Analytical post column detection

Both on-column and post-column detection schemes have been developed for radionuclide detection for CE. The most common type used is an on-column configuration, which yields detection limits in the 10 M range for isotopes such as P. Isotachophoretic separations of C were among the first examples of online capillary radionuclide detection, performed by Kaniansky et al. The associated instmment uses 300-p,m ID fluorinated ethylene-propylene copolymer capillary tubing, and the separation eluent flows directly into a plastic scintillator cell between two PMTs. The scintillation events are detected coincidentally between the two PMTs, such that only if both PMTs receive an input within a short time will they register the count as signal. This kind of coincidence detection ensures that nonscintillation photons that come from outside the detection cell and only hit one PMT are not counted. This system exhibits a detection limit of 16 Bq for analytes, with a detector efficiency of 13-15%. 

In the second case monitoring is performed on the column outlet with the main objective of identifying when the individual fractions elute. Post column detection is a very importemt part of analytical chromatographs where sophisticated diode array detectors may be utilized, and base line correction and peak area calculations are used for concentration determination of individual fractions. In contrast, in process chromatography, post column detection tends to be used purely qualitatively and may even be absent. 

Chromatographic methods including thin-layer, hplc, and gc methods have been developed. In addition to developments ia the types of columns and eluents for hplc appHcations, a significant amount of work has been done ia the kiads of detectioa methods for the vitamin. These detectioa methods iaclude direct detectioa by uv, fluoresceace after post-column reduction of the quiaone to the hydroquinone, and electrochemical detection. Quantitative gc methods have been developed for the vitamin but have found limited appHcations. However, gc methods coupled with highly sensitive detection methods such as gc/ms do represent a powerful analytical tool (20). 

In liquid chromatography, in contrast to gas chromatography [see Section 9.2(2)], derivatives are almost invariably prepared to enhance the response of a particular detector to the substance of analytical interest. For example, with compounds lacking an ultraviolet chromophore in the 254 nm region but having a reactive functional group, derivatisation provides a means of introducing into the molecule a chromophore suitable for its detection. Derivative preparation can be carried out either prior to the separation (pre-column derivatisation) or afterwards (post-column derivatisation). The most commonly used techniques are pre-column off-line and post-column on-line derivatisation. 

At this point, the anaiyte may not be amenabie to UV, FL, or EC detection. In this case, the best course of action may be to choose LC/MS (see Section 4.2). However, one other option is to use a pre- " or post-coiumn derivatization step to increase the detectabiiity of the anaiyte with respect to FL or UV. Fluorescent or UV labels are available for carboxylic acids," amines, phenols, and thiols. The decision to use pre- or post-column derivatization is predicated upon the functionality of the analyte available for derivatization and the rate and extent of the reaction between each derivatizing agent and the analyte. 

The increased use of IV-methyl carbamate insecticides in agriculture demands the development of selective and sensitive analytical procedures to determine trace level residues of these compounds in crops and other food products. HPLC is the technique most widely used to circumvent heat sensitivity of these pesticides. However, HPLC with UV detection lacks the selectivity and sensitivity needed for their analysis. In the late 1970s and early 1980s, HPLC using post-column hydrolysis and derivatization was developed and refined with fluorescence detection to overcome these problems. The technique relies on the post-column hydrolysis of the carbamate moiety to methylamine with subsequent derivatization to a fluorescent isoindole product. This technique is currently the most widely used HPLC method for the determination of carbamates in water" and in fruits and vegetables." " ... 

Lovdahl, M. J. and Pietrzyk, D. J., Anion-exchange separation and determination of bisphosphonates and related analytes by post-column indirect fluorescence detection,. Chromatogr A, 850, 143, 1999. 

Aromatic and sulfur-containing amino acids were separated by HPLC, and subjected to post-column UV irradiation before electrochemical detection with GCE vs AgCl/Ag electrodes. The analytes showed different behavior during lamp off and on periods. Thus, for example, tyrosine (46) and tryptophan (47) showed inherent electrochemical response at +0.80 V, but none at +0.60 V however, on turning on the UV lamp they showed sensitive response at both potentials126. 

RP-HPLC-vis measurements were performed in an ODS column (250 X 2 mm i.d. particle size 5 jum). A 10 X 2 mm column filled with lead(IV) oxide was employed for the post-column oxidation of the analytes. The method of post-column oxidation allowed the simultaneous detection of each dye and metabolite in the visible range. Dyes were eluted with a two-phase step gradient programme mobile phase 1 (0.05 M aqueous ammonium... 

A fraction collector and a post-column derivatization system were included (Figure 2.1) for a comprehensive and multi-purpose instrument. However, the fraction collector is needed only when collecting components from the effluent, and is generally not included in an analytical system. The post-column derivatization system is connected only when required for the selective and sensitive detection of specially targeted compounds. Usually, most compounds are directly detected by an on-line spectroscopic or other detector. 

Like dissolves like is the basic concept for the selection of solvents in the eluent for liquid chromatography. Controlling the solubility of analytes is the key to success. If the selected solvent or mixture of solvents does not interfere with detection, it is a good eluent. The selection of a suitable solvent for low-wavelength absorption detection and post-column derivatization detection is important to obtain highly sensitive detection. The selection of a volatile solvent is the key for preparative-scale liquid chromatography and for mass spectro-metric detection. 

It is used in IC systems when the amperometric process confers selectivity to the determination of the analytes. The operative modes employed in the amperometric techniques for detection in flow systems include those at (1) constant potential, where the current is measured in continuous mode, (2) at pulsed potential with sampling of the current at dehned periods of time (pulsed amperometry, PAD), or (3) at pulsed potential with integration of the current at defined periods of time (integrated pulsed amperometry, IPAD). Amperometric techniques are successfully employed for the determination of carbohydrates, catecholamines, phenols, cyanide, iodide, amines, etc., even if, for optimal detection, it is often required to change the mobile-phase conditions. This is the case of the detection of biogenic amines separated by cation-exchange in acidic eluent and detected by IPAD at the Au electrode after the post-column addition of a pH modiher (NaOH) [262]. 

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