LC-MS分析饲料中的己烯雌酚

己烯雌酚(diethylstilbestrol)是一种非甾体激素,曾被广泛用于促进动物生长,增加瘦肉率,提高饲料转化率。从而其药物原形或代谢产物不可避免地残留于食品动物的器官、组织或通过排泄物进入外界环境成为环境雌激素,通过食物链进入消费者体内或通过人与污染的外界环境的接触而在体内发挥雌激素的效果而影响消费者健康。2002年农业部第176号公告和第193号公告中规定禁止使用己烯雌酚,但由于己烯雌酚作为畜禽增重剂效果好、经济回报高,仍有一些不法饲养业主受利益驱使而违法使用。本实验建立了高效液相色谱-质谱联用法(LC-MS)测定饲料中己烯雌酚,旨在为政府从源头上监控饲料中己烯雌酚的违法使用提供准确可靠的检测方法。     

高效液相色谱-串联质谱法同时测定动物肝脏中玉米赤霉醇及其类似物残留量

采用高效液相色谱-串联质谱方法(HPLC-MS/MS)同时测定了动物肝脏中6种玉米赤霉醇及其类似物的残留量。酶解后的样品采用乙醚提取,经液-液分配(LLP)和HLB固相萃取(SPE)柱净化后,采用HPLC-MS/MS电喷雾电离(ESI-),多反应监测(MRM)模式检测,基质匹配外标曲线定量。按照欧盟法规2002/657/EC的要求进行验证。在添加浓度1～4μg/kg范围内,方法回收率在70%～90%之间;相对标准偏差小于20%。6种玉米赤霉醇及其类似物的判断限(CCα)在0.17～0.31μg/kg之间,检测能力(CCβ)在0.26～0.42μg/kg之间。本方法对其它动物源食品中玉米赤霉醇及其类似物残留的检测具有适用性。     

高效液相色谱–串联质谱内标法测定动物源性食品中玉米赤霉烯酮及其代谢产物

建立高效液相色谱–串联质谱法(HPLC–MS/MS)测定动物源性食品中玉米赤霉烯酮及其5种代谢产物(α-玉米赤霉烯醇、β-玉米赤霉烯醇、α-玉米赤霉醇、β-玉米赤霉醇、玉米赤霉酮)残留量。在样品中加入4种同位素内标(13C18–玉米赤霉烯酮,D7–α-玉米赤霉烯醇,D7–β-玉米赤霉烯醇,D5–α-玉米赤霉醇)后,经β-葡萄糖苷酶/硫酸酯酶酶解,用叔丁基甲基醚萃取,取上清液氮吹至近干后用三氯甲烷复溶,再用氢氧化钠溶液反向萃取,以HLB固相萃取柱净化后,用HPLC–MS/MS检测。结果表明,玉米赤霉烯酮及其代谢产物在1.0~100.0μg/L范围内线性关系良好,相关系数均大于0.996,方法的检出限为0.04~0.13μg/kg,定量限为0.11~0.43μg/kg。在1.0、4.0、10.0μg/kg三种加标浓度水平下,回收率为77.7%~105.5%,测定结果的相对标准偏差为4.8%~9.8%(n=6)。该方法准确、可靠,灵敏度高,适用于动物源性食品中玉米赤霉烯酮及其代谢产物的定量分析。     

复合免疫亲和柱净化-液相色谱-串联质谱法测定动物源食品中6种黄曲霉毒素和6种玉米赤霉醇类真菌毒素残留量

建立了动物源食品(猪肉、鱼肉、猪肝)中6种黄曲霉毒素(AFB1、AFB2、AFG1、AFG2、AFM1和 AFM2)和6种玉米赤霉醇类真菌毒素(α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮和玉米赤霉烯酮)残留量的复合免疫亲和柱净化-高效液相色谱-串联质谱(HPLC-MS/ MS)检测方法。样品经β-葡萄糖苷酸/硫酸酯复合酶酶解后,用甲醇-乙腈(20∶80, V/ V)提取,提取液经玻璃纤维滤纸过滤,滤液用PBS 溶液稀释,复合免疫亲和柱富集和净化后,采用 HPLC-MS/ MS 法分析。12种目标分析物中 AFB2和 AFG2的线性范围为0.03~6.0μg/ L,其余目标分析物的线性范围为0.05~20μg/ L,线性相关系数均大于0.999,检出限在0.01~0.03μg/ kg 范围内,定量限在0.04~0.09μg/ kg 范围内。分别以0.5,1.0和5.0μg/ kg 添加浓度水平进行方法学验证,平均回收率为73.6%~98.4%,相对标准偏差(RSD)为1.9%~11.2%。本方法简便、灵敏,能够满足动物源食品中痕量黄曲霉毒素和玉米赤霉醇类真菌毒素残留的测定要求。     

