金丝桃素的提取纯化工艺研究

用乙醇从贯叶连翘中提取金丝桃素并对其进行纯化,考察了金丝桃素的工艺流程.用乙醚作为除杂剂,用乙醇热浸贯叶连翘,后浓缩浸提液为浸膏;用热水溶解浸膏,乙酸乙酯萃取,浓缩萃取液为浸膏(粗提物).然后再用乙醇溶解,利用树脂柱层析分离纯化金丝桃素,减压浓缩洗脫液,干燥,即得金丝桃素产品.经HPLC测定,结果表明:该产品中金丝桃素的含量高达1.2% ;该工艺不但有效改善了金丝桃素产品易吸湿、易发粘的缺点,而且具有操作简单,经济易行的特点.     

大孔吸附树脂分离纯化贯叶连翘中的金丝桃素

以贯叶连翘粗提浸膏为材料,考察六种大孔吸附树脂分离纯化金丝桃素的性能。采用静态、动态吸附方法筛选树脂,以中压分离方法确定分离纯化的条件。结果表明:HZ-801树脂以其高吸附率、高洗脱率成为优选的分离填料,其最佳分离纯化条件为:进样液质量浓度为0.1125 g/m L,进样速度为5.0 m L/min、洗脱速度为20 m L/min,0~95%乙醇溶液梯度洗脱,金丝桃素的纯度为79.11%。HZ-801可以较好分离纯化金丝桃素,纯化工艺具有较大实践性及参考价值。     

高效液相色谱法测定贯叶金丝桃粉剂中金丝桃素的含量

为了有效控制贯叶金丝桃粉剂的质量,试验采用高效液相色谱(HPLC)法,流动相为乙腈-水-乙酸铵(三者比例为50∶40∶10;乙酸铵浓度为0.30 mol/L,pH值为3.75),流速为1 mL/min,检测波长为588 nm,测定贯叶金丝桃粉剂中金丝桃素的含量。结果表明:金丝桃素的检测浓度在10～80μg/mL范围内与峰面积的线性关系良好(r=0.999 6),平均回收率为98.3%,RSD=1.14%。说明该方法简便、准确、快速、重复性好,可用于贯叶金丝桃粉剂的质量控制。     

金丝桃素提取、分离方法的研究

目的:研究贯叶连翘中有效成分金丝桃素提取方法。方法:应用有机溶剂溶解法、薄层色谱法、紫外检测、高效液相色谱(HPLC)等方法探讨贯叶连翘中金丝桃素提取方法。结果:用石油醚除叶绿素、花青素,以丙酮为溶剂在60℃水浴中提取贯叶连翘中金丝桃素和酮类衍生物,在真空度为0.07MPa、温度为60℃水浴条件下浓缩。后用100-200目硅胶分离金丝桃素,钢瓶冻干,得含金丝桃素为1.08%深红色产品。     

贯叶连翘中金丝桃素的薄层色谱检测

目的:建立贯叶连翘中金丝桃素含量的测定方法。方法:以硅胶GF254为固定相,以石油醚-乙酸乙酯-甲醇(2:4:1)为展开剂,在588nm波长的条件下进行单波长薄层色谱扫描来测定贯叶连翘中金丝桃素含量。结果 :金丝桃素在0.15-7.5μg范围内呈线性关系,(R=0.99987),平均回收率97.92%,RSD=1.82(n=5)。结论:该方法操作简便、分离效果好、价廉,可用于贯叶连翘中金丝桃素的含量测定及质量控制。     

乌腺金丝桃中金丝桃素超声法提取工艺研究

为了筛选乌腺金丝桃中金丝桃素超声提取最佳工艺,试验采用高效液相色谱(HPLC)法测定金丝桃素的含量,并以金丝桃素提取率为评价指标,通过正交试验优化提取工艺。结果表明:超声提取金丝桃素的影响因素顺序为,乙醇浓度温度超声时间料液比;最佳提取工艺条件为,样品15倍量的70%乙醇在超声温度40℃条件下提取60 min,提取2次,此时金丝桃素提取率可达到94.66%。说明超声法提取乌腺金丝桃中金丝桃素的得率较高、工艺可行,具有一定的推广意义。     

