Monitoring cell culture media degradation using surface enhanced Raman scattering (SERS) spectroscopy

The quality of the cell culture media used in biopharmaceutical manufacturing is a crucial factor affecting bioprocess performance and the quality of the final product. Due to their complex composition these media are inherently unstable, and significant compositional variations can occur particularly when in the prepared liquid state. For example photo-degradation of cell culture media can have adverse effects on cell viability and thus process performance. There is therefore, from quality control, quality assurance and process management view points, an urgent demand for the development of rapid and inexpensive tools for the stability monitoring of these complex mixtures. Spectroscopic methods, based on fluorescence or Raman measurements, have now become viable alternatives to more time-consuming and expensive (on a unit analysis cost) chromatographic and/or mass spectrometry based methods for routine analysis of media. Here we demonstrate the application of surface enhanced Raman scattering (SERS) spectroscopy for the simple, fast, analysis of cell culture media degradation. Once stringent reproducibility controls are implemented, chemometric data analysis methods can then be used to rapidly monitor the compositional changes in chemically defined media. SERS shows clearly that even when media are stored at low temperature (2–8 °C) and in the dark, significant chemical changes occur, particularly with regard to cysteine/cystine concentration.     

An investigation of the surface enhanced Raman scattering (SERS) from a new substrate of silver-modified silver electrode by magnetron sputtering

'Pure' silver nanoparticles on silver electrode were prepared by magnetron sputtering. The silver-modified silver electrode has good stability and the silver nanoparticles on silver electrode have homogeneous size distribution. Compared with the silver colloid modified silver electrode, there were no any extraneous component ions on the electrode, for the modified silver nanoparticles are prepared by magnetron sputtering. Synchronously, we obtained much higher quality SERS spectra of adenine molecules on the silver electrode modified by magnetron sputtering (SEMMS), and the study of the adsorption behavior of adenine on the silver-modified silver electrode by surface enhanced Raman scattering (SERS) indicated that the silver-modified silver electrode was highly efficient substrates for SERS investigation. From the rich information on the SEMMS obtained from high-quality potential-dependent SERS, we may deduce the adsorption behavior of adenine and the probable SERS mechanism in the process. The probable reasons are given.     

Detection of volatile organic compounds using surface enhanced Raman spectroscopy substrates mounted on a thermoelectric cooler

Preliminary results for a volatile organic compound (VOC) sensor based on surface enhanced Raman spectroscopy (SERS) are described. The sensor is comprised of a SERS substrate mounted on a thermoelectric cooler (TEC). The SERS substrate is chemically modified with a thiol coating that prevents oxidation of the roughened silver surface and attracts the analyte of interest to the SERS surface. Using this sensor, detection of chlorinated solvents, aromatic compounds, and methyl t-butyl ether (MTBE) is demonstrated.     

Surface-enhanced Raman scattering (SERS) for probing internal cellular structure and dynamics

Surface-enhanced Raman scattering (SERS) provides vibrational information about molecules that are located within several nanometers of the surface of a metallic nanoparticle. This review describes the various challenges and successes of applying SERS inside living cells in order to gain information about the internal structure and dynamic processes occurring in the intracellular matrix. In particular, the challenges associated with the introduction of metal nanoparticles into cells are described, as well as the complexity of interpreting SERS spectra from within complex biological environments. Strategies for understanding and improving the specificity of SERS in vivo are also presented.

Semi-quantitative trace analysis of nuclear fast red by surface enhanced resonance Raman scattering

