酸水解法从葛根中提取分离葛根素和大豆苷元

本文报道了葛根黄酮的提取精制方法,以６０％乙醇为溶剂,６０℃温浸６ｈ的优化工艺条件下,采用逆流萃取法从葛根中提取葛根黄酮,其收得率为１９．２８％,含量５１．０５％。葛根黄酮提取物采用５％盐酸水解４ｈ,酸水解液用乙酸乙酯萃取放置析出葛根素,乙酸乙酯相经水洗、浓缩和重结晶可得大豆苷元。采用酸水解法可以将葛根黄酮中的葛根素及大豆苷元衍生物水解成葛根素和大豆苷元,有利于葛根素、大豆苷元的分离及产率的提高;该方法从葛根中提取分离葛根素和大豆苷元具有操作简便、产品纯度和产率高、成本低的特点。葛根素和大豆苷元产率分别为１．３６％和０．４５％,纯度为９８．３２％和９１．２５％。     

超声提取和超声水解法从野葛根中提取纯化葛根素和大豆苷元

采用超声法从野葛根中提取葛根异黄酮,再将提取物在盐酸水溶液中超声水解结合有机溶剂萃取法从野葛根中分离纯化葛根素和大豆苷元。葛根素收得率为1．2％,纯度为97．8％;大豆苷元收得率为0．5％,纯度为98．2％。超声法从野葛根中提取分离葛根异黄酮活性成分葛根素和大豆苷元具有省时、节能、提取率和产品纯度高的优点。     

太白山野葛根中异黄酮和葛根素含量随季节的变化

为了考察太白山野葛根中异黄酮及葛根素含量随季节的变化。采集一年12个月份的野葛根,采用紫外分光光度法测定其中异黄酮含量;以7高效薄层色谱扫描法测定野葛根中游离葛根素及总葛根素含量。结果表明：三月份异黄酮含量最高,为18.53%;四月份游离葛根素及总葛根素含量最高,分别为7.45%和8.93%。对不同季节太白山野葛根中异黄酮、游离葛根素及总葛根素进行定量分析,为这一野生资源的合理采收和开发利用提供了实验依据。     

RP-HPLC法测定不同产地红枣中甜菜碱的含量

目的:通过对影响甜菜碱提取和测定的符种因素进行了研究,建立了一种准确、快速的测定不同产地红枣中甜菜碱的方法.方法:样品采用超声波甲醇提取的方法,得到的样品采用反相C18柱进行分离,色谱条件为:流动相为50mmol/L pH 4.5的KH2PO4溶液;192nm波长测定,流速0.7ml/min.结果:总分析时间:4.5min.甜菜碱平均保留时间2.698min.甜菜碱在5.0～25.0mg/ml线性关系良好(r2=0.9911),平均回收率为96.87%.结论:该法测定红枣中甜菜碱含量简便、快速、准确,重现性好.     

6种晋产野生葛根中葛根素、大豆苷和总黄酮含量比较

采用高效液相色谱法对山西省5地6处野生葛根中主要有效成分葛根素、大豆苷及总黄酮含量进行了分析,并对HPLC法的测定条件(流动相和洗脱程序)进行了优化。结果表明,6种晋产野生葛根中主要有效成分含量差异显著,葛根素的含量为1.46%~4.48%,大豆苷的含量为0.11%~0.62%,总黄酮的含量为11.19%~26.14%。葛根素、总黄酮含量与纬度高低呈正相关。晋南绛县么里镇小北山与夏县泗交镇太宽河自然保护区的野生葛根中葛根素含量低于《中国药典》规定的2.4%,而晋北灵丘县的野生葛根中葛根素含量则高达4.48%。     

茉莉酸甲酯和ABA对野葛毛状根中异黄酮含量的影响

10～ 10 0 μmol·L-1茉莉酸甲酯 (MJ)可提高野葛毛状根培养液中葛根素和大豆甙元的水平 ,促进毛状根内总异黄酮含量的增加。而用 0 .5～ 2 .0mg·L-1ABA处理后 ,无论对野葛毛状根还是培养液中葛根素、大豆甙元的含量仅略有提高 ,但对总异黄酮的合成与分泌等则有显著的促进。 10 0 μmol·L-1MJ和 1mg·L-1ABA处理 2 4～ 72h ,可促进毛状根内葛根素和培养液中总异黄酮的水平 ,以处理 4 8h的增加量最大     

