激光镊子拉曼光谱法优化红法夫酵母合成虾青素条件

用激光镊子拉曼光谱法优化红法夫酵母生产虾青素的条件。首先,将红法夫酵母细胞拉曼光谱与虾青素标准品溶液拉曼光谱进行对比,找出定量虾青素的拉曼特征峰;然后对照红法夫酵母生长曲线与不同时间点红法夫酵母细胞内虾青素含量曲线,进行发酵时间的优化;最后将红法夫酵母分别用不同氮源、碳源培养基培养,对比其细胞的虾青素拉曼特征峰强度,最终优化培养基。由实验可得,适宜定量虾青素的拉曼特征峰是1520cm-1峰;最佳发酵时间为72h;最佳优化培养基氮源含硫酸铵4g/L+硝酸钾4g/L,碳源含蔗糖60g/L。从上述结果可知,用激光镊子拉曼光谱法对红法夫酵母合成虾青素的条件进行优化,充分发挥了其操作简便、耗时短、样品用量少、对样品无损伤等优势,使所得结果更准确可信。因此,激光镊子拉曼光谱法是红法夫酵母细胞合成虾青素条件优化的理想选择。     

光镊拉曼光谱法分析红法夫酵母内虾青素含量

建立一种用激光镊子拉曼光谱法快速定量红法夫酵母细胞内虾青素含量的方法。测定不同浓度虾青素标准品溶液的拉曼光谱,取其1 520cm-1峰峰高绘制虾青素标准曲线;取不同氮源、碳源的红法夫酵母细胞,一部分用激光镊子拉曼光谱法测定,一部分用紫外-可见分光光度法测定;最后分析两者间的相关性。结果表明虾青素标准曲线的相关系数到达0.998 3。在单位质量红法夫酵母虾青素含量与单位体积红法夫酵母发酵液虾青素产量方面,对比激光镊子拉曼光谱法与紫外-可见分光光度法,两种方法所得数据具有良好的相关性,其相关系数分别达到0.917 7和0.905 4,表明激光镊子拉曼光谱法能够达到紫外-可见分光光度法的测定效果,是定量分析红法夫酵母细胞内虾青素的更有效方法。     

共振拉曼光谱在体测量类胡萝卜素含量

类胡萝卜素是人体内重要的抗氧化剂,能够消除体内的自由基和其它有害氧分。研究表明类胡萝卜素含量与癌症、心血管疾病、老化等疾病发病率呈反比关系。目前检测方法为采用血清液相色谱检测方法,不能做到在体无损伤、实时测量。文章介绍了一种用于测量类胡萝卜素的新技术——共振拉曼光谱技术,具有在体无损伤、灵敏度高和实时检测等特点。该技术利用强度远小于美国ANSI Z136.1-2000标准的473 nm激光激发拇指中类胡萝卜素,产生强荧光和叠加在其上的弱共振拉曼光,通过测量拉曼散射光强度在体评估类胡萝卜素的含量。同时,利用组织通透技术,改善了测量信噪比。测量了不同饮食习惯志愿者的类胡萝卜素含量,说明其含量与其摄入量成正比。该技术在临床应用和科学研究等领域具有重要意义。     

拉曼光谱法结合薄层层析分析红酵母色素

利用薄层层析色谱法分离红酵母色素,结果显示,红酵母细胞能合成至少三种色素,即β-胡萝卜素、红酵母红素、圆酵母红素;采集三种色素的拉曼光谱,光谱数据经过背景扣除、基线校正、三点平滑等方法预处理,统计不同色素的平均光谱,结果表明三种色素的CC拉曼位移不同,并且β-胡萝卜素的拉曼位移最多,红酵母红素和圆酵母红素的含量较多;定量分析色素特征峰高比值,各色素峰高比值差异不大,峰高比值能用作参数,为深入研究活体细胞内色素的相对含量提供参考。以上结果表明,拉曼光谱法结合薄层层析能够分析红酵母色素,可以提供红酵母色素的丰富信息,是研究色素的有效方法。     

