山姜素与人血清白蛋白相互作用的荧光光谱法研究

利用荧光光谱法和紫外-可见光谱法研究了山姜素与人血清白蛋白(HSA)之间的相互作用。证实了山姜素对HSA的荧光猝灭为动态猝灭过程,并测定了不同温度下的猝灭常数; 根据Fōrster非辐射能量转移理论,计算出山姜素在蛋白质中的结合位置与色氨酸残基间的距离为4.05 nm; 由求得的热力学参数,推断了山姜素与HSA之间主要靠疏水作用力结合;用三维荧光光谱及同步荧光光谱技术探讨了山姜素对HSA构象的影响。     

Separation and determination of alpinetin and cardamonin in Alpinia katsumadai Hayata by flow injection-micellar electrokinetic chromatography

A simple, rapid, and accurate method for the separation and determination of alpinetin and cardamonin in Alpinia katsumadai Hayata was developed by combination of flow injection (FI)-micellar electrokinetic chromatography (MEKC) for the first time. The analysis was carried out using an unmodified fused-silica capillary (50 μm i.d.; total length 13.6 cm; effective length 10.3 cm) and direct ultraviolet (UV) detection at 214 nm. The sample throughput was 11-24 samples per hour using the background electrolyte (BGE) containing 4 mM sodium borate-8 mM NaH₂PO₄ (pH 8.1)-8 mM sodium dodecyl sulfate (SDS)-19% (v/v) ethanol. The repeatabilities (n = 4) reached relative standard deviation values (R.S.D.) of 3.0% and 2.5% for the peak areas and 2.5% and 3.1% for peak heights of alpinetin and cardamonin, respectively. Regression equations revealed linear relationships (r²: 0.9993-0.9994) between the peak area of each analyte and the concentration. Recoveries were in the range 90-92% and 99-105% for alpinetin and cardamonin, respectively.     

中草药有效成分山姜素的固体表面室温磷光法研究

本文选择滤纸作基质，以ＬｉＡｃ作重原子微扰剂，建立了测定中草药中痕量有效成分山姜素的滤纸表面室温磷光法。该法取样量少（２μＬ），线性范围宽（２．９～５８０ｎｇ／斑），灵敏度高（检出限０．４０ｎｇ／斑），操作简便、快速。     

山姜素与脱氧核糖核酸的相互识别研究

应用荧光光谱法及紫外-可见光谱法研究了生理条件下(pH 7.4)山姜素(ALP)与DNA分子之间的相互识别。考察了不同温度下(25,32和39 ℃),DNA对山姜素荧光猝灭情况。实验发现, DNA能猝灭山姜素的内源性荧光,随着温度的升高,荧光猝灭常数K_SV逐渐减小(K_SV分别为3.288×103, 2.923×103和2.467×103 L·mol-1),并且DNA对山姜素的猝灭速率常数K_q要大于药物小分子与生物大分子之间的最大扩散所控制的碰撞猝灭常数,得出DNA对山姜素的荧光猝灭是单一的静态猝灭过程。DNA与山姜素相互作用紫外-可见光谱显示,DNA不能使得山姜素的吸收峰发生减色效应和红移现象,而山姜素也不能使溴化乙锭-DNA体系的荧光强度及最大荧光峰位置发生变化,即山姜素不和溴化乙锭竞争与DNA的结合位点。DNA热变性实验发现,解链DNA对山姜素的荧光猝灭程度要大于正常DNA的猝灭程度,由此推断山姜素与DNA不存在嵌插作用。同时,I-离子效应和盐效应表明,山姜素与DNA之间主要以沟槽模式相结合。     

