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91.
本文采用Mannich反应合成黄曲霉毒素B1(AFB1)人工抗原,免疫小鼠制备AFB1单克隆抗体。采用直接搅拌法将异硫氰酸荧光素(FITC)标记AFB1抗体,经Sephadex-50凝胶柱纯化,制得FITC-AFB1荧光标记抗体,分析其免疫学特性,从而建立了一种快速灵敏的直接竞争荧光免疫分析方法(FIA)以用于检测中药材中的AFB1含量。结果表明,AFB1-FITC标记抗体的结合比率为4.19。通过对检测体系多项影响因素的筛选优化,FIA检测方法的标准曲线方程为I=33.45 log C+25.55,R=0.9913,线性检测范围1~100 ng/m L,检测限0.69ng/m L,回收率90.4%~106.6%。该方法具有操作简单、快速灵敏、特异性高等特点,可用于中药材中AFB1的分析测定。 相似文献
92.
A new method based on solid-support reaction is described to realize fluorescent derivatization of proteins at concentrations as low as 10−8 M. A simple, low-cost homemade capillary C18 cartridge was fabricated as the solid-support reactor. Using bovine serum albumin (BSA) as a test protein, we demonstrated that the protein can be captured by this reactor and then labeled by fluorescein isothiocyanate (FITC, isomer I) on solid-support. Unwanted fluorescent intruder (excrescent FITC and products of secondary reactions) were removed from target easily. The analysis by nano-HPLC with laser-induced fluorescence (LIF) detection was described. The effect of reaction conditions on the derivatization has been evaluated and discussed. The use of the solid-support reactor allows easy handling of as little as 8.5 pmol of BSA. A fraction from weak anion-exchange chromatography (WAX) of human liver extract was used as an illustrative example of application to real samples. 相似文献
93.
在λex/λem=405/547 nm,于缓冲溶液和表面活性剂存在的情况下,荧光素和曙红Y能够发生有效能量转移,而牛血清白蛋白(BSA)的加入使得曙红Y荧光猝灭,该体系可用于微量蛋白质的测定。系统探讨了荧光素-曙红Y能量转移体系发生荧光猝灭的条件,最佳条件为:2.0 mL pH=3.8的B-R缓冲溶液,0.4 mL 0.05%曲拉通X-100,1.5 mL 1.0×10-4mol/L的荧光素水溶液,2.0 mL 1.0×10-4mol/L的曙红Y水溶液,最佳实验时间为溶液配制完成静置15 min后60 min内,最佳加入顺序为pH=3.8缓冲溶液+荧光素+曙红Y+曲拉通+蛋白质标准溶液或样品。在优化的实验条件下,蛋白质含量在0~2.0μg/mL范围内与荧光猝灭强度呈良好的线性关系。检出限为6.6 ng/mL;测定样品的相对标准偏差(RSD)在±5%以内;样品加标回收率为90.4%~95.3%。该法可用于人血清、牛奶中蛋白质含量的测定。 相似文献
94.
银二氧化硅核壳( Ag@ SiO2)可使异硫氰酸荧光素酯(FITC)产生金属增强室温磷光(MERTP).当所研究体系加入赖氨酸时,其MERTP猝灭.据此,建立了检测赖氨酸的新方法.在FITC和Ag@ Si02的加入量之比为1∶1.5,pH- 8.0时,本方法检测赖氨酸的检出限为0.270 μmol/L;线性范围为1.50~90.5 μmol/L,人二合成样品加标回收率为97.0%~108.0%;相对标准偏差为2.1%(n=9).本方法用于检测赖氨葡锌颗粒和赖氨酸磷酸氢钙片中的赖氨酸,结果满意. 相似文献
95.
