Relaxant activity of raspberry (Rubus idaeus) leaf extract in guinea-pig ileum in vitro

Tea made from the leaves of Rubus idaeus L. (raspberry) has been used for centuries as a folk medicine to treat wounds, diarrhoea, colic pain and as a uterine relaxant. Extracts of dried raspberry leaves prepared with different solvents, (n-hexane, ethyl acetate, chloroform and methanol) were tested in vitro for relaxant activity on transmurally stimulated guinea-pig ileum. The methanol (MeOH) extract exhibited the largest response and also indicated that the active compounds are of a relatively polar nature. Hence the bulk of the leaves were extracted with methanol and the dried extract fractionated on a silica gel column, eluting with chloroform, mixtures of chloroform and methanol and finally methanol. Each fraction was examined by thin layer chromatography and tested for relaxant activity in an in vitro transmurally stimulated guinea-pig ileum preparation. The fractions eluted with chloroform (CHCl(3)) lacked relaxant activity. Samples eluted with CHCl(3)/MeOH (95:5) had moderate relaxant activity, while a second distinctive peak of activity eluted with a more polar solvent mixture (CHCl(3)/MeOH 50:50) provided strong dose dependent responses. Evidence was obtained that there are at least two components of raspberry leaf extract which exhibit relaxant activity in an in vitro gastrointestinal preparation.     

液相色谱-质谱串联法测定清血内消丸中马兜铃酸A含量

目的:建立清血内消丸中马兜铃酸A的限量检查法.方法:采用1％碳酸钠溶液多次加热溶解样品,用三氯甲烷多次提取,弃去三氯甲烷层,碱水层用稀盐酸调pH,加三氯甲烷提取多次,收集合并三氯甲烷层,蒸干,残渣加甲醇溶解,精密吸取1 mL,置20 mL量瓶,加乙腈-水(40∶60)稀释至刻度,以水(0.1％甲酸+5 mmol· L-1甲酸铵)/(％)和乙腈(含5％水,0.1％甲酸+5 mmol· L-1甲酸铵)/(％)为流动相梯度洗脱,应用高效液相色谱/电喷雾质谱多离子反应监测模式(MRM)测定.结果:马兜铃酸A在11.21 ～299.0 pg进样量与马兜铃酸A峰面积呈良好的线性关系(r=0.999 5),检测限0.1μg·L-1,定测限0.3 μg·L-1,平均回收率119.20％,RSD4.0％(n=9).结论:方法准确,快速,简便,重复性好,干扰少,特异性好,能满足马兜铃酸A的痕量残留的检测要求.     

Biological activity of Spatoglossum asperum: a brown alga

Chloroform and methanol fractions of an ethanol extract of Spatoglossum asperum showed antifungal activity against the highly destructive plant pathogen, Macrophomina phaseolina while the n-hexane fraction showed activity against Rhizoctonia solani and Fusarium solani. n-Hexane and methanol fractions also showed nematicidal activity against the plant parasitic nematode, Meloidogyne javanica. Column chromatography of the ethanol extract of seaweed yielded oily fractions named SA-I to SA-VI. GC-MS data of these oily fractions showed the presence of various fatty acid esters. Some of these oily fractions exhibited strong antifungal and nematicidal activities. Oily fractions of S. asperum also showed hypolipidaemic activity in normal, triton-induced and high fat diet-induced hyperlipidaemic rats without producing any ill effect on cardiac and liver enzymes.     

Constituents of the root wood of Austroplenckia populnea var. ovata

The root wood of Austroplenckia populnea var. ovata was extracted successively with chloroform and methanol. Lapachol and dehydro-beta-lapachone were isolated from the chloroform extract, and euonine, alatusinine, wilfordine, 2-O-deacetyleuonine (1), 7-O-deacetyleuonine (2), and austronine (3) from the methanol extract. The structures of the new compounds 1-3 were elucidated by spectroscopic data interpretation. Lapachol, dehydro-beta-lapachone, euonine, alatusinine, and wilfordine are known compounds that are newly identified from root wood of Austroplenckia populnea.     

