Analytical methods cleanup/purification

Cleanup by solid-phase extraction has also been widely employed since it is a simple, fairly inexpensive, and easy-to-perform procedure for purification of the crude extract. The use of disposable solid-phase extraction columns is currently part of most, if not all, modern analytical methods for the determination of anthelminthics in biological matrices at residue levels. Both normal-phase columns based on silica (333-335, 340, 367, 372), alumina (346, 373-375), or aminopropyl (339, 365, 370) materials, and reversed-phase columns based on Ci8 (319, 323, 324, 328, 344, 346, 347, 349-351, 357-359, 364, 367) and cyclohexyl (329, 332, 360) sorbents have been described in analytical applications. 

By nature, analytical results are variable. The science of trace analysis (analysis at parts per million or below levels) is not as precise as most layman and many scientists view it to be (Rogers, 1986). Once devised, analytical methods are like life-forms, subject to evolution. Natural selection is mediated by analytical chemists, which ensures that only the fittest analytical mcthod,s survive. Therefore, an analytical method must be fit for purpose. In order to be able to determine trace levels of OPs and CMs in the environment and biological, food, and feed samples, it is necessary to follow a series of operations. Note that most of the advancements and improvements in the analytical methods have occurred by making necessary changes and improvements in various. steps used in analytical methods. The papers by Sawyer (1988) and Seiber (1988) clearly demonstrate this point. The various steps in an analytical method are (i) extraction of the sample, (ii) cleanup and purification... 

Two general approaches have been used to overcome matrix effects (1) partial purification of the analyte prior to analysis by immunoassay ( cleanup methods) and (2) the use of a matrix blank when preparing the calibration curve. Both options are widely used, but each has its individual limitations. 

Liquid chromatography cleanup on a LiChrosorb Diol column has been further proposed for the offline purification of chloramphenicol residues from bovine muscle and eggs (32). An online approach based on reversed-phase principles has also been described for isolation of chloramphenicol residues from swine kidney by an automated column switching system (63). Use of a protein exclusion column (Hisep) has been also suggested in an online trace-enrichment method for the determination of chloramphenicol in animal tissues (52). By employing a column-switching system, all chloramphenicol that eluted from the protein exclusion column was trapped at the entry of a 5 m Supelcosil LC-18 preconcentration column, to be subsequently back-flashed into the analytical column. 

Preliminary extraction of 5-HIAA may be used as an