免疫法制备液相色谱生物样品的研究——血中沙丁胺醇的分析

 用动物免疫法制备了免疫亲和柱纯化水溶性的沙丁胺醇血浆样品。琥珀酸酐交联沙丁胺醇和牛血清白蛋白获得抗原免疫家兔抗沙丁胺醇抗体——免疫球蛋白。琼脂糖Ｓｅｐｈａｒｏｓｅ４Ｂ与抗体交联制成免疫球蛋白亲和柱。对高效液相色谱法测定中的一般提取方法和固相小柱提取法作了比较，后者具有内源性杂质干扰少的优点，是生物样品预处理的一种有效的方法。     

莱克多巴胺、沙丁胺醇与克伦特罗三合一免疫亲和柱的制备及应用

将莱克多巴胺抗体和克伦特罗抗体偶联到溴化氰活化的琼脂糖凝胶上,制成免疫亲和柱,对其柱性能进行测试,并优化了使用条件,结合高效液相色谱-串联质谱对实际饲料样品进行检测。结果表明:该免疫亲和柱对莱克多巴胺、沙丁胺醇、克伦特罗的柱容量依次为49,27,24 ng/mL胶,保质期为5个月,最佳洗脱液为甲醇,对3种物质的回收率为78.9%~96.5%,RSD均小于1.5%,检出限为0.1~1 ng/g。该法可同时检测饲料中3种瘦肉精的含量。     

一种孔雀石绿免疫亲和柱及其制备方法和用途

<正>申请公布号:CN104174185A申请公布日:2014.12.03申请人:北京美正生物科技有限公司摘要本发明涉及一种孔雀石绿免疫亲和柱制备方法及其用途。该免疫亲和纯化柱利用蛋白G偶联到琼脂糖凝胶上载体,然后用抗孔雀石绿的抗体与琼脂糖上的蛋白G偶联。再利用交联剂对结合孔雀石绿抗体的蛋白G–琼脂糖凝胶载体进行交联。用交联后的载体制备免疫亲和柱。该纯化柱     

尿样中克仑特罗免疫亲和色谱-气/质联用检测方法的研究

以单抗为配基制备克仑特罗免疫亲和柱,建立了简单、快速、特异的样品净化技术,柱容量达到100 ng。偶联抗体量与柱容量呈线性关系,线性方程为:y=236.36x 6.6214,R2=0.9461。测定尿样中的克仑特罗,不需前处理直接用亲和柱提纯后以气质联用(GC/MS)方法检测。空白猪尿添加浓度为205、01、00 ng时,回收率分别为85%±1.3,85.9%±1.3,81.5%±2.7。同时以NY/XQ421-2003标准中净化方法为对照,回收率分别为113%±5.6,94.5%±10,78.6%±1.7,偶联非特异抗体的对照组亲和柱对样品保留值为7.03%。数据显示:免疫亲和柱作为提纯方法,与传统固相萃取提纯方法相比平行性好,提纯后样本杂质少。因此免疫亲和柱是净化样品的一个有效方法。     

饲料中伏马毒素的检测方法

摘要本发明一种饲料中伏马毒素的检测方法，免疫亲和柱一荧光仪检测法的原理为：将饲料样品与提取液混合、均匀、过滤，然后将滤液通过键合有伏马毒素特殊抗体的分离柱。用淋洗液通过分离柱，将伏马毒素从抗体上分离下来。最后，将淋洗液注入荧光计进行测定。     

粮食中呕吐毒素含量的免疫亲和柱净化超高效液相色谱法快速测定

建立了免疫亲和柱净化/超高效液相色谱法快速测定粮食中呕吐毒素(DON)的检测方法.样品经纯水提取后,用免疫亲和柱净化、浓缩,Waters Acquity UPLC BEH C18(2.1 mm×50 mm,1.7 μm)色谱柱分离,以甲醇-水(10:90)为流动相,流速为0.1 mL/min,进样量1μL,用Acqui...     

免标型沙丁胺醇免疫电化学传感器

构建了以茜素为探针,二氧化钛掺杂乙炔黑和壳聚糖复合材料为信号放大平台的新型免标型沙丁胺醇免疫传感器。于修饰电极的表面电沉积金纳米粒子,实现沙丁胺醇抗体的固定并进一步增大电流响应。采用扫描电子显微镜(SEM)对材料进行表征,循环伏安法(CV)和电化学阻抗谱图(EIS)对修饰电极的构建过程及传感器的性能进行电化学表征。采用差分脉冲伏安法(DPV)检测茜素在修饰电极表面的峰电流值,不同浓度的沙丁胺醇(SAL)抗原与抗体发生免疫亲和反应后,该峰电流值随着沙丁胺醇浓度的增大而减小,并呈线性关系,据此建立峰电流值与沙丁胺醇浓度的关系并实现了对沙丁胺醇的定量检测。探究了缓冲溶液p H值、孵化温度和孵化时间对免疫传感器的影响。在最佳条件下,该免疫传感器对沙丁胺醇的线性响应范围为1.0~100μg/L,检出限(LOD)为0.67μg/L(3σ/k)。该免疫传感器具有良好的重现性、选择性和稳定性,已成功用于猪饲料和猪肉样品中沙丁胺醇的检测,加标回收率分别为100.2%~102.4%(相对标准偏差为1.9%~4.7%)和97.5%~103.3%(相对标准偏差为2.1%~3.5%)。     

