表面增强拉曼光谱法快速鉴别非法染色的关黄柏药材

目的采用表面增强拉曼光谱法(SERS)对非法染色的关黄柏进行快速检测。方法采用原位还原法制备的银胶纸作为SERS基底,擦拭经乙醇溶液润湿的关黄柏,银胶纸的擦拭区立即进行SERS检测。本文先后对硝酸银的浓度、银胶纸的增强效果及稳定性等因素进行考察和优化。结果成功鉴别经0.01 mg·m L-1金胺O染料染色的关黄柏。结论表面增强拉曼光谱法可实现对染色黄药材的快速、简单、灵敏和无损的鉴别。     

基于整体柱的表面增强拉曼光谱法检测非法染色青黛

目的 建立基于整体柱的表面增强拉曼光谱(surface enhanced Raman spectroscopy, SERS)法快速检测非法染色的青黛。 方法 采用混合溶剂乙醇-水(1:1,V/V)对染色青黛中的染料进行提取,利用整体柱中网状孔隙有助于银纳米颗粒在空间上耦合的特性,将样品溶液和银胶溶液混匀后滴加于整体柱上进行SERS光谱采集。 结果 此法可得到青黛中染色掺伪量低至500 μg/kg的孔雀石绿SERS信号。 结论 该方法具有快速、简便、专属性强等优点,可运用于中药材青黛非法染色的快速检测。     

表面增强拉曼光谱法检测微量添加物质的研究

目的 快速、简便、准确地检测中成药与食品中的微量非法添加物质.方法 用表面增强拉曼光谱法对市场上添加了微量物质的中成药及食品进行检测.结果与结论 该方法快速、简便、准确,检测结果与用传统方法检测的结果相吻合.     

薄层色谱-表面增强拉曼光谱法快速检测染色掺伪的西红花

目的 建立一种薄层色谱-表面增强拉曼光谱法对染色掺伪的西红花进行检测。方法 将乙醇溶液润湿的西红花按压在表面喷有银溶胶的薄层板上,然后对按压区域进行表面增强拉曼散射（SERS）检测。依次对润湿剂的浓度、银溶胶的喷洒时间和喷洒量等因素进行优化。结果 成功检测经金胺O、新品红、柠檬黄和胭脂红4种常见染料染色的西红花药材。结论 薄层色谱-表面增强拉曼光谱法可实现对染色西红花简单、快速、灵敏的检测要求,并满足现场快速检测的需求。     

表面增强拉曼光谱法对盐酸四环素注射液的定量分析

目的 利用表面增强拉曼光谱(SERS)法对注射用盐酸四环素进行定量分析.方法 利用便携式拉曼光谱仪测量盐酸四环素注射液的拉曼特征峰,并对峰位进行了归属和指认.结果 盐酸四环素的拉曼特征峰主要位于1278、1311、1450、1617cm-1.建立了盐酸四环素SERS特征峰强与浓度的函数关系,通过线性拟合,得到拟合方程:y=191.95x-4842,R2=0.9976,进而为盐酸四环素的定量分析奠定基础.结论 ERS具有快速、简便、准确等优点,有望在注射用盐酸四环素的鉴定和检测中使用.     

利用表面增强拉曼光谱技术检测硫酸阿托品

摘 要 目的：建立硫酸阿托品的表面增强拉曼光谱（SERS）技术检测方法。方法: 通过理论计算与试验检测相结合,对硫酸阿托品分子的拉曼峰进行归属,并选取1 002 cm-1拉曼峰作为特征峰,开展定量分析。结果: 硫酸阿托品水溶液中的检测限约为0.5 μgSymbolWC@ml^-1;1 002 cm-1处特征峰强度随水溶液浓度变化关系的线性范围在1~8 μg·ml^-1,r=0.983 9;平均回收率为103.3%,RSD为4.5%（n=9）。同时,对该方法的批次间稳定性进行了测试,相对标准偏差为5.7%。另外,对含有硫酸阿托品的中药苍术掺杂样品进行检测,依然能检测到1 002 cm-1处的特征峰。结论: 该方法具有快速、准确、无损、操作简便等优点,在硫酸阿托品检测方面具有很好的应用潜力。     

