GMA-EDMA-TiO2扩张床介质合成的改进及其在复合干扰素分离纯化中的应用

通过合成条件的改进,得到了一种性能更好的扩张床介质GMA-EDMA-Lipo-TiO2(GELT),其平均密度为1.25g/ml,孔容为0.564ml/g,平均孔径为15.6nm,比表面积为127.58m2/g,渗磨圆球率为95%。研究表明GELT在氢氧化钠溶液是稳定的。同时讨论了甲苯致孔剂对孔结构的影响及二氧化钛添加量对湿真密度的影响;将GELT与二乙胺偶联得到阴离子树脂(GELT-DEAE),在静态和动态条件下,GELT-DEAE吸附量分别88mgBSA/ml、65mgBSA/ml。将GELT-DEAE应用于扩张床直接从发酵液中分离复合干扰素,一步活性收率为75%,后经SPSepharoseFF离子交换层析和CellufastGel01凝胶层析进一步纯化。最后得到的成品其比活为6.82×108IU/mg,与商品干复津(Infergen)的比活相当,纯度为95%。     

高效液相色谱-蒸发光散射检测法测定蛋黄卵磷脂的含量

建立了蛋黄磷脂中卵磷脂（即磷脂酰胆碱,PC)的高效液相色谱-蒸发光散射检测（HPLC-ELSD）的测定方法。以Nov a-Pak Silica 60A硅胶柱(3.9 mm i.d.×150 mm,4 μm)为分离柱,正己烷-异丙醇-3%冰醋酸水溶液(体积比为35∶65 ∶8)为流动相，等度洗脱,流速1.0 mL/min,柱温30 ℃。蒸发光散射检测器漂移管温度50 ℃,雾化气(空气)压力350 kPa。在上述条件下测得PC在0.16~1.61 g/L范围内线性关系良好(r2=0.9979),检测限为0.64 μg,方法的精密度为3.2%(n=5),回收率为98.2%~128.2%。将该方法用于实际样品的测定,获得了令人满意的结果。该方 法预处理简单,分析速度快,可用于蛋黄磷脂中卵磷脂的测定。     

离子交换色谱法分离纯化鸡卵黄免疫球蛋白

建立了高效、经济、大规模获得鸡卵黄免疫球蛋白(IgY)的生产方法。在对传统的水稀释法改良的基础上,结合聚乙二醇沉淀与离子交换色谱进行IgY的分离纯化。结果显示,用8倍无菌水稀释蛋黄液,用0.1 mol/L HCl调节pH为5.2,在4 ℃下静置8 h,于5000×g力离心可得上清粗IgY液,经测定回收率可达93.47%。然后用6%聚乙二醇沉淀后经DEAE-Toyopearl 650 M离子交换纯化,最佳的纯化条件: 0.05 mol/L磷酸盐缓冲液(PBS, pH 7)平衡上样,0.075 mol/L PBS(pH 7)洗脱。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析结果显示所得的IgY的纯度为95.02%,活性保持率高达73.77%。本研究弥补了传统分离方法不能同时达到高纯度和高回收率的缺点,且可用于大规模生产。     

磷脂的分离纯化及高效毛细管电泳分析

采用溶剂提取和柱色谱法分离纯化市售大豆粉末磷脂(卵磷脂含量14.05%),得到高纯度的卵磷脂产品(纯度92.80%)。重点建立了磷脂的胶束电动毛细管色谱(MECC)分离分析方法。以分离度和峰面积为优化指标,对表面活性剂及其浓度、电泳缓冲液pH、有机改性剂及其含量、缓冲液浓度、温度等条件进行优化,确定了最优化电泳条件:电泳缓冲液为35 mmol/L脱氧胆酸钠-1 mmol/L 硼砂缓冲液/正丙醇(体积比为57∶43)(pH 8.30),柱温44 ℃,操作电压25 kV,检测波长200 nm;内加法定性磷脂组分;外标法定量卵磷脂。结果表明,MECC法能有效分离5种磷脂组分;0.1～1 g/L的质量浓度范围内卵磷脂的线性关系良好(r=0.9990),平均回收率为98.0%,日内、日间精密度分别为1.36%和3.27%,定性结果与薄层色谱法、红外光谱法的定性结果相符。     

用流动注射分析法测定强碱性阴离子交换树脂的交换容量

建立了快速测定微量氯离子的FIA分光光度法和快速测定强碱性阴离子交换树脂交换容量的FIA法。本方法将离子交换微型柱导入树脂交换容量的测定中 ,不但获得了很好的精度 (RSD <0 .9% ) ,而且使测定效率提高了近 30倍。优选实验得到的最佳测定条件为再生液流速 0 .5mL/min ,再生剂用量 :333mL/ g,再生剂浓度 :3.0 % (W /V) ;样品液流速 :1 .5mL/min ;树脂交换容量测定温度 :2 5℃± 5℃。     