应用光镊子拉曼光谱鉴别阴道毛滴虫和口腔毛滴虫的研究

应用激光镊子拉曼光谱技术比较分析阴道毛滴虫和口腔毛滴虫的拉曼吸收峰,寻找具有分类学鉴别价值的特征拉曼峰。通过光镊随机俘获单个大小一致、有活力的虫体,记录其拉曼光谱,收集数据并做主成分分析。在937cm-1、1002cm-1和1446cm-1谱峰的强度和谱型上有差异,根据1002cm-1峰的信号强度,并结合937cm-1和1002cm-1峰强度比值(I937/I1002)以及1002cm-1和1446cm-1峰强度比值(I1002/I1446)可以作为区分阴道毛滴虫和口腔毛滴虫的量化指标。该法是简便、快捷、有效地鉴别两种人体毛滴虫的新方法。     

激光拉曼光谱在类胡萝卜素研究中的应用

本文简述了类胡萝卜素的组成、特性及功能;激光拉曼光谱在类胡萝卜素浓度测量中的应用。类胡萝卜素作为一种广泛存在的天然色素,在预防人类的某些疾病,尤其是癌症方面有着比较重要的作用。拉曼光谱技术作为一种快速、无损、安全、所需样品少、可在体测量的分析测试手段而被广泛的应用,而类胡萝卜素以其独特的链式结构又比较容易被激发出强的拉曼信号,随着对类胡萝卜素各种功能的研究和相关产品的开发应用,激光拉曼光谱技术也必在此领域中有广阔的发展空间。     

拉曼光谱分析人类单个血小板的类胡萝卜素

应用拉曼光镊对人的单个血小板细胞(Plt)进行拉曼光谱收集与分析.拉曼信号显示,人类血液成分中,血小板的类胡萝卜素浓度最高.虽然类胡萝卜素在人类血小板中普遍存在,但是单个血小板之间差异极显著,且与血小板的脂类物质、蛋白质含量没有相关性.血小板激活过程引起蛋白质拉曼光谱的变化,而类胡萝卜素基本没有变化;常温(25℃)保存...     

人体单红血细胞的拉曼光谱分析

对人体红血细胞进行了拉曼光谱谱线测定,并对谱线进行了初步指认和分析,由此得出红血细胞的基本特征,为疾病诊断提供初步的实验依据.     

不同物态及单个杆菌芽孢中的2,6-吡啶二羧酸的拉曼光谱特征与结构分析

利用激光镊子拉曼系统(LTRS)测定了2,6-吡啶二羧酸(DPA)及其钙盐(Ca-DPA)的固体(粉末与晶体)和水溶液及单个细菌芽孢的光谱并进行了对比分析.不同物态的 DPA 及 Ca-DPA 的拉曼光谱有很大区别.分析导致光谱差异的结构上的原因可能有:(1)DPA 晶体产生的散射要大于 DPA 粉末所产生的散射,DPA 晶体的拉曼光谱所反映的 DPA 的特征信息比 DPA 粉末更为精确;(2)晶体状态的 DPA及Ca-DPA 中可能有水分子的存在,其与水分子间的相互作用是很强;(3)钙离子的存在影响吡啶环两边的羧基,从而使吡啶环产生一定的几何形变,羧基中的羟基也受到破坏;(4) DPA与Ca-DPA 溶液中主要存在的是DPA-2,在 Cla-DPA 溶液中钙离子主要是对吡啶环结构的稳定性产生影响.在测定单个杆菌芽孢内 DPA 的研究中发现,芽孢体内的 DPA 是以 Ca-DPA 晶体的形式存在的.     

南极海洋芳香烃低温降解微生物的拉曼光谱分析

激光镊子拉曼光谱技术可以实现在自然状态下对单个细胞或细胞器较长时间的观察研究.应用激光镊子拉曼光谱技术实时观察南极微生物低温降解芳香烃过程中单个南极细菌的细胞生长和胞内生物大分子的动态变化过程,收集、分析其拉曼光谱,结果发现:单细胞的拉曼光谱反映了其细胞内部的生命物质组成,南极动球菌 NJ41 和希瓦氏菌 NJ49 生...     