表面增强拉曼光谱法检测保健品中添加的微量西布曲明

摘 要 目的： 对保健品中添加的西布曲明进行快速检测。方法: 用便携式拉曼光谱仪对市场上几种添加了微量西布曲明的保健品进行快速检测,实验条件：激发波长: 785 nm±0.5 nm,光谱覆盖范围: 175～3 100 cm^-1,信号采集时间20 s,光谱分辨率: 8 cm^-1。结果: 研究表明, 表面增强拉曼光谱法可快速检测出添加于保健品中的微量西布曲明,检测结果与传统方法检测的结果相吻合。结论:该方法快速、简便、无损,可作为检测保健品中非法添加的微量西布曲明的快检方法。     

表面增强拉曼光谱法快速鉴别失活白色念珠菌

目的 建立表面增强拉曼光谱（surface-enhanced Raman spectroscopy,SERS）法快速判别失活白色念珠菌。方法 利用热、甲醛、杀菌药（两性霉素B）分别灭活白色念珠菌,然后利用SERS法对失活前后的白色念珠菌进行光谱考察比较。结果 白色念珠菌灭活后的SERS光谱显示出相似的特征峰,且与失活前的光谱有明显差异。结论 该实验方法快速、简便,实现了对失活白色念珠菌的快速检测,有望成为快速鉴别更多病原微生物失活状态的有利工具。     

红花注射液表面增强拉曼光谱研究

摘 要 目的： 研究红花的表面增强拉曼光谱,利用表面增强拉曼光谱技术对红花注射液进行快速有效的鉴别。 方法: 通过对红花注射液图谱与相应对照药材标准图谱的比较分析,实现对红花注射液的快速鉴别。结果:研究表明,红花的几个特征峰在表面增强拉曼光谱中得到了很明显的增强,表面增强拉曼光谱可很好地识别红花注射液。结论: 该方法简单、快速、可靠、专属性强,可以作为鉴别红花及红花注射液的方法。     

盐酸二甲双胍的表面增强拉曼光谱研究

目的:研究盐酸二甲双胍在不同激发光波长下的固体拉曼光谱以及不同酸碱度条件下的表面增强拉曼光谱,对比固体谱,分析盐酸二甲双胍表面增强拉曼光谱中的分子振动模式.方法:用2种不同的激发光波长对盐酸二甲双胍固体粉末及不同pH 条件下盐酸二甲双胍溶液的表面增强拉曼散射(SERS)进行考察.结果:体系的pH对盐酸二甲双胍的增强效应有较大的影响,在碱性条件下,盐酸二甲双胍的SERS效应最强,而其他两种pH条件下均不明显.另外,激发光波长对盐酸二甲双胍的SERS效应也有较大影响.结论:本方法简单、快速、专属性强,可以作为分析盐酸二甲双胍分子结构信息的方法.     

青风藤中挥发油成分的分析

目的 建立气相色谱-质谱联用(GC-MS)方法分析青风藤中的挥发油成分。方法 采用水蒸气蒸馏法提取青风藤挥发油,运用GC-MS技术,结合数据库检索对其挥发油进行分离鉴定。对青风藤中的挥发油进行分析。结果 实验共鉴定出80个化合物,通过与软件中的质谱标准谱图库进行比较,获得匹配的有63个化合物,主要成分为脂肪酸、甾醇、烯烃以及少量的醛类等物质,含量最多的为十八烷酸和十六烷酸。结论 该实验丰富了青风藤的物质基础研究,为青风藤质量控制提供了更多的参考依据。     

Rapid separation and characterization of active flavonolignans of Silybum marianum by ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry

Ultra-performance liquid chromatography (UPLC) interfaced with the electrospray ionization (ESI) tandem mass spectrometer (MSⁿ) was developed for the simultaneous determination of silychristins A (1) and B (2), silydianin (3), silybins A (4) and B (5), and isosilybins A (6) and B (7), major bioactive flavonolignans in silymarin, a herbal remedy derived from the milk thistle Silybum marianum. In this study, the seven major active flavonolignans including the diastereomers 1/2, 4/5, and 6/7 were completely separated using UPLC with an ACQUITY UPLC C₁₈ column and a MeOH/water/formic acid mobile phase system. The collision-induced dissociation (CID) MSⁿ spectra of these flavonolignans were studied systematically using ESI-MS. The results with the present methodology show that UPLC–MSⁿ can be useful for general screening of active natural products from plant extracts and for the specific quality control of silymarin.     

