Determination of contents of eight alkaloids in fruits of Macleaya cordata (Willd) R. Br. from different habitats and antioxidant activities of extracts

A reliable ultrasound-assisted extraction （UAE） method combined with HPLC-UV for quantification of eight active alkaloids in fruits of Macleaya cordata （Willd） R. Br. was developed. The optimization conditions of UAE were obtained by using Box-Behnken design of response surface methodology. Chromatography was carried out using a Kromasil C18 column by gradient elution with 0.1% phosphoric acid aqueous solution for HPLC-UV. All calibration curves showed good linear correlation coefficients （R^2〉0.999 6） and recoveries （from 97.3% to 104.9%） were acceptable. 1,1-diphenyl-2-picrylhydrazyl （DPPH） method was employed to test the antioxidant activity of the extract from the samples. The proposed method was successfully applied to quantifying eight components in nine samples of M.cordata, and significant variations of alkaloid contents and antioxidant aetivity of the samples from different habitats were demonstrated. It presents a powerful proof for the selection of the best sources to extract eight kinds of alkaloids.

Stability of Sanguinarine and Chelerythrine in Macleaya cordata (Willd.) R. Br.

[Objectives]To explore the stability of sanguinarine and chelerythrine in Macleaya cordata(Willd.)R.Br.[Methods]The solubility and stability of sanguinarine and chelerythrine in seven solvents were measured by HPLC.Besides,the effects of water quality,light source,oxidant,temperature,and pH on stability were investigated.[Results]The solubility and stability of sanguinarine and chelerythrine in methanol and ethanol are good;the stability of sanguinarine and chelerythrine in distilled water and rainwater is not affected by light and is very stable,but they are unstable in tap water whether they are protected from light or not;oxidants have a great influence on the stability of sanguinarine and chelerythrine;sanguinarine and chelerythrine are stable at room temperature lower than 54℃;sanguinarine is stable in pH 2.5-7.0,while chelerythrine is stable in pH 2.5-8.0.[Conclusions]The good stability of sanguinarine and chelerythrine under specific conditions shows that they have broad development prospects and value.

Influence of extraction solvents on antioxidant and antimicrobial activities of the pulp and seed of Anisophyllea laurina R.Br.ex Sabine fruits

Objective:To evaluate the influence of extraction solvents on antioxidant and antimicrobial activities of the pulp and seed of Anisophyllea laurina R.Br.ex Sabine fruits.Methods:The antibacterial activities of pulp and seed extracts were tested by using disk diffusion method against eight bacterial strains and three fungal strains.Total phenolic,flavonoid,monomeric anthocyanin and tannin contents,and antioxidant activities were determined by spectrometric methods.Results:The antioxidant analysis of pulp extract revealed the strong radical scavenging capacity and total phenolic content(4 329.66 mg of gallic acid/100 g),while seed extract showed the high antioxidant activity and total tannin content(5 326.78 mg catechin equivalent/100 g).Antibacterial and antifungal activities of methanol and ethanol extracts exhibited potent growth inhibitory activity against Aeromonas hydrophila,Bacillus subtilis,Escherichia coli O157:H7,Pseudomonas aeruginosa,Salmonella Typhimurium and Staphylococcus aureus ATCC 6538 with minimum inhibitory concentration values ranged from 125 to 250 μg/mL.However,seed extract had the strongest potential activity against Aspergillus niger and Candida albicans with minimum inhibitory concentration value of 500 μg/mL compared to pulp extract.Conclusions:Our results therefore demonstrated that ethanol and methanol extractions were more efficient in extracting antioxidants and bioactive compound in pulp and seed.These results support that these plant extracts can be used for the treatment of bacterial infections.

Optimization of Extraction Technology of Pigments from Petals of Matthiola incana(L.)R.Br.of Different Colors

[ Objective] The paper aimed to study the optimum extraction technology of pigments from petals of Matthiola incana（ L. ） R. Br. of differ- ent colors. [Method] Petal pigments of Aida variety （white violet）, Francesco variety （ red violet） and Arabella variety （ purple violet） were ana- lyzed by colorimetric reaction method and ultraviolet-visible spectrum method, and the optimum extraction technology of flavonoid compounds from petals of them was determined by single factor experiment and orthogonal experiment. [ Result] The optimum extraction conditions of white violet were as follows： with 95% ethanol ＋ 5% HCI as solvent, solid-liquid ratio （ 1 ： 40）, extraction time （3 h）, extraction temperature （65 ℃ ）. The opti- mum extraction conditions of red violet and purple violet were as follows： with acetone as solvent, solid-liquid ratio （1：40）, extraction time （3 h）, extraction temperature （65 ℃）. [ Conclusion] The study provided theoretical foundation for further development and exploitation of pigments from M. incana（ L. ） R. Br.

