Raman Spectroscopy for the Undergraduate Teaching Laboratory: Quantification of Ethanol Concentration in Consumer Alcoholic Beverages and Qualitative Identification of Marine Diesels Using a Miniature Raman Spectrometer

Raman spectroscopy has steadily gained popularity as a powerful tool in both the analytical lab and the undergraduate classroom. The technique is attractive because it allows for rapid, nondestructive qualitative or quantitative analyses of many analytes with little or no sample preparation requirements. The introduction of less expensive, smaller, and more powerful diode laser excitation sources and the recent availability of rugged, red‐sensitive, charge‐coupled device–based miniature modular spectrometers has prompted the integration of Raman spectroscopy into the undergraduate curriculum. We have evaluated the analytical utility of a small, portable Raman instrument for the qualitative and quantitative analyses of two “real” samples. The experiments in this paper were designed to be used as a laboratory component for undergraduate education and include the quantification of ethanol in consumer alcoholic beverages and the qualitative identification of marine diesel fuels that had been spilled on surface waters. In the case of the liquor samples, the ethanol concentration in colorless, odorless alcoholic beverages could be determined very rapidly, but colored and heavily scented liquors proved more difficult and required pretreatment with activated carbon to remove fluorescence that masked the Raman signal. Similarly, a high‐intensity fluorescence background was observed to mask characteristic Raman bands of the diesel fuels. Some reduction in the intensity of the fluorescence was observed after carbon pretreatment of the fuels. The set of undergraduate experiments described in this paper treat the concepts of quantitative and qualitative analysis using portable instrumentation, instrumental calibration by the standard addition and external curve methods, and method development for the analysis of real consumer and environmental samples.     

拉曼光谱定性和定量检测青蒿素研究

青蒿素是从中药青蒿中提取的含有过氧基团的倍半萜内酯药物,具有良好的抗疟特性,是治疗疟疾的特效药。运用激光拉曼光谱分析了100～3 500 cm-1光谱范围内青蒿素的声子振动特性。指出位于724 cm-1的拉曼峰为与青蒿素中过氧基团直接相关的一个特性声子振动模式,可用于检测过氧桥键的存在。位于1 734 cm-1的拉曼峰为与青蒿素中内酯基团直接相关的一个特性声子振动模式,可用于进一步检测分析青蒿素。由于这两个特征拉曼峰对应于青蒿素分子中特征化学键的振动,而且在实验上较容易观察分析,因而它们可以很好的用于拉曼光谱法快速初步定性检测青蒿素。同时,通过分析比较不同纯度青蒿素样品中724和1 734 cm-1处特征拉曼峰的平均散射信号强度比,拉曼光谱法可以用于定量检测青蒿素样品的纯度。与常用的高效液相色谱法相比,拉曼光谱法更快速方便,检测精度更高,而且可以检测青蒿素样品纯度的均匀性。拉曼光谱法定性和定量检测青蒿素纯度的功能对分析检测中药青蒿的品质也有重要意义。     

Combined Laser-Induced Breakdown with Raman Spectroscopy: Historical Technology Development and Recent Applications

Abstract: A modern trend in the development of laser-based analytical instruments is to integrate more functions in a single device, which could supply complementary information and improve the capability to identify different components. This review presents historical development of the combined analysis by laser-induced breakdown spectroscopy (LIBS) and Raman Spectroscopy, which briefly describes basic instrumental principles and technological aspects of the hyphenated technique, as well as the requirements in the design of interactive instruments, including laser systems, optical systems for laser beam delivery, emission collection devices, and spectrum measurement systems. Various configurations of the combined instrumentation have been designed and used depending on the different applications, such as pigment analysis in the cultural heritage field, detection of explosives, analysis of geological samples, and applications in future planetary missions. In addition, the feasibility, applicability, and prospective aspects of the hybrid technology are discussed in detail.     