湘西产野葛不同部位中总黄酮及葛根素的含量测定

本文采用UV、TLC法对野葛不同部位总黄酮及葛根素的含量进行了测定,结果野葛块根比茎叶中黄酮和葛根素含量都高,其黄酮含量为4.12%,是叶的4.24倍;葛根素含量为2.43%,是叶的243倍。     

HPLC法测定不同品种不同产地菊芋叶片中绿原酸的含量

为建立同时测定大丰地区3个菊芋品种及大庆地区2个菊芋品种叶片绿原酸含量的方法,采用高效液相色谱法对不同品种不同产地菊芋叶片中绿原酸含量进行测定。结果表明:1)在检测条件流动相为1%磷酸溶液、乙腈(87∶13)、流速1 mL/min和检测波长327 nm时,绿原酸在0.01～0.1 mg/mL呈良好的线性关系,精密度良好,平均回收率为98.21%,RSD=0.68%;2)大丰地区南芋1号、青芋2号、南芋10号叶片中绿原酸含量分别为干重的0.431%、0.040%和0.933%;3)大庆地区南芋1号、南芋9号叶片中绿原酸含量分别为干重的1.245%、2.139%。研究结果表明菊芋叶片中绿原酸含量在不同品种不同生态区域间有差异,HPLC方法简便、快速、准确,可用于菊芋叶片中绿原酸含量的测定。     

UHPLC法测定不同产地野菊花中4种有机酸和蒙花苷含量

建立采用UHPLC同时测定野菊花中绿原酸、3,4-二咖啡酰奎尼酸、3,5-二咖啡酰奎尼酸、4,5-二咖啡酰奎尼酸和蒙花苷含量的分析方法。UHPLC分析条件:Agilent ZORBAX RH C18色谱柱(50 mm×2.1 mm,1.8μm),检测波长326 nm,流动相为乙腈-0.1%磷酸水进行梯度洗脱,流速0.4 mL/min,柱温25℃。实验结果显示绿原酸、3,4-二咖啡酰奎尼酸、3,5-二咖啡酰奎尼酸、4,5-二咖啡酰奎尼酸、蒙花苷分别在1.20~24.00、1.16~23.20、2.23~44.60、1.75~35.00、2.25~45.00μg/mL范围内线性关系良好(r0.9998),平均回收率分别为98.08%、98.42%、97.75%、98.27%、98.36%;RSD分别为0.38%、1.50%、0.77%、0.81%、0.62%。UHPLC法分析速度快、高效、重复性好、结果准确简便,可用于野菊花中四种有机酸和蒙花苷含量的测定。     

RP-HPLC法测定不同产地及不同部位马蓝中六种活性成分的含量

建立了RP-HPLC法测定马蓝中色胺酮、2-苯并噁唑酮、靛玉红,靛蓝、阿克苷和角胡麻苷六种活性成分含量的方法,并对不同产地及不同部位药材含量进行了分析。采用ZORBAX SB-C18色谱柱(4.6×250 mm,5μm),以乙腈和水为流动相梯度洗脱,流速为1 m L/min;检测波长280 nm;柱温30℃。马蓝中六种化学成分分离良好,各成分含量与峰面积在测定范围内均呈良好的线性关系(r≥0.999 1),加样回收率分别为100.25%、99.93%、100.46%、99.94%、99.92%、100.02%,RSD值分别为1.46、0.13、0.52、0.09、0.16、0.40。该方法简便快捷、重现性好,可为南板蓝根药材的品质评价和质量控制标准提供科学依据。     

In vitro isoflavonoid production in callus from different organs of Pueraria lobata (Wild.) Ohwi

Pueraria lobata (Wild.) Ohwi is a medicinal plant producing large amounts of isoflavonoid glycosides. Here, the ability of in vitro callus cultures to synthesize isoflavonoids was tested. Callus cultures have been initiated from different explants of in vitro germinated plants using modified MS medium. Roots, leaves and stem segments were the best sources of callus tissue. The isoflavonoid profile and content was determined by means of chromatographic methods. Callus from all organs contained isoflavonoid aglycones: genistein and daidzein and daidzein glycosides: daidzin, puerarin and 3'-methoxypuerarin. The differences between each kind of explant were observed in both the total amount of isoflavonoids and in the proportion of individual compounds. The highest content was in root callus, followed by leaf- and stem callus.     

