试析乌腺金丝桃中提取分离金丝桃素的相关工艺

根据研究可知,乌腺金丝桃素具有很大的药用价值,将其有机应用,则可以有效抗击癌症、病毒以及抑郁,同时还可以止血消炎,国内外专家加强了对该项物质的研究,研究更科学、有效的提取工艺,满足临床药用需求.本文首先对金丝桃素进行简单介绍,在此基础上对乌腺金丝桃素提取工艺进行研究,希望可以促进金丝桃素提取工艺技术的提升,促进有效应用.     

不同方法除鞣质后贯叶金丝桃提取物中金丝桃素的含量变化

目的 考察用不同方法检测除鞣质后金丝桃素的含量变化和收率,建立贯叶金丝桃良好的除鞣质方法。方法采用明胶沉淀法、改良明胶沉淀法及碱性醇沉法除鞣质后,用高效液相色谱（HPLC）法测定贯叶金丝桃提取物中金丝桃素含量变化和收率;色谱柱为Phenomenex—C18柱（250mm×4．6mm,5μm）,流动相为甲醇-0．006mol／LNa214PO4（7：1V／V,H3P04调至pH=6．5）,流速为1．0mL／min,柱温30℃,检测波长590nm,外标法计算。结果金丝桃素的进样量线性范围为0．0194～0．7760μg（r=0．9999）,平均回收率为100．25％。RSD：1．29％（n=5）;明胶沉淀法、改良明胶沉淀法和碱性醇沉法除鞣质后金丝桃素含量分别为0．092％,0．098％和0．093％,收率分别为70．15％,85．21％和89．16％。结论改良明胶沉淀法具有鞣质去除完全、金丝桃素含量较高、损失较少的优点,且方法简便易行．可应用于贯叶金丝桃提取物除鞣质处理。     

具有抗抑郁作用的金丝桃草制剂在美国热销

金丝桃草(Hypericumperforatum)又称贯叶连翘,因其花期正值西方的圣约翰日,所以在国外惯称圣约翰草(St John'sWort)。金丝桃草提取物中主要含金丝桃素(hypericin)、伪金丝桃素(pseudohypericin)、儿茶精及黄酮类化合物,具有较强的广谱抗病毒作用。最近美、德两国的研究表明金丝     

贯叶金丝桃止痛酊的制备及质量控制

目的：研制贯叶金丝桃止痛酊，并建立其质量标准。方法：用70％乙醇提取贯叶金丝桃，配以丁香油、薄荷油及氮酮制成酊剂，测定pH值及鉴别，并用可见紫外分光光度法测定含量。结果：该制剂1mL含总金丝桃素0．27mg，平均加样回收率100．06％。结论：本制剂制备工艺简单，质量控制方法简便、可靠。     

山地虎耳草质量控制方法研究

目的 建立山地虎耳草的质量控制方法。方法 采用薄层色谱法对山地虎耳草中金丝桃苷进行定性鉴别；采用高效液相色谱法测定山地虎耳草中金丝桃苷和当药醇苷含量。结果 金丝桃苷薄层色谱鉴别斑点清晰，特征明显，专属性强，可行性良好，12批样品均显示有金丝桃苷的斑点；高效液相色谱法测定金丝桃苷和当药醇苷2种成分分别在0.124 6～1.494 6μg(r=0.999 9)、0.045 3～0.543 6μg(r=0.999 9)范围内与其峰面积积分值均呈良好的线性关系。结论 该文建立的质量控制方法简便、准确、可行，可用于山地虎耳草的质量控制。     

乌腺金丝桃药理作用研究进展

乌腺金丝桃为滕黄科金丝桃属植物的干燥全草,具有通乳消痈、止血镇痛的功效,临床上用于治疗乳腺炎和功能性子宫出血等病。研究显示乌腺金丝桃主要含有黄酮类、挥发油、间苯三酚衍生物类成分,其中黄酮类成分应用广泛;药理实验证明其有抗心律失常、抗心肌缺血、抗肺动脉高压、抗炎镇痛、抗抑郁症、抗病毒以及抗肿瘤等作用,对乌腺金丝桃的药理作用的研究进展进行综述,以期为新药开发与临床应用提供参考。     

