高效液相色谱法测定蛹虫草中腺苷和虫草素

在蛹虫草中腺苷和虫草素含量的反相高效液相色谱分析中,所用色谱柱为WatersNOVA-PAK C18(3.9 mm×300 mm,4μm),流动相为0.01 mol.L-1KH2PO4-K2HPO4缓冲溶液(pH 6.86)+1%(体积分数)四氢呋喃,流速为1.0 mL.min-1。紫外检测的波长为260 nm,峰面积与腺苷浓度和虫草素浓度之间的线性范围依次为1.27~50.5 mg.L-1和3.23~129.0 mg.L-1,其检出限依次为0.18 ng和0.25 ng。在腺苷及虫草素的浓度水平依次为5.05 mg.L-1和12.9 mg.L-1时测试方法的精密度,求算得RSD(n=7)值依次为0.094%和0.792%。用标准加入法试验方法的回收率,求得两者的平均回收率依次为103.2%和98.5%。     

蛹虫草及其培养残基中腺苷和虫草素含量的快速测定

建立了以氰基正相色谱柱反相条件下快速分析腺苷和虫草素的方法。以微波辅助提取蛹虫草及其培养残基样品,每次提取1.5 min,共提取2次。采用Eclipse XDB-CN色谱柱测定提取液中腺苷和虫草素的含量,以甲醇-水(7:93, v/v)为流动相进行等度洗脱,检测波长260 nm,并考察和分析了影响分离性能的流动相组成和pH值。结果腺苷和虫草素在4.5 min内实现完全分离且无基质干扰。腺苷和虫草素在线性范围内线性关系良好,线性相关系数r2分别为0.9998和0.9995,定量限(以10倍信噪比计)分别为0.21 mg/L(腺苷)和0.083 mg/L(虫草素)。该方法日内和日间精密度的相对标准偏差(RSD)小于2%;腺苷和虫草素的平均加标回收率为93.8%～102.9%, RSD值不大于3.62%(n=5)。本方法简便、快速、准确、成本低,可用于冬虫夏草、蛹虫草子实体、虫草培养残基及虫草制剂中腺苷和虫草素含量的快速测定。     

富硒蛹虫草的抗氧化活性研究

采用现代分离技术从富硒蛹虫草中提取硒多糖,然后以苯酚-硫酸法、电感耦合等离子体质谱（ICP—MS）法对硒多糖中多糖含量和硒含量进行测定;在Fenton体系的最佳实验条件下,研究了硒多糖对羟自由基的清除作用及硒多糖对支撑磷脂双层膜（Supported Bilayer Lipid Membranes,s—BLM）的保护作用。结果表明：硒多糖清除羟自由基的效果比与之相同浓度的无机硒、多糖都要明显。其中硒多糖对羟自由基的最高清除率可达38．02％,硒多糖具有很强的抗脂质过氧化、保护支撑磷脂双层膜的作用。     

蛹虫草中硒的赋存形态及蛋白硒分析

为研究蛹虫草中硒的赋存形态,并进一步探究蛹虫草硒蛋白组分中硒含量的分布,以蛹虫草子实体为研究对象,用DAN荧光法测定了硒含量,用连续提取法进行了蛋白硒分析。结果表明,蛹虫草最高硒含量为89.486mg·kg-1,有机硒占85.96%;蛹虫草中蛋白硒、多糖硒和核酸硒分别占有机硒的71.4%~77.1%、19.7%~32.3%和0.138%~2.727%;在硒蛋白组分中虫草盐溶性蛋白、水溶性蛋白、醇溶性蛋白和碱溶性蛋白结合硒分别占总蛋白硒的55.13%~56.80%、20.47%~24.60%、15.30%~16.49%和2.99%~3.35%。蛹虫草具有很强的富硒能力;在蛹虫草子实体中,有机硒主要与蛋白质、多糖、核酸等生物大分子结合,其中蛋白质结合硒是硒的主要赋存形态;在蛋白质组分中,又以盐溶性蛋白质结合硒量最多。     

富硒蛹虫草试样中硒的形态分析

本文采用连续浸提法研究了富硒蛹虫草中硒的赋存形态。结果表明,碱溶态>盐溶态>水溶态>醇溶态>残渣态。可见,碱浸取是获取植物蛋白的有效途径,在富硒蛹虫草试样中硒主要以硒蛋白的形式存在。     