单克隆抗体亲和柱分离荧光法测定花生和玉米中黄曲霉毒素B1的研究

本文建立了用单克隆抗体亲和柱分离荧光法测定花生和玉米中黄曲霉毒Ｂ１的方法，用６０％甲醇提取样品中的ＡＦＴ，经氢氧化铁沉淀除去大部分杂质，通过亲和柱将ＡＴＦＢ１进一步与杂质分离，甲醇洗脱后加溴处理，测定荧光强度，与标准比较定量。     

猪肉中β-受体激动剂AlphaLISA检测方法的建立

建立了快速检测猪肉中克伦特罗、沙丁胺醇、溴布特罗和特布他林4种β-受体激动剂的光激化学发光纳米均相时间分辨荧光免疫(Alpha LISA)分析方法。样品经乙酸铵和β-葡萄糖醛酸酶/硫酸酯酶酶解提取过柱,浓缩至干后,用缓冲溶液定容。样品与生物素化抗原、抗体、供体微珠和受体微珠进行酶联免疫,Alpha LISA进行检测,基质标准曲线定量。结果表明:沙丁胺醇、克伦特罗、溴布特罗和特布他林在0.1~500ng/m L范围内呈良好线性,相关系数均大于0.98,沙丁胺醇与克伦特罗、溴布特罗和特布他林的交叉反应率分别为143.3%,179.2%和95.6%,检出限为0.03~0.08 ng/m L。在0.5,10,20μg/kg 3个添加水平下,4种化合物的平均回收率为82.5%~115.9%,相对标准偏差(RSD)均小于12.0%。该方法操作简单,快速准确,适用于猪肉中β-受体激动剂的筛选与检测。     

免疫亲和柱净化、在线电化学衍生化高效液相色谱法检测花生中的黄曲霉毒素

 采用免疫亲和柱净化、在线电化学衍生化高效液相色谱法测定了花生中黄曲霉毒素（AFT）B 1,B2,G1和G2。以体积分数为80%的甲醇提取样品中的AFT,经免疫亲和柱净化洗脱 后,以Kobra Cell装置在线衍生,反相HPLC分离定量。4种毒素的分离在13 min内完成,检 出限均达到0.1 μg/kg。5次测定花生样品的RSD值为9.2%～15%;样品添加标样0.5 ～9.0 μg/kg,回收率为74.8%～97.3%。     

二氧化碳和丙烯直接合成甲基丙烯酸的NiPMo/TiO2催化剂的研究

用溶胶-凝胶法以磷钼酸(MPA)的镍盐溶液水解钛酸四丁酯制备了NiPMo/TiO2催化剂.使用ICP、 XRD、 TG-DTA、 IR、 TPD-MS和微反应技术研究了催化剂的化学组成、热稳定性、化学吸附性质和催化反应性能.杂多钼酸盐与TiO2通过O2-在TiO2表面发生了键合.在623 K下,杂多阴离子仍保持原有的Keggin结构.CO2在Lewis酸位Ni(Ⅱ)和Lewis碱位Ni-O-Mo的桥氧协同作用下生成CO2卧式吸附态Ni(Ⅱ)←O-(CO)←(O--Ni).丙烯有多种吸附态在催化剂上吸附.在563 K、 1 MPa和空速1500 h-1的反应条件下,丙烯的摩尔转化率为3.2%,产物MAA选择性为95%.     

Synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives

Different approaches for the synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives from simple amino acids have been investigated. A library of 33 precursors for the preparation of N-hydroxy pyrazinones was obtained in moderate to good yields.     

Toward new camptothecins. Part 6: Synthesis of crucial ketones and their use in Friedländer reaction

In the context of the preparation of camptothecin and luotonin A analogs, the synthesis of some key keto-precursors and their use in Friedländer condensation are described. This paper also focuses on the stability of these keto intermediates and emphasizes the major differences between indolizinones and pyrroloquinazolinones series. Noteworthy is also the report of some original structures isolated as by-products of some experiments.     