Antibacterial potential from Indian Suregada angustifolia

Phytochemical analysis of the leaves from Indian Suregada angustifolia (Baill. ex Muell. Arg.) Airy Shaw (Euphorbiaceae) resulted in the isolation and identification of six known compounds, viz. friedelin, epi-friedelinol, n-octacosanol, alpha-amyrin, beta-sitosterol and beta-sitosterol-3-beta-D-glucopyranoside. Aqueous (room temperature, boiled and autoclaved) and various solvent (methanol, chloroform and hexane) extracts of leaves were tested against 12 human pathogenic bacteria by the agar well-diffusion method. Aqueous extracts did not express any activity. Antibacterial activity was recorded in the order of methanol, hexane and chloroform extracts. Maximum activity recorded against Staphylococcus aureus (skin infections) in methanol and hexane extracts and moderate activity recorded against diarrhoea causing bacteria, Vibrio vulnificus (hexane extract) and Vibrio cholerae (chloroform extract).     

防已黄芪汤的抗氧化活性研究

目的测定防己黄芪汤(防己、黄芪、白术、甘草)中甲醇提取物和三氯甲烷提取物的抗氧化活性。方法采用2,2-二苯基-1-苦味基肼自由基清除(DPPH)实验、还原力实验、羟基自由基清除实验对其抗氧化活性进行研究。结果清除DPPH活性研究表明,甲醇提取物与三氯甲烷提取物的抗氧化能力较弱。还原能力实验证明三氯甲烷提取物的抗氧化能力只略低于甲醇提取物。清除羟基自由基实验结果则表明两种提取物均具有很高的清除羟基自由基能力。结论试验结果表明,甲醇提取物及三氯甲烷提取物具有很好的清除羟基自由基能力。     

Synergistic effect of methanol extract of Abies webbiana leaves on sleeping time induced by standard sedatives in mice and anti-inflammatory activity of extracts in rats

During the determination of LD50 values of extracts of Abies webbiana, it was observed that the methanol extract (MEAW) produces sedation of animals. This led to investigation of the effect of MEAW on sleeping time in mice. When various doses of the methanol extract (100, 150, and 200 mg/kg body weight) were administered alone, no hypnotic activity was observed. However, these exhibited significant synergistic effects (P < 0.001) at those dose levels in mice when administered prior to the administration of standard sedatives (pentobarbitone sodium: 50 mg/kg and diazepam: 6 mg/kg, respectively). In addition anti-inflammatory effects of methanol, chloroform, and petroleum ether extracts of Abies webbiana leaves in rats were performed to assess scientific validity of the medicinal claim of Indian folk medicine. The effects of leaf extracts (methanol, chloroform, and petroleum ether) against inflammation were studied by carrageenan-induced paw edema model in rats. The methanol extract (400 mg/kg p.o.) of leaves of Abies webbiana showed the best significant anti-inflammatory activity as compared to that of diclofenac sodium (150 mg/kg p.o.). The LD50 values of methanol, chloroform, and petroleum ether extracts were found to be 986, 1387, and > 3200 mg/kg, respectively. Thus, the therapeutic index of methanol extract may be favorable to open a new vista on combination therapy of hypnotics and may also against inflammation.     

黄药子的肝脏毒性研究

目的:考察黄药子总甲醇提取物不同极性部位的肝脏毒性。方法:灌胃给药10d后测定大鼠的直接胆红素(direct bilirubin,DBil),谷-丙转氨酶(glutmic-pyruvic transaminase,GPT);称量肝重量并计算肝指数;观察大鼠外观、行为及饮食的变化;对肝脏组织进行光镜及电镜观察。结果:甲醇总提取物及其氯仿部位对大鼠的GPT、肝指数、外观、行为、饮食及肝脏的组织形态有显著的影响。结论:氯仿部位是黄药子甲醇总提取物的肝脏毒性部位。     

海南龙血树抗肿瘤新用途研究

目的:筛选海南龙血树的抗肿瘤活性部位.方法:分别用溶剂萃取法和大孔吸附树脂柱层析法对海南龙血树的95%乙醇提取物进行分段处理,利用MTT法测定各提取部位体外抗肿瘤活性.结果:氯仿提取物、乙酸乙酯提取物和正丁醇提取物对体外肿瘤细胞表现出较强的抑制活性,甲醇洗脱物显示了弱活性,水提取、水洗脱物和50%甲醇-水洗脱物则没有活性.在各提取物中氯仿提取物活性最强.结论:氯仿提取部位是海南龙血树主要的体外抗肿瘤活性部位.     

Evaluation of anti-tussive activity of Chamaedorea tepejilote

Aim of the study

Evaluate the potential inhibition of cough in a mouse model induced by sulfur dioxide gas of methanol and chloroform extracts of aerial parts of Chamaedorea tepejilote.