The dye nuclear fast red has been detected and determined semi-quantitatively by means of surface enhanced resonance Raman scattering (SERRS) and surface enhanced Raman scattering (SERS), using laser exciting wavelengths of 514.5 and 632.8 nm, respectively, by employing a citrate-reduced silver colloid. A good linear correlation is observed for the dependence of the intensities of the SERRS bands at 989 cm⁻¹ (R=0.9897) and 1278 cm⁻¹ (R=0.9872) on dye concentration over the range 10⁻⁹ to 10⁻⁷ M, when using an exciting wavelength of 514.5 nm. At dye concentrations above 10⁻⁷ M, the concentration dependence of the SERRS signals is non-linear. This is almost certainly due to the coverage of the colloidal silver particles being in excess of a full monolayer of the dye. A linear correlation is also observed for the dependence of the intensities of the SERS bands at 989 cm⁻¹ (R=0.9739) and 1278 cm⁻¹ (R=0.9838) on the dye concentration over the range 10⁻⁸ to 10⁻⁶ M when using an exciting wavelength of 632.8 nm. Strong fluorescence prevented collection of resonance Raman scattering (RRS) spectra from powdered samples or aqueous solutions of the dye using an exciting wavelength of 514.5 nm, but weak bands were observed in the spectra obtained from both powdered and aqueous samples of the dye using an exciting wavelength of 632.8 nm. A study of the pH dependence of SERRS/SERS and UV–VIS absorption spectra revealed the presence of different ionisation states of the dye. The limits of detection for nuclear fast red by SERRS (514.5 nm), SERS (632.8 nm) and visible spectroscopy (535 nm) are 9, 89 and 1000 ng ml⁻¹, respectively.     

A simple approach for ultrasensitive detection of bisphenols by multiplexed surface-enhanced Raman scattering

Bisphenol A (BPA) is well known for its use in plastic manufacture and thermal paper production despite its risk of health toxicity as an endocrine disruptor in humans. Since the publication of new legislation regarding the use of BPA, manufacturers have begun to replace BPA with other phenolic molecules such as bisphenol F (BPF) and bisphenol B (BPB), but there are no guarantees regarding the health safety of these compounds at this time. In this context, a very simple, cheap and fast surface-enhanced Raman scattering (SERS) method was developed for the sensitive detection of these molecules in spiked tap water solutions. Silver nanoparticles were used as SERS substrates. An original strategy was employed to circumvent the issue of the affinity of bisphenols for metallic surfaces and the silver nanoparticles surface was functionalized using pyridine in order to improve again the sensitivity of the detection. Semi-quantitative detections were performed in tap water solutions at a concentrations range from 0.25 to 20 μg L⁻¹ for BPA and BPB and from 5 to 100 μg L⁻¹ for BPF. Moreover, a feasibility study for performing a multiplex-SERS detection of these molecules was also performed before successfully implementing the developed SERS method on real samples.     

Effect of layer structures of gold nanoparticle films on surface enhanced Raman scattering

Using a method of collecting nanoparticles at a water/hexane interface in a close-packed monolayer film and transferring such films onto a solid substrate, three-dimensional multilayer films of nanoparticles were formed. The packed nanoparticles were gold nanospheres (NS) with a 26 nm diameter or gold nanorods (NR) with a 31 nm diameter and 74 nm length. We investigated variations in the surface enhanced Raman scattering (SERS) intensities from such nanoparticle films as the layer compositions were changed. The films stacked with NR layers generated much higher SERS intensity than those of NS layers. The SERS intensities from both kinds of films increased as the number of layers were increased. However, when the NR layer and NS layer were stacked alternately, SERS intensity varied in a zigzag fashion. It was found that the structure of top layer plays a distinguishable role in generating strong SERS enhancement while the lower layers contribute to SERS with less dependency on structures. Interlayer coupling as well as intralayer coupling was considered in order to explain the observations.     

胶体分形聚集体分维数与随时间变化的SERS强度关系研究

由于胶体聚集的动力学过程可用分形理论描述,从而对胶体动力学生长机制有了新的理解。Lin等研究了金胶、硅胶及聚乙烯乳胶的直接光散射后认为,胶体的两种生长机制DLCA和RLCA聚集体具有不依赖于胶体个性的普适分数维。对这一结果用不同方法对不同胶体进行验证十分必要。我们曾研究了吡啶吸附于AgBr胶体随时间变化的SERS规律,本文进一     