金丝桃素的研究进展

金丝桃素是贯叶连翘中最具生物活性的物质,具有抗抑郁、抗肿瘤、抗病毒、止血消炎及光动活性等。在德国,将金丝桃素与蛋白结合,用于治疗艾滋病及甲、乙型肝炎;在美国作为食品添加剂已被广泛应用。金丝桃素具有多种药理活性,目前已成为国内外研究开发的热点。现将近几年金丝桃素的研究进展阐述如下。1金丝桃素的化学结构及特性金丝桃素名为4,4',5,5',7,7'-六羟基-2,2-二甲基-中位-萘并二蒽酮,分子式C30H16O8,属于二蒽酮类化合物,是一种蓝黑色针状结晶体,其熔点是320℃,易溶于吡啶和其它有机胺类,呈橙红色并带红色荧光,可溶于碱水溶液中。pH…     

RP-HPLC测定贯叶连翘中的金丝桃素含量

采用反相高效液相色谱法测定了贯叶连翘提取物金丝桃素浸膏中金丝桃素的含量。用Kromasil C18(5μm,250×4.6 mm)色谱柱为固定相,乙腈-水(140∶10)为流动相,流速为1 mL/min,检测波长为284 nm。结果金丝桃素的含量为0.914%,平均回收率为96.76%,日内精密度RSD为2.67%,日间精密度RSD为3.65%。     

紫花苜蓿黄酮的提取与纯化工艺初探

为探讨紫花苜蓿(Medicago sativeL.)黄酮的提取与纯化工艺,采用不同乙醇浓度和不同pH进行提取,然后用不同来源和型号的大孔树脂进行纯化,以得出苜蓿黄酮的最佳提取和纯化方法.结果表明:30％乙醇、水浸提(pH为7.5)对紫花苜蓿黄酮的浸出率最高,分别为0.46％和0.49％,pH在9.0时所得提取物黄酮含量最高为1.61％;pH为2.2～2.4时,对紫花苜蓿黄酮有最好的沉淀效果,所得沉淀物中黄酮含量为9.80％;9种型号的大孔树脂中DS-17对乙醇提取物中紫花苜蓿黄酮有较好的分离效果,所得提取物黄酮为5.87％;在大孔树脂对碱溶酸沉提取物中黄酮的分离中,2组试验均以DM130的效果最好,所得分离物黄酮含量最高分别为24.25％和29.35％.     

益肾清列口服液的质量标准研究

目的:建立益肾清列口服液的质量标准。方法:采用TLC法对益肾清列口服液中白术、白芍、菟丝子定性鉴别;采用HPLC法对其中菟丝子中主要有效成分金丝桃苷进行含量测定。结果:方中各药TLC鉴别法分离好、专属性强、重现性好、阴性无干扰;HPLC法中,以金丝桃苷浓度为横坐标、相应峰面积为纵坐标得回归方程为Y=1.782×10~7X-3 736.7 (r=0.999 9),金丝桃苷浓度在0.005 15～0.103 00 mg/mL范围内线性关系良好,平均加样回收率为97.9%,RSD为2.1%(n=6)。结论:该研究建立的定性定量方法分离好、专属性强、结果准确、重现性好,可有效控制益肾清列口服液质量。     

