苦橙叶精油化学成分及抗炎作用研究

为探究苦橙叶精油的抗炎作用。实验采用气相-质谱联用法(GC-MS)分析精油成分,并建立脂多糖(LPS)诱导RAW 264.7细胞炎症模型,用Griess法检测一氧化氮(NO)含量评价其体外抗炎作用,随后进一步通过巴豆油致小鼠耳肿胀模型和鸡蛋清致小鼠足肿胀模型评价其体内抗炎作用。结果表明苦橙叶精油成分以酯类、醇类物质为主;25μg/mL浓度能显著抑制RAW 264.7细胞NO的释放;中浓度苦橙叶精油能明显减轻小鼠耳肿胀程度;低、中、高浓度苦橙叶精油均对小鼠足肿胀模型有炎症缓解作用,并于肿胀前期呈浓度依赖性。以上实验证明苦橙叶精油在体外和体内具有一定抗炎作用。     

代代花挥发油的超临界萃取及其在烟草中的应用

为了开发新的天然烟用香料,采用超临界流体萃取技术萃取代代花并优化了萃取工艺,GC-MS对挥发油成分进行了分离鉴定,并对其进行了卷烟加香试验.结果表明:①超临界萃取代代花的最佳工艺为:萃取压力40 MPa、萃取温度50℃萃取时间15 min、改性剂为20%乙醇;②代代花挥发油中共鉴定出45种挥发性成分,主要成分为柠檬烯、...     

农杆菌介导佛手遗传转化主要影响因素的研究

采用根癌农杆菌介导的佛手叶盘转化法,在建立转海藻糖合酶基因(TPS)佛手体系过程中,对影响农杆菌转化频率的各种因素进行了研究。结果表明,佛手叶盘需在黑暗条件下MT培养基上预培养2-3d,与农杆菌共培养3d较合适;农杆菌菌液浓度OD600约为0.6-0.8,感染时间20min;抑制农杆菌生长的抗生素浓度以头孢霉素(Cef)250mgL-1和羧苄青霉素(Cb)250mgL-1且延迟筛选时间4d最好;共培养基中添加100μmol/L乙酰丁香酮(AS)和400mgL-1半胱氨酸(L-Cys)对佛手遗传转化有明显的促进作用。经GUS报告基因和PCR技术检测,初步证实TPS基因已整合到佛手基因组中,且GUS报告基因瞬时表达率为5.9%。     

A Study on the Chemical Components of the Essentialoi from Peel of Citrus medica L. var. muhensis W. D. et Y.

The chemical components of the essential oil from peel of Citrus medica L. var. muliensis W. D. et Y. has been studied by means of the GC-MS-DS, the retention index of eapillarly gas chromatography and the Authentic sample addition process. Twenty-nine constituents has been identified from sixty-four seperated peaks. The main constituents is d-limonene (68.2%), geranial (9.5%), neral (5.37%), nerol (2.72%), β-ocimen (2.47%) and (+)-Carvone.     

Selectivity and potential interference from phenolic compounds in chemiluminescence methods for the determination of synephrine

Three recently reported chemiluminescence methods (based on reactions with alkaline luminol and hexacyanoferrate(III); acidic cerium(IV) and rhodamine B; and acidic permanganate with polyphosphates) for the determination of synephrine were re‐evaluated in terms of their selectivity towards this analyte in comparison to other phenolic compounds. A fourth reagent system, acidic soluble manganese(IV) and formaldehyde, was also examined. Each set of reagents was sensitive towards synephrine (limits of detection were 3 × 10^?9, 5 × 10^?8, 1 × 10^?8 and 1 × 10^?8 mol/L, respectively) but also responded with numerous other phenolic compounds, including some that are present in citrus fruit extracts, dietary supplements and/or biological fluids. It is therefore recommended that the determination of synephrine in these matrices should incorporate physical separation of sample components (e.g. chromatography or electrophoresis). In more general terms, this study illustrates that accurate percentage recoveries for an analyte in spiked samples (without validation against another analytical method) are insufficient to confirm the analytical utility of new flow‐injection analysis (FIA) procedures. Copyright © 2008 John Wiley & Sons, Ltd.     