刺五加叶中金丝桃苷的薄层鉴别和含量测定

目的：以金丝桃苷为指标成分,建立刺五加叶的薄层定性和含量测定的方法。方法：采用薄层色谱法对刺五加叶中金丝桃苷进行定性鉴别,并用高效液相色谱法进行含量测定,色谱柱：Kromasil 100-5C18色谱柱（4.6mm×250mm）;流动相：甲醇-0.1％冰醋酸水溶液(35：65);流速：0.8mL/min;检测波长：360nm;柱温：30℃;进样量：10μL。结果：刺五加叶金丝桃苷的薄层鉴别特征明显,金丝桃苷在0.13～0.78μg范围内具有良好线性关系,r=0.9994,平均回收率为98.17％,RSD为1.02％（n=5）。结论：此方法简便、快速、准确,可用于刺五加叶的质量控制。     

紫外分光光度法测定贯叶金丝桃胶囊中金丝桃素的含量

目的：建立制剂贯叶金丝桃胶囊中金丝桃素的含量测定方法。方法：采用紫外分光光度法，在590nm波长处检测。结果：金丝桃素在3.29—16.49mg.L^-1(r＝0．9999)范围内吸收值与其浓度呈良好的线性关系，方法平均回收率99．54％，RSD为0.44％(n＝6)。结论：该方法简便、快速、准确，可作为本制剂的质量控制方法。     

金丝桃素合酶基因的快速克隆及功能鉴定

金丝桃素合酶能催化大黄素生成金丝桃素,根据已发表的金丝桃素合酶的基因序列,设计了6对引物,通过连续重叠PCR快速克隆得到了金丝桃素合酶基因hyp-1。构建含hyp-1基因的原核表达载体pET32ahyp,将该载体导入大肠杆菌进行诱导表达。SDS-PAGE结果表明,hyp-1基因在大肠杆菌中获得表达;Western blot结果表明,重组的Hyp-1蛋白具有特异的免疫活性,表明hyp-1基因在E.coli中得到了表达。酶促反应表明,Hyp-1确实能催化大黄素形成金丝桃素,上述结果表明,本研究克隆的hyp-1基因是金丝桃素合酶基因,具备催化大黄素形成金丝桃素的能力,从而为通过合成生物学技术制备金丝桃素奠定了物质基础。     

金丝桃素提取物稳定性考察

目的考察金丝桃素提取物的稳定性。方法以金丝桃素和伪金丝桃素含量变化为指标,考察光照、温度、抗坏血酸等因素对金丝桃素提取物短、长期稳定性的影响。结果短期内,金丝桃素提取物在常温避光及加抗坏血酸条件下较稳定;光照能使提取物中金丝桃素、伪金丝桃素含量短时间内升高,然后开始下降;在高温条件下,提取物稳定性差。8周内,提取物在常温光照、常温避光及加抗坏血酸条件下金丝桃素、伪金丝桃素含量均有不同程度地下降,在低温避光条件下比较稳定。结论金丝桃素提取物应低温、避光保存。     

Influence of the developmental stage on the (proto)-hypericin and (proto)pseudohypericin levels of Hypericum plants from Crete

The contents of (pseudo)hypericin and their immediate precursors were studied in wild populations of various Hypericum species on the island of Crete, Greece. Therefore, the aerial parts of wild grown H. perforatum, H. triquentrifolium, H. empetrifolium and H. perfoliatum shoots were collected throughout the island and the quantitative variations in (proto)hypericin and (proto)pseudohypericin examined. The plant material was harvested at different stages of the life cycle of the species and the contents in the above-mentioned compounds analyzed discriminating between flowers/fruits and leaves/petioles. HPLC analysis of hypericin, pseudohypericin and their immediate precursors, protohypericin and protopseudohypericin, revealed great differences in the contents of the compounds in dependence on the developmental stage of the plants. In all examined species the highest concentrations of hypericin were found during blossoming whereas the lowest concentrations were present during ripening of the fruits. H. perforatum and H. triquentrifolium show much higher hypericin levels in flowers/fruits compared to leaves/petioles, whereas the species H. empetrifolium and H. perfoliatum show similar concentrations of total hypericins in both flowers/fruits and leaves/petioles. In the different species the levels of (proto)hypericin and (proto)pseudohypericin varied, but in almost all samples from flowers/fruits and leaves/petioles the ratio of (proto)hypericin to (proto)pseudohypericin was higher than one. When the total amount of hypericins per entire aerial part of a plant was calculated for all developmental stages, we found that H. perforatum contained the highest amount of hypericin. This in combination with the comparatively high concentration of hypericins in flowers/fruits and in leaves/petioles in this species, as well as the high ratio of (proto)hypericin to (proto)pseudohypericin, especially during the developmental stage of blossoming, encourages us to think about the possibility of cultivating Hypericum perforatum in Crete as a medicinal plant in the future.     