结合DAD光谱及单级质谱快速鉴定壮阳类中药制剂中的非法添加物

目的建立快速鉴定补肾壮阳类中药制剂中非法添加物的方法。方法通过HPLC-DAD和LC-MS(单级质谱)分析,利用保留时间、DAD光谱和准分子离子峰及其同位素丰度比4个方面的信息,检测10批市售中药制剂中的非法添加物。结果采用所建立的方法检测出3批中药制剂中含有枸橼酸西地那非和(或)他达拉非。结论本文建立的方法结果准确可靠,可用于基层单位的快速筛查,同时可为建立中药制剂中非法添加化学药物的技术标准提供依据。     

Detection of monofluoroacetate in Palicourea and Amorimia species

Numerous plant species worldwide including Palicourea marcgravii and Tanaecium bilabiatum in Brazil cause sudden death and are known to contain monofluoroacetate (MFA). Other species in Brazil including some species traditionally assigned to Mascagnia but now properly called Amorimia species and other Palicourea species are reported to cause sudden death in livestock and are suspected to contain MFA due to the similarity of clinical signs. In this study, an HPLC-APCI-MS method to detect and quantify MFA was developed and was used to investigate plant material from field collections and/or herbarium specimens of Mascagnia, Amorimia, and Palicourea species suspected of causing sudden death. MFA was detected in Amorimia amazonica, Amorimia camporum, Amorimia exotropica, Amorimia pubiflora, Amorimia rigida, and Amorimia septentrionalis as well as Palicourea aeneofusca. MFA concentrations differ greatly between Palicourea species and Amorimia species, which may explain the incidence of poisoning and the amount of plant material required to cause sudden death between these taxa.     

Diverse role of microbially bioconverted product of cabbage (Brassicaoleracea) by Pseudomonassyringe pv. T1 on inhibiting Candida species

This study was carried out to evaluate the anticandidal effects of bioconverted product, obtained from the microbial conversion of cabbage (Brassicaoleracea) by a bacterial strain Pseudomonassyringe pv. T1 (Ps-T1) against various isolates of Candida species. The diameters of zones of inhibition of bioconverted product of cabbage (10 μl, corresponding to 500 μg/disc) against Candidaalbicans KACC 30003 and 30062, Candidageochares KACC 30061, Candidasaitoana KACC 41238 and Candidaglabrata P00368 were found between 10 ± 1 and 16 ± 0.8 mm. The bioconverted product was tested for the minimum inhibitory and minimum fungicidal concentration values against the tested pathogens which were found in the range of 125-500 and 125-500 μg/ml, respectively. On the viable counts of the tested fungal pathogens, the bioconverted product showed a remarkable anticandidal effect. Also the study of using scanning electron microscopy on the morphology of C.albicans KACC 30062 revealed potential detrimental effect of bioconverted product at MIC concentration. The results of this study suggest that bioconverted product of cabbage by Ps-T1 holds potential therapeutic value and medicinal significance to control Candida species.     

Microbial conversion and anticandidal effects of bioconverted product of cabbage (Brassica oleracea) by Pectobacterium carotovorum pv. carotovorum 21

This study was undertaken to examine the anticandidal effects of microbially bioconverted product of cabbage, obtained from the microbial conversion of cabbage (Brassica oleracea var. capitata) by a bacterial strain Pectobacterium carotovorum pv. carotovorum 21 (Pcc 21) against various isolates of Candida species including a clinical isolate. The bioconverted product (10 μl, corresponding to 500 μg/disc) displayed potential anticandidal effect against Candida albicans KACC 30062, Candida geochares KACC 30061, Candida albicans KACC 30003, Candida saitoana KACC 41238 and Candida glabrata P00368 (clinical isolate) as a diameter of zones of inhibition, found in the range of 14 ± 0.9 to 19 ± 1.1 mm. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of bioconverted product against the tested isolates were found in the range of 62.5–250 and 125–250 μg/ml, respectively. Also the bioconverted product had remarkable anticandidal effect on the viable counts of the tested Candida isolates. Further, scanning electron microscopic study revealed potential detrimental effect of bioconverted product on the morphology of C. albicans KACC 30062 at MIC concentration. All these findings together indicate that bioconverted product of cabbage has potential therapeutic value of medicinal significance to control Candida species including clinical isolates.     