A NEW GUANIDINE DERIVATIVE FROM ASTRAGALUS COMPLANATUS R.Br.

A new guanidine derivative,complanatin has been isolated from the seeds of Chinese traditional medicine Astragalus complanatus R.Br..Its structure was established on the basis of spectral data and X-ray crystallographic analysis to be N-[3-carboxypropyl]-N-[3-methyl-2-butenyl]guanidine.

Inhibitory Effect of Flavonoid Glycosides from Chlorophytum comosum on Nasopharyngeal Carcinoma 5-8F Cells and Its Mechanism

[Objectives]To study the inhibitory activity of two flavonoid glycosides isolated from Chlorophytum comosum Laxum R.Br on human nasopharyngeal carcinoma(NPC)cell line 5-8F in vitro and its mechanism.[Methods]The flavonoid glycosides were isolated and purified from the ethanol alcoholic extract of the roots of Liliaceae plant Chlorophytum comosum by silica gel column chromatography,macroporous resin column chromatography,Sephadex LH-20,and reverse column chromatography(ODS).The inhibitory activity of flavonoid glycosides on human nasopharyngeal carcinoma cells was analyzed by CCK-8 method,and the potential mechanism was preliminarily analyzed by molecular docking.[Results]Two flavonoid glycosides were identified as isovitexin 2″-0-rhamnoside and 7-2″-di-O-β-glucopyranosylisovitexin.Two flavonoid glycosides showed promising inhibitory effect on human nasopharyngeal carcinoma cell line 5-8F,with IC_(50) values of 24.8 and 27.5μmol/L,respectively.Molecular docking results showed that the potential targets of two flavonoid glycosides include CyclinD1,Bcl-2β-Catenin,ILK,TGF-β,in addition,two glycosides showed higher predicted binding affinity towards CyclinD1,which verifies the cytotoxicity of the two compounds on human nasopharyngeal carcinoma cell line 5-8F in vitro.[Conclusions]Two flavonoid glycosides are the active molecules in Chlorophytum comosum that can inhibit the proliferation of human nasopharyngeal carcinoma cells,and have the potential to be used in the research and development of anti nasopharyngeal carcinoma drugs.

博落回叶与小果博落回叶中4种生物碱的含量比较

目的比较博落回叶与小果博落回叶中4种生物碱含量的差异。方法 采用反相高效液相色谱法对博落回叶与小果博落回叶中4种生物碱———原阿片碱、别隐品碱、血根碱、白屈菜红碱的含量进行比较研究。结果在博落回叶中原阿片碱、别隐品碱、血根碱、白屈菜红碱的含量分别为0.48%～2.00%、0.36%～1.72%、0.18%～0.81%、0.07%～0.33%,小果博落回叶中分别为0.01%～0.13%、0.07%～0.48%、0.24%～0.75%、0.35%～1.24%。结论博落回叶中原阿片碱与别隐品碱含量较高;小果博落回叶中血根碱与白屈菜红碱含量较高。

博落回根的化学成分研究

目的:研究博落回根的化学成分。方法:采用硅胶柱色谱、重结晶、半制备液相色谱等进行分离纯化,根据波谱数据进行结构鉴定。结果:从博落回根中分离得到7个化合物,通过质谱、核磁等波谱手段确定了结构,分别鉴定为:6-氰基二氢白屈菜红碱(1)、6-氰基二氢白屈菜黄碱(2)、二氢白屈菜玉红碱(3)、6-甲氧基去甲白屈菜红碱(4)、二氢血根碱(5)、6-丙酮基二氢血根碱(6)、豆甾醇(7)。结论:其中,化合物1、2为新的天然化合物,化合物4首次从该植物中分离得到。

手参属植物化学成分及药理活性研究进展

兰科手参属植物手参、西南手参、短距手参与峨眉手参等为珍稀药用植物,具有较高的临床药用价值,近年来逐渐受到国内外学者的关注。综述手参属植物化学成分和药理活性的研究进展,其中化学成分包括二氢类、菲类、苷类、苯丙素类、异戊二烯类、脂肪酸衍生物等多种类型;药理活性包括抗氧化、抗过敏、促进祖细胞增殖、抑制乙型肝炎病毒表面抗原等作用,以期为今后对该属植物进一步研究和开发利用提供参考。

