Ultraviolet and visible molecular spectroscopyProbably the first physical method used in analytical chemistry was based on the quality of the color in colored solution. The first things we observe regarding colored solutions are their hue，or color，and the colors depth，or intensity.These observations led to the technique historically called colorimetry;the color of the solution could identify species(qualitative analysis) while the intensity of the color could identify the concentration of the species present(quantitative analysis).This technique was the first use of what we now understand to be absorption spectroscopy for chemical analysis .When white light passes through a solution and emerges as red light，we say that the solution is red.What has actually happened is that the solution has allowed the red component of white light to pass through，whereas it has absorbed the complementary color，yellow and blue. The more concentrated the sample solution，the more yellow and blue light is absorbed and the more intensely red the solution appears to the eye. For a long time，experimental work made use of the human eye as the detector to measure the hue and intensity of colors in solutions. However，even the best analyst can have difficulty comparing the intensity of two colors with slightly different hues，and there are of course people who are color-blind and cannot see certain colors.Instruments have been developed to perform the measurements more accurately and reliably than the human eye. While the human eye can only detect visible light，this chapter will focus on both the ultraviolet(UV) and the visible (Vis) portions of the spectrum. The wavelength range of UV radiation starts at the blue end of visible light(about 400nm) and ends at approximately 200nm for spectrometers operated in air. The radiation has sufficient energy to excite valence electrons in many atoms and molecules;consequently，UV radiation is involved with electronic excitation. Visible light，considered to be light with wavelengths from 800 to 400nm，acts in the same way as UV light. It is also considered part of the electronic excitation region. For this reason we find commercial spectroscopic instrumentation often operates with wavelengths between 800 and 200nm.Spectrometers of this type are called UV/Visible(or UV/Vis) spectrometers.The vacuum UV region of the spectrum extends below 200 nm to the X-ray region of the spectrum, at100 A. It has called the vacuum UV region because oxygen，water vapor，and other molecules in air absorb UV radiation below 200nm，so the spectrometer light path3 must be free of air to observe wavelengths200nm.The instrument must be evacuated(kept under vacuum) or purged with an appropriate non-UV absorbing gas such as helium for this region to be used. Vacuum UV radiation is also involved in electronic excitation but the spectrometers are specialized and not commonly found in undergraduate or routine analytical laboratories. For our purposes the term UV will mean radiation between 200nm and 400nm，unless stated otherwise.The interaction of UV and visible radiation with matter can provide qualitative identification of molecules and polyatomic species，including ions and complexes. Structural information about molecules and polyatomic species，especially organic molecules,can be acquired. This qualitative information is usually obtained by observing the UV/Vis spectrum,the absorption of UV and visible radiation as a function of wavelength by molecules.The shape and intensity of UV/Vis absorption bands are related to the electronic structure of the absorbing species.The molecule is often dissolved in a solvent to acquire the spectrum . We will look at how we can use the absorption maximum of a chromophore and a set of guidelines to predict the position of the absorption maximum in a specific molecule.We will also consider how the solvent affects the spectrum of some molecules. As a reminder,the transitions that give rise to UV/Vis absorption by organic molecules are the n*,*，and n*，transitions. Some terms need to be defined. A chromophore is a group of atoms(part of a molecule)that gives rise to an electronic absorption. An auxochrome is a substituent that contains unshared (nonbonding) electron pairs，such as OH，NH，and halogens. An auxochrome attached to a chromophore with electrons shifts the absorption maximum to longer wavelengths. A shift to longer wavelengths is called a bathochromic shift or red shift.A shift to shorter wavelengths is called a hypsochromic shift or blue shift. An increase in the intensity of absorption band(that is，an increase in max)is called hyperchromism;a decrease in intensity is called hypochromism. These shifts in wavelength and intensity come about as a result of the structure of the entire molecule or as a result of interaction between the solute molecules and the solvent molecules. As described at the beginning of the chapter，the types of compounds that absorb UV radiation are those with nonbonded electrons(n electrons) and conjugated double bond systems (electrons) such as aromatic compounds and conjugated olefins. Unfortunately，such compounds absorb over similar wavelength ranges，and the absorption spectra overlap considerably. As a first step in qualitative analysis，it is necessary to purify the sample to eliminate absorption bands due to impurities. Even when pure，however，the spectra are often broad and frequently without fine structure. For these reasons，UV absorption is much less useful for the qualitative identification of functional groups or particular molecules than analytical methods such as MS，IR，and NMR. UV absorption is rarely used for organic structural elucidation today in modern laboratories because of the ease of use and power of NMR，IR，and MS. UV and visible absorption spectrometry is a powerful tool for quantitative analysis. It is used in chemical research，biochemistry，chemical analysis，and industrial processing.Quantitative analysis is based on the relationship between the degree of absorption and the concentration of the absorbing material. Mathematically，it is described for many chemical systems by Beers Law A=abc. The term applied to quantitative absorption spectrometry by measuring intensity ratios is spectrophotometry. The use of spectrophotometry in the visible region of the spectrum used to be referred to as colorimetry. UV/Vis spectrophotometry is a widely used spectroscopic technique. It has found use everywhere in the world for research，clinical analysis，environmental analysis，and many other applications. Some typical applications of UV absorption spectroscopy include the determination of(I)the concentrations of phenol，nonionic surfactants，sulfate，sulfide，phosphates，fluoride，nitrate，a variety of metal ions，and other chemicals in drinking water in environmental testing;(II)natural products,such an steroids or chlorophyll;(III)dyestuff materials;(IV)vitamins，proteins，DNA，and enzymes in biochemisty. Spectrophotometry in the UV region of the spectrum is used for the direct measurement of many organic compounds，especially those with aromatic rings and conjugated multiple bonds. There are also colorless inorganic species that absorb in the UV. A good example is the nitrate ion，NO3-.A rapid screening method for nitrate in drinking water is performed by measuring the absorbance of the water at 220 nm and at 275 nm. Nitrate ion absorbs at 220 nm but not at 275 nm;the measurement at 275 nm is to check for interfering organic compounds that may be present. Spectrophotometic analysis in the visible region can be used whenever the sample is colored. Many materials are inherently colored without chemical reaction(e.g.，inorganic ions such as dichromate，permanganate，cupric ion，and ferricion) and need no further chemical reaction to form colored compounds. Colored organic compounds，such as dyestuffs，are also naturally colored. Solution of such materials can be analyzed directly.The majority of metal and nonmetal ions，however，are colorless. The presence of these ions in a sample solution can be determined by first reacting the ion with an organic reagent to form a strongly absorbing species. If the product of the reaction is colored，absorbance can be measured in the visible region;alternatively，the product formed may be colorless but absorb in the UV. The majority of spectrophotometric determinations result in an increase in absorbance (darker color if visible) as the concentration of the analyte increases. Quantitative analysis by absorption spectrophotometry requires that the samples be free from particulates，that is，free from turbidity. The reason for this is that particles can scatter light. If light is scattered by the sample away from the detector，it is interpreted as an absorbance.The absorbance will be erroneously high if the sample is turbid. We can make use of the scattering of light to characterize samples as discussed in Section 5. 7，but particulates must be avoided for accurate absorbance measurements. Quantitative analysis by absorption spectrophotometry generally requires the preparation of a calibration curve，using the same conditions of pH，reagents added，and so on for all of the standards，samples，and blanks. It is critical to hav