Metabolic profiling and biological capacity of Pieris brassicae fed with kale (Brassica oleracea L. var. acephala)

Phenolic and organic acid profiles of aqueous extracts from Pieris brassicae material and the host kale (Brassica oleracea L. var. acephala) leaves were determined by HPLC/UV–DAD/MSⁿ-ESI and HPLC–UV, respectively. The identified phenolics included acylated and nonacylated flavonoid glycosides, hydroxycinnamic acyl gentiobiosides, and sulphate phenolics. Kale exhibited the highest content (11 g/kg lyophilized extract), while no phenolics were identified in the butterflies or exuviae. Nine different organic acids were characterized in the materials, with kale showing the highest amount (112 g/kg lyophilized extract). With the exception of the exuviae extract, the rest were screened for bioactivity. Using spectrophotometric microassays, all exhibited antiradical capacity against DPPH and NO in a concentration-dependent way, whereas only kale and excrement extracts were active against superoxide. All displayed activity on intestinal smooth muscle, albeit with distinct relaxation–contraction profiles. Larvae and butterfly extracts were more efficacious for intestinal relaxation than was kale extract, whereas excrement extract evoked only contractions, thus evidencing their different compositions. Collectively, these results show that P. brassicae sequesters and metabolizes kale’s phenolic compounds. Moreover, the extract’s bioactivities suggest that they may constitute an interesting source of bioactive compounds whose complex chemical structures preclude either synthesis or isolation.     

Evidence of Methylobacterium spp. and Hyphomicrobium sp. in azaspiracid toxin contaminated mussel tissues and assessment of the effect of azaspiracid on their growth

A flagellar protein belonging to the genus Methylobacterium or Agrobacterium was previously observed by proteomics in azaspiracids (AZA) toxic mussels. Here, we report the isolation of two different Methylobacterium spp. (NTx1 and Tx1) from non-toxic and AZA toxic mussels, respectively, which when co-cultured with AZA exhibited significantly different growth responses - isolate Tx1 growth rate was enhanced, whereas growth of isolate NTx1 was adversely affected, compared to non-AZA supplemented control cultures. A Hyphomicrobium sp. (Tx2) also isolated from the toxic mussels achieved greater cell density in AZAs supplemented cultures.     

Toxicity assessment of crude and partially purified extracts of marine Synechocystis and Synechococcus cyanobacterial strains in marine invertebrates

Among the Cyanoprokaryota, the genera Synechocystis and Synechococcus have rarely been studied with respect to potential toxicity. This is particularly true with marine environments where studies about the toxicity of cyanobacteria are restricted to filamentous forms at the warmer temperate and tropical regions and also to filamentous forms at cold seas such as the Baltic Sea. In this study, we describe the effects of cyanobacterial strains of the Synechocystis and Synechococcus genera isolated from the marine coast of Portugal, on marine invertebrates. Crude and partially purified extracts at a concentration of 100 mg/ml of freeze-dried material of the marine strains were tested for acute toxicity in nauplii of the brine shrimp Artemia salina, in the rotifer Brachionus plicatillis and in embryos of the sea urchin Paracentrotus lividus and the mussel Mytilus galloprovincialis. The cyanobacterial extracts, especially the crude extract, had an impact on A. salina nauplii. No significant toxic effects were registered against the rotifer. A negative impact of all strains was recorded on the embryonic development of the sea urchin, with toxic effects resulting in an inhibition of embryogenesis or development of smaller larvae. To the mussel embryos, the effects of cyanobacterial extracts resulted in a complete inhibition of embryogenesis. The results of all assays indicate that Synechocystis and Synechococcus marine strains contained toxic compounds to marine invertebrates.