Reassessment of the toxicity of Hypericum perforatum (St John's wort) for cattle

OBJECTIVE: To investigate the clinical effect of administering sufficient Hypericum perforatum to cattle to deliver quadruple the reported oral toxic dose. ANIMALS: Thirty-six yearling Hereford (n = 18) and Angus (n = 18) steers. DESIGN: A series of six experiments was conducted, each using 12 animals in a 2 x 2 factorial design, with two breeds of cattle (Hereford, Angus) and two dose levels of hypericin, 1.5 mg/kg (treated group) and 0 mg/kg (control group). Each set of 12 steers was used in duplicate experiments, with all animals alternated between treated and control groups. PROCEDURES: Treated groups received finely milled H. perforatum administered orally in gelatin capsules to provide 1.5 mg hypericin/kg body weight. All cattle were then exposed to direct sunlight for 5 h per day for 5 successive days. Rectal temperatures were measured immediately before and at the end of each sunlight exposure session. Rectal temperature above 40 degrees C, together with some other clinical sign of hypericin poisoning, was considered indicative of intoxication. RESULTS: No animals developed a rectal temperature above 40 degrees C or other clinical signs of hypericin poisoning. CONCLUSIONS: While the reported bovine oral toxic dose of 3 g dried plant/kg body weight, for flowering stage, presumed narrow leaved biotype, H. perforatum, is probably correct, the corresponding dose for hypericin of 0.37 mg/kg is incorrect. Based on its known concentration in this plant the toxic dose of hypericin for partially pigmented Hereford-cross cattle is estimated at about 10.5 mg/kg body weight and more than this for fully pigmented cattle. This would imply that cattle of the former type should be about three and a half times better protected against H. perforatum toxicity than are unpigmented, wool protected, Merino sheep. Cattle, particularly if fully pigmented, may have a role in grazing management to control H. perforatum.     

Sunlight associated hyperthermia as a consistent and rapidly developing clinical sign in sheep intoxicated by St John's wort (Hypericum perforatum)

OBJECTIVE: To assess the usefulness of rectal temperature responses in Australian bred Merino sheep, following the oral administration of Hypericum perforatum (St John's wort), as an early indicator of Hypericum intolerance. DESIGN: Thirty-three Merino ewes were divided into three groups of 11. Each group was dosed with finely ground, dried, flowering growth stage H perforatum plant material at either 5.7, 4.0, or 2.85 g dry plant per kg live weight. This corresponded to 5.3, 3.7 and 2.65 mg hypericin per kg live weight, respectively. PROCEDURE: The sheep were dosed with a plant slurry by stomach tube and then exposed to bright sunlight for up to 5 h per day over successive days. Their clinical responses were observed and rectal temperature measured. RESULTS: Ingestion of H perforatum followed by exposure to bright sunlight frequently resulted in clinical signs attributable to skin irritation and central nervous effects, including an inappropriate increase in body temperature. A decrease in H perforatum ingestion from 5.7 to 2.85 g dry plant per kg live weight and a corresponding decrease in hypericin ingestion from 5.3 to 2.65 mg per kg live weight, was associated with a decrease in the severity of the clinical signs, including the severity of the hyperthermia. CONCLUSIONS: The rectal temperature rise in affected sheep is a reliable indicator of the early development of an adverse clinical effect. There appears to be an absolute requirement for exposure to bright sunlight before any effects of H perforatum will develop. A single dose of H perforatum remains potentially effective for up to 4 days. In the small group of Merino sheep tested a tolerance level for H perforatum, eaten at the flowering stage, of < 1% (plant wet weight) of body weight and a tolerance level for hypericin of < 2.65 mg per kg live weight, were demonstrated.     

The effect of shade, shearing and wool type in the protection of Merino sheep from Hypericum perforatum (St John's wort) poisoning