CoFe2O4自形成磁性液体场致结构化对磁化的影响

The Langevin paramagnetic theory can’t describe the relation between magnetization of ferrofluids and applied magnetic field. The structuralization of ferrofluids, which is considered the main influence factor of the magnetization, is regarded. The part of magnetization works is deposited when the structure is forming. This action influences the magnetization of ferrofluids directly or indirectly. On the base of the “compressing” model, the Langevin function that usually describes the magnetization of ferrofluid is modified, and a well-fitted curve is obtained. An equation of the relation between the equivalent volume fraction after being “compressed” and the intensity of magnetic field is discovered, which approximately describes the process of magnetization. The relation between the approximate initial susceptibility and the volume fraction can be obtained from modified formula.     

Highly regioselective Buchwald–Hartwig amination at C-2 of 2,4-dichloropyridine enabling a novel approach to 2,4-bisanilinopyridine (BAPyd) libraries

The highly regioselective Buchwald–Hartwig amination at C-2 of the cheap and readily accessible reagent, 2,4-dichloropyridine with a range of anilines and heterocyclic amines is described. This new methodology is robust and provides a facile access to 4-chloro-N-phenylpyridin-2-amines on 0.25 mol scale. These intermediates undergo a further Buchwald–Hartwig amination at higher temperature to enable rapid exploration of the chemical space at C-4 and to provide a library of 2,4-bisaminopyridines.     

KMnO4-mediated oxidative CN bond cleavage of tertiary amines: Synthesis of amides and sulfonamides

KMnO₄-mediated oxidative CN bond cleavage of tertiary amines producing secondary amine was introduced, which was trapped by electrophiles (acyl chloride and sulfonyl chloride) to form amides and sulfonamides. The reaction could take place at mild condition, tolerating a wide range of function groups and affording products in moderate to excellent yields.     

Chemistry of heterocyclic compounds at the A. E. Favorsky Irkutsk Institute of Chemistry, Siberian Branch of the Russian Academy of Sciences, over 50 years (Review)

The review contains a concise historical account and information on the most significant researches undertaken by the staff at the A. E. Favorsky Irkutsk Institute of Chemistry, Siberian Branch of the Russian Academy of Sciences on the Chemistry of Heterocyclic Compounds. Dedicated to Academician of the Russian Academy of Sciences B. A. Trofimov on his 70th jubilee. Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 10, pp. 1443–1502, October, 2008.     

A general and convenient synthesis of 4-(tosylmethyl)semicarbazones and their use in amidoalkylation of hydrogen,heteroatom, and carbon nucleophiles

A general synthesis of previously unknown semicarbazone-based α-amidoalkylating reagents, 4-(tosylmethyl)semicarbazones, has been developed. The synthesis involved three-component condensation of semicarbazones of aliphatic or aromatic aldehydes with the same or other aldehydes and p-toluenesulfinic acid. The scope and limitations of this reaction were investigated. The compounds obtained were demonstrated to be an efficient α-(4-semicarbazono)alkylating agents. They were reacted with H- (sodium borohydride), O- (sodium methylate), S- (sodium phenylthiolate), N- (pyrrolidine, sodium succinimide), P- (trialkyl phosphites), and C-nucleophiles (sodium diethyl malonate) to give the corresponding products of the tosyl group substitution, 4-substituted semicarbazones, including analogues of nitrofurazone. Among the prepared compounds tested in vitro for antibacterial and antifungal activity, three nitrofuryl-containing semicarbazones exhibited high biological activities with minimum inhibitory concentration (MIC) values of 8–32 μg/mL.     

Sulfur-directed metal-free and regiospecific methyl C(sp3)–H imidation of thioanisoles

Regiospecific and direct imidation of the methyl C(sp³)–H bond of thioanisoles is realized under mild and metal-free conditions with N-fluorobis(benzenesulfonyl)imide as an oxidant and nitrogen source. Proposed mechanism suggests that thionium ion intermediates and a Pummerer-type reaction are involved. The imidation has advantages such as high step-economy, excellent functionality tolerance, and regiospecificity, giving structurally diverse imidation products.     

Synthesis of novel chiral bidentate hydroxyalkyl-N-heterocyclic carbene ligands and their application in palladium-catalyzed Mizoroki–Heck couplings and asymmetric addition of diethylzinc to benzaldehyde

A small library of new chiral bidentate hydroxyalkyl-imidazolium salts 1 is conveniently synthesized on multi-gram scale from inexpensive and commercially available chiral pool amino acids. The corresponding carbenes, generated by deprotonation of imidazolium salts 1, in combination with palladium(II) chloride were tested in the Mizoroki–Heck coupling reaction. The most significant results in terms of yields and reactivities were achieved with low catalyst loading. The catalytic activities of these imidazolium salts were also investigated in the asymmetric addition of diethylzinc to benzaldehyde. The use of MgO nanoparticles as an additive in conjunction with these ligands played a crucial role in increasing the efficiency of these reactions.