Although a definitive causative role for drugs could be identified in only five patients, in 52 (60%) patients drugs were a contributing factor (potassium supplements 24, ACE inhibitors 16, nonsteroidal antiinflammatory drugs 12). Thirty-five of the 86 (41%) patients died during their hospital admission. Nineteen of the 35 deaths occurred within three days of the hyperkalaemia being recorded. A normal plasma potassium was eventually documented in 50 of the 86 patients. Of the remaining 36 patients, 25 (69%) subsequently died. In general the treatment of patients with hyperkalaemia focused on identifying and treating the underlying cause. Hyperkalaemia must always be considered seriously and regard given to the overall clinical status of the patient, with particular attention to drug therapy, renal and cardiac function, acid base status and the possibility of sepsis.     

Changes in angiopoietin expression in glomeruli involved in glomerulosclerosis in rats with daunorubicin-induced nephrosis

AIM: To study the potential pathological role of endogenous angiopoietins in daunorubicin-induced progressive glomerulosclerosis in rats. METHODS: Seventy male Wistar rats were allocated randomly into a daunorubicin group (DRB; n=40) or a control group (n=30). The rats in the DRB group were injected with DRB (15 mg/kg), in their tails. Subsequently, at intervals of 1, 2, 4, 6, 8, and 12 weeks, 5 male Wistar rats in each group were chosen randomly for 24 h urinary protein quantitative measurements (24 h UPQM), and determination of plasma tumor necrosis factor alpha (TNF-alpha), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) levels. Kidney sections were examined by electron microscopy, Periodic Acid Schiff (PAS) staining, immunohistochemical staining and in situ hybridization histochemistry. RESULTS: As glomerulosclerosis progressed in the DRB group, expression of Ang1 mRNA and protein in glomeruli decreased and expression of TNF-alpha protein, Ang2 mRNA and protein in glomeruli increased. Expression of Ang1 mRNA and protein in glomeruli were negatively correlated with 24 h UPQM, Fn protein expression, and mean area of extracellular matrix (MAECM). In comparison, expression of Ang2 mRNA and protein in glomeruli were positively correlated with 24 h UPQM, Fn protein expression and MAECM; furthermore, there was a positive correlation between plasma Ang2 and 24 h UPQM. Plasma TNF-alpha and expression of TNF-alpha in glomeruli were positively correlated with expression of Ang2 mRNA and protein in glomeruli. There was a negative correlation between Ang1 protein expression and Ang2 protein expression in glomeruli. CONCLUSION: During DRB-induced glomerulosclerosis, podocyte injury led to a shift in the balance of Ang1 and Ang2 in glomeruli. Increased TNF-alpha in plasma and glomeruli may upregulate Ang2 expression in glomeruli. Elevated Ang2 in both plasma and glomeruli may mediate protein permeability through the glomerular filtration barrier. Moreover, local expression of Ang2 may facilitate the progress of glomerulosclerosis by upregulating a component expression of extracellular matrix.