OBJECTIVE: To investigate the roles of shade, fleece length and wool type in the protection of sheep from Hypericum perforatum poisoning. ANIMALS: Adult Merino ewes of superfine, fine and medium wool type. DESIGN: Seventy sheep were divided into seven equal groups. During late spring and summer a series of successive, replicate experiments was conducted, each using one group and lasting 5 days. The sheep carried 14 to 24 weeks wool growth. In each experiment the treatments tested were Hypericum +, sunlight + (n = 7); Hypericum +, sun - (n = 1); Hypericum -, sun + (n = 1); Hypericum -, sun - (n = 1). Next, 24 sheep in two equal groups were used in experiments of similar design to the above. Each group consisted of nine recently (1 to 3 weeks previously) shorn and three wool covered (25 to 26 weeks growth) sheep. The treatments tested were Hypericum +, sunlight +, fleece - (n = 9); Hypericum +, sun -, fleece + (n = 1); Hypericum -, sun +, fleece + (n = 1); Hypericum -, sun -, fleece + (n = 1). PROCEDURES: Finely milled Hypericum was administered by gavage to provide 3 mg hypericin / kg body weight. Sheep were sheltered from direct sunlight or were exposed for 5 h per day for 4 successive post-treatment days. Rectal temperatures were measured immediately before and at the end of each sunlight exposure session. Rectal temperature above 40 degrees C was considered indicative of hypericin poisoning. RESULTS: After Hypericum treatment hypericin poisoning was displayed by 26.5% of woolled sheep that were exposed to sunlight, but by none of those that were fully shaded. In similarly treated but recently shorn sheep 94% displayed hypericin poisoning when exposed to sunlight. In the wool covered group the percentages of poisoned animals based on wool type were: superfine 14%, fine 28.5%, medium 33.3%. In the recently shorn group the percentage for all three approached 100%. CONCLUSIONS: A majority of Merinos with at least 14 weeks wool growth will not be poisoned by a single oral dose of 3 mg hypericin /kg, but because hypericin persists in the blood circulation for several days this safe dose will be lowered by continuous daily ingestion. Sheep with access to substantial areas of shade could safely ingest much greater amounts of hypericin. Wool removal greatly increases the risk of poisoning. Superfine Merinos with a wool cover should be able to ingest more hypericin than comparable, medium wool types, without any increased risk of poisoning. The ability of ruminant livestock to safely ingest Hypericum is probably determined more by the amount of skin protection they have against incident sunlight than by differences in hypericin metabolism and excretion capacity.     

贯叶连翘引种驯化过程中生理生化特性、活性物质含量及抗氧化能力的变化

为了探明贯叶连翘引种驯化过程中生长适应能力以及活性物质的变化,以野生和栽培盛花期植株为材料,分别对生理生化特性、活性物质含量以及抗氧化能力进行了测定与分析,结果表明,栽培贯叶连翘的净光合速率、蒸腾速率、气孔导度、叶绿素含量和根系活力均高于野生植株,尤其是其盛花期和地上部分干重分别较野生植株提前了7 d和提高了16.67%。总黄酮和金丝桃素含量在栽培和野生植株同一器官中无显著差异;总酚类在野生花中的含量显著高于栽培花,而叶中的含量则相反;野生和栽培植株各器官中,总黄酮和酚类化合物含量在叶和花中的含量显著大于茎,而金丝桃素的含量表现为花>叶>茎。野生和栽培各器官抗氧化能力为叶>花>茎,其中,野生和栽培叶、茎提取液的抗氧化能力无显著差异,而栽培花的抗氧化能力显著大于野生花。以上研究结果表明,贯叶连翘野生转栽培后,不仅具有较好的生长适应能力,而且主要活性物质的积累量及抗氧化能力并未显著降低。该研究结果将对贯叶连翘种植栽培以及保护野生资源具有重要的参考价值和实践意义。     

Recent plant poisoning in ruminants of northern and eastern Germany. Communication from the practice for the practice]

It is reported of botanical intoxications in 1379 ruminants (dairy cattle, beef cattle and ewes) in north-eastern Germany. The intoxications of these animals were caused by consumption of the following plants: meadow saffron (Colchicum autumnale L.), cowbane (Cicuta virosa L.), sweet clover (Melilotus alba MED.), sheep's sorrel (Rumex acetosella L.), bracken (Pteridium aquilinum KUHN) and St. John's wort (Hypericum perforatum L.). In all the five described cases animals died as a result of plant intoxications. The authors come to the conclusion that periodical inspections of the meadows are necessary, checking the plants in cooperation with veterinary surgeons.     