SE-HPLC-ICP-MS联用技术在富硒蛹虫草硒蛋白形态分析中的应用研究

从富硒培养的虫草中提取可溶性蛋白质, 经过(NH4)2SO4分级盐析, 采用ICP-MS对不同盐析组分中蛋白及硒分布进行测定, 结果表明, 富硒蛹虫草的可溶性硒含量占总硒含量的69.3%以上, 在所得的各级蛋白组分中, 除A组分(0~30%饱和度的盐析)外, 其它各组分均存在含硒蛋白, 其硒含量高低顺序为B组分(30%~50%饱和度的盐析)>C组分(50%~75%饱和度的盐析)>D组分(75%~100%饱和度的盐析)>A组分. 采用SE-HPLC-ICP-MS联机技术对样品中的硒蛋白进行分离检测, 结果表明, 其分子量分布在114000和1000之间, 且含量各异. 进一步研究结果表明, 硒在虫草体内参与了蛋白的合成, 这对于以蛹虫草为载体将无机硒有机化具有重要意义.     

海红果红色素的理化性质

食用色素是食品添加剂的重要组成部分,应用广泛。许多合成色素有慢性毒性及致癌作用,天然色素的研究和开发日益引起人们的重视。海红果属蔷薇科属西府海棠(Malus-micro-malus Makino)类,为落叶乔木果树,在陕西、内蒙古、山西等地广泛种植。果实成熟时呈刺红色,     

HPLC-ELSD法测定蛹虫草中6种游离氨基酸的含量

建立了反相高效液相色谱-蒸发光散射检测(HPLC-ELSD)直接测定6种未衍生游离氨基酸(脯氨酸Pro、缬氨酸Val、蛋氨酸Met、异亮氨酸Ile、亮氨酸Leu、苯丙氨酸Phe)含量的分析方法。采用VenusilMP-C18色谱柱,以甲醇和水为流动相进行线性梯度洗脱,流速为0.8mL·min-1,漂移管温度40℃,氮气流速2.5L·min-1;优化了流动相的洗脱条件。结果表明,在优化实验条件下,6种氨基酸峰面积的对数值与氨基酸质量浓度的对数值呈良好线性,检出限为10.0~15.0mg·mL-1,方法精密度及稳定性良好。采用该法对微波法提取的蛹虫草子实体中的氨基酸进行测定,待检蛹虫草样本中不含缬氨酸(Val),其余5种氨基酸的总量为16.00mg/g,约占蛹虫草总质量的1.60%,其中脯氨酸(Pro)(占0.72%)和苯氨酸(Phe)(占0.56%)的含量较高。已检出的5种Pro、Met、Ile、Leu、Phe含量分别为7.20、0.88、0.69、1.65、5.58mg/g。     

成都市城市绿地土壤理化性质研究

对成都市绿地土壤的理化性质进行了研究,并与郊区的土壤作了比较.结果显示,该市绿地土壤的pH明显高于市郊,而有机质含量和总硫含量则低于市郊.     

La(NO3)3对蛹虫草活性物质含量的影响EI北大核心CSCD

蛹虫草是一种著名的药用和食用真菌,研究者们常在培养基中添加亚硒酸钠、Fe SO4等无机物来提高蛹虫草活性物质的含量。本文采用高效液相色谱法和紫外分光光度法测定了La(NO3)3处理后蛹虫草活性物质的含量,探究La(NO3)3对蛹虫草活性物质含量的影响。结果表明,在实验浓度范围内,10,50和200 mg·L-1这3个浓度对蛹虫草的活性物质含量影响较大。La(NO3)3浓度为10 mg·L-1时,对喷司他丁、虫草酸和多糖合成具有促进作用,其含量分别为空白组的6.2倍、2.4倍和1.3倍,La(NO3)3对蛹虫草的腺苷和N6-2-羟乙基腺苷合成具有抑制作用,其含量仅为空白组的24.29%和16.87%。La(NO3)3浓度为50 mg·L-1时,对虫草素合成具有显著促进作用,其含量为空白组的8.2倍,La(NO3)3对多糖合成具有抑制作用,其含量仅为空白组的73.14%。La(NO3)3浓度为200 mg·L-1时,对腺苷和N6-2-羟乙基腺苷合成具有促进作用,其含量分别为空白组的1.5倍和2.6倍。La(NO3)3对麦角甾醇和蛋白质的含量未产生显著影响(P>0.05)。     

Assembly of layer-by-layer films of superoxide dismutase and gold nanorods:A third generation biosensor for superoxide anion

Based on the layer-by-layer self-assembly of positively charged cetyltrimethylammonium bromide (CTAB) wrapped gold na- norods (AuNRs) and negatively charged superoxide dismutase (SOD) from their aqueous solutions on cysteine modified gold electrode (Cys/Au), a third generation electrochemical biosensor ((SOD/AuNRs)2/Cys/Au) for superoxide anion (O2·-) was developed. The two layers assembly of SOD/AuNRs can significantly enhance the direct electron transfer between SOD and the electrode. The functional enzym...     