动态表面增强拉曼光谱在敌瘟磷快速定量分析中的应用

动态表面增强拉曼光谱是在干态与湿态表面增强拉曼光谱(SERS)检测的基础上发展而来的,不仅具有极好的信号增强,还具有良好的重复性与稳定性。提出了一种基于动态SERS与多元分析方法的敌瘟磷快速定量分析方法。实验中,首先测量100,50,10,5,1,0.5和0.1 mg·L-1敌瘟磷动态SERS谱图,并使用多项式校正方法去除光谱基线漂移。然后,处理后的全范围(600~1 800 cm-1)与特征范围(674~713,890~1 195,1 341~1 399和1 549~1 612 cm-1)光谱分别利用支持向量机回归(SVR)构建定量模型,实现对敌瘟磷的定量分析。同时,实验还评估了主成分分析(PCA)对定量分析结果的影响。实验结果表明特征范围光谱所建立的模型预测误差较小,而数据经过PCA处理后预测误差得到进一步下降。最优回归模型是由特征范围光谱经PCA处理后所构建的模型(RMSECV=0.065 7 mg·L-1),模型能够准确地预测敌瘟磷溶液浓度。为了测试实际检测中的效果,该方法被用来对苹果表面的敌瘟磷残留进行检测,并通过气相色谱法进行验证。结果表明该方法对于同一样本多次检测值波动较小,且检测均值与气相色谱检测值相差较小,相对误差最大仅为5.13%。此外,动态SERS检测可在2 min内完成,且后续数据处理也可在数秒内完成,同时整个过程的试剂消耗仅在2 μL左右。因此,所提出的方法在敌瘟磷快速准确检测具有极大优势。     

拉曼光谱在多组分定量分析中方法学验证的探讨

拉曼光谱在中国药典2010版第一次被收录于“附录XIX L拉曼光谱法指导原则”中,但该原则对拉曼多组分定量分析的方法学验证未作论述,对多组分复杂体系中采用拉曼数学模型定量分析时的方法学验证进行独创性的探索和研究,分别从仪器性能测试、线性、定量限、准确度等方面进行方法学验证研究;另外由于拉曼光谱定量数学模型样品为复杂成分样品,无须像传统方法提供空白样品和进行破坏实验等专属性实验验证。通过研究探讨,提出了拉曼多组分定量分析的方法学验证内容,为方法学验证的完善和标准的制定提供了基础和依据。     

基于等离激元增强拉曼光谱的便携式快速检测仪设计与实现

用于食品安全、环境污染、毒品、化学战剂检测等的现场快速检测要求检测设备便携、快速和准确。目前的常规实验室检测方法虽然能够实现准确检测,但是其实时性较差,无法满足现场快速检测的要求。因此,设计开发了基于等离激元增强拉曼光谱的便携式快速检测仪,能够实现对滥用添加剂、违禁食品添加剂、农药残余、毒品、化学战剂和环境水污染等大量有害物质的现场快速定性检测。该检测仪基于ARM嵌入式系统开发,编写了其操作交互界面和底层驱动程序,实现了拉曼光谱数据自动标定和特征谱图快速识别等算法。在仪器中搭建了检测物质拉曼光谱的标谱数据库,数据库根据检测科目进行分类,每种科目包含该类别的多种物质,每种物质均包含其高中低多种浓度的标准品和样本谱图。设计实现了大类科目的辨识比对算法和GPU硬件加速算法,实现了对某一类科目的所有物质的快速比对,与传统的纯CPU算法实现相比较,在辨识速度上提高了20倍以上。通过市场购买的功能饮料、违规葡萄酒与果汁等实际样品对所制备的样机仪器进行测试,测试结果符合预期,具有良好的灵敏度与重现性,满足了现场快速检测的要求。     