Material and methods

The antitussive effect was examined using the Ogyhara method. The extracts and theobromine were orally administered to mice. Each extract was administered to one group (400 mg/Kg) and theobromine (2 mg), and negative control received PVP. The frequency of cough was measured at 60, 90 min after the administration of chloroform extract.

Results

The water extract of C. tepejilote had no effect on the pharmacological model used in this study. The effects of the methanol and chloroform extracts of this plant on sulfur-dioxide-induced cough in experimental animals show activity dose-dependently in the range between 100 to 500 mg/Kg. At doses of 400 mg/kg, the methanol and chloroform extracts inhibited cough by 41.72% and 49.1%, respectively; these results are comparable to the effect produced by the antitussive agent theobromine (53.4% inhibition).

Conclusions

Both extracts exhibited significant antitussive activity, and the effect of the chloroform extract was dose dependent. The antitussive activity of this extract was comparable to that of theobromine. The effect of chloroform extract on the trachea was also tested; it did not relax pre-contractions in guinea pig trachealis induced KCl, carbachol or histamine.     

Which extractant should be used for the screening and isolation of antimicrobial components from plants?

Freeze dried and finely ground leaves of two plants with known antimicrobial activity, Anthocleista grandiflora and Combretum erythrophyllum were extracted with acetone, ethanol, methanol, methylenedichloride, methanol/chloroform/water and water at a 1 to 10 ratio in each case. The quantity and diversity of compounds extracted, number of inhibitors extracted, rate of extraction, toxicity in a bioassay, ease of removal of solvent and biological hazard were evaluated for each extractant. An arbitrary scoring system was developed to evaluate the above parameters for the different extractants. Acetone gave the best results with these plants with an arbitrary value of 102 followed by methanol/chloroform/water (81), methylene dichloride (79), methanol (71), ethanol (58) and water (47). Four five minute sequential extractions of very finely ground A. grandiflora shaking at a high rate extracted 97% of the total antimicrobial activity.     

The chloroform fraction of Solanum nigrum suppresses nitric oxide and tumor necrosis factor-α in LPS-stimulated mouse peritoneal macrophages through inhibition of p38, JNK and ERK1/2

Solanum nigrum L., commonly known as black nightshade, is used worldwide for the treatment of skin and mucosal ulcers, liver cirrhosis and edema. We aimed to determine the anti-inflammatory active fraction of S. nigrum by serial extractions. S. nigrum was first extracted with methanol, then fractionated with chloroform and water. The effects of S. nigrum fractions, diosgenin and α-solanine on LPS/interferon-gamma-induced nitric oxide (NO) and inducible NO synthase (iNOS), or LPS-induced tumor necrosis factor-α (TNF-α) and interleukin (IL)-6, in mouse peritoneal macrophages were determined. Western blotting analysis was used to detect LPS-induced phosphorylation of p38, JNK and ERK1/2. The chloroform fraction of S. nigrum was cytotoxic in a time and concentration dependent manner; however, the methanol and water fractions were not. The chloroform fraction reduced NO through inhibition of iNOS synthesis and inhibited TNF-α and IL-6 at the level of protein secretion; the methanol and water fractions showed a weak or no effect. The chloroform fraction also suppressed p38, JNK and ERK1/2. Diosgenin and α-solanine were cytotoxic at a high concentration. In particular, diosgenin was able to inhibit TNF-α and IL-6, but both compounds did not affect LPS-induced iNOS expression. These results indicate that the anti-inflammatory compounds of S. nigrum exist preferentially in the nonpolar fraction, ruling out the possibility that diosgenin and α-solanine are the likely candidates. The inhibition of iNOS, TNF-α and IL-6 by the chloroform fraction may be partly due to the suppression of p38, JNK and ERK1/2. Further study is required to identify the active compounds of S. nigrum.     

Evaluation of the analgesic and topical anti-inflammatory effects of Hypericum reflexum L. fil

The present study investigates the analgesic and topical anti-inflammatory effects of the infusion, methanol extract and fractions of the aerial part in blossom of Hypericum reflexum L. fil. in mice. The acetic acid-induced writhing test, formalin test, tail flick test and the tetradecanoylphorbol acetate (TPA)-induced ear inflammation model in mice were used to determine these effects. Our findings show that oral administration of all extracts tested from this species significantly inhibit acetic acid-induced writhing in mice. Only the methanol extract and chloroform fraction were significantly active in both phases of formalin-induced pain and in the tail flick assays, suggesting that they may have central analgesic properties. On the other hand, the topical treatment of methanol extract, butanol and chloroform fractions of this species significantly reduced the TPA-induced ear oedema. In conclusion, the results indicate analgesic and topical anti-inflammatory activities in mice for the Hypericum species studied.     