野葛内生真菌的分离、鉴定及体外细胞毒活性

目的:药用植物内生真菌是一类重要的微生物资源,其代谢产物有着广泛的生物学活性。该研究拟从野葛(Pueraria lobata(Willd.)Ohwi)中分离有细胞毒活性的内生真菌。方法:组织块改良法分离内生真菌;细胞毒活性测定采用Alamar blue法;结合形态学和分子生物学方法进行菌种鉴定。结果:获得的34株内生真菌中菌株KLBMP f0027对HepG2、HO-8910、NCI-H460、SGC-7901等细胞株均有显著活性,ID50分别为437.4、460.0、542.5、771.2。而且活性产物相对稳定,对温度、pH变化及蛋白酶不敏感。代谢过程研究显示菌株KLBMP f0027的活性产物合成与细胞生长属于部分耦联关系类型。形态学和ITS-rDNA序列分析鉴定菌株为Aspergillus ostianus strain KLBMP f0027。结论:野葛内生真菌A.ostianus strain KLBMP f0027可作为细胞毒活性候选菌株进行深入研究。     

Regulation of isoflavone production in hydroponically grown Pueraria montana (kudzu) by cork pieces, XAD-4, and methyl jasmonate

A mini-hydroponic growing system was employed for seedlings of kudzu vine (Pueraria montana) and contents of isoflavones (daidzein, genistein, daidzin, genistin, and puerarin) from shoot and root parts of seedlings were analyzed quantitatively. In addition, exogenous cork pieces, polymeric adsorbent, XAD-4, and universal elicitor, methyl jasmonate (MeJA), were used to regulate the production of these isoflavones. It was shown that cork pieces up-regulate the production of daidzein and genistein up to seven- and eight-fold greater than the levels obtained for control roots. In contrast, levels of glucosyl conjugates, daidzin and genistin, decrease up to five- and eight-fold, respectively. Cork treatment also induces the excretion of the root isoflavone constituents into the growth medium. Minimal levels of isoflavones are absorbed by the cork pieces. XAD-4 stimulates the production of glucosyl conjugates, daidzin and genistin, in root parts about 1.5-fold greater than that obtained in control roots. These are the highest amounts of daidzin and genistin that are observed (5.101 and 6.759 mg g⁻¹ dry weight, respectively). In contrast to these two adsorbents, MeJA increases the accumulation of isoflavones in shoot rather than in root parts of seedlings, about three- to four-fold over control levels, with the exception of genistein. These studies reveal new observations on the regulation of isoflavone production in hydroponically grown Pueraria montana plants by two adsorbents (cork pieces and XAD-4) and MeJA elicitor.     

Ultrasonication assistance increases the efficiency of isoflavones extraction from kudzu (Pueraria lobata Ohwi) roots waste

This study assesses the use of ultrasonication to improve the extraction process of classical solvent extraction methods for extracting isoflavones from the kudzu roots waste. The kudzu roots waste was produced after squeezing fresh kudzu roots to make juice. The effects of extraction time, extraction temperature, ultrasonic power, and ethanol concentration in ethanol/water mixtures were investigated. The extraction yield was found to increase with extraction time and temperature. The application of ultrasonication-assisted extraction (UAE) increased the extraction yield of water/ethanol mixture (20:80) at 25°C 3 fold. A maximum amount (7.28 g) of isoflavone was obtained from 100 g of dried kudzu roots waste by UAE with water/ethanol mixture (20:80) for 6 h at 80°C. Combining the use of ultrasonication with conventional vacuum evaporation method also reduced the concentration time for extracts from 45 to 24 min.     