Organic-solvent-free extraction of carotenoids from yeast Rhodotorula glutinis by application of ultrasound under pressure

The extraction of Rhodotorula glutinis carotenoids by ultrasound under pressure (manosonication) in an aqueous medium has been demonstrated. The influence of treatment time, pressure, and ultrasound amplitude on R. glutinis inactivation and on the extraction of carotenoids was evaluated, and the obtained data were described mathematically. The extraction yields were lineal functions of those three parameters, whereas inactivation responded to a more complex equation. Under optimum treatment conditions, 82% of carotenoid content was recovered. Extraction of carotenoids in an aqueous medium was attributed to the capacity of ultrasound for cell disruption and emulsification. Cavitation caused the rupture of cell envelopes and the subsequent formation of small droplets of carotenoids surrounded by the phospholipids of the cytoplasmic membrane that would stabilize the emulsion. Analysis of the dispersed particle size of the extracts demonstrated that a fine, homogeneous emulsion was formed after treatment (average size: 230 nm; polydispersity <0.22). This research describes an innovative green process for extracting carotenoids from fresh biomass of R. glutinis in which only two unit operations are required: ultrasonic treatment, followed by a centrifugation step to discard cell debris. The extract obtained thanks to this procedure is rich in carotenoids (25 mg/L) and could be directly incorporated as a pigment in foods, beverages, and diet supplements; it can also be utilized as an ingredient in drugs or cosmetics.     

Resonance Raman detection of carotenoid antioxidants in living human tissues

We have used resonance Raman scattering as a novel noninvasive optical technology to measure carotenoid antioxidants in living human tissues of healthy volunteers. By use of blue-green laser excitation, clearly distinguishable carotenoid Raman spectra superimposed on a fluorescence background are obtained. The Raman spectra are obtained within less than a minute, and the required laser light exposure levels are well within safety standards. Our technique can be used for rapid screening of carotenoid levels in large populations and may have applications for assessing antioxidant status and the risk for diseases related to oxidative stress.     

In vivo Raman spectroscopic analysis of the influence of UV radiation on carotenoid antioxidant substance degradation of the human skin

It is well known from the literature that carotenoid antioxidant substances decrease after the irradiation of the skin with UV light. Available literature has shown no information about the response time and total dynamics of degradation of carotenoid antioxidants after UV irradiation. The measurements were made with the HPLC method, which is time-consuming and does not give relevant information about the dynamics of degradation of carotenoids in the skin after UV irradiation. With the introduction of new noninvasive spectroscopic methods, it became possible to measure in vivo the behavior of carotenoid antioxidant substances, such as betacarotene and lycopene in the skin after UV light exposure. In the present study, the resonance Raman spectroscopy method was used as a fast and noninvasive optical method to measure the dynamics of degradation of beta-carotene and lycopene in living human skin after UV exposure. It was found that the beta-carotene and lycopene concentration in the skin does not decrease immediately after UV irradiation. There is a time delay, which varies from 30 up to 90 min for beta-carotene and from 0 up to 30 min for lycopene. A strong nonlinear correlation between the individual antioxidant level of volunteers and the magnitude of destruction of antioxidants in the skin was found.     

Photobleaching as a method of increasing the accuracy in measuring carotenoid concentration in human skin by Raman spectroscopy

It was shown experimentally that the effect of photobleaching in noninvasive measurement of the Raman spectra of light used in determining the carotenoid concentration in human skin can be used to increase measurement accuracy. Increased accuracy occurs as a consequence of a decrease in the measurable Raman spectra of the wideband fluorescent background intensity when a sample is irradiated by laser radiation. Furthermore, it was demonstrated that, for the spectra of skin from nine volunteers, the fluorescent background intensity can be decreased on average by a factor of 1.4, which leads to an increase in the signal-to-noise ratio for Raman lines of skin carotenoids by a factor of 1.2 on average. The kinetics of photobleaching of humans can be described by biexponential decay with a correlation coefficient close to unity, which agrees with the presented theoretical calculations.     

