不同产地博落回各部位血根碱与白屈菜红碱含量分析

采用HPLC法同时测定不同产地博落回各部位血根碱、白屈菜红碱含量.采用Welch Material XB-C18分析色谱柱(4.6×250 mm,5μm),以(A)体积分数为0.1%冰乙酸水溶液-(B)乙腈梯度洗脱(0~24 min,B的体积分数为10%~46%)为流动相,流速为1 m L/min,柱温25℃,检测波长270 nm.结果在一定质量浓度范围内血根碱、白屈菜红碱含量测定方法的线性关系良好,相关系数分别为0.999 7、0.999 3;平均回收率分别为95.6%、95.9%,RSD分别为1.06%、0.6%(n=3).该方法操作简便、准确、灵敏度高,适合于博落回中血根碱、白屈菜红碱含量的测定,不同部位两种生物碱含量均存在明显差异,不同采集地点的样品之间两种生物碱含量也存在一定差异.     

博落回生物碱的超声提取动力学模型

超声法提取博落回果荚中主要生物碱,并采用HPLC法对提取液进行生物碱含量测定,以此来研究博落回果荚中主要生物碱在超声条件下提取的动力学特征.获得了提取液中四种主要生物碱(原阿片碱,别隐品碱,血根碱和白屈菜红碱)的含量,并根据等温动力学方程图解法确定了提取过程的反应级数.求得了不同温度下四种主要生物碱的反应速率常数k,进一步依据速率常数k求得了四种生物碱的活化能.结果表明,超声提取博落回果荚中主要生物碱的过程符合二级动力学方程的模型.     

HPLC法测定白屈菜不同器官多种生物碱的质量比

采用高效液相色谱法分析白屈菜不同器官生物碱的种类及质量比.取白屈菜不同器官粉碎,以甲醇∶水溶液(4∶1,V∶V)热回流提取,提取液蒸干后用50%甲醇溶液溶解,滤过后采用HPLC分析.采用Diamonsil C18色谱柱,流动相为乙腈-0.1%三乙胺水溶液(24∶76,三乙胺溶液用磷酸调至p H至3),流速为1.0 m L/min,柱温为30℃,检测波长为290 nm.白屈菜碱、白屈菜红碱、血根碱、小檗碱、黄连碱分离良好,各成分质量浓度与峰面积在测定范围内均呈良好的线性关系(r0.999 6),重现性良好,5种成分的RSD值(n=5)均小于2.0%;平均加样回收率(n=5)为97.78%~99.63%.白屈菜不同器官所含生物碱的种类有所差异,根、茎及果实中所含生物碱的种类较多,质量比较高.明确了白屈菜碱各器官所含主要生物碱种类,为白屈菜的合理开发和利用奠定了基础;所建立的白屈菜多种生物碱同时测定的HPLC方法稳定可靠,简便易行,可用于白屈菜药材及其制剂的质量评价.     

具有生物活性的山梗碱类生物碱的合成研究

山梗碱类生物碱多为2,6位上各有一侧链的哌啶类化合物。本文报道了山梗酮碱(Ⅱ)、去甲山梗酮碱(Ⅲ)、山梗醇碱(Ⅳ)、去甲山梗醇碱(Ⅴ)、2,6-二丙酮基哌啶(Ⅵ),N-甲基-2,6-二丙酮基哌啶(Ⅶ)及2-丙酮基-6-苯甲酰甲基哌啶(Ⅷ)的合成。以苯基锂为催化剂使2-甲基-6-丙酰甲基吡啶的缩酮(Ⅺ)与苯甲醛反应时,缩酮被破坏而得到2-甲基-6-(2,2-二苯基丁基)吡啶(ⅩⅥ)。以氨基钠作为催化剂时,苯甲醛转变为苯甲酰胺。Ⅳ和Ⅴ对呼吸中枢有较强的兴奋作用。     