博落回内生放线菌的分离及活性菌株的鉴定

目的:从博落回Macleaya cordata(Willd.)R.Br根部筛选具有抑菌活性的放线菌,并对其进行种属鉴定。方法:采用平板涂布法分离皖西大别山野生植物博落回根部的内生放线菌;采用琼脂块法初步研究博落回内生放线菌的抑菌活性;通过形态特征、生理生化特性及分子生物手段鉴定菌种。结果:从博落回根部共分离筛选得到内生放线菌15株,有6株放线菌对供试菌株表现出抑菌活性,占总分离菌株的40%,其中菌株BL7对金黄色葡萄球菌与枯草芽孢杆菌的拮抗作用明显,其抑菌圈直径分别为29、20 mm。根据菌株BL7的形态学特征、生理生化特征并结合16S r DNA系统发育分析将其初步鉴定为茂原链霉菌Streptomyces mobaraensis。结论:链霉菌属放线菌在医药上具有广阔的应用前景。而茂原链霉菌Streptomyces mobaraensis首次从博落回植株中筛选得到,具有进一步研究开发的价值。

一测多评法测定博落回果实中原阿片碱、别隐品碱、血根碱和白屈菜红碱的含量

目的建立一测多评法测定博落回中原阿片碱、别隐品碱、血根碱和白屈菜红碱的含量。方法以博落回果实为研究对象,建立别隐品碱、血根碱、白屈菜红碱对原阿片碱的相对校正因子,利用该相对校正因子计算别隐品碱、血根碱、白屈菜红碱的含量,实现一测多评;同时采用外标法测定药材中4种生物碱的含量,比较两种方法的差异,验证一测多评法的准确性和可行性。结果 9批博落回果实中4个生物碱的含量可以用一测多评法进行测定,其计算值与实测值间无显著差异。结论建立的一测多评法同步测定博落回果实中的原阿片碱、别隐品碱、血根碱、白屈菜红碱是可行的、准确的。

博落回中生物碱的超声提取和分离

运用超声波辅助提取技术,通过L9(3)4正交试验考察了乙醇浓度(A)、液固比(B)和超声时间(C)3个因素对博落回生物碱提取的影响,同时还研究了D101大孔树脂对粗提液中生物碱的吸附和分离效果.结果表明最佳提取工艺为A3B1C3,即用9倍量的90%的乙醇超声提取45 min.D101大孔树脂能从博落回的pH2～3的盐酸提取液中以游离碱形式全部吸附其生物碱,分别采用50%乙醇和丙酮洗脱能有效地将极性和弱极性生物碱分离开来.

博落回果荚中生物碱的研究

目的:研究博落回果荚的化学成分。方法:采用硅胶柱层析以及制备高效液相色谱法进行分离纯化,并根据波谱学数据鉴定化合物的结构。结果:从博落回果荚的乙醇提取物中分离纯化得到8个生物碱,分别鉴定为:6-氰基二氢血根碱(1)、6-丙酮基二氢血根碱(2)、6-丙酮基二氢白屈菜红碱(3)、白屈菜红碱(4)、二氢白屈菜红碱(5)、13,14-去氢-N-甲基黄连碱(6)、N-甲基氢化小檗碱(7)、小檗碱(8)。结论:其中,化合物1、6为新的天然产物,化合物7为首次在博落回属植物中报道。

HPLC法同时测定博落回不同部位原阿片碱、别隐品碱的含量

目的采用HPLC法同时检测博落回不同部位原阿片碱、别隐品碱含量。方法采用Luna-C18色谱柱(150 mm×4.6 mm,5μm),以0.1%磷酸加三乙胺缓冲溶液(pH 3.5)-乙腈(78∶22)为流动相,流速为0.8 mL/min,柱温25℃,检测波长284 nm。结果原阿片碱、别隐品碱含量测定方法的线性关系良好,线性范围和相关系数分别为4.2～52μg/mL(r=0.999 4),6.9～86μg/mL(r=0.999 9);平均回收率分别为99.2%、99.4%;RSD分别为0.9%、1.8%(n=6)。结论该方法操作简便、准确、灵敏度高,适合于博落回中原阿片碱、别隐品碱含量的同时测定,不同部位两种生物碱均存在一定的差异。

燃油各组分在咪唑盐类离子液体中溶解性的研究

利用微波法合成了不同烷基取代的溴化1-烷基-3-甲基咪唑盐类离子液体[RMIm]Br(R=E,B,H;E表示乙基,B表示1-丁基,H表示1-庚基),并对其进行了红外光谱表征,用平衡法测定了燃油各基本组分的溶解性.结果表明:饱和烷烃在离子液体中几乎不溶,苯系物在离子液体中有一定的溶解度,而含S和含N化合物在离子液体中有较大的溶解度;升高温度有助于燃油各组分在离子液体中的溶解;离子液体的含水量对含S和含N化合物的溶解度影响较大.