复合SERS基底的构建及性质

通过自组装方法以对巯基苯胺(PATP)为偶联分子, 在石英基片上构筑了多种形貌的银钠米粒子单层结构和三明治结构. 研究了组装膜在不同激发线下表面增强拉曼散射(SERS)的增强差异. 研究结果表明, 单层基底和三明治基底中偶联分子的SERS信号因银纳米粒子间的电磁场耦合而显著增强, 且在三明治结构中增强更加明显. 对复合SERS基底增强因子进行计算可知, 复合SERS基底的表面等离子体共振(SPR)峰与激发线的匹配程度越好, 其增强因子越大. 在三明治结构中更易发生PATP分子转变为对巯基偶氮苯(DMAB)分子的激光诱导催化偶联反应. 另外, 该激光诱导催化偶联反应与激发波长密切相关.     

Surface-enhanced Raman scattering for perchlorate detection using cystamine-modified gold nanoparticles

Perchlorate (ClO₄⁻) has recently emerged as a widespread environmental contaminant found in groundwater and surface water, and there is a great need for rapid detection and monitoring of this contaminant. This study presents a new technique using cystamine-modified gold nanoparticles as a substrate for surface-enhanced Raman scattering (SERS) detection of perchlorate at low concentrations. A detection limit of 5 × 10⁻⁶ M (0.5 mg/L) has been achieved using this method without sample preconcentration. This result was attributed to a strong plasmon enhancement by gold metal surfaces and the electrostatic attraction of ClO₄⁻ onto positively charged, cystamine-modified gold nanoparticles at a low pH. The methodology also was found to be reproducible, quantitative, and not susceptible to significant interference from the presence of anions such as sulfate, phosphate, nitrate and chloride at concentrations <1 mM, making it potentially suitable for rapid screening and routine analysis of perchlorate in environmental samples.     

Full-range analysis of ambient volatile organic compounds by a new trapping method and gas chromatography/mass spectrometry

This study investigated the feasibility of analyzing a full range of ambient volatile organic compounds (VOCs) from C(3) to C(12) using gas chromatograph mass spectrometry (GC/MS) coupled with thermal desorption. Two columns were used: a PLOT column separated compounds lighter than C(6) and a DB-1 column separated C(6)-C(12) compounds. An innovative heart-cut technique based on the Deans switch was configured to combine the two column outflows at the ends of the columns before entering the MS. To prevent the resolved peaks from re-converging after combining, two techniques were attempted (hold-up vs. back-flush) to achieve the intended "delayed" elution of heavier components. Thus, the resulting chromatogram covering the full range of VOCs is a combination of two separate elutions, with the heavier section following the lighter section. With the hold-up method, band-broadening inevitably occurred for the delayed C(6)-C(7) DB-1 compounds while the light compounds eluted from the PLOT column. This broadening problem resulted in peak tailing that was largely alleviated by adding a re-focusing stage while the DB-1 compounds were back-flushed, and this modified technique is referred to as the back-flush method. With this modification, the separation of the C(6)-C(7) compounds improved dramatically, as revealed by the decrease in peak asymmetry (As) and increase in resolution. Linearity and precision for these peaks also improved, yielding R(2) and RSD values better than 0.9990 and 2.8%, respectively.     

Sampling volatile organic compounds using a modified solid phase microextraction device

Headspace solid phase microextraction (headspace SPME) has been demonstrated to be an excellent solvent-free sampling method. One of the major factors contributing to the success of headspace SPME is the concentrating effect of the fiber coating toward organic compounds. The affinity of the fiber coating toward very volatile analytes, such as chloromethane, may, however, not be large enough for detection at the parts per trillion concentration level. Static headspace analysis, on the other hand, is very effective for these very volatile compounds. As analyte volatility decreases, the sensitivity of static headspace analysis drops. The complementary nature of these two sampling methods can be exploited by combining the SPME device with a gastight syringe. The sensitivity of the new sampling device is better than that of SPME for very volatile compounds or that of static headspace analysis for less volatile compounds. This new method can sample a wide range of compounds from chloromethane (b.p. −24°C) to bromoform (b.p. 149°C) with estimated limits of detection at the low parts per trillion level.     