收山虎的化学成分

从收山虎药用根和皮中分离出九个化合物。确定其中八种结构为:廿六碳脂肪酸,β-谷甾醇,L-细辛素,白藓碱,N-去甲基白屈菜红碱,木兰素,Arnottianamide和11-Acetonyldihydrochelerythrine;后者过去被人认为是在提取过程中产生的丙酮加成物,而本实验结果表明它应该是天然存在的。另有一种生物碱可能是苯并菲啶生物碱。     

HPLC法同时测定白屈菜中白屈菜碱和白屈菜红碱

采用高效液相色谱法建立白屈菜药材中含白屈菜碱和白屈菜红碱量的测定方法,色谱柱选用依利特Hypersil ODS2(4.6 mm×200 mm,5μm),以乙腈-0.01 mol/L磷酸二氢钾水溶液(38∶62,磷酸二氢钾水溶液用磷酸调pH=3)为流动相,流速为1.0 mL/min,检测波长为282 nm,柱温为35℃,进样量20μL.结果表明白屈菜碱和白屈菜红碱均在0.25~1.50 mg范围内与峰面积线性关系良好,平均加样回收率白屈菜碱和白屈菜红碱分别为(n=6)为98.72%、97.34%,RSD=0.65%、0.86%.该方法灵敏、准确、重复性好.     

丽江紫乌头中的二萜生物碱

 对丽江紫乌头中的二萜生物碱进行研究.利用反复硅胶柱层析,重结晶等分离手段从中分离鉴定了6个二萜生物碱:查斯曼宁(chasmanineⅠ),塔拉萨敏(talatizamine Ⅱ),草乌甲素(crassicauline A Ⅲ),丽江乌头碱(foresaconitine Ⅳ),acoforestinine(Ⅴ),滇乌碱(yunaconitine Ⅵ).其中化合物Ⅰ、Ⅲ、Ⅳ、Ⅴ为首次从该种植物中分离得到.     

采用UPLC-ESI-MS快速测定博落回各器官中王要生物碱的含量

采用超高效液相色谱-电喷雾质谱联用(UPLC-ESI-MS)技术,对中药博落回各器官中的主要生物碱成分进行系统分析.样品经超声萃取后,通过UPLC-ESI-MS快速测定了8种生物碱在根、茎、叶和果荚中的含量,获得相应化合物的保留时间、相对分子质量和离子碎片等信息,并从博落回果荚中分离、鉴定隐品碱.在此基础上,建立种快速、灵敏、准确测定博落回中多种生物碱成分的新方法.研究结果表明:这8种生物碱含量在一定范围内呈线性关系,相关系数为0.992 5-0.998 3,平均加标回收率为96.6％～104.0％,相对标准偏差均小于4.3％,检出限为0.123～1.270 μg/L.     

猪殃殃酚类成分研究

采用正相、反相硅胶柱色谱、葡聚糖凝胶柱、半制备高效液相色谱(HPLC)对猪殃殃氯仿和乙酸乙酯部位进行了化学成分的研究,分离得到了9个酚类化合物,分别为：对苯二甲酸二丁酯(1),原儿茶酸丁酯(2),原儿茶酸乙酯(3),二氢咖啡酸乙酯(4),顺式咖啡酸乙酯(5),没食子酸甲酯(6),7-羟基-6-甲氧基色酮(7),1-丙烯酸-3-(3,4-二羟苯基)戊酯(8),1-(3,4-二羟基苯)丙酮(9).这9个化合物都是首次从该属植物中分离得到.     

吴茱萸化学成分研究

作者对中药吴茱萸Evodia rutaecarpa(Juss.)Benth.的化学成分作了较为系统的研究,分离鉴定了六个化合物,分别为吴茱萸碱(evod iam ine 1)、吴茱萸次碱(rutaecarpine 2)、14-甲酰基二氢吴茱萸次碱(14-formyld ihydrorutaecarpine 3)、1-甲基-2-[(6Z,9Z)6,9-十五二烯]-4(1H)-喹诺酮{1-methy-2-[(6Z,9Z)-6,9-pentadecad ienyl]-4(1H)-quinolone 4}、二氢吴茱萸新碱(d ihydroevocarpine 5)、β-谷甾醇(-βsitosterol 6).其中主要有效成分吴茱萸次碱的收率比其它文献[1,2]报道的高出几十倍,作者提供了一种简单快速、收率高的吴茱萸次碱提取分离方法,从而为大规模分离生产吴茱萸次碱提供一个科学的方法参考.     