Near-field Raman spectroscopy of biological nanomaterials by in situ laser-induced synthesis of tip-enhanced Raman spectroscopy tips

We report a new approach in tip-enhanced Raman spectroscopy (TERS) in which TERS-active tips with enhancement factors of ～10(-5)× can be rapidly (1-3 min) produced in situ by laser-induced synthesis of silver nanoparticles at the tip apex. The technique minimizes the risks of tip contamination and damage during handling and provides in situ feedback control, which allows the prediction of the tip performance. We show that TERS tips produced by this technique enable the measurement of spatially resolved TERS spectra of self-assembled peptide nanotubes with a spatial resolution of ～20 nm.     

茶氨酸拉曼光谱分析

采用显微共焦拉曼光谱技术测试研究了L-茶氨酸,得到了L-茶氨酸的拉曼谱图,发现在250～1 700和2 800～3 000cm-1能够观测到明显的拉曼信号;通过对谱峰进行归属和分析,得到了不同波数范围内的特征振动模式;其中,在321,900,938,1 153,1 312,1 358,1 454和1 647cm-1找到8个较强的拉曼信号,可作为L-茶氨酸的特征峰。结果表明:拉曼光谱可能成为一种直接、准确和快速的检测茶氨酸的方法。     

Raman analysis of synthetic eritadenine

Eritadenine, 2(R),3(R)‐dihydroxy‐4‐(9‐adenyl)‐butyric acid, is a cholesterol‐reducing compound naturally occurring in the shitake mushroom (Lentinus edodes). To identify the unknown Raman spectrum of this compound, pure synthetic eritadenine was examined and the vibrational modes were assigned by following the synthesis pathway. This was accomplished by comparing the known spectra of the starting compounds adenine and D ‐ribose with the spectra of a synthesis intermediate, methyl 5‐(6‐Aminopurin‐9H‐9‐yl)‐2,3‐O‐isopropylidene‐5‐deoxy‐β‐D ‐ribofuranoside (MAIR) and eritadenine. In the Raman spectrum of eritadenine, a distinctive vibrational mode at 773 cm⁻¹ was detected and ascribed to vibrations in the carbon chain, ν(C C). A Raman line that arose at 1212 cm⁻¹, both in the Raman spectrum of MAIR and eritadenine, was also assigned to ν(C C). Additional Raman lines detected at 1526 and at 1583 cm⁻¹ in the Raman spectrum of MAIR and eritadenine were assigned to ν(N C) and a deformation of the purine ring structure. In these cases the vibrational modes are due to the linkage between adenine and the ribofuranoside moiety for MAIR, and between adenine and the carbon chain for eritadenine. This link is also the cause for the disappearance of adenine specific Raman lines in the spectrum of both MAIR and eritadenine. Several vibrations observed in the spectrum of D ‐ribose were not observed in the Raman spectrum of eritadenine due to the absence of the ribose ring structure. In the Raman spectrum of MAIR some of the D ‐ribose specific Raman lines disappeared due to the introduction of methyl and isopropylidene moieties to the ribose unit. With the approach presented in this study the so far unknown Raman spectrum of eritadenine could be successfully identified and is presented here for the first time. Copyright © 2008 John Wiley & Sons, Ltd.     

Raman analysis of defects in n-type 4H-SiC

This paper employs micro-Raman technique for detailed analysis of the defects (both inside and outside) in bulk 4H-SiC. The main peaks of the first-order Raman spectrum obtained in the centre of defect agree well with those of perfect bulk 4H-SiC, which indicate that there is no parasitic polytype in the round pit and the hexagonal defect. Four electronic Raman scattering peaks from nitrogen defect levels are observed in the round pit (395\,cm$^{-1}$, 526\,cm$^{-1}$, 572\,cm$^{-1}$, and 635\,cm$^{-1})$, but cannot be found in the spectra of hexagonal defect. The theoretical analysis of the longitudinal optical plasmon--phonon coupled mode line shape indicates the nonuniformity of nitrogen distribution between the hexagonal defect and the outer area in 4H-SiC. The second-order Raman features of the defects in bulk 4H-SiC are well-defined using the selection rules for second-order scattering in wurtzite structure and compared with that in the free defect zone.