Enhancement of the detection sensitivity for volatile organic compounds by using an annular type photoionization detector and a pre-concentration system

Photoionization detector (PID) was developed for a sensitive on-site detection of trace amounts of volatile organic compounds (VOCs) based on an annular type ion collection electrode assembly. An ion collector with an annular geometry could detect more stable ion signals in the PID system when compared to the other types of ion collectors when an UV lamp of 10.6 eV was used as an ionization source. In order to enhance the detection sensitivity, a pre-concentration system, which was developed by adopting a ceramic heater packed with rod shaped molecular sieves, was adopted for a detection of VOCs. The adopted ceramic heaters had a resistance of 10-20 Ohm, and the temperature of the heater was optimized by controlling the heating time of the resistor. The enhancement of the detection sensitivity was found to be 8-10 times with the PID system when compared to the signals measured without a pre-concentrator. The overall detection sensitivity of the developed PID system was estimated as 10 ppb or better.     

Detection of hexavalent chromium using gold/4-(2-mercaptoethyl) pyridinium surface enhanced Raman scattering-active capture matrices

In this communication, the fabrication of SERS-active capture matrices for the detection of hexavalent chromium is described. The amine groups of amine-modified magnetic microparticles were used to immobilize gold colloidal particles. Once immobilized, the gold was reacted with 4-(2-mercaptoethyl) pyridinium (MEP) hydrochloride to form a self-assembled monolayer (SAM). The MEP SAM exhibits great selectivity for hexavalent chromium. It was shown that calibration curves could be generated by ratioing MEP peaks that increased in intensity upon complexation with chromate with a peak that did not change. Flow experiments, using Au/MEP capture matrices held in place by a magnet, showed instantaneous response to changes in chromate concentration.     

表面增强拉曼散射强度与金纳米粒子粒径关系

表面增强拉曼散射（SurfaceenhancedRamanscattering,简称SERS）的增强机理主要分为两类’‘－‘’：电磁增强和化学增强．通常SERS活性表面的获得需要粗糙化．MOSkovitS最先提出,可将粗糙化的SERS活性表面模型化为平整金属基底上排列的金属胶体粒子“’．这样的模型与实际体系比较符合,同时给理论处理提供了便利．在这一模型基础之上,人们提出了一系列SERS电磁增强的理论计算方法’‘－“．在这些理论计算中,大多包含有SERS强度与粒径关系的结果．粒径对SERS强度的影响体现在两方面：1）SERS与粗糙度有关,粒径可视为粗…     

基于表面增强拉曼光谱技术对医疗美容产品中正壬酸香草酰胺的快速检测

以银纳米线为拉曼基底,运用表面增强拉曼光谱技术(SERS)建立了对发热剂中正壬酸香草酰胺的检测方法。采用简便有效的两步滴加多元醇法制备了具有SERS活性的银纳米线,利用扫描电镜和紫外-可见光谱仪对银纳米线进行了表征。对正壬酸香草酰胺进行了SERS研究并对正壬酸香草酰胺的SERS谱带进行了归属。正壬酸香草酰胺的质量浓度在1～1.0×10^-8mg/L范围内与其在1588 cm^-1处的SERS特征峰强度有良好的线性关系,方法的最低检出浓度可达0.66 pg/L。对样品进行前处理后,运用加标回收法考察其回收率。该方法可以用于发热剂中正壬酸香草酰胺的检测。     

电化学粗糙银电极表面上共吸附体系的SERS研究

自1974年发现表面增强喇曼散射(SERS)光谱以来,人们不但对其进行实验和理论上的探讨,还开展了应用方面的研究,如用于痕量分析、催化和腐蚀等,并已开始用于研究物质的吸附状态以及多物种共存体系。本文通过对吡啶、苯甲酸共存体系的SERS谱及其随外加电位变化的实验研究,运用镜像场等理论探讨了在电化学粗糙银电极表面上吡啶和苯甲酸共存体系中的吸附状态。