乔木茵芋中黄酮和生物碱类化合物及其镇痛活性研究

 对乔木茵芋进行化学成分研究.采用硅胶柱色谱、葡聚糖凝胶LH-20柱色谱、RP-Csub>18柱色谱等方法分离化学成分,¹H NMR、¹C NMR等方法进行结构鉴定,从中分离鉴定了5个黄酮类化合物:槲皮素(1),槲皮苷(2),山奈酚-3,7-葡萄糖苷(3),枸橘苷(4),橙皮苷(5);1个生物碱类化合物:1-甲基-2-苯基-4-喹啉酮(6).所有化合物均为首次从该种植物中分离得到.并采用小鼠醋酸扭体法和热板法检测乔木茵芋总提取物及乔木茵芋总生物碱提取物的镇痛活性.结果显示乔木茵芋总提物和乔木茵芋总生物碱提取物均可明显抑制醋酸所致小鼠扭体反应,此外乔木茵芋总生物碱提取物还可明显提高小鼠对热刺激的痛阈值,具有较好的镇痛活性.     

近无柄金丝桃中的口山酮类成分

 在细胞毒性试验结果指导下,同步对近无柄金丝桃(Hypericum subsessile N.Robson)全株的有效部位进行化学成分分离纯化,得6个口山酮类化合物,通过理化数据测定及波谱数据分析,鉴定了结构,分别是:2,3-二甲氧基口山酮(1),1,3-二羟基-5,6-二甲氧基口山酮(2),Toxyloxanthone B(3),Kielcorin(4),Candensin D(Hypericorin)(5)和Subalatin(6).化合物(1)～(6)均为首次从该植物中分离得到;同时,细胞毒试验发现,近无柄金丝桃的氯仿及乙酸乙酯提取物有一定细胞毒活性,其IC₅₀分别为8.8,14.6μg/mL.     

Diversity and high nitrogenase activity of endophytic diazotrophs isolated from Oryza rufipogon Griff.

Diversity and nitrogenase activity of endophytic diazotrophs colonized in the wild rice Oryza rufipogon Griff grown in Boluo, Huilai County in Guangdong Province and Lingshui County in Hainan Province were studied. Thirty-seven isolates obtained from Oryza rufipogon were identified as putative endophytic nitrogen-fixing bacteria by ARA （acetylene reduction assay） test and further confirmed by PCR amplification of nifH gene fragments. All obtained strains have ARA activity and the same sized nifH gene fragments. Above the similarity level of 80%, the obtained isolates were assigned as Group Ⅰ to Ⅷ by the clustering of IS-PCR fingerprints. The SDS-PAGE whole-cell protein patterns were similar to those of IS-PCR fingerprints. Components and contents of fatty acid methyl esters （FAMEs） were used to differentiate the representative strains （Ls13, Ls8, BL1, BL12, HL6, Ls4） from Group Ⅰ to Group Ⅵ. The six representative strains showed significant difference in contents and components of cellular fatty acid methyl ester. 16S rDNA sequencing analysis showed that strains of Group Ⅰ to Ⅶ were located in Enterobacteraceae （y-proteobacteria）. Strains of Group Ⅰ and Group Ⅱ were closely related to Klebsiella sp.; Strain Ls8 of Group Ⅱ was a little far away from the genus of Pantoea （homology level 96% with Pantoea agglomerans）, which may represent a new species or genus in Enterobacteraceae; Strains of Groups Ⅳ and Ⅴ belonged to different Enterobacter sp.; Strain Ls4 and Ls 9 representing Group Ⅵ were close to Citrobacter amalonaticus with 98% sequence similarity; Strain Ls15 of Group Ⅶ showed 98% sequence identity with Pantoea sp.; Strains of Group Ⅷ were assigned to the genus Ideonella （β-proteobacteria）. Based on the above results, endophytic diazotrophs isolated from O. rufipogon showed great diversity and some diazotrophs showed high nitrogenase activity with 42.52 μmol/mL. h C2H4. Inoculation to rice tests indicated that the isolated endophytic diazotrophs significantly promoted the rice growth.     