A study on some macrocyclic bicopper(Ⅱ) complexes as the model compounds of superoxide dismutase

24-Membered macrocyclic bicopper(Ⅱ) complexes wth bridging ligand imidzao-late,SCN-,N3-and H2O respectively were synthesized by condensation of diacetylpyridine with 3-oxapentane-1,5-diamine.As the model compounds of superoxide dismutase (SOD),the relationship between their SOD activities and properties of bridging ligands and redox potentials of copper(Ⅱ) were studied.     

Detection of synergistic effect of superoxide dismutase and jujubosides on scavenging superoxide anion radical by capillary electrophoresis

A new capillary electrophoresis method was developed to study the synergistic effect of superoxide dismutase and jujuboside A or B on scavenging superoxide anion radical in serum matrix respectively, in which superoxide anion radical was generated from pyrogallol autoxidation. The electrophoresis conditions, and the factors affecting the productive rate of purpurogallin, such as pyrogallol autoxidation product and the activity of superoxide dismutase, were optimized. Under optimal conditions, the content of superoxide dismutase in Gibco newborn calf serum was 7.06 mg/L, RSD was 2.01% and the average recovery was 98.4%. The values of IC₅₀ for jujuboside A and B in the serum matrix were 157.67 and 31.60 mg/L respectively, and they both had synergy on scavenging superoxide anion radical with superoxide dismutase, but there was no the dose‐dependency on this synergy.     

Oxazolidine copper complexes: Synthesis,characterization and superoxide dismutase activity of copper(II) complexes with oxazolidine ligands derived from hydroxyquinoline carboxaldehyde

This work describes the synthesis of copper(II) complexes, their identification using spectroscopic and electrochemical methods, structural characterization with X‐ray diffraction and density functional theory calculations (DFT), as well as their catalytic activity mimicking that of superoxide dismutase. Structural analysis revealed the formation of complexes with cyclization of ligands L1 and L2 with the formation of a heterocycle, 1,3‐oxazolidine. The DFT calculations confirmed this trend by stabilizing with lower energy. In addition, evaluations of mimetic enzymatic activity of complexes C1, C2, C3 and C4 revealed promising IC₅₀ values compared to other results seen in the literature (IC₅₀ = 0.22, 0.328, 0.55 and 0.92). Also, calf thymus DNA interactive experiments using UV–visible spectroscopy were conducted in the presence of the copper(II) complexes.     

Microcalorimetric studies on the dismutation of superoxide anion catalyzed by superoxide dismutase

Ｓｕｐｅｒｏｘｉｄｅｄｉｓｍｕｔａｓｅ(ＳＯＤ,ＥＣ1.15.1.1),ｗｈｉｃｈｗａｓｆｏｕｎｄａｎｄｉｓｏｌａｔｅｄｆｒｏｍｂｏｖｉｎｅｅｒｙｔｈｒｏｃｙｔｅｓｂｙＭｃＣｏｒｄａｎｄＦｒｉｄｏｖｉｃｈｉｎ1969[1],ｉｓａｎｉｍｐｏｒｔａｎｔｍｅｍｂｅｒｏｆｔｈｅｆａｍｉｌｙｏｆｂｉｏｌｏｇｉｃａｌａｎｔｉｏｘｉｄａｎｔｓｔｒｅｓｓｅｎｚｙｍｅｓ.Ｔｈｉｓｅｎｚｙｍｅｈａｓｂｅｅｎｄｅｔｅｃｔｅｄｉｎａｗｉｄｅｒａｎｇｅｏｆｌｉｖｉｎｇｔｈｉｎｇｓａｎｄｈａｓｂｅｅｎｉｍｐｌｉｃａｔｅｄｉｎｔｈｅｉｎｔｅｒ…     

绿豆萌发过程中SOD的分离纯化及性质研究

制备不同萌发时期绿豆SOD(超氧化物歧化酶)粗酶液,筛选出酶比活力最高的时期,并运用热变性、硫酸铵分级、DEAE-Sepharose FF阴离子交换层析和Sephacryl S-100 HR分子筛层析分离纯化并作对比考察,提纯的SOD进行了理化性质研究。结果表明,绿豆种子萌发第3天时SOD的比活力最高,经纯化获得的SOD酶比活力为2256.4 U/mg,纯化倍数为3.9。绿豆中SOD为Cu/Zn SOD和Mn SOD,分子量为68KD,热稳定性高,在pH5-8条件下稳定,对高浓度SDS具有抗性。EDTA可以一定程度上抑制SOD活性,H2O2可以完全抑制SOD活性。     