免疫亲和柱净化-高效液相色谱法同时检测鸡蛋中6种玉米赤霉醇类化合物残留量

建立了鸡蛋中6种玉米赤霉醇类化合物(α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮和玉米赤霉烯酮)残留量的免疫亲和柱净化-高效液相色谱检测方法。样品酶解后用叔丁基甲醚提取、氢氧化钠反萃取,经免疫亲和柱富集和净化后,采用高效液相色谱-紫外检测器进行测定。色谱柱: Agilent Eclipse XDB-C18(150 mm×4.6 mm, 3.5 μm);流动相: 甲醇-乙腈-水(50:15:35, v/v/v);流速: 1.0 mL/min;检测波长: 270 nm。结果表明,6种目标物在0.01～0.2 mg/L范围内线性关系良好,相关系数(r)≥0.9998,检出限(LOD,S/N≥3)为1.0 μg/kg,平均回收率为73.2%～95.7%,相对标准偏差小于8%。该方法灵敏度高、重现性好,适用于鸡蛋样品中痕量玉米赤霉醇类药物残留的测定。     

免疫亲和固相萃取-超高效合相色谱-串联质谱法同时测定牛奶中6种玉米赤霉醇类化合物残留

建立了免疫亲和固相萃取（IAC-SPE）-超高效合相色谱-串联质谱（UPC²-MS/MS）同时测定牛奶中α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉烯酮和玉米赤霉酮残留的分析方法。样品用去离子水稀释,经IAC-SPE富集净化后,采用Waters ACQUITY UPC² Torus 2-PIC色谱柱（50 mm×3.0 mm,1.7 μm）分离,以超临界CO₂和0.1%（v/v）甲酸甲醇溶液为流动相,经梯度洗脱后在ESI^-模式下检测。经过稀释离心的牛奶样品采用免疫亲和柱净化后没有明显的基质效应,6种目标化合物在1~200 ng/mL范围内线性关系良好,相关系数（r²）≥0.9957;6种目标化合物在3个加标水平下的平均回收率为75.9%~106.5%,日内和日间精密度均≤11.4%。该法专属性好,操作简便,有机溶剂使用量小,与已有的样品测定方法比较更绿色环保,可用于牛奶中α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮和玉米赤霉烯酮的残留检测。     

快速高分离液相色谱-串联质谱法测定植物组织中雌激素玉米赤霉醇类化合物

应用快速高分离液相色谱-串联质谱仪(RRLC-MS/MS),建立了植物组织中玉米赤霉醇类化合物(α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮、玉米赤霉烯酮)的检测方法.植物组织样品采用乙腈提取,碱性水溶液反萃取,经混合阴离子(MAX)固相萃取柱进行净化和富集后,用RRLC-MS/MS检测,多反应监测(MRM)模式下进行定性与定量分析.结果表明:玉米赤霉醇类化合物在0～20 μg/kg的线性范围内均具有良好的线性关系,方法检出限(LOD)为0.5 μg/kg,定量限(LOQ)为1.0 μg/kg,6种玉米赤霉醇类化合物的平均回收率为75.8%～105.4%,相对标准偏差为2.4%～12.1%.本方法可用于植物组织中玉米赤霉醇类化合物含量的测定.     

液相色谱-串联质谱法测定牛尿中多种非类固醇同化激素残留量

玉米赤霉醇(Zearalanol,ZER)和二苯乙烯类激素(Stilbenes)属人工合成非类固醇同化性激素,为农业部公布《食品动物禁用的兽药及其他化合物清单》中禁用兽药[1].     

固相萃取-超高效液相色谱-串联质谱法检测粮食及其制品中的玉米赤霉烯酮类真菌毒素

建立了粮食及其制品中6种玉米赤霉烯酮类物质(α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇、β-玉米赤霉烯醇、玉米赤霉酮和玉米赤霉烯酮)的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。样品用84%(体积分数)乙腈水溶液提取,通过ENVI-Carb石墨化炭黑(GCB)固相萃取柱进行富集净化,用6 mL二氯甲烷-甲醇(7:3, v/v)溶液洗脱,采用UPLC-MS/MS进行测定。在ACQUITY UPLCTM BEH C18反相柱上分离,梯度洗脱,流动相为水和乙腈;质谱采集模式为电喷雾负离子多反应监测模式。以α-玉米赤霉烯酮-d4为内标,6种目标物的线性范围为0.1～50 μg/L,相关系数(R2)大于0.99,检出限为0.1～0.2 μg/kg, 3个不同水平的加标平均回收率为79.9%～104.0%,相对标准偏差不大于10%。应用该方法对北京市的粮食及相关产品进行了分析,结果发现玉米赤霉烯酮的检出率最高,含量为0.42～220.7 μg/kg;此外还检出了α-和β-玉米赤霉烯醇。该方法具有操作简单、灵敏度高、重现性好等特点,符合食品样品中痕量污染物的检测要求。     

玉米赤霉醇人工抗原合成及其多克隆抗体的制备

对玉米赤霉醇(ZER)16-OH进行改造,设计合成了模拟玉米赤霉醇特征结构的半抗原———ZER-16-羧丙基丁醚。用混合酸酐法将其与载体蛋白BSA连接,用于免疫新西兰大白兔,制备的抗血清效价达1∶102 400。基于活性酯法合成的包被抗原建立了CI-ELISA检测方法,该法对玉米赤霉醇检测的线性范围为18.39~1744.70 ng/mL,检出限为8.61 ng/mL。     

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%.     