金瓜和白瓜花叶病毒病病原的初步鉴定

崇明是上海出口蔬菜生产基地之一,也是国家级的绿色食品园区之一,具有生产绿色蔬菜商品的生态环境和资源优势。特色出口蔬菜金瓜(Cucurbita pepo L.var.ovifera)、白瓜(Cucumis melo L.var.conomon,又名盐渍菜瓜)1997～1999统计病毒病发病率为23％,被感染的金瓜、白瓜田     

西双版纳黄瓜Cs-Psy1基因的序列特征与表达分析

西双版纳黄瓜是我国特有的果肉橙黄色的黄瓜变种资源,不同种质间的β-胡萝卜素含量差异明显。PSY是胡萝卜素生物合成途径中的第1个限速酶。本文以西双版纳黄瓜为试材,分别克隆西双版纳黄瓜八氢番茄红素合成酶(Cs-PSY1)的DNA和c DNA序列,结果显示,DNA长2797 bp,包含5个内含子和6个外显子,c DNA序列长1385 bp,编码421个氨基酸。Psy1推测的氨基酸序列包含该家族的2个特征序列,保守性很高。该蛋白为不稳定蛋白,无明显疏水区,未预测到跨膜结构;系统进化分析结果显示,西双版纳黄瓜的Cs-PSY1蛋白与甜瓜的同源性较高;与栽培黄瓜深度测序材料"9930"和"GY14"的序列进行比较分析,结合115份黄瓜重测序结果,共发现5个SNP,其中2个位于起始密码子上游27 bp处和971 bp处,3个位于内含子区域。其中SNP4在重测序的19份西双版纳黄瓜中的突变率为100%,在96份栽培黄瓜中的特异性为5.3%。转录因子结合位点预测结果显示,在普通栽培黄瓜该位点处存在一个CTAG motif,在西双版纳黄瓜中该位点突变后则不存在该motif。利用实时荧光定量PCR技术分析Cs-Psy1的表达量变化趋势,结果表明,在黄瓜不同果实发育时期,该基因的表达量均呈现先上升后下降的趋势,在西双版纳黄瓜中表达量变化的差异明显,在授粉后50 d达到最大值,是果实发育初期表达量的8倍多,是同时期普通黄瓜的4倍多,而普通黄瓜表达量的总体变化相对平缓。西双版纳黄瓜果实内果皮的表达水平明显高于中果皮,最高相差约5倍,普通黄瓜差异不明显。从上述研究结果推测Psy1基因可能影响西双版纳黄瓜的β-胡萝卜素积累。     

Simultaneous determination of flavonoids and phenylethanoids in the flowers of Verbascum densiflorum and V. phlomoides by high‐performance liquid chromatography

Introduction – Mullein (Verbascum) flowers are highly valued herbal drugs used in the treatment of inflammation, asthma, spasmodic coughs and other respiratory tract diseases. Their phenolic constituents are considered to be responsible for the anti‐inflammatory and antimicrobial activity of the herb. However, knowledge about the contents of phenolics in flowers is limited and no HPLC method for their analysis is available. Objective – To develop and validate an RP‐HPLC‐UV method for the simultaneous determination of eight flavonoids and two phenylethanoids in the flowers of Verbascum densiflorum and V. phlomoides. Methodology – HPLC separation was accomplished on a C₁₈ Lichrosphere 100 column (5 µm, 250 mm × 4.6 mm, i.d.) with an acetonitrile gradient elution using aqueous 0.5% (w/v) orthophosphoric acid solution containing 1% (v/v) tetrahydrofurane. Results – All the calibration curves showed good linear correlation coefficients (r > 0.997) over the wide test ranges. The relative standard deviation of the method was less than 3.4% for intra‐ and inter‐day assays, and the average recoveries were between 93.5 and 101.9%. High sensitivity was demonstrated with detection limits of 0.062–0.083 µg/mL for flavonoid aglycones, 0.156–0.336 µg/mL for flavonoid glycosides and 0.390–0.555 µg/mL for phenylethanoids. The flower samples of V. phlomoides were found to contain high levels of diosmin and tamarixetin 7‐rutinoside (2.327–2.392% of dry weight), whereas verbascoside (0.688–0.742% of dry weight) and luteolin 7‐glucoside (0.204–0.279% of dry weight) dominated in the V. densiflorum flower. Conclusion – The HPLC method established is appropriate for the quality assurance and the differentiation of V. phlomoides and V. densiflorum samples. Copyright © 2009 John Wiley & Sons, Ltd.     