野葛离体培养生产异黄酮类化合物研究进展

野葛含有大量的异黄酮类化合物,其中葛根素是葛属植物特有药用成分。该文综述了野葛细胞和器官培养生产异黄酮类化合物的研究现状以及各种影响葛根素等异黄酮类化合物产生的因素,并评述葛根素的应用前景。     

Molecular Surveillance for Lymphoproliferative Disease Virus in Wild Turkeys (Meleagris gallopavo) from the Eastern United States

Lymphoproliferative disease virus (LPDV) is a poorly understood, oncogenic avian retrovirus of domestic turkeys that has historically been restricted to Europe and Israel. However, a recent study reported LPDV in multiple wild turkey diagnostic cases from throughout the eastern United States of America (USA). To better understand the distribution of LPDV in the eastern USA, we surveyed 1,164 reportedly asymptomatic hunter-harvested wild turkeys from 17 states for the presence of LPDV proviral DNA by PCR. In total, 564/1,164 (47%) turkeys were positive for LPDV. Wild turkeys from each state had a relatively high prevalence of LPDV, although statewide prevalence varied from 26 to 83%. Phylogenetic analysis revealed two major clades of LPDV in the USA, although one was at a low frequency suggesting restricted transmission, as well as significant clustering by state of isolation. To determine the best tissue to target for diagnostic purposes, liver, spleen, and bone marrow were tested from a subset of 15 hunter-harvested wild turkeys and 20 wild turkey diagnostic cases. Overall, bone marrow provided the highest level of detection for both hunter-harvested turkeys and diagnostic cases. The sensitivity of LPDV detection between tissues was not significantly different for diagnostic cases, but was for hunter-harvested birds. These results indicate that LPDV infection is common and widespread in wild turkey populations throughout the eastern USA, even without overt signs of disease.     

RP-HPLC法同时测定十种五加属植物叶中三萜Chiisanoside和Chiisaogenin

建立反相高效液相色谱法(RP-HPLC)测定chiisanoside和chiisanogenin的定量分析方法,利用最优色谱条件对十种中韩五加属植物叶中的三萜chiisanoside和chiisanogenin进行定量分析.研究结果表明,最佳色谱条件为:ODS-C18色谱柱(250mm×4.6mm,5μm);流动相:乙腈-水(55:45);检测波长:205 nm;柱温:40℃;流速:1.0 mL/min.Chiisanoside和chiisanogenin线性范围和回归方程分别为16.5～ 181.5 μg/mL,Y=5498356X-9738(r=0.9995)和4.88～78.0 μg/mL,Y=5753131X-8289 (r=0.9979),加样回收率分别为98.87％(标准偏差为1.14％)和98.83％(标准偏差为0.72％).     

Ethylene Production by the Kudzu Strains of Pseudomonas syringae pv. phaseolicola Causing Halo Blight in Pueraria lobata (Willd) Ohwi

Significant amounts of ethylene was produced by Pseudomonassolanacearum (all strains), P. syringae pv. phaseolicola (Kudzustrains isolated from Pueraria lobata) and Erwinia rhapontici(2 strains out of 22) out of 24 species, 3 subspecies and 38pathovars of plant pathogenic bacteria tested in yeast extract-peptonebroth. The bean strains of P. syringae pv. phaseolicola causinghalo blight in kindney bean plants did not produce ethylene.The Kudzu strains produced ethylene at a rate of 7 to 100?10^–9nl cell^–1 h^–1, which was 500 to 1,000 times higherthan that of P. solanacearum and several times higher than thatof Penicillium digitatum, the most potent ethylene producerknown among microorganisms. The presence of living cells was essential for ethylene productionby the Kudzu strains. The bacterium effectively produced ethylenefrom amino acids such as glutamate, aspartate and their amides.Although glucose and succinate were also good substrates forethylene biosynthesis, the rate of ethylene production was significantlysmaller than that with glutamate. Methionine, which is knownas the precursor of ethylene in plants, had no effect on ethyleneproduction by the bacterium. 1-Aminocyclopropane-1-carboxylicacid (ACC) also had no effect on ethylene production, and therewas not enough ACC in the bacterial cells to account for thehigh rate of ethylene production. Ethylene production from glutamatewas inhibited by n-propylgallate and EDTA, but not by aminoethoxyvinylglycine.These results indicate that ACC is not involved as an intermediatein the process of ethylene biosynthesis by the bacterium, suggestingthe presence of a pathway different from that of plant tissues. (Received September 4, 1984; Accepted October 27, 1984)