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.     

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.     

Selective syntheses of 2H-1,3-oxazines and 1H-pyrrol-3(2H)-ones via temperature-dependent Rh(II)-carbenoid-mediated 2H-azirine-ring expansion

The Rh(II)-catalyzed reaction of 2-carbonyl-substituted 2H-azirines with ethyl 2-cyano-2-diazoacetate or 2-diazo-3,3,3-trifluoropropionate provides an easy access to 2H-1,3-oxazines and 1H-pyrrol-3(2H)-ones. These compounds can be selectively prepared from the same starting material using temperature as the only varied parameter. The 2-azabuta-1,3-diene intermediate, a common precursor for both heterocyclic products, isomerizes into 2H-1,3-oxazine under kinetic control, while 1H-pyrrol-3(2H)-one is the sole product of the reaction at elevated temperatures. According to DFT-calculations a one-atom oxazine ring contraction involving ring-opening to a 2-azabuta-1,3-diene intermediate, followed by a 1,5- and 1,2-prototropic shift leads to the consecutive formation of imidoylketene and azomethine ylide, which then further undergo cyclization to the pyrrole derivative.     

Copper catalyzed/mediated synthetic methodology for ebselen and related isoselenazolones

Scope of the copper catalyzed/mediated selenium-nitrogen coupling reaction has been studied for the synthesis of isoselenazolones. It is noticed that the 2-chloro, 2-bromo-, and 2-iodo-aryl amides substrates can be exploited in the selenium-nitrogen coupling reaction by employing 25-100 mol % of CuI/1,10-phenanthroline (L) and potassium carbonate as a base in DMF. Furthermore, electron rich 2-chloro-arylamides also underwent selenium-nitrogen coupling reaction to give biologically important selenium-nitrogen heterocycles. Also, copper-catalyzed selenium-nitrogen coupling reaction has been meticulously applied for the synthesis of diaryl diselenides having methoxy, amine, and amide functionality from respective aryl iodides in the presence of stoichiometric amount of succinimide as an external Se-N coupling partner.     

Knoevenagel condensation catalyzed by novel Nmm-based ionic liquids in water

A series of novel N-methyl morpholine (Nmm) based ionic liquids with 1,2-propanediol group were synthesized and used as catalysts for Knoevenagel condensation at room temperature in water. Under the effect of the catalyst, various aldehydes or aliphatic ketones could react with a wide range of activated methylene compounds well, including malononitrile, alkyl cyanoacetate, cyanoacetamide, β-diketone, barbituric acid, 2-arylacetonitrile and thiazolidinedione. Furthermore, most of the products could be separated just by filtrating and washing with water. Additionally, the catalyst is recyclable and applicable for the large-scale synthesis.     

Construction of polyheterocyclic spirotetrahydrothiophene derivatives via sulfa-Michael/aldol cascade reaction

A series of polyheterocyclic spirotetrahydrothiophene derivatives were obtained in moderate to excellent yields via a catalyst-free sulfa-Michael/aldol cascade reaction of chalcones 1 and commercially available 1,4-dithiane-2,5-diol 2 under mild conditions. We also present the first asymmetric sulfa-Michael/aldol cascade reaction of chalcones 1 and commercially available 1,4-dithiane-2,5-diol 2 with moderate to good enantioselectivities catalyzed by readily available chiral phase-transfer catalysts (PTCs).     

Suzuki-Miyaura reactions of the soluble guanylate cyclase inhibitor NS2028: a non-product specific route to C-8 substituted analogues

Both soluble guanylate cyclase (sGC) inhibitors ODQ 1 and NS2028 2 are synthesized via improved protocols. In the former case treating 3,4-dihydroquinoxalin-2(1H)-one oxime 8, which can be prepared in two steps from 1,2-benzenediamine, with 1,1′-carbonyldiimidazole (CDI) gives the dihydro-ODQ 10 that in the presence of KMnO₄ oxidises to give ODQ 1 in an overall yield of 46% starting from 1,2-benzenediamine. In the latter case, the synthesis affords NS2028 2 from 2-amino-4-bromophenol 3 in three steps with an overall yield of 85% and avoids the need for chromatography. Furthermore, Suzuki-Miyaura reaction conditions are described that enable the preparation of 8-aryl and 8-heteroaryl derivatives of NS2028 directly from NS2028 2. Finally, demethylation of the 8-(methoxyphenyl) substituted analogues afforded the 8-(hydroxyphenyl) derivatives 40-42. All new products are fully characterised.