博落回鲜叶中生物碱类化学成分的分离与结构鉴定

目的:研究博落回叶的化学成分。方法:采用硅胶柱色谱以及制备液相色谱法进行分离纯化,并根据波谱数据鉴定化合物结构。结果:从博落回新鲜叶的乙醇提取物中分离纯化得到14个生物碱,分别鉴定为:原阿片碱(1)、别隐品碱(2)、血根碱(3)、白屈菜红碱(4)、去甲基白屈菜红碱(5)、去氢紫堇碱(6)、N-甲基四氢黄连碱(7)、黄柏碱(8)、6-甲氧基去甲基血根碱(9)、6-氰基二氢血根碱(10)、6-氰基二氢白屈菜红碱(11)、6-丙酮基二氢血根碱(12)、二氢血根碱(13)、二氢白屈菜红碱(14)。结论:其中化合物5~9为首次在博落回属植物中分离得到。

沙苑子黄酮对CCl_4及D-氨基半乳糖致急性肝损伤的保护作用

目的探讨沙苑子黄酮(FAC)对肝损伤的保护作用。方法采用0.1%CCl4(10mL/kg)、D-氨基半乳糖(D-Gal,700mg/kg)分别ip造成小鼠急性肝损伤模型,并ig不同剂量的FAC,测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性,观察肝组织病理改变;用CCl4损伤原代培养大鼠肝细胞,测定细胞培养液ALT、AST活性,MTT法测定肝细胞增殖活性。结果FAC能显著降低小鼠CCl4和D-Gal损伤原代培养大鼠肝细胞模型ALT和AST活性(P<0.01),病理学观察显示,FAC明显减轻肝组织损伤程度(P<0.01)。FAC显著提高肝细胞活性,促进肝细胞增殖。结论FAC具有显著的保肝作用。

博落回不同部位提取物对大肠菌群的抑菌作用研究

研究了博落回提取物对大肠菌群的抑制作用。本实验以大肠杆菌、产气肠杆菌、阴沟肠杆菌为指示菌,采用牛津杯法筛选对三种指示菌有最佳抑制作用的博落回组织粗提物;对抑菌效果好的粗提物依次用石油醚、乙酸乙酯、正丁醇进行萃取,采用滤纸片法筛选出抑菌活性最高的萃取物,并测定其最小抑制浓度和最小杀菌浓度。实验结果表明博落回不同部位粗提物有抑制指示菌的活性,其中以博落回根部提取物的抑菌活性最佳;根部萃取物抑菌实验结果表明正丁醇萃取物对三种指示菌均有较好的抑制效果,抑菌圈直径均在12 mm以上,远高于粗提物,对大肠杆菌,其最小抑制浓度和最小杀菌浓度分别为400、600μg/m L,对产气肠杆菌与阴沟肠杆菌,其最小抑制浓度均为500μg/m L,最小杀菌浓度均为700μg/m L。博落回根部提取物对上述三种指示菌均有抑制作用,尤其以正丁醇萃取物的抑菌效果最佳。

基于HPLC-Q-TOF/MS技术鉴定博落回叶中化学成分

目的快速获取博落回叶中的生物碱组分信息。方法应用HPLC-Q-TOF/MS技术鉴定博落回叶的化学成分。结果从博落回叶的甲醇提取物中鉴定了19个化合物,包括17个生物碱（3个苯并菲啶类、1个二氢苯并菲啶类、2个苄基异喹啉类、2个原小檗碱类、5个四氢原小檗碱类、3个普罗托品类和1个氮甲基-四氢原小檗碱类生物碱）、2个黄酮类化合物（1个黄酮苷和1个槲皮素）。结论化合物1~6,8首次在博落回中报道。

基于质谱示踪研究博落回根的化学成分

目的:研究博落回根的化学成分。方法:采用LC-MS确定分离目标,运用硅胶柱层析以及制备液相色谱法进行分离纯化,并根据波谱数据鉴定化合物结构。结果:从博落回根的乙醇提取物中分离纯化得到8个化合物,分别鉴定为:6-hydroxymethyldihydrosanguinarine(1)、7-去甲基二氢白屈菜红碱(2)、四氢非洲防己胺(3)、四氢小檗红碱(4)、hunnemannine(5)、白藜芦醇(6)、对羟基苯甲醛(7)、2,2-二甲氧基乙酸(8)。结论:8个化合物均首次在博落回属中分离得到。