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1.
天麻特异DNA序列的克隆及其在天麻鉴定中的应用 总被引:1,自引:0,他引:1
应用改进的RAPD方法测定了名贵中药材天麻基因组DNA指纹图谱;通过选择和回收各种天麻种群共有和优良种群特有的DNA片段,加以克隆、测序和生物信息学分析,证明其中5个DNA序列是未报道的,已被美国基因数据库收录,并运用高效液相色谱技术测定了天麻样本的有效成分天麻素含量。运用PCR技术研究了这些DNA序列在9个天麻种群中的分布及其与天麻素含量的关系。结果表明这5个DNA序列在这些天麻种群中的分布各不相同,其中DNA序列1是所研究的全部天麻种群共有、而其伪品没有的特异DNA分子标记;DNA序列2可能与天麻的天麻素含量高有关。这些DNA标记序列可用于天麻的真伪鉴别、品种鉴定和优选优育等。 相似文献
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鳖甲消痔胶囊中中药化学成分的测定 总被引:2,自引:0,他引:2
孙芳 《武汉生物工程学院学报》2007,(3)
采用薄层色谱法(thin-layer chromatography,TLC)对药方中的土大黄、地榆、槐角和栀子进行定性鉴别,并用高效液相色谱法(high performance liquid chromatography,HPLC)测定胶囊中盐酸小檗碱的含量。结果表明,TLC法可用于土大黄、地榆、槐角和栀子的定性鉴别;盐酸小檗碱对照品在0.1~1.0μg范围内呈线性关系,r=0.999 4;加样回收率为97.80%,RSD为1.40%。本方法结果准确,灵敏度高,重复性好,能够有效地控制鳖甲消痔胶囊的质量。 相似文献
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《现代生物医学进展》2016,(34)
目的:探讨天麻熄风胶囊的DNA条形码鉴定方法与价值。方法:2015年1月到11月,选择3批天麻熄风胶囊(批号20150918、200150919、20151019)进行显微鉴定与DNA条形码鉴定,选择的基因序列为天麻素线粒体A基因,进行基因测序与遗传分析。结果:显微鉴定显示天麻细胞无色,呈现长卵形、长椭圆形或类圆形,遇碘液显棕色;蔓荆子非腺毛2-3个细胞,顶端细胞基部稍粗;甘草纤维成束、壁厚,周围薄壁细胞可形成晶纤维;半夏椭圆形细胞存在于粘液细胞中。3批天麻熄风胶囊的天麻素线粒体A基因特定序列中,A含量为27.9%、C为24.9%、G为18.3%、T为28.9%,G-T含量明显高于G/C含量(43.2%)(P0.05)。遗传分析显示20150918批次的种间最小K-2P距离为0.006,种间最大K-2P距离为0.241;20150919批次种间最小K-2P距离为0.008,最大K-2P距离为0.074;20151019批次的种间最小K-2P距离为0.008,种间最大K-2P距离为0.133。结论:天麻熄风胶囊成分复杂,显微鉴定技术存在一定的缺陷,DNA条形码鉴定能有效反映胶囊成分,且能很好判定药物批次的种内与种间变异,具有很好的鉴别价值。 相似文献
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首次采用高效液相色谱法对中药复方止咳清肺胶囊中的有效成分—芥子碱硫氰酸盐进行了含量测定研究,由此建立了该复方的HPLC法质量分析方法。 相似文献
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目的:天麻具有许多药理作用和重要开发价值,通过探索天麻的未知化学成分和鉴定西藏天麻种群的品质,为深入研究、开发和利用西藏优良天麻种群提供科学技术支持。方法:本研究采用HLPC-MS(液质联用)技术测定西藏天麻的化学成分;采用高效液相色谱技术测定西藏6个天麻种群的生化指纹图谱,利用天麻素峰面积、数学公式和SPSS软件,计算和比较西藏6个天麻种群的平均天麻素含量和各种化学成分分离峰的总面。结果:从天麻块茎中发现了33个未报道的化学物质;西藏天麻的生化指纹图谱具有7个较大的共有特征分离峰,分别位于1.854、2.759、7.279、7.591、8.500、9.557、10.753min;根据生化指纹图谱特征可将它们分为三个主要类型;凡是带有一型生化指纹图谱的天麻个体和种群往往含有更高的天麻素和更大的分离峰总面积,品质更优良。结论:西藏天麻种群3和种群6以一型生化指纹图谱为主,其天麻素含量最高,分离峰总面积最大,品质最优良,具有重要研究、开发和保护价值。本研究成果对于天麻的化学成分分析与鉴定、天麻种群和品质鉴定与评价以及西藏天麻的品种选优、资源保护与利用具有重要指导意义和科学价值。 相似文献
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采用浸苗法将野生天麻总DNA导入马铃薯试管苗,对筛选得到的转化植株进行蛋白及药用成份的分析。结果显示:(1)在200株转化的马铃薯中有21株的紫外扫描图谱与正常对照组有显著差异,且在220nm有明显吸收峰。(2)5株经PCR扩增出野生天麻抗真菌蛋白(GAFP)基因。(3)转基因马铃薯与正常马铃薯的蛋白表达有明显差异,并且在转基因马铃薯中有一条与野生天麻抗真菌蛋白(GAFP)相同的条带。而正常马铃薯中无此条带。(4)通过薄层层析法检测出3株转基因马铃薯表达野生天麻的有效药用成份天麻素。说明采用浸苗法进行外源总DNA导入是可行的。 相似文献
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为选择一种准确快捷的方法测定银耳多糖的单糖组成,对薄层色谱法(TLC)、气相色谱法(GC)、高效液相色谱法(HPLC)三种色谱方法进行比较。结果表明,前两种方法的测定结果均不理想,而HPLC法,操作简便,灵敏度高,分离效果好,信息完整。测定结果为由葡萄糖、甘露糖、葡萄糖醛酸、木糖、岩藻糖组成,其摩尔比为0.24∶1.00∶0.06∶0.29∶0.25。HPLC法对酸性杂多糖组成糖分析是一种比较理想的选择。 相似文献
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通过薄层色谱(TLC)与高效液相色谱(HPLC)联用技术鉴定了暗纹东方鲀(Takifugu obscurus)肌肉中存在肌肽和谷胱甘肽,同时利用高效液相色谱法测定了肌肽和谷胱甘肽(GSH)的含量。薄层层析采用的展开剂为正丁醇:乙酸:水(4∶2∶1),层析板为硅胶板。高效液相色谱利用Kromasil C18反相柱分析,流动相为10%的纯乙腈和90%含有0.05%三氟乙酸的超纯水。结果表明:通过薄层色谱和高效液相色谱鉴定了暗纹东方鲀肌肉中肌肽和谷胱甘肽,其中暗纹东方鲀肌肉中肌肽含量约213μg/g(鲜重),还原性谷胱甘肽含量约211μg/g(鲜重)。本法样品无需衍生,操作简便,适合于暗纹东方鲀肌肉中肌肽和谷胱甘肽的测定。本文为生物体内肌肽和谷胱甘肽的研究提供借鉴意义。 相似文献