Functional characterization and mapping of two MADS box genes from peach （Prunus persica）

Previously an AGAMOUS gene homologue PpMADS4 and a FRUITFULL gene homologue PpMADS6 were isolated from peach （Prunus persica）, and both genes were shown to express in the developing floral and fruits. To gain insight into their function, the two genes were constitutively expressed in Arabidopsis thaliana and their effects on plant growth and floral organ development were studied in this work. The transgenic plants all displayed early flowering and conversion of inflorescence to floral meristem. However, the two genes had different effects on the floral organ structures in A. thaliana. The transgenic plants overexpressing PpMADS4 displayed homeotic conversion of floral organs, and particularly the perianth abscission was inhibited. The plants overexpressing PpMADS6 showed early flowering, produced higher number of carpels, petals, and stamens than nontransgenic plants, and pod shatter was prevented; significantly, the transgenic plants yielded more than one siliques from a single flower. A SSR molecular marker was developed for PpMADS4, and it was then assigned into the G5 linkage group of Prunus sp. Both PpMADS4 and PpMADS6 genes were located at the same region in the G5 linkage group. Our results showed the potential application of these two MADS box genes for crop and fruit tree improvement.     

红树白骨壤果实中芳香脂类化学成分研究

【目的】研究红树白骨壤(Avicennia mavina)果实中芳香酯类化合物。【方法】采用柱色谱、凝胶层析和高效液相色谱分离技术,从白骨壤果实中分离芳香酯类单体化合物,运用理化和波谱分析方法鉴定其化学结构。【结果】从白骨壤果实中分离获得7个芳香酯类化合物,分别鉴定为甲基(4-氯-4[N-羟氨基]苯基)醋酸酯(1)、4-(2-羟基乙氧基)苯甲酸甲酯(2)、邻苯二甲酸二异丙酯(3)、咖啡酸甲酯(4)、原儿茶酸甲酯(5)、邻苯二甲酸二(2-乙基)己酯(6)、邻苯二甲酸二丁酯(7)。【结论】化合物1~7均是首次从该种海洋植物中分离得到,其中化合物1和2为新天然产物。     

黄根脂溶性成分的GC-MS分析

采用气相色谱-质谱联用技术（GC-MS）,分析并鉴定黄根的脂溶性化学成分。从黄根挥发油中分离出74个成分,并鉴定了其中的36个成分。黄根挥发油主要成分为十八碳6烯酸（7.52%）,棕榈酸（6.34%）,Phosphetane,2,2,3,4,4-pentamethyl-1-phenoxy-,1-oxide （6.05%）等。从黄根石油醚提取物中分离出64个成分,并鉴定了其中的37个成分。黄根石油醚提取物主要成分为反式油酸（18.85%）,棕榈酸（17.66%）,硬脂酸（4.49%）等。利用GC-MS分析鉴定黄根脂溶性成分,具有快速、稳定、重复性好的特点,可用于黄根药材成分的分析鉴定。     

海芒果可培养细菌的分离鉴定及其抑制海洋鱼类致病菌活性研究

【目的】研究红树植物海芒果Cerbera manghas L.及其根际土壤可培养细菌多样性及其代谢产物抑制鱼类致病菌的生物活性。【方法】采用纯培养法分离并基于16SrRNA基因序列的系统发育分析法对分离得到的细菌进行多样性研究,采用滤纸片法研究其抑制海洋鱼类致病菌的生物活性。【结果】从海芒果及其根际土壤中共分离得到15株可培养细菌,采用16SrRNA基因序列的系统发育分析后发现,这些菌分属于2个大的细菌发育类群:厚壁菌门(Firmicutes,占比为53%)和放线菌门(Actinobacteria,占比为47%),其中4株为芽孢杆菌属,6株为链霉菌属。活性研究发现,所有测试菌株对3种海洋鱼类致病弧菌均有一定的抑制作用,其中VT86(Kitasatospora paranensis),V139-2(Streptomyces luteireticuli),V140-2(Streptomyces jiujiangensis),VT19(Streptomyces orinoci),VT113(Streptomyces niveiscabiei)这5株菌的抑菌效果较好,以VT86(K.paranensis)抑菌活性最好。【结论】红树植物海芒果及其根际土壤中的细菌丰富多样,部分细菌具有抑制海洋鱼类致病菌生长的生物活性,具有开发成新型抑制鱼类致病菌药物的潜力。     