Phytochemical analysis of nine Hypericum L. species from Serbia and the F.Y.R. Macedonia

The methanol extracts of the aerial parts of nine Hypericum species (H. barbatum, H. hirsutum, H. linarioides, H. maculatum, H. olympicum, H. perforatum, H. richeri, H. rumeliacum and H. tetrapterum), collected on different locations in Serbia and the F.Y.R. Macedonia, were obtained by accelerated solvent extraction (ASE) and analyzed for the content of four constituents (hyperoside, quercitrin, hyperforin and hypericin) by LC-MS/ MS. All studied extracts contained the characteristic four constituents, but their contents varied between different species and locations. The content of hypericin in H. barbatum was significantly higher (3.9 times) than that in H. perforatum.     

Flavonoid and dianthranoid levels of the St.-John's-wort flowering tops

Dried flowering tops of Hypericum perforatum L. (16 batches) and H. maculatum (3 batches) were studied according to the harvest period. The mean levels of the flavonoid and dianthranoid compounds were respectively: total flavonoids 3.20 and 3.92%, hyperoside 0.87 and 2. 12%, rutin 0.54 and 0.02%, isoquercitrin 0.48 and 0.75%; total dianthranoids 0.14 and 0.13%, hypericin 0.04 and 0.02%, pseudohypericin 0.08 and 0.11%. The flavonoid and dianthranoid levels were higher in young flowering tops of H. perforatum. 3 commercial batches were also examined for a comparative study. Pharmacopoeial specifications are proposed for a revision of the monograph "St.-John's-wort".     

Final report on the safety assessment of Hypericum perforatum extract and Hypericum perforatum oil

Hypericum Perforatum Extract is an extract of the capsules, flowers, leaves, and stem heads of Hypericum perforatum, commonly called St. John's Wort. Hypericum Perforatum Oil is the fixed oil from H. perforatum. Techniques for preparing Hypericum Perforatum Extract include crushing in stabilized olive oil, gentle maceration over a period of weeks, followed by dehydration and filtration. Propylene Glycol and Butylene Glycol extractions were also reported. The following components have variously been reported to be found in H. perforatum: hypericin, naphtodianthrones, flavonoids, terpene and sesquiterpene oils, phenylpropanes, biflavones, tannins, xanthones, phloroglucinols, and essential oils. Hypericum Perforatum Extract is used in over 50 cosmetic formulations and Hypericum Perforatum Oil in just over 10, both across a wide range of product types. Acute toxicity studies using rats, guinea pigs, and mice indicate that the extract is relatively nontoxic. Animals fed H. perforatum flowers for 2 weeks showed significant signs of toxicity, including erythema, edema of the portion of the body exposed to light, alopecia, and changes in blood chemistry. In a chronic study, rats fed H. perforatum gained less weight than control animals. Mixtures containing the extract and the oil were not irritants or sensitizers in animals. Because of the presence of hypericin, H. perforatum is a primary photosensitizer. In clinical tests, a single oral administration of Hypericum extract resulted in hypericin appearing in the blood. With long-term dosing, a steady-state level in blood was reached after 14 days. The polyphenol fraction of H. perforatum had immunostimulating activity, whereas the lipophilic portion had immunosuppressing properties. Mixtures of the extract and the oil produced minimal or no ocular irritation in rabbit eyes. Mutagenic activity in an Ames test was attributed to flavonols in one study and to quercitin in another, but other genotoxicity assays were negative. No carcinogenicity or reproductive and developmental toxicity data were available. A mixture of the extract and the oil was not irritating in clinical studies. Adverse reactions to Hypericum extract in the clinical treatment of depression include skin reddening and itching, dizziness, constipation, fatigue, anxiety, and tiredness. Absent any basis for concluding that data on one member of a botanical ingredient group can be extrapolated to another in a group, or to the same ingredient extracted differently, these data were not considered sufficient to assess the safety of these ingredients. Additional data needs include current concentration of use data; function in cosmetics; photosensitization and phototoxicity data using visible light; gross pathology and histopathology in skin and other major organ systems associated with repeated dermal exposures; dermal reproductive/developmental toxicity data; human skin irritation and sensitization data using the oil; and ocular irritation data, if available. Until these data are available, it is concluded that the available data are insufficient to support the safety of these ingredients in cosmetic formulations.     