Synthesis,characterization, and superoxide dismutase activity of two new copper(II) complexes of benzoylpyridine 4-phenylsemicarbazone

Two mononuclear complexes containing copper(II) and 2-benzoylpyridine 4-phenylsemicarbazone (BPS) and pseudohalides, [Cu(BPS)(N₃)] (1) and [Cu(BPS)(NCS)(H₂O)]NO₃ (2) have been synthesized and characterized by UV-Vis, Fast atom bombardment, electron paramagnetic resonance, and infrared spectroscopy. Crystal structures of these two complexes have been resolved by using single crystal X-ray studies. Complex 1 crystallizes in the triclinic lattice with space group P1 and is a distorted square planar geometry. Complex 2 also crystallizes in the triclinic lattice with space group P1 but is a distorted square pyramidal geometry with N₃O₂ chromophore. Both the complexes have g _∥ > g _┴ > 2.0023 and a G value less than 4, consistent with a d _{x ²? y ²} ground state. Superoxide dismutase activities have also been examined.     

铜锌超氧化物歧化酶的金属重组酶的活性部位与组氨酸的相互作用

应用核磁共振和电子自旋共振技术研究了铜锌超氧化物歧化酶（ＣｕＺｎＳＯＤ）的金属重组酶与水溶液中游离组氨酸（Ｈｉｓ）的相互作用。发现在水溶液中ＣｕＺｎＳＯＤ的金属重组酶活性中心的金属可与加入的Ｈｉｓ发生作用而被部分地诱导出来，与Ｈｉｓ形成络合物。     

Age-dependent dichotomous effect of superoxide dismutase Ala16Val polymorphism on oxidized LDL levels

We investigated the association between superoxide dismutase (SOD) Ala16Val polymorphism and the levels of oxidized LDL lipoprotein-C (ox-LDL-C) in two age-different Greek cohorts. Four hundred fifteen middle-aged (n=147 females: 43.2+/-13 years, n=268 males: 43.3+/-14 years) Caucasian Greek subjects consisted the middle aged cohort. One hundred seventy five elderly (n=88 females: 79.9+/-4 years; n=87 males: 80.6+/-4 years) were selected from the elderly cohort. Genotype data were obtained for all of them. Multiple linear regression analysis, stratified by gender and adjusted for age, smoking habits and body mass index as covariates, showed higher ox-LDL-C levels for the middle aged men with the Val/Val genotype, compared to the other allele (Ala/Ala and Ala/Val) carriers (65.9+/-25.7 vs. 55.7+/-20.5 mg/dl; standardized beta coefficient=0.192, P=0.012). On the contrary, elderly women with the Val/Val genotype occurred with lower ox-LDL-C levels compared to the Ala/Ala or Ala/Val genotype (74.2+/-22.1 vs. 86.5+/-26.6 mg/dl; standardized beta coefficient= -0.269, P=0.015). The same trend was also recorded in elderly men, however without reaching statistical significance (standardized beta coefficient= -0.187, P=0.077). Moreover, elderly men and women with the Ala/Ala or Ala/Val genotype presented higher triglycerides levels compared to Val/Val (women: 145.2+/-68.7 vs. 114.3+/- 34.3 mg/dl, P= 0.027; men: 147.8+/-72.4 vs. 103.7 +/-38.0 mg/dl, P=0.002). Additionally, middle aged men with the Val/Val genotype had higher HDL-C levels compared to the Ala allele carriers. The results suggest that SOD Ala16Val polymorphism is an age-dependent modulator of ox-LDL-C levels in middle-aged men and elderly women.     

高效凝胶渗透色谱法研究超氧化物歧化酶的分子量

本文采用凝胶渗透色谱仪，对修饰后的牛血ＳＯＤ进行测定。仪器：美国ＷａｔｅｒｓＡＬＣ／ＧＰＣ２４４型高效液相色谱仪，ＳｈｏｄｅｘＰ－８２葡聚精为高分子标样。采用紫外和示差折光双检测器进行测定。所得数据与经典的凝胶电泳法完全一致，为此类高分子材料的Ｍｎ、ＭＷ和Ｄ值的测定，建立了准确、快速的测定方法。