Postcolumn photochemical reactions

Postcolumn photochemical reactions are another approach to the detection problem. High-intensity UV light, generally provided by a Hg or Zn lamp, photolyzes the HPLC effluent, which passes through a Teflon (47) or quartz tube. The photolysis reaction determines the nature of the subsequent detection. If the compound has a UV chromophore, such as an aromatic ring, and an ionizable heteroatom, such as chlorine, then the products of the reaction can be detected conductometrically. Busch et al. (48) have examined more than 40 environmental pollutants for applicability to detection with photolysis and conductance detection. Haeberer and Scott (49) found the photoconductivity approach superior to precolumn derivatization for the determination of nitrosoamines in water and waste water. The primary limitation of this detection approach results from the inability to use mobile phases that contain ionic modifiers, that is, buffers and... 

Figure 27.18 Common configuration for postcolumn reactors with electrochemical analysis. (A) LC-chemical reaction-EC. Postcolumn addition of a chemical reagent (for example, Cu2+ or an enzyme). (B) LC-enzyme-LC. Electrochemical detection following postcolumn reaction with an immobilized enzyme or other catalyst (for example, dehydrogenase or choline esterase). (C) LC-EC-EC. Electrochemical generation of a derivatizing reagent. The response at the second electrode is proportional to analyte concentration (for example, production of Br2 for detection of thioethers). (D) LC-EC-EC. Electrochemical derivatization of an analyte. In this case a compound of a more favorable redox potential is produced and detected at the second electrode (for example, detection of reduced disulfides by the catalytic oxidation of Hg). (E) LC-hv-EC. Photochemical reaction of an analyte to produce a species that is electrochemically active (for example, detection of nitro compounds and phenylalanine). Various combinations of these five arrangements have also been used. [Reprinted with permission from Bioanalytical Systems, Inc.]...

Applicability of the conductivity detector can be extended by chemical derivatization or by the use of postcolumn photochemical reactions [78]. The use of a photochemical reaction detector, also known as a photoconductivity detector, can also be very selective. Only certain organic compounds such as trinitroglycerin, chloramphenicol, and hydrochlorothiazide will undergo photolytic decomposition to produce ionic species. 

Electrochemical detection is inherently a chemical rather than a physical technique (such as ultraviolet, infrared, fluorescence, or refractive index). It is, therefore, not surprising to hnd that many imaginative postcolumn reactions have been coupled to LC-EC. These include photochemical reactions, enzymatic reactions, halogenation reactions, and Biuret reactions. In each case, the purpose is to enhance selectivity and therefore improve limits of detection. While simplicity is sacrihced with such schemes, there are many published methods that have been quite successful. 

Postcolumn detection by reaction uses a chemical or photochemical reaction to generate a detectable product species (P) from the analyte (A) by addition of a reagent or photons ... 

Many LC methods involving the use of UV [29,55-57,80,81,84,104,106,107], diode-array [30,52,60,65,67-69,72,73,108], photoconductivity [109], Fourier-transform infrared spectrometry [79], and, after postcolumn photochemical reaction, fluorescence [110-112], detectors have been developed for analyzing fungicides and herbicides in water samples. 

Hydrogen peroxide [26-33] and analytes converted to hydrogen peroxide via either an enzymatic [34-52] or a photochemical postcolumn reaction [53-55]... 

Postcolumn reactions in LCEC can take several forms, most of which are illustrated in Figure 27.18. These include transformation of the analyte by chemical, electrochemical, photochemical, and enzymatic methods. 

Fluorescence spectroscopy plays an important function in modern food analysis as can be seen from its wide use in the determination of numerous food components, contaminants, additives, and adulterants. This technique has made available very sensitive and selective methods that satisfy the requirements of food analysis, which are usually very complex, taking into account the large number of species to be determined, frequently at very low concentrations, and the wide variety of foodstuffs available. Initially, the use of fluorescence spectroscopy in food analysis was limited to the determination of species with intrinsic fluorescence (e.g., vitamins, aflatoxins, and some polycyclic aromatic hydrocarbons (PAHs)), but now it is widely applied to nonfluorescent species, using several physicochemical means such as chemical or photochemical derivatization reactions. Numerous techniques involve fluorescence detection in liquid chromatography (LC), frequently using pre- or postcolumn derivatization. In addition to conventional fluorime-try, which is commonly chosen for this purpose, other fluorimetric techniques such as laser-induced... 

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