一种危险液体混合物的拉曼光谱定性定量识别方法

危险液体混合物的拉曼光谱定性定量分析一直是现场应用难点,为解决该问题,分析了多种物质混合后拉曼光谱的峰位、峰值、峰型变化情况,选取拉曼光谱关键特征峰进行数学简化,构建了从混合物物质成分到混合物拉曼光谱的映射关系,该映射关系描述多种物质成分混合的混合物拉曼特征峰响应只和混合物中各成分本身拉曼特征峰响应以及各物质成分混合比例有关,各物质成分按混合比例贡献拉曼特征谱峰,共同形成最终的混合物拉曼光谱。由该映射关系求逆,可实现从采集到的混合物拉曼光谱计算出各物质成分的混合比例。基于此,设计了危险液体混合物成分定性定量识别方法,主要方法步骤包括,首先进行拉曼光谱数据采集,然后进行拉曼光谱数据处理并获得拉曼特征峰,再进行测试样品与数据谱库标准品的正反向特征峰匹配,如果正反向特征峰匹配系数都比较高,在满足一定阈值条件下,可认定测试样品是某种纯净物,如果不是纯净物,则进入混合物分析,通过拉曼光谱特征峰反向匹配系数筛选,确定混合物成分构成,混合物成分确定后再进行混合物成分比例计算,最终实现危险液体混合物定性定量分析。实验部分,选定丙酮、甲苯、三氯甲烷、乙醇及其混合物进行实验验证,当混合物样品是丙酮、乙醇两种成分按3∶7比例混合时,经拉曼光谱识别方法计算,混合成分计算值是丙酮占比0.245 7,乙醇占比0.706 0;当混合物样品是甲苯、三氯甲烷两种成分按3∶7比例混合时,经拉曼光谱识别方法计算,混合成分计算值是甲苯占比0.323 4,三氯甲烷占比0.763 0;当混合物样品是丙酮、甲苯、乙醇三种成分按4∶3∶3比例混合时,经拉曼光谱识别方法计算,混合成分计算值是丙酮占比0.795 9、甲苯占比0.303 5、乙醇占比0.287 5,实验结果表明,当危险液体混合物成分是两种或三种成分混合时,混合成分计算值基本和实际值吻合,应用危险液体混合物的拉曼光谱定性定量识别方法,可较准确的从拉曼混合光谱中解析出各混合物成分以及各成分在混合物中的比例,可以判断混合物每个拉曼特征谱峰都来自于哪个成分或哪些成分拉曼特征谱峰的混合,谱图解析结果良好,对危险液体混合物现场分析鉴别有较大应用价值。     

一种便携式快速等离激元增强拉曼光谱检测前处理仪的研制与在食品安全中的应用

便携式拉曼光谱仪在现场快速分析中往往由于现场样品比较复杂,需要处理后才能采用光谱仪进行测定,因此一定程度上限制它的使用。常规实验室前处理虽然行之有效,但是过程繁琐费时,不适合现场使用。本课题组开发了一种便携式快速PERS前处理仪器,具有便携、功耗低、操作简便,提取时间短以及重复性好等特点,能满足现场快速检测的前处理需求。该仪器由超声波提取单元、加热抽气挥发单元、尾气净化与吸收单元以及控制系统四个功能模块组成,并且设有LED控制面板和报警装置。整个工作流程主要包括超声提取、液液萃取和溶剂加热抽气挥发三个环节。以三种实际样品(辣椒粉、辣椒油、豆瓣酱)中的违禁添加剂罗丹明B检测为例,结果表明：与常规实验室前处理方法相比,两种前处理方法对于PERS谱图影响不大,但是检测时间缩短一半。另外,重复性实验检测结果的相对标准偏差(relative standard deviation, RSD)小于±5%。     

New Developments and Applications of Handheld Raman,Mid-Infrared,and Near-Infrared Spectrometers

Abstract: Recently, miniaturization of Raman, mid-infrared (IR), and near-infrared (NIR) spectrometers has made substantial progress. Though mid-infrared systems are based exclusively on attenuated total reflection (ATR) measurements, near-infrared spectrometers operate in the diffuse reflection or transmission mode. The reduction in size, however, must not be accompanied by deterioration in measurement performance, and portable instrumentation will only have a real impact on quality and process control if Raman, IR, and NIR spectra of comparable quality to laboratory spectrometers can be obtained.