一种在多糖分离纯化过程中新的脱蛋白方法

目的　寻找一种在多糖 (香菇多糖 )分离纯化过程中适宜的脱蛋白方法。方法　以正交试验法为主。结果　通过正交试验法找到了一种在多糖 (香菇多糖 )分离纯化过程中新的脱蛋白方法。 5因素 3水平正交试验的结果表明 ,影响脱蛋白率 ( DPR)的主要因素是 B(正丁醇 )、次主要因素是 D(乙醇 ) ,而因素 C(甲醇 )和 E(氯化钠 )的影响则较小 ;影响 DPR的主次顺序为 B>D>A(氯仿 ) >E>C。 DPR随着正丁醇浓度的增加而变化很大 ,开始缓慢上升 ,然后迅速降低 ;随着乙醇浓度的增大 ,DPR逐渐降低 ;而当氯仿浓度增大时 ,DPR则开始上升 ,然后下降 ;当甲醇浓度增大时 ,DPR则变化很小。脱蛋白的最佳条件是 A2 B2 C1 D1 E2 ,即在氯仿 4 0 %、正丁醇 10 %、甲醇 2 %、乙醇4 0 %、氯化钠 0 .0 1%的条件下 ,杂蛋白的去除率最高。验证性试验结果表明 ,在此条件下 ,杂蛋白的去除率达99.98 %。通过反复试验 ,还找到了适用于工业化生产的包括香菇原料的蒸煮、煮沸样品的浓缩、粗多糖的沉淀和脱色、粗多糖的干燥等在内的原料预处理的最佳工艺。结论　这种新的脱蛋白方法具有工艺简单、无需贵重设备、加工成本低、容易放大等特点 ,特别适合于多糖类产品的精制或其他需要去除杂蛋白的单元操作     

In vitro antiproliferative and antioxidant activities and total phenolic contents of the extracts of Melastoma malabathricum leaves

The present study aims to determine the in vitro antiproliferative and antioxidant activities of various extracts from the leaves of Melastoma malabathricum using various established in vitro assays. The aqueous extract inhibited the proliferation of Caov-3 and HL-60 cell lines, while the chloroform extract exhibited antiproliferative activity against the Caov-3, HL-60, and CEM-SS cell lines. The methanol extract demonstrated antiproliferative activity against more cell lines, including the MCF-7, HeLa, Caov-3, HL-60, CEM-SS, and MDA-MB-231 cancer cell lines. Interestingly, all extracts did not inhibit the proliferation of 3T3 cells, thus indicating their noncytotoxic properties. Unlike the chloroform extracts, the aqueous and methanol extracts of M malabathricum (20, 100, and 500 μg/ml) produced high antioxidant activity for the superoxide scavenging assay with only the 500 μg/ml aqueous and methanol extracts exhibited high activity for the 2,2-diphenyl -1-picrylhydrazyl radical scavenging assay. The total phenolic content recorded for the aqueous, methanol, and chloroform extracts were 3344.2 ± 19.1, 3055.1 ± 8.7, and 92.5 ± 7.3 mg/100 g of gallic acid, respectively. The M malabathricum leaves possessed potential antiproliferative and antioxidant activities that could be attributed to its high content of phenolic compounds.     

Antienterobacterial activity of Hemidesmus indicus R. Br. root extract

The antienterobacterial activity of the chloroform and methanol extracts of Hemidesmus indicus root was demonstrated using a variety of methods and different enterobacterial strains. Although the constituents were similar in the chloroform extract (CHI) and the fatty substance separated (ME1) from the methanol extract (MHI), ME1 was found to be more effective than CHI as evident from the disc diffusion method. ME1 was found to be more active than MHI, followed by CHI. This may be due to the inefficient diffusion of CHI into the medium. In a modified agar well diffusion and swab method the activity of the extract against different strains was observed in a single plate. The extracts inhibited growth in a dose dependent manner; both MHI and CHI were most effective against S. flexneri, least effective against S. dysenterie and moderately effective against the other strains. The presence of antimicrobial trace elements such as copper and zinc, along with other active constituents may contribute to the antienterobacterial activity of Hemidesmus indicus root.     

