Ultraviolet/visible spectroscopy detector

Ultraviolet/visible spectroscopy is one of the most common methods for online monitoring of analytes from a chromatographic column. This type of detector is fast (many data points can be taken every second) and sensitive (parts per million concentration levels can register a significant response). When two or more compounds elute through the flow cell at the same time, the UV signal is typically the superposition of the signals from each... 

The phenomenon of fluorescence has been synonymous with ultraviolet (UV) and visible spectroscopy rather than near-infrared (near-IR) spectroscopy from the beginning of the subject. This fact is evidenced in definitive texts which also provide useful background information for this volume (see, e.g., Refs. 1-6). Consequently, our understanding of the many molecular phenomena which can be studied with fluorescence techniques, e.g., excimer formation, energy transfer, diffusion, and rotation, is based on measurements made in the UV/visible. Historically, this emphasis was undoubtedly due to the spectral response of the eye and the availability of suitable sources and detectors for the UV/visible in contrast to the lack of equivalent instrumentation for the IR. Nevertheless, there are a few notable exceptions to the prevalence of UV/visible techniques in fluorescence such as the near-IR study of chlorophyll(7) and singlet oxygen,<8) which have been ongoing for some years. 

The use of ultraviolet (UV) spectroscopy for on-line analysis is a relatively recent development. Previously, on-line analysis in the UV-visible (UV-vis) region of the electromagnetic spectrum was limited to visible light applications such as color measurement, or chemical concentration measurements made with filter photometers. Three advances of the past two decades have propelled UV spectroscopy into the realm of on-line measurement and opened up a variety of new applications for both on-line UV and visible spectroscopy. These advances are high-quality UV-grade optical fiber, sensitive and affordable array detectors, and chemometrics. 

Ultraviolet-visible (UV-Vis) spectrophotometric detectors are used to monitor chromatographic separations. However, this type of detection offers very little specificity. Element specific detectors are much more useful and important. Atomic absorption spectrometry (AAS), inductively coupled plasma-atomic emission spectroscopy (ICPAES) and inductively coupled plasma-mass spectrometry (ICP-MS) are often used in current studies. The highest sensitivity is achieved by graphite furnace-AAS and ICP-MS. The former is used off-line while the latter is coupled to the chromatographic column and is used on-line . 

Analysis is an integral part of research, clinical, and industrial laboratory methodology. The determination of the components of a substance or the sample in question can be qualitative, quantitative, or both. Techniques that are available to the analyst for such determinations are abundant. In absorption spectroscopy, the molecular absorption properties of the analyte are measured with laboratory instruments that function as detectors. Those that provide absorbance readings over the ultraviolet-visible (UV-vis) light spectrum are commonly used in high-performance liquid chromatography (HPLC). The above method is sufficiently sensitive for quantitative analysis and it has a broader application than other modes of detection. 

Apparatus. Ultraviolet, visible and near infrared spectra were recorded with a Cary 17 spectrophotometer, y spectroscopy was carried out with a Ge-Li detector and a Zoomax (Sein) multichannel analyzer. pH measurements were taken with an Aries 10000 (Tacussel) potentiometer, a spectroscopy was carried with a solid state a detector and a (Intertechnique) multichannel analyzer. 

Once the transient species has been formed, it has to be monitored by some form of kinetic spectroscopy, typically with ultraviolet-visible absorption or emission, infrared (time-resolved infrared or TRIR) (74), or resonance Raman (time-resolved resonance Raman or TR3) (80) methods of detection. The transient is usually tracked by a probe beam at a single characteristic frequency, thereby giving direct access to the kinetic dimension. Spectra can then be built up point by point, if necessary, with an appropriate change of probe frequency for each point, although improvements in the sensitivity of multichannel detectors may be expected to lead increasingly to the replacement of the laborious point-by-point method by full two-dimensional methods of spectroscopic assay (that is, with both spectral and kinetic dimensions). 

Unlike IR spectroscopy where nowadays FT instrumentation is solely used, in Raman spectroscopy both conventional dispersive and FT techniques have their applications, the choice being governed by several factors. The two techniques differ significantly in several performance criteria, and neither one is best for all applications. Contemporary dispersive Raman spectrometers are often equipped with silicon-based charge coupled device (CCD) multichannel detector systems, and laser sources with operating wavelength in the ultraviolet, visible or near-infrared region are employed. In FT Raman spectroscopy, the excitation is provided exclusively by near-infrared lasers (1064 nm or 780 nm). 

In this regard, various analytical methods have been developed and their potential for the VOO authentication has been evaluated. Thus, spectroscopic tools, such as FT-near-infrared spectroscopy, FT-Raman spectroscopy, fluorescence and ultraviolet-visible detectors, and chromatographic techniques have been widely used in the field of VOO authentication [43-48],... 

Initially, simple methods such as ultraviolet-visible (UV-Vis), fluorescence or infrared (IR) spectroscopy were proposed in order to estimate the total amount of antioxidants in various food samples. However, coupled methods such as gas chromatography-mass spectrometry (GC-MS), high performance liquid chromatography with ultraviolet-visible (HPLC-UV-Vis) or nuclear magnetic resonance detector (HPLC-NMR) and liquid chromatography-mass spectrometry (LC-MS) are employed more to quantify individual tocols or carotens from various corn-based food samples. In this chapter all these methods of analysis will be briefly described. 

Four basic types of elution are used in HPLC, namely, the isocratic system, the basic gradient system, the inert system and the advanced gradient system (see Figure 1.1). The most commonly used detectors are those based on spectroscopy in the region 185-400 nm, visible-ultraviolet (UV) spectroscopy in the region 185-900 nm, post-column derivatisation with fluorescence detection (see next), conductivity [7] and multiple wavelength UV detectors using a diode array system detector (see next). Other types of detectors available are those based on electrochemical principles, refractive index, differential viscosity, and mass detection [8]. 

The scope of ultraviolet and visible spectrophotometry can be further extended when combined with a chromatographic separation step such as HPLC. The development of rapid-scanning detectors based on the linear photodiode array permits spectra to be acquired during the elution of peaks. Computer-aided manipulation of these spectra has led to new strategies for the examination of chromatographic peak homogeneity, based on classical techniques in spectroscopy. The use of microcomputers enables the development of archive retrieval methods for spectral characterisation (A. F. Fell etal, J. Chromat., 1984, 316, 423-440). 

Over the past decade, there has been considerable development in imaging type detectors for the measurement of ultraviolet (UV) and visible light. These new detectors have attracted the interest of a number of analytical spectroscopists. For absorption spectroscopy, analytical chemists have traditionally used such instruments as the photometer, which uses a narrow-band light source (for example the 254 nm emission line from a low pressure Hg lamp or a continuous source with a filter), a sample cell and a photomultiplier tube (FMT) as the detector. While useful for many specific applications, the single-wavelength photometer cannot determine multiple sample components simultaneously or provide a general absorbance characterization of the system. When information at multiple wavelengths is desired,... 

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