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F2-Isoprostanes in plasma and urine are generally determined by labor-intensive methods requiring sample purification by solid-phase extraction and thin-layer chromatography (TLC). A streamlined and more sensitive method for the measurement of esterified plasma F2-isoprostanes was developed by replacing these steps with high-performance liquid chromatography (HPLC) using an amino column with a hexane/2-propanol gradient. Pentafluorobenzyl esters of F2-isoprostanes were prepared and purified by HPLC, silylated, and then analyzed by gas chromatography (GC) with negative chemical ionization mass spectroscopy (NCI-MS). This method permits analysis with lower plasma volumes (100 microL) and greater sensitivity (to 10 pg; allowing detection to 50 pg/mL) than provided by other methods. Urinary F2-isoprostanes can also be efficiently quantified by this method, with 8-iso-PGF2alpha being identified as a major isomer. With this procedure, esterified plasma F2-isoprostanes were found to be 8.3-fold higher in an end-stage renal failure patient on hemodialysis and urinary 8-iso-PGF2alpha was 7.1-fold higher in a cigarette smoker than respective control subjects. This method, particularly the substitution of the TLC step common to other methods with HPLC, results in a more sensitive and reproducible assay. 相似文献
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József Téren János Varga Zsuzsanna Hamari Edit Rinyu Ferenc Kevei 《Mycopathologia》1996,134(3):171-176
One hundred and fifty-seven strains belonging to Aspergillus section Nigri were tested for ochratoxin A production using three different methods: a relatively new immunochemical method based on an enzyme-linked immunosorbent assay (ELISA), thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The monoclonal antibody-based ELISA technique was successfully used to screen for low levels of ochratoxin A in the black Aspergilli without concentrating the culture filtrates. The results were confirmed by TLC and HPLC analysis and chemical derivatization. These latter methods required concentrated filtrates. Ochratoxin A was detected in the culture filtrates of five of the 12 A. carbonarius strains, none of the 45 A.japonicus strains and three of the 100 isolates in the A. niger aggregate (A. foetidus, A. awamori and A. niger).Abbreviations ELISA
enzyme-linked immunosorbent assay
- HPLC
high-performance liquid chromatography
- OA
ochratoxin A
- TLC
thin-layer chromatography 相似文献
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A simple and rapid method for the analysis of lipids on a thin layer chromatography (TLC) plate by matrix-assisted secondary ion mass spectrometry (SI-MS) is reported. Analysis was performed without elution of the sample from the TLC plate. Mass spectra obtained by this method are free from interference due to the TLC plate absorbent and reagents used for the detection of the spots. About 1 micrograms of lipids applied on a TLC plate can be analyzed by this method. On scanning the plate, mass chromatograms of each lipid were obtained based on its migration distance along the plate. 相似文献
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Oztunc A Onal A Erturk S 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2002,774(2):149-155