不同收获期贯叶连翘花中抗氧化能力、主要活性物质变化及挥发性组分分离鉴定

为了探明不同收获期(花蕾期、盛花期、结果期)贯叶连翘花中抗氧化能力、总黄酮、酚类和金丝桃素含量等的变化,以栽培花器官为实验材料,采用HPLC和GC-MS等方法对70%乙醇提取液进行测定与分析。结果表明,花蕾期和盛花期贯叶连翘花提取液自由基抑制率(inhibition percentage, I%)和铁离子还原/氧化能力(ferric reducing/antioxidant power, FRAP)均显著高于结果期,花蕾期和盛花期二者之间无显著差异;总黄酮和酚类化合物以及金丝桃素含量均呈现为盛花期>花蕾期>结果期,且在P<0.05水平下达到显著差异;盛花期花中分离鉴定得到37种挥发性化学组分,其中,主要成分有1,1-二乙氧基-乙烷(19.26%)、1-十六醇(17.85%)、β-衣兰烯(10.71%)、(Z, Z)-9,12-十八碳二烯酰氯(8.42%)、十六烷酸乙酯(8.40%)、叶绿醇(5.79%)、石竹烯氧化物(4.56%)等。以上研究结果表明,花蕾期至盛花期采集贯叶连翘花器官较佳,提取液抗氧化能力较强,主要活性物质含量较高,挥发性成分较为丰富,该研究结果将对贯叶连翘生产、大面积种植栽培具有重要的参考价值和实践意义。     

金丝桃素的提取分离及其药理活性研究现状

金丝桃素为贯叶连翘中抗病毒的活性单体,属二蒽酮类。植物化学和药理学研究发现,金丝桃素具有抗抑郁、抗肿瘤、抗病毒、止血消炎以及光动活性等特点。目前关于金丝桃素的研究,国内多处于药理临床研究阶段,主要集中于肿瘤的辅助治疗和抗病毒研究方面,国外现已将其用于抑郁症、艾滋病等的临床治疗。鉴于金丝桃素突出的抗病毒作用,以及作为癌症的光化学治疗的一种有前途的内源性光敏剂,文章对金丝桃素的分离提取及其多种药理作用做了系统阐述,以期为金丝桃素的基础研究和临床应用提供借鉴和参考。     

盐酸恩诺沙星平衡水溶解度的测定

利用水平摇床充分溶解盐酸恩诺沙星,在10h以内得到pH5．3的稳定的饱和溶液,用紫外分光光度计测定室温下盐酸恩诺沙星的平衡水溶解度,研究pH对盐酸恩诺沙星溶解度的影响,为测定盐酸恩诺沙星溶解度提供一种简单易行、科学准确的检测方法,为其生态风险评价及相关研究提供数据。结果表明,当盐酸恩诺沙星饱和水溶液pH分别为2．8、4．0、5．3、7．0、8．2和9．9时,室温下溶解度分别为1．161g／L、10．637g／L、19．770g／L、4．198g／L、1．874g／L和16．108g／L;盐酸恩诺沙星水溶液在pH5．3时溶解度最大,溶液中的恩诺沙星绝大部分以阳离子形式存在,小部分以两性离子形式存在;溶解度次最大时PH为9．9,此时的恩诺沙星几乎完全以阴离子形式存在于溶液中。     

桑枝粉对水中Cr(Ⅵ)离子的吸附研究

桑枝粉富含纤维素和木质素,具有吸附去除废水中重金属离子的功能。采用桑枝粉吸附去除水中Cr(Ⅵ)离子,考察pH值、吸附时间、桑枝粉用量、Cr(Ⅵ)离子溶液初始浓度等因素对吸附效果的影响,测定吸附等温线,对等温吸附规律和吸附动力学过程作数学模拟。结果表明,桑枝粉对水中Cr(Ⅵ)离子有很好的吸附效果。吸附量随着pH值的增大而减小,随着吸附时间的延长而增大,吸附平衡时间为6 h;桑枝粉用量增加,吸附去除率提高;Cr(Ⅵ)离子溶液初始浓度增大,吸附去除率下降。适宜的吸附条件为:桑枝粉用量5 g/L,pH值1.5,温度25℃,吸附时间6 h,Cr(Ⅵ)离子溶液初始浓度为10 mg/L。桑枝粉对Cr(Ⅵ)离子的等温吸附规律符合Freundlich、Langmuir和Temkin模式,吸附呈单分子层形式,吸附性能良好,吸附易于进行。吸附动力学过程可用准一级和准二级动力学模式进行模拟。