Simultaneous quantification of five triterpenoid saponins in Clematis L. spp. by high-performance liquid chromatography with evaporative light scattering detection

A simple and accurate method involving high-performance liquid chromatography with evaporative light scattering detection was developed for the simultaneous determination of five triterpenoid saponin components in Clematis L. spp. for the first time. The analysis was performed on a Zorbax SB-C(18) column and gradient elution with acetonitrile and water with 0.1% formic acid was utilised. All the calibration curves exhibited good linear characteristics with correlation coefficients in the range from 0.9979 to 0.9997. The limits of detection and limits of quantification were less than 0.152 and 0.506 microg, respectively. The overall recoveries for the five analytes were between 91.3 and 99.5%. A total of 10 samples from Clematis L. spp. were analysed under optimised conditions and the chemical profiles provided information for the identification of botanical origin, the development of new medicinal resources and chemotaxonomic investigation.     

Secretory Cavity Development and Its Relationship with the Accumulation of Essential Oil in Fruits of Citrus medica L. var. sarcodactylis （Noot.） Swingle

The developmental types of secretory cavities in Citrus remain controversial. The relationship between secretory cavity development and the accumulation of essential oil in fruits of Citrus species is also unknown. In order to develop better insights into these problems, histological, histochemical, and cytochemical methods were used to investigate secretory cavity development and the accumulation of essential oll at different developmental stages of fruits of Citrus medica L. var. sarcodactylis （Noot.） Swingle. The results indicate that the secretory cavity of the variety seemed to originate from an epidermal cell and a subepldermal cell. These two cells underwent successive divisions, resulting In the formation of two parts： （Ⅰ） a conical cap; and （Ⅱ） a globular gland. The formation of the lumen was schlzolysigenous. Regular changes in the size of vacuoles and the accumulation of essential oil were revealed during the process of secretory cavity development. In addition, when fruits were a light yellow or golden color, the structure of secretory cavities was well developed and the content of essential oil in a single fruit reached a maximum. It would be most appropriate to collect the fruit as a medicinal material at this time.     

紫花曼陀罗幼苗中紫色异细胞分布特征及其形态建成影响条件的初步研究

应用植物显微技术观察了幼苗期紫花曼陀罗（Datura stramonium L.var.tatula Torrey）中紫色异细胞的分布及不同温度与光照下培养的紫花曼陀罗幼苗茎中紫色异细胞的形态和分布差异。结果：（1）紫花曼陀罗中紫色异细胞分布在茎与叶地上器官中,根中则不存在。（2）强光和高温下培养的紫花曼陀罗幼苗茎中紫色异细胞的颜色明显加深、透明度下降,细胞间隙减小;而弱光和低温下其形态特征表现与上述相反。（3）强光处理下紫色异细胞横切面积、纵切长度以及数量均显著增加（P<0.05）;弱光下该异细胞横切面积、纵切长度均显著减少（P<0.05）,但数量显著增加（P<0.05）。（4）高温处理下紫色异细胞横切面积、纵切长度以及数量均显著增加（P<0.05）;低温下该异细胞仅数量显著增加（P<0.05）,其横切面积、纵切长度虽减小但变化差异性不显著（P<0.05）。结论：紫花曼陀罗中紫色异细胞的形态建成受光照和温度影响,曼陀罗（Datura stramonium L.）变种紫花曼陀罗（D.stramonium L.var.tatula Torrey）各营养器官中进化出紫色异细胞对其更好地适应光照和温度等生态环境变化有积极作用。     

南方铁杉和长苞铁杉技叶的挥发油成分

南方铁杉〔Ｔｓｕｇａｃｈｉｎｅｎｓｉｓ (Ｆｒａｎｃｈ .)Ｐｒｉｔｚ.ｖａｒ.ｔｃｈｅｋｉａｎｇｅｎｓｉｓ (Ｆｌｏｕｓ)ＣｈｅｎｇｅｔＬ .Ｋ .Ｆｕ〕和长苞铁杉 (ＴｓｕｇａｌｏｎｇｉｂｒａｃｔｅａｔａＣｈｅｎｇ)均为我国特有种 ,国家重点保护植物[1 ,2 ] ,主要分布于长江以南各省区。二者不仅是高级材用树种 ,而且是很好的造林及观赏植物 ,并有一定的药用价值。作者在野外调查中发现 ,民间用铁杉的枝叶治疗关节炎和胃病 ,为进一步探讨其药用成分和价值 ,对南方铁杉和长苞铁杉的叶及幼枝的挥发油成分进行了分析。1　材料和方法1.1　…     

A qualitative and quantitative HPTLC densitometry method for the analysis of cannabinoids in Cannabis sativa L.