贯叶金丝桃药材中萘骈二蒽酮类成分的分光光度法和HPLC法定量分析

目的:建立测定贯叶金丝桃药材中萘骈二蒽酮类成分的紫外-可见分光光度法和HPLC法,比较两者测定结果的差异与定量可靠性。方法:紫外-可见分光光度法以金丝桃素为指标性成分,在590nm处测定。HPLC法以DiamonsilC18柱(150mm×4.6mm,5μm为色谱柱,甲醇-10mmol/L CH3COONH4水溶液(pH值为6.2)(90∶10)为流动相,流速1.0ml/min,检测波长590nm,柱温30℃。经紫外光谱与质谱确认其中主要色谱峰成分为伪金丝桃素和金丝桃素,并以伪金丝桃素、金丝桃素为对照品进行定量,萘骈二蒽酮大类成分含量以伪金丝桃素、金丝桃素含量之和来表征。结果:紫外-可见分光光度法定量金丝桃素在2.44～19.52μg/ml范围内线性关系良好(r=0.999 6,n=3),测得贯叶金丝桃药材中萘骈二蒽酮类成分以金丝桃素计为(0.990 2±0.006 0)μg/mg。HPLC法定量伪金丝桃素在0.402～8.032μg/ml范围内线性关系良好(r=0.999 5,n=3)),定量金丝桃素在0.488～9.760μg/ml范围内线性关系良好(r=0.999 7,n=3)),测得贯叶金丝桃药材中伪金丝桃素和金丝桃素含量分别为(0.387 2±0.001 4)、(0.220 2±0.000 7)μg/mg,两者的总量为(0.605 5±0.001 2)μg/mg。结论:对贯叶金丝桃药材中萘骈二蒽酮类成分的定量分析,紫外-可见分光光度法和HPLC法测定结果差异明显;即使化学结构与光谱特性相似的大类成分,其紫外-可见分光光度法的定量准确性仍需要验证与评价。     

不同采收期及药用部位的长柱金丝桃中有效成分的含量变化

目的 考察长柱金丝桃药用部位、采收期对金丝桃苷和金丝桃素的含量变化.方法 采用HPLC法,色谱柱为Agilent Zorbax Extend-C18 (250 mm×4.6 mm,5 μm),金丝桃苷的流动相为乙腈-0.4％磷酸梯度洗脱,柱温为室温,检测波长360 nm,流速1.0mL·min-1;金丝桃素流动相为甲醇-0.1 mol·L-1磷酸二氢钠(90∶10),柱温为室温,流速为1.0 mL·min-1,检测波长590nm.结果 长柱金丝桃花中金丝桃苷及金丝桃素的含量最高;长柱金丝桃在5月中旬其有效成分含量达到最高,后又逐渐降低.结论 长柱金丝桃中金丝桃苷和金丝桃素的含量随着采收时间的不同而不同,而且不同器官中含量差异显著.     

Screening for the Presence of Hypericins in Southern Brazilian Species of Hypericum

Eight species of Hypericum (H. brasiliense, H. caprifoliatum, H. carinatum, H. connatum, H. cordatum, H. myrianthum, H. piriai and H. polyanthemum) growing in southern Brazil were analyzed by TLC and HPLC for the presence of hypericin and pseudohypericin. Although these polycyclic quinones have been identified in some Hypericum species, they were not detectable in the presently assessed samples. The chemotaxonomy of the taxon is briefly discussed.     

Biochemical activities of extracts from Hypericum perforatum L. 5th communication: dopamine-beta-hydroxylase-product quantification by HPLC and inhibition by hypericins and flavonoids

Extracts from the herb "St. John's wort" (Hypericum perforatum L.) exhibit beneficial effects on patients suffering from mental depressions. Lack of catecholamine neurotransmitters may be one biochemical mechanism for this problem under discussion. It has been recently reported that alcoholic extracts from Hypericum perforatum inhibit dopamine-beta-hydroxylase (D-beta-H) with an I50 of 0.1 mumol/l on the basis of total hypericin content and with an I50 of 21 mumol/l with pure commercial hypericin. As test system polarographic determination of oxygen uptake with tyramine as a substrate analogue was used. In the present paper the quantification of the enzymatic activity and the potential influence of inhibitors are reported using dopamine as substrate and product (noradrenaline) quantification by HPLC. With this test system it could be shown that D-beta-H is strongly inhibited by pseudohypericin (I50 = approx. 3 mumol/l) and hypericin (I50 = approx. 5 mumol/l), whereas the I50-values of various flavonoids (quercitrin, isoquercitrin, hyperoside, rutin, quercetin, amentoflavone, kaempferol) are in the range of 50 mumol/l or higher.