In the present communication, a short overview on the building principles of novel handheld systems will be provided and the results of qualitative and quantitative analyses of selected liquid and solid sample systems obtained with these Raman, Fourier transform infrared (FTIR), and NIR spectrometers will be evaluated in terms of their comparability with laboratory instruments and their suitability for on-site and field measurements.     

纳米银粒子表面增强拉曼光谱在宠物饲料中三聚氰胺的速测研究

研究将制备的纳米银粒子作为表面增强拉曼光谱的增强试剂,实现对宠物饲料中三聚氰胺的快速定性定量分析。以709与1542cm-1处拉曼位移作为定性依据,以1149cm-1峰强度作为归一化标准,在1.0～10.0mg·kg-1浓度范围实现定量计算,检测限0.5mg·kg-1。研究发现,纳米银粒子对三聚氰胺具有较强的拉曼增强效应,特征峰信号强度与纳米银粒子加入时间,溶液涡旋强度有很大的关系,同时也受提取溶剂的种类、提取方式及样品加入量的影响。本方法中每个样品5min内完成分析,与现有方法相比,快速简便,对宠物饲料中三聚氰胺实现现场、快速检测。     

Quantitative Analysis of Amoxicillin Residues in Foods by Surface-Enhanced Raman Spectroscopy

Amoxicillin is one of the most widely used antibiotics for food-producing animals and human beings. Amoxicillin residue in food-producing animals has become a growing concern for consumers because it has been proven to show potential carcinogenic, teratogenic, and mutagenic effects at a high-dose level in clinical use. So there is an urgency to develop a rapid, simple, and accurate detection method for amoxicillin residue in animal foods. In this study, surface-enhanced Raman spectroscopy (SERS) coupled with gold nanoparticles was used for the rapid analysis of amoxicillin. Density functional theory (DFT) calculations were conducted with Gaussian 03 at the B3LYP level using the 6-31G(d) basis set to analyze the assignment of vibrations. Results showed that the theoretical Raman spectra of amoxicillin were completely consistent with its experimental spectra. Gold nanoparticles used as a SERS substrate can significantly increase spectra signals of amoxicillin, and acidic pH can improve its characteristic peaks' shape and SERS sensitivity. Under optimum pH (pH 2), the detection limit could reach the level of 1 µg/mL in case of distilled water as solvent. Quantitative analysis of amoxicillin residues in foods revealed that the SERS technique with gold nanoparticles was sensitive and of a good stability and linear correlation. It was well suited for rapid analysis of amoxicillin residue in a great deal of food samples.     

激光拉曼光谱法测定不同采收期连翘叶中连翘苷的含量

采用激光拉曼光谱内标法对不同采收期连翘叶中连翘苷的含量进行测定。分别选取连翘苷甲醇溶液的拉曼光谱中连翘苷呋喃环上C-H对称伸缩振动(2844 cm-1处强峰)和溶剂甲醇中CH3-O反对称伸缩振动(2975 cm-1)作为定量峰和内标参比峰。以两拉曼峰峰强的比值组成相对强度,绘制标准曲线,并对线性和加样回收率进行考察。在1.1×10-5mol.L-1~3.5×10-4mol.L-1(5.9 mg.mL-1~189.1 mg.mL-1)范围内,连翘苷的浓度与相对峰强的线性关系良好,r=0.9980,检出限为5.2×10-6mol.L-1.通过精密度、重复性和加样回收率测定,结果证明该法具有良好的精密度、重复性和准确性。对不同采收期的连翘叶样品进行了测定,结果表明该法简便快速、高效灵敏,可用于其含量测定。拉曼光谱内标法用于连翘苷的定量分析具有操作简便和无需添加其它试剂等优点,可以用于药物的含量测定。     