海南草豆蔻及其主要活性组分、金属元素含量的谱学测定

采用红外光谱、紫外光谱、荧光光谱及微波消解电感耦合等离子体质谱法对海南白沙和霸王岭两个产地的草豆蔻进行了分析。其乙醇提取物的红外光谱都具有1 051,1 390,2 976及3 300cm~(-1)左右表征黄酮类成分的特征吸收峰,二阶红外导数谱图显示了在2 977.72及2 899.94有相似吸收峰,在1 922.36及1 650.87cm~(-1)有显著差异吸收峰,可以鉴定区分海南不同产地的草豆蔻。利用紫外光谱法表征了两个不同产地草豆蔻的特征谱图,并根据有关的吸收原理,建立了测定草豆蔻植物中主要组分山姜素与豆蔻明含量的方法,获得了良好的分析精密度(0.42%~0.29%)和回收率(84.28%~117.41%)。测得白沙草豆蔻中山姜素和豆蔻明的含量分别为4.23%和3.83%,而霸王岭的分别为3.72%和3.34%。荧光测定结果表明,不同产地的草豆蔻乙醇提取物具有相似的荧光发射及激发光谱谱图,荧光强度的差别也再次证明白沙草豆蔻乙醇提取物的浓度大于霸王岭的。利用微波消解电感耦合等离子体质谱法分析了草豆蔻中的K,Ca,Mg,Na,Al,Fe,Mn,Cu,Zn等十七种金属元素及其含量。该研究为定性、定量地鉴定区别不同产地或不同种植批次的草豆蔻及测定其中主要活性组分的含量建立了简单可行的方法;根据实验测得的金属元素种类及含量的结果,可为进一步开发利用草豆蔻及提高食品安全意识提供更好的理论依据。     

Spectrometric study of the interaction between Alpinetin and bovine serum albumin using chemometrics approaches

The binding interaction of Alpinetin (APT) with bovine serum albumin (BSA) was studied by fluorescence, UV-visible and synchronous fluorescence spectroscopy (SFS) under simulated physiological conditions. The measured complex spectra were resolved by multivariate curve resolution-alternating least squares (MCR-ALS), yielding a host of data and information, which otherwise would have been impossible to obtain. The extracted profiles corresponded to the spectra of the single species in the APT/BSA mixture. In addition, the presence of the APT-BSA complex was demonstrated, and it was shown that the associated quenching of the fluorescence from the BSA protein resulted from the formation of APT-BSA complex via a static mechanism. The binding constant (K_a(ave) = 2.34 × 10⁶ L mol⁻¹) and the number of sites (n = 1) were obtained by fluorescence methods as were the thermodynamic parameters (ΔH⁰, ΔS⁰ and ΔG⁰). This work suggested that the principal binding between APT to BSA was facilitated by hydrophobic interactions. The thermodynamic parameters for APT were compared to those from the structurally similar Chrysin and Wogonin molecules. It appeared that the entropy parameters were relatively more affected by the small structural changes. SFS from the interaction of BSA and APT showed that the ligand affected the conformation of BSA. The competitive interaction of APT and site makers with BSA indicated site I as the binding area of APT in BSA.     

微波辅助萃取多物理场耦合模型的构建及验证

以姜科类植物草豆蔻中山姜素为模拟对象, 基于微波辅助萃取(MAE)过程中微波场、 温度场及浓度场等物理场之间的耦合关系, 利用Comsol软件构建了MAE多物理场耦合模型. 模拟了不同萃取时间、 微波功率下萃取体系的电磁场分布、 温度分布以及山姜素扩散分布; 通过比较萃取液温度和山姜素浓度的模拟值与实验值, 结合误差分析对模型进行了验证, 以同类植物砂仁中的异槲皮苷为萃取目标验证了模型的适用性. 结果表明, 微波功率越大, 电磁场强度越强, 萃取液温度越高, 山姜素扩散越明显, 越有利于萃取; MAE萃取山姜素和异槲皮苷的萃取液温度模拟值与实验值之间的相对均方根误差(RRMSE)分别在1.9%~4.5%和1.9%~2.8%之间, 其浓度RRMSE分别在1.7%~3.2%及1.6%~4.1%之间, 均小于5.0%, 表明建立的模型准确、 可靠, 且适用性良好. 该模型综合考虑了MAE过程中的多个物理场及其耦合关系, 可为深入研究其萃取机理提供参考.