大孔树脂分离纯化宁夏枸杞总黄酮的研究

考察了NKA-9、S-8、XDA-1、AB-8、HPD-100、HPD-600 6种大孔吸附树脂对宁夏枸杞总黄酮的吸附和解吸性能,筛选出XDA-1树脂的效果最佳;考察了pH值、样液浓度、流速等对XDA-1树脂静态吸附效果的影响;并进行了动态吸附试验,确定出XDA-1树脂动态吸附枸杞总黄酮的最佳条件:样品液浓度为0.25mg/mL,控制流速为0.5mL/min,样品液pH 5;最佳洗脱条件:洗脱液为80%的乙醇水溶液,用量为35mL (5倍柱床体积).在此条件下,枸杞总黄酮含量从27.2%提高到79.8%,回收率为87.4%,表明XDA-1树脂可用于宁夏枸杞总黄酮的分离纯化.     

大孔阳离子交换树脂分离纯化Zwittermicin A

以抑菌圈为指标,通过离子交换法对双效菌素Zwittermicin A进行了分离纯化,并研究其抑菌活性。首先通过静态法比较D151、D113、D001-cc 3种大孔阳离子交换树脂吸附效果,结果表明,D151对ZwittermicinA分离纯化效果最好。利用单因素轮换法,确定了ZwittermicinA的最佳纯化工艺为:上柱样品的最佳pH=7;第1次冲洗液NH4H2PO4的最适浓度为5mmol/L;第2次冲洗液CH3COONH4的最适浓度为30mmol/L;1mol/L氨水洗脱液的最少用量为12.7mL/cm3树脂;Zwittermicin A的总收率达90%以上。利用红外光谱仪对D151树脂的吸附前后进行了分析。     

离子交换树脂层析法分离木糖醇结晶母液

研究了001系列离子交换树脂抗衡阳离子型式、树脂交联度、树脂粒径等因素对木糖醇结晶母液分离为糖组分和糖醇组分的影响.筛选出了具有较好分离效果的 001×8(Ca)(0.15mm～0.30mm)树脂.对柱温、柱长径比、进料量、进料浓度、洗脱速度、洗脱液pH等操作因素进行了分离条件优化.在优化条件下,对未糖醇结晶母液进行分离,得到去除糖组分的糖醇组分回收率为92.3%,完成了木糖醇结晶母液的阶段性分离纯化任务.     

两种氢氧化铁胶体纯化新方法

针对在Fe(OH)3胶体纯化过程中,难以在短时间内获得一定纯度的Fe(OH)3胶体的问题,探究了两种Fe(OH)3胶体纯化的新方法,冷热交替渗析法和混合离子交换树脂法,并且对两种方法的最佳纯化条件及其各自的优劣进行探索。实验结果表明:使用冷热交替渗析法,60°C热水浴渗析35 min后冷水浴渗析15 min,胶体的电导率可达560μS?cm-1;而混合离子交换树脂法,当阴阳离子交换树脂的总质量为10 g,且质量比为1:1,粗胶体的体积为45 m L时,胶体的电导率可降至42μS?cm-1。两种方法各有优劣,在教学实践过程中可根据实际情况灵活选择。     

盐析色层

色层分离中应用离子交换树脂作为固定相以分离或纯化各种物质的方法,简便而有效,且能分离性质非常相近的物质。近来已成为一个化学的单元操作而获得广泛的应用。离子交换树脂在色层分离中作为固定相而加以应用的可概括为三:(1)离子交换色层(ion exchange chromatogra-phy),是以离子交换树脂为固定相,用电解质溶液(酸、碱或中性盐)为淋洗剂,分离二种或更多种的电解质混合物,如碱金属、希土元素、卤素离子及超铀元素等。(2)离子排斥的方法(ion exclusion),是以离子交换树脂为固定相,用水为淋洗剂,分离电解质与非电解质的混合物。这种方法广泛地用以纯化各种有机物质如葡萄糖、甘油等。(3)盐析色层(salting-out chromatography),是以离子交换树脂为固定相,不同浓度的盐溶液为淋洗剂,分离二种或多种的水溶性非电解质混合物。     

α-Amino acid derived enaminones and their application in the synthesis of N-protected methyl 5-substituted-4-hydroxypyrrole-3-carboxylates and other heterocycles

A new and simple synthesis of novel N-protected methyl 5-substituted-4-hydroxypyrrole-3-carboxylates, which exist in equilibrium with their 4-oxo tautomers, has been developed in two steps starting from N-protected α-amino acids. The key intermediates are enaminones, which can also be isolated, characterized, and used for the construction of other functionalized heterocycles, before they spontaneously decompose to pyrrole products. 4-Hydroxypyrroles are prone to partial aerial oxidation but can be efficiently alkylated or reduced to stable polysubstituted pyrrolidine derivatives.     