Using 7,7,8,8-tetracyanoquinodimethane (TCNQ) as a new derivatization reagent for HPLC and TLC, novel methods are described to detect secondary amine-bearing antidepressants (paroxetine, desipramine, fluoxetine, nortriptyline, maprotiline). The HPLC method is sensitive enough to detect these drugs in plasma at therapeutic levels whereas the latter has potential to detect them in overdose or forensic cases. The methods are based on purple chromogens formed by the displacement reaction of the drugs with TCNQ. The resulting chromogens are directly separated by either reversed-phase HPLC on a C(18) column or TLC on silicagel plates. For HPLC, acetonitrile-water (60:40) was used as mobile phase, with detection at 567 nm and separation in 40 min. For TLC, three developing solvent systems were used. By HPLC, 36 ng ml(-1) spiked plasma concentration of the drugs gave easily detectable signals whereas by TLC, detection limits varied mostly between 240 and 480 ng ml(-1). The HPLC method was applied to real plasma samples. The methods described are simple and very selective; some metabolites of these antidepressants and a vast number of drugs do not interfere with detection. 相似文献
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Oligosaccharides were analyzed by a combination of high-performance liquid chromatography (HPLC) and mass spectrometry (MS). First, oligosaccharides labeled with 2-aminopyridine were studied to see if they could be analyzed by MS under the conditions used for separation by HPLC. Pyridylamino (PA)-oligosaccharides could be analyzed under these conditions, although the mass spectra were affected. Then, liquid chromatography-mass spectrometry was used to analyze a PA-oligosaccharide mixture derived from human immunoglobulin G. The PA-oligosaccharides were separated on a reversed-phase column and mass-analyzed directly. The observed molecular weights were close to or identical to those expected from the structures, which were estimated from the elution position on HPLC. This method is rapid and simple, as the mass spectrometer can give the accurate molecular weight of each PA-oligosaccharide in one chromatography run, even if the HPLC separation is incomplete. This method can be used to extend the so-called two-dimensional mapping of PA-oligosaccharides. The structure can be studied in greater detail by tandem MS. 相似文献
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Thin layer chromatography (TLC) methods for identifying and quantifying deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone
in grain samples were compared to immunoassay (ELISA) and high performance liquid chromatography (HPLC) methods to determine
the reliability of the less expensive TLC. There was a very good agreement between levels of DON measured by TLC and competitive-direct
ELISA, and between levels of fumonisin B1 measured by TLC and HPLC, over a wide range of concentrations. Correlation coefficients
(Pearson's) were 0.978, 0.914 and 0.953 for DON in maize, DON in wheat and FB1 in maize respectively. A lower correlation
coefficient (r = 0.672) was obtained when zearalenone was quantified by TLC and HPLC. Possible reasons for this are discussed.
A cost comparison of the various methods revealed that TLC was the least expensive for sample analysis. It is recommended
that researchers choose which analytical method to use based upon individual considerations of cost and precision.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献