Screening for the antidepressant activity of some species of Hypericum from South Brazil

The crude methanol extracts of three species of the genus Hypericum (H. brasiliense, H. caprifoliatum and H. myrianthum) native to South Brazil were evaluated for the antidepressant activity according to the forced swimming test, a classical animal model for antidepressant drug screening. Among the species studied, only H. caprifoliatum showed activity. Thus, this plant was fractionated with solvents in increasing polarity (petroleum ether; petroleum ether: chloroform (1:1); chloroform and methanol). The petroleum ether fraction was the only one which demonstrated antidepressant activity at a dose of 270 mg/kg/day (i.p.). The chemical analyses showed that this fraction is rich in phenolic compounds, mainly of the phloroglucinol type.     

刺山柑果实抗炎活性部位与致敏部位的分离

目的从刺山柑果实的5个化学部位中分离出抗炎活性部位以及致敏部位。方法采用蛋清法所致的小鼠关节炎模型以及局部皮肤过敏模型2个实验进行研究。结果刺山柑果实95%乙醇提取物的乙酸乙酯部位、甲醇部位及水部位对小鼠踝关节肿胀抑制有显著的抗炎作用(P<0.01),而氯仿部位、乙酸乙酯部位及水部位对小鼠皮肤均呈现不同程度的致敏性。结论乙酸乙酯部位具有显著的抗炎活性和中度致敏性,甲醇部位有显著的抗炎活性且无致敏性。     

Effect of Plantago major on cell proliferation in vitro

Plantago major (Plantaginaceae) is popularly used to treat tumors, infections and as a blood purifier. Aqueous, methanol, chloroform and hexane extracts of the aerial parts (leaves and seeds) were added to CD(1) mice bone marrow and spleen cultures incubated at 37 degrees C for 72h, and also added to Escherichia coli, Bacillus subtilis and Candida albicans cultures, while methanol extract dilutions were added to HTC-15, OVCAR, UISO and KB cell line cultures. Doses of 0.4 and 0.2 mg/mL of aqueous and methanol extracts increased the bone marrow cell concentration by 2.70- and 3.15-fold, respectively, and increased the spleen cell concentration by 3.38- and 6.39-fold, respectively (p < 0.001). Aqueous extract inhibited Bacillus subtilis growth from 78 to 21%; hexane extract inhibited the growth of Escherichia coli, and methanol and chloroform extracts weakly inhibited the growth of Bacillus subtilis and Escherichia coli, respectively. Methanol extract (1 microg/mL) decreased the UISO and OVCAR cell concentrations to 59 and 82%, respectively. Data demonstrate for the first time that Plantago major has hematopoietic activity in vitro.     

Inhibition of Helicobacter pylori-induced inflammation in human gastric epithelial AGS cells by Phyllanthus urinaria extracts

AIM OF THE STUDY: Helicobacter pylori is linked to a majority of peptic ulcers and to some types of gastric cancer, and its resistance to antibiotic treatment is now found worldwide. This study is aimed at evaluating the antimicrobial activity of Phyllanthus urinaria Linnea (Euphorbiaceae), chloroform (PUC) and methanol (PUM) extracts, and its eight isolates on H. pylori-infected human gastric epithelial AGS cells. MATERIALS AND METHODS: The in vitro anti-bacterial activity of P. urinaria chloroform (PUC) and methanol (PUM) extracts, and its eight isolates were determined. Additional experiments were also performed to know the PUC and PUM ability to inhibit the H. pylori adhesion to and invasion of AGS cells, in addition to the effect of PUC on NF-kappaB activity as well as IL-8 synthesis during H. pylori infection of AGS cells. RESULTS: The results revealed that crude extracts PUC and PUM showed potent antimicrobial activity against H. pylori than pure isolates. On the other hand, in vitroH. pylori-infection model revealed that the inhibition of bacterial adhesion and invasion to AGS cells has dramatically reduced by treatment of extract PUC, while PUM has the same moderate effect. Furthermore, H. pylori-induced nuclear factor (NF)-kappaB activation, and the subsequent release of interleukin (IL)-8 in AGS cells were also inhibited by the extract PUC. CONCLUSIONS: These results open the possibility of considering P. urinaria a chemopreventive agent for peptic ulcer or gastric cancer, but this bioactivity should be confirmed in vivo in the future.