Bertrand Schweitzer-Chaput Massaba Keita Thierry Milcent Sandrine Ongeri Benoit Crousse 《Tetrahedron》2012,68(35):7028-7034
A series of fluorinated N-aminoaziridines have been synthesized by the PhI(OAc)2-mediated aziridination procedure. The reaction was carried out with various protected hydrazides and fluorinated alkenes. The reaction was extended to alkenes bearing an amino acid and the ring opening of the CF3–N-aminoaziridines has been investigated. 相似文献
96.
Abstract The absorption, excitation and fluorescence spectra of the fluorescein dianion (FL2?) in N, N-Dimethylformamide (DMF)/water (H2O) and Dimethylsulphoxide (DMSO)/H2O solvent mixtures have been investigated. It is found that the absorption λmax and emission maxima EX, nax are both hypsochromic shifted when the H2O content in the solvent mixtures increases. However, the shoulder peaks λs remain constant at 483. 5nm within the range of H2O mole fraction, x= 0 to 0. 518 in a DMF/H2O solvent mixture and at 484. 4nm within the range, x=0 to 0. 304, in a DMSO/H2O mixture. Further increases in H2O content, cause a hypsochromic shift in λs. The molar energies for the electronic transition from the ground state (S0) to S1, the first excited singlet state, (ET(1)) and to S2, the 2nd excited singlet state, (ET(2)) of FL2?in the solvent mixtures are also plotted against x. Linear straight lines and intersection points are observed at x=0. 73 for ET(1) and at x=0. 51 for ET(2) in the DMF/H2O mixture and at x=0. 71 for ET(1) and at x=0. 31 &; 0. 69 for ET(2) in the DMSO/H2O mixture. Hydrogen-bonding stabilization effects are used to explain the above observations. The variation in relative fluorescence quantum yields of S1 and S2 of FL2? with x in the aprotic solvent-H2O mixtures to FL2? in pure aprotic solvent are determined and discussed. 相似文献
97.
We have investigated the properties of cellulose diacetate in solution by using laser light scattering. The cellulose diacetate molecules can form micelles and micellar clusters in acetone besides the individual chains. As the concentration increases, the average hydrodynamic radius (Rh) linearly increases, whereas the ratio of gyration radius to hydrodynamic radins 〈Rg〉/〈Rh〉 linearly decreases. It indicates that the micelles associate and form micellar clusters due to the intermolecular interactions. 相似文献
98.
99.
研究了头孢羟氨苄和头孢拉定对荧光素的荧光猝灭反应及相互作用机理。因形成荷移络合物,头孢羟氨苄和头孢拉定对荧光素的荧光强度有很强的猝灭作用。采用紫外-可见光谱、红外光谱结合量子化学计算分析了荷移反应过程,揭示了头孢菌素类药物的成键规律和反应机理。在此基础上建立了测定头孢羟氨苄和头孢拉定含量的荧光光谱法。络合物的激发波长和发射波长分别为483, 517和519 nm,头孢羟氨苄和头孢拉定浓度分别在0.3~13.5 mg·L-1和0.1~1.2 mg·L-1范围内为线性关系,相关系数分别是r=0.999 7和r=0.999 3。应用该方法测定了头孢羟氨苄和头孢拉定药物制剂含量, 回收率分别在99.63%~99.91%和99.71%~100.08%之间。测定结果与文献值基本吻合。 相似文献
100.
研究了荧光素(Fluorescein)与阴离子表面活性剂(Anioni surfactant)十二烷基苯磺酸钠(Sodiun dodecyl benzenesulfonate)、十二烷基磺酸钠(Sodium laurylsulfonate)、十二烷基硫酸钠(Sodiumdodecyl sulphate)的荧光性质,探讨了实验条件对荧光素与阴离子表面活性剂荧光性质的影响。研究结果表明,在室温下,pH 7.3-7.7 Na2HPO4-KH2PO4缓冲溶液中,体系中加入十二烷基苯磺酸钠、十二烷基磺酸钠后荧光强度增强,加入十二烷基硫酸钠后荧光强度减弱。建立了荧光光度法测定十二烷基苯磺酸钠的新方法。 相似文献