甘蔗糖蜜总糖中提高蔗糖纯度的菌株分离鉴定及生物纯化性能

从甘蔗糖厂周边环境采集污泥、土壤及污水样品,通过甘蔗糖蜜纯化发酵试验筛选提高甘蔗糖蜜总糖中蔗糖纯度的菌株,并对菌株进行形态观察、生理生化特征比较和26SrDNA基因序列同源性分析鉴定。结果筛选获得1株提高甘蔗糖蜜总糖中蔗糖纯度的菌株m-6。菌株m-6纯化发酵10h,甘蔗糖蜜中葡萄糖、果糖的去除比例分别为96.5%、100%,蔗糖在糖蜜总糖中的纯度由初始的55.6%提高至85.7%,菌株m-6对甘蔗糖蜜有良好生物纯化作用。菌株m-6属于库德毕赤酵母(Pichia kudriavzevii)。     

灰毛豆Tephrosia purpurea中查尔酮类化合物的分离鉴定

【目的】为探讨灰毛豆属植物Tephrosia purpurea化学成分中的查尔酮类化合物,对其枝、叶部位进行化学成分分离及结构解析。【方法】采用乙醇提取、溶剂萃取、柱层析分离方法得到多种灰毛豆属T.purpurea化学成分,并用波谱分析手段对其进行结构表征。【结果】从灰毛豆属T.purpurea中共分离鉴定出11个查尔酮类化合物,分别为elatadihydrochalcone(1),spinochalcone A(2),3′,5′-diisopentenyl-2′,4′-dihydroxychalcone(3),candidachalcone(4),O-methylpongamol(5),(+)-tephrosone(6),spinochalcone C(7),epoxyobovatachalcone(8),tunicatachalcone(9),crassichalone(10),(S)-elatadihydrochalcone(11)。【结论】以上化合物均为查尔酮,且均属首次从该植物中分离得到。     

Microcalorimetric investigation of effect of berberine alkaloids from Coptis chinensis Franch on intestinal diagnostic flora growth

The inhibitory effect of three berberine alkaloids (BAs) from Coptis chinensis Franch, a traditional Chinese medicinal (TCM) herb, on Bifidobacterium adolescentis growth was investigated by microcalorimetry. The power-time curves of B. adolescentis with and without BAs were acquired, meanwhile the extent and duration of inhibitory effect on the metabolism were evaluated by the growth rate constant (k), half inhibitory ratio (IC₅₀), maximum heat-output power (P _max), peak time of maximum heat-output power (t _p) and total heat production (Q _t). k, P _max and Q _t decreased, and t _p was prolonged with the increase of BAs concentration. The IC₅₀ of BAs is 806 μg/mL for berberine, 341 μg/mL for coptisine and 236 μg/mL for palmatine. The sequence of antimicrobial activity of BAs is berberine < coptisine < palmatine. Combined with previous studies, it could be shown that the sequences of antimicrobial activity of BAs on both Bacillus shigae and Escherichia coli are berberine > coptisine > palmatine. The structure-function relationship of BAs indicates that the functional group methylenedioxy or methoxyl at C2 and C3 might be the major group inducing the activities of BAs on E. coli and B. adolescentis. Meanwhile, the substituent groups at C2, C3, C9 and C10 almost have equal effect on B. shigae. Supported by the National Natural Science Foundation of China (Grant Nos. 30625042 and 30873385) and Key Projects of National Science & Technology Pillar Program of China in the 11th Five-Year Period (Grant No. 2007BAI40B05)