Sorbent selectivity, HPLC development

An on-line concentration, isolation, and Hquid chromatographic separation method for the analysis of trace organics in natural waters has been described (63). Concentration and isolation are accompHshed with two precolumns connected in series the first acts as a filter for removal of interferences the second actually concentrates target solutes. The technique is appHcable even if no selective sorbent is available for the specific analyte of interest. Detection limits of less than 0.1 ppb were achieved for polar herbicides (qv) in the chlorotriazine and phenylurea classes. A novel method for deterrnination of tetracyclines in animal tissues and fluids was developed with sample extraction and cleanup based on tendency of tetracyclines to chelate with divalent metal ions (64). The metal chelate affinity precolumn was connected on-line to reversed-phase hplc column, and detection limits for several different tetracyclines in a variety of matrices were in the 10—50 ppb range. 

Selected data published by Patsias and Papadopoulou-Mourkidou [114] illustrate sorption s dependence on sample volume (Figure 2.36). Their research pursues development of an automated online SPE-HPLC methodology for analysis of substituted anilines and phenols. Recovery (%) was measured for numerous compounds on various polymeric sorbents, but the only data presented here are those in which a styrene-divinylbenzene polymeric sorbent was used for analysis of aniline, phenol, 4-nitroaniline, and 4-nitrophenol. Aqueous sample volumes of 5, 10, 25, 50, 75, 100, 125, and 150 mL were acidified to pH 3 before SPE. 

The selection of starting conditions is the first step in HPLC method development. Instead of starting with arbitrary conditions or conditions derived from the chromatographer s experience or intuition, the software can predict suitable initial conditions for reversed-phase methods from analyte structures and properties of the sorbent/solvent system [1-3]. If structures are known, the theoretical approach has the potential to save time and effort, since in this way the experimental method development process will start under the theoretically predicted optimum conditions. 

Eurther improvements can be expected on each of the different steps of isolation, cleanup, chromatography, and detection systems for determination of VK in biological samples. TLC and direct-phase HPLC as chromatographic methods will be reduced to a minimum. Development of new materials for solid phase extraction (e.g., Oasis, a new polymeric SPE sorbent) and improved materials for reversed-phase HPLC, including smaller, narrow-bore or capillary columns, should bring advantage to RP-HPLC, especially in connection with MS detection, but also with fluorescence and electrochemical detection and help to obtain smaller sample volumes and/or shorter analysis times. However, the greatest advance, in our opinion, will be in the field of detection instruments—in particular, in mass selective detection instruments. 

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