Specific intramolecular aromatic CH insertion of diazosulfonamides

The chemoselectivity in the intramolecular CH insertion of various diazosulfonamides has been experimentally studied. The results reveal that the aliphatic 1,4-, 1,5-, or 1,6-C(sp³)?H insertions of diazosulfonamides are not accessible, while the aromatic 1,5-C(sp²)?H insertion can be realized specifically by adjusting the diazo-adjacent group. In addition, the general chemoselectivities in the intramolecular CH insertions of diazosulfonyl compounds are summarized. Generally, diazosulfones undergo both aromatic 1,5-C(sp²)?H and aliphatic 1,5- and 1,6-C(sp³)?H insertions, while diazosulfonates undergo aliphatic 1,5- and 1,6-C(sp³)?H insertions. However, diazosulfonamides only undergo aromatic 1,5-C(sp²)?H insertion.     

N-Heterocyclic carbene-palladacyclic complexes: synthesis,characterization and their applications in the C-N coupling and α-arylation of ketones using aryl chlorides

N-Heterocyclic carbene-palladacyclic complexes 3 were successfully achieved in a one-pot procedure under mild conditions. The structure of 3a was unambiguously confirmed by X-ray single crystal diffraction and it was an active catalyst in the Buchwald-Hartwig amination and α-arylation of ketones even at very low catalyst loadings (0.01?mol%).     

Iodine-mediated oxidative Pictet-Spengler reaction using terminal alkyne as the 2-oxoaldehyde surrogate for the synthesis of 1-aroyl-β-carbolines and fused-nitrogen heterocycles

An efficient iodine-mediated oxidative Pictet-Spengler reaction in dimethyl sulphoxide (DMSO) using terminal alkynes as the 2-oxoaldehyde surrogate for the synthesis of aryl (9H-pyrido[3,4-b]indol-1-yl)methanones is described. The scope of the protocol includes the total synthesis of Fascaplysin, Eudistomins Y₁ and Y₂. The methodology is extended for preparing pyrrolo[1,2-a]-quinoxaline and indolo[1,5-a]quinoxaline derivatives. The utility of 1-aroyl-β-carbolines was demonstrated by performing palladium-catalyzed β-carboline directed ortho-C(sp2)-H functionalization of the phenyl ring with thiomethyl (SMe) group using DMSO as source and for accessing 4-aryl-canthin-6-ones.     

Facile synthesis of bicyclic 1-arylpyrazol-5-ones

In this Letter, we described a facile method for constructing fused bicyclic 1-arylpyrazol-5-one ring system. We employed various methylene-containing carboxylic acids as the substrates and proved that the pyrazolone ring closure requires activated methylene group in intermediate II. Accordingly, a series of structurally diversified, fused bicyclic 1-arylpyrazol-5-ones was prepared in moderate to high yields using the requisite substrates.     

Synthesis of pyrrolo[1,2-a]pyrazines and pyrazino[1,2-a]indoles by Curtius reaction in Morita-Baylis-Hillman derivatives

Synthesis of substituted pyrrolo[1,2-a]pyrazines and pyrazino[1,2-a]indoles from the Morita-Baylis-Hillman derivatives of acrylates via saponification followed by Curtius reaction is described.     

正丁胺对激光染料荧光谱影响的研究

用正丁胺作为碳源,采用射频辉光放电制备碳膜,选用激光染料R6G和聚乙二醇混合液作为蒸气源,采用单源热蒸发,在蒸发室与染料同时沉积得到混合膜,用拉曼光谱和红外光谱分析了碳膜的结构和键合方式,分析表明:碳膜中存在胺基团和氢原子.混合膜的荧光谱测量结果表明,认为正丁胺对染料荧光谱的影响是因为胺基和氢原子的存在.     

Cu0-catalysed 1,3-dipolar cycloadditions of α-amino acid derived N,N-cyclic azomethine imines to ynones

A series of 20 CuAIAC reactions between eight 4-acylamino substituted pyrazolidine-3-one-1-azomethine imines and four terminal ynones were performed using Cu⁰ as catalyst. The corresponding fluorescent cycloadducts were obtained in very high yields upon simple workup. Thus, Cu-metal turned out to be a better catalyst than Cu^I in terms of yield and ease of isolation. Availability of azomethine imines, mild reaction conditions, and simple workup enable a “click” access to libraries of densely substituted 2,3-dihydro-1H,5H-pyrazolo[1,2-a]pyrazol-1-ones. Reactivity of differently substituted dipoles was evaluated experimentally and by quantum chemical methods (DFT).     

Synthesis and antioxidative/anti-inflammatory activity of novel fullerene–polyamine conjugates

(E)-4-(Fullerenopyrrolidin-1-yl)-3-methylbut-2-enoic acid and its corresponding succinimidyl ester, readily obtained through Prato-type modification of C₆₀, were used for the selective N-acylation of polyamines. The thus obtained conjugates were evaluated for their antioxidative and anti-inflammatory activity and their cytotoxicity was determined. Members of this family of compounds showed interesting anti-lipid peroxidation, anti-lipoxygenase and anti-inflammatory activity and comparable cytocompatibility to spermidine.