拉曼光谱在线测量技术在聚合物合成与加工中的应用研究进展

聚合物的合成与加工过程影响着材料的聚合度、结晶度、取向度以及共混组分的均匀性,最终影响聚合物产品的性能。因此对聚合物合成与加工过程中材料的实时状态进行监控至关重要。拉曼光谱在线测量技术凭借其无损检测、采样简单、可靠性高,对试样的外形和物态没有任何特定要求,且能在高温、高压、有毒的环境中直接进行在线测量等特点,在高分子材料合成与加工领域有了越来越广泛的应用。将此在线测量技术应用到高分子合成与加工过程中,实时测量高分子材料的化学组成和物理性质,并将测量数据用于控制合成与加工过程,可以提高高分子材料的性能。本文选择聚合反应转化率、聚合产物粒径及其分布、聚合物结晶度、聚合物取向度、多相共混聚合物组分含量等五个反映聚合物合成或加工过程状态的参量,概述了拉曼光谱在线测量上述参量的相关研究进展,并对拉曼光谱在线测量技术在聚合物合成及加工中的应用发展进行了展望。     

红外和拉曼光谱的煤灰矿物组成研究

研究煤灰中矿物质的性质通常从矿物组成的表征入手。为了分析两种高硅铝煤灰的矿物成分,采用傅里叶变换红外光谱(FTIR)、拉曼光谱和X射线衍射(XRD)技术对煤灰样进行了测试和综合表征,将FTIR和拉曼光谱的分析结果与XRD进行了比较。FTIR结果表明,在1 100~1 000 cm-1范围内高硅铝煤灰出现最强的特征峰,例如石英峰(1 089 cm-1)和偏高岭石峰(1 042 cm-1),它们都归属于Si-O伸缩振动。对原始红外谱图进行二阶导数处理后,可获得重叠峰的峰位,有助于更完整的解析矿物吸收峰,从而获得更丰富的矿物组成信息。煤灰中硬石膏的红外和拉曼光谱发现,在1 157,1 126和674 cm-1的拉曼光谱峰与在1 151,1 120和678 cm-1的红外光谱峰振动模式分别相同且峰位接近,还存在一些完全不同的拉曼光谱与红外光谱峰,表明这两种光谱存在互补性。尽管煤灰中锐钛矿含量很低,但由于Ti-O的极化率很高,因此拉曼光谱显示锐钛矿的144 cm-1峰远远强于石英的461 cm-1峰。XRD结果表明,煤灰中主要存在石英、云母、赤铁矿、硬石膏和未知的无定形相矿物,FTIR和拉曼光谱综合分析的结果表明除了这些矿物,还存在偏高岭石、无定形氧化硅、长石、方解石和锐钛矿等。在定性分析方面,将FTIR和拉曼光谱结合起来比XRD单独获得的矿物组成信息更为详细。     

Validation of Raman and FTIR Spectroscopy Methods in Forensic Analysis of Questioned Documents

Abstract

Paper is of great interest to the examiner of questioned documents and forensic chemists because its examination can often answer questions concerning the identity of paper, its source or origin, and its date of manufacture. This information is often used to determine whether a document is authentic or fraudulent. Raman and infrared spectroscopy are two complementary spectroscopic techniques that can produce fast and efficient analysis of the paper of questioned documents. Considering the applicability of quantitative methods of analysis and the importance of the fields in which they are used, method validation is fundamental to the quality of the final results. In this study, a validation of infrared and Raman spectroscopy methods used to differentiate paper samples is described.     

激光诱导击穿光谱对污染鱼体内重金属元素分布与含量的分析

环境污染可造成生物体的中毒、病变及死亡。生物体各组织对污染物质的吸收、积累情况是不同的。生物体各组织对污染物质重金属元素的吸收是生物医学中的一项重要研究。采用激光诱导击穿光谱(LIBS)方法对受污染的鱼体各组织进行了重金属元素定量分析。实验中得到了一组探测鱼类组织元素成份的LIBS最佳条件,拟合了Pb和Ba元素的定标曲线并通过外标法测定了重金属含量。实验结果表明,鱼体中肝脏及口腮等部位有重金属积累,而在鱼肉中重金属含量极低。所提出的方法可用于生物体受污染影响的评估研究, 在生物医学领域有重要推广价值。     