Introduction – Cannabis and cannabinoid based medicines are currently under serious investigation for legitimate development as medicinal agents, necessitating new low‐cost, high‐throughput analytical methods for quality control. Objective – The goal of this study was to develop and validate, according to ICH guidelines, a simple rapid HPTLC method for the quantification of Δ⁹‐tetrahydrocannabinol (Δ⁹‐THC) and qualitative analysis of other main neutral cannabinoids found in cannabis. Methodology – The method was developed and validated with the use of pure cannabinoid reference standards and two medicinal cannabis cultivars. Accuracy was determined by comparing results obtained from the HTPLC method with those obtained from a validated HPLC method. Results – Δ⁹‐THC gives linear calibration curves in the range of 50–500 ng at 206 nm with a linear regression of y = 11.858x + 125.99 and r² = 0.9968. Conclusion – Results have shown that the HPTLC method is reproducible and accurate for the quantification of Δ⁹‐THC in cannabis. The method is also useful for the qualitative screening of the main neutral cannabinoids found in cannabis cultivars. Copyright © 2009 John Wiley & Sons, Ltd.     

A study by cryo-scanning electron microscopy of the initiation and early development of an inflorescence in glasshouse celery (Apium graveolens L. var. dulce (Miller) Pers.) grown under inductive conditions

The morphology of flower initiation and early development in glasshouse celery (Apium graveolens L. var. dulce (Miller) Pers.) cv. Celebrity was studied by means of apical dissections and cryo-scanning electron microscopy. Easily recognisable morphological features were used to define seven stages in the early development of the inflorescence. A highly significant linear regression was established between the logarithm of the apical diameter (measured diametrically across the apical dome between the two most recently initiated leaf or inflorescence primordia) and these discrete floral stages. There was no strong evidence that either the origin or the slope of the regression varied with different combinations of temperature (viz. 10°C or 14°C) and daylength (viz. natural, short or long) which were conducive for the initiation and development of an inflorescence. It is suggested that both apical diameter and floral stage may be used as parameters for assessing the influence of environmental factors such as temperature and daylength on the floral development of glasshouse celery.     

Comparison of three different extraction methods and HPLC determination of the anthraquinones aloe‐emodine,emodine, rheine,chrysophanol and physcione in the bark of Rhamnus alpinus L. (Rhamnaceae)

Introduction – Rhamnus alpinus L. (Rhamnaceae), a traditional plants in the flora of the Abruzzo region, is known to contain active anthraquinone secondary metabolites. However, the content of anthraquinones varies among R. alpinus samples depending on collection season and site. Thus, using simple, reliable and accurate analytical methods for the determination of anthraquinones in R. alpinus extracts allows comparative study of different methods of extraction. Objective – After a partial validation of an HPLC method for the simultaneous determination of five anthraquinones, aloe‐emodine, rheine, emodine, chrysophanol and physcione, in the bark of R. alpinus, we compared three different methods of extraction. Methodology – Anthraquinones were extracted from the bark of R. alpinus using different techniques (methanol maceration, ultrasonic and supercritical CO₂ extraction). Separation and quantification of anthraquinones were accomplished using a reversed‐phase C₁₈ column with the mobile phase of H₂O–methanol (40 : 60, v/v, 1% formic acid) at a wavelength of 254 nm. The qualitative analyses were also achieved at wavelength of 435 nm. Results – All calibration curves were linear over the concentration range tested (10–200 mM) with the determination coefficients ≥0.991. The detection limits (S/N = 3) were 5 mM for each analytes. All five anthraquinones were found in the samples tested at concentrations reported in experimental data. Conclusion – The described HPLC method and optimised extraction procedure are simple, accurate and selective for separation and quantification of anthraquinones in the bark of R. alpinus and allow evaluation of the best extraction procedure between the tested assays. Copyright © 2009 John Wiley & Sons, Ltd.