活性污泥中抗性细菌筛选及拉曼光谱检测

抗生素制药废水是污水处理中比较难处理的废水。抗生素与微生物作用的机理研究对于解决抗生素废水处理问题具有重要意义。拉曼光谱作为一种对样品无损,非侵入型,并且对水不敏感的检测手段,已越来越多地用于生物、医学的研究。从普通污水处理反应器的活性污泥里通过涂布法和平板划线法分离得到了4株对头孢他啶具有抗性的菌株,研究了其对头孢他啶的降解效果,降解率最高达45.44%。采用16S rRNA基因测序进行菌种鉴定,得到这些菌种分别属于气单胞菌属,芽孢杆菌属和红球菌属。对筛选出的细菌进行不同条件下拉曼光谱测定,探讨了细菌拉曼光谱的测试条件,结果显示CaF₂是测定生物样品拉曼光谱时效果最好的基底。通过拉曼光谱技术和主成分分析法对菌种进行了差异分析,提取了三个主成分,累积荷载达到83.9%。其中,第一主成分主要包括 748,1 003和1 126 cm-1等比较明显的峰位,第二主成分包括1 661和1 448 cm-1,第三主成分只有855 cm-1。本研究利用拉曼光谱对活性污泥筛选的抗性菌株进行生物大分子的差异表征,通过拉曼光谱对抗性菌种进行了较好的区分,与基因测序的结果相吻合,证明了拉曼光谱用于抗生素废水生物处理作用机理研究的可行性及优越性。     

Recent Applications of Infrared and Raman Spectroscopy in Art Forensics: A Brief Overview

Abstract

A brief overview on the recent applications of infrared and Raman spectroscopy in the field of art forensics is presented.     

Applications of Raman Spectroscopy in Agricultural Products and Food Analysis: A Review

Abstract

Raman spectroscopy has been gaining popularity as an analytical tool due to advances in development of Raman spectrometry and the power of personal computers. Due to to its narrow and highly resolved bands, Raman spectroscopy allows for nondestructive extraction of chemical and physical information about samples and aids in rapid on-line analysis without any special sample preparation. In this review, Raman spectroscopic techniques such as dispersive Raman spectroscopy, Fourier transform Raman spectroscopy, surface-enhanced Raman spectroscopy, and spatially offset Raman spectroscopy are briefly introduced. In addition, applications of Raman spectroscopy are explored, within various fields of agricultural products and food, including fruits and vegetables, crops, meat and dairy products, oil, as well as beverages. In addition, some discussion on the importance of Raman spectroscopy as fundamental and applied research of agricultural products and food is provided.     

显微激光拉曼定量分析CO_2气体碳同位素组成方法研究

制备了一系列不同比例的~(12)CO_2/N_2和~(13)CO_2/N_2混合物,对样品进行显微激光拉曼测试分析后发现气体拉曼特征峰峰面积比与其摩尔分数比成正比例关系,拟合方程的斜率被认为是拉曼量化因子F_(12CO_2)和F_(13CO_2)。用气相组分中只含有~(12)CO_2和N_2的流体包裹体验证了当F_(12CO_2),为1.163 49时,根据气体的拉曼特征峰峰面积比能估算出其摩尔分数比。由线性拟合后的方程斜率得出F_(13CO_2)和F_(12CO_2)分别为1.610 86和1.163 49,它们的比率F_(13CO_2)/F_(12CO_2)是1.3845。在确定稳定同位素分子的拉曼参数和实验条件基础上,CO_2气体碳同位素摩尔分数比C_(12)/C_(13)可根据A_(12Co_2)/A_(13CO_2)(拉曼峰峰面积比)和F_(13CO_2)/F_(12CO_2)的乘积求出。此外,用已知摩尔分数比(C_(12)/C_(13))的人造包裹体验证了此方法具有一定的可行性,可以建立起定量分析CO_2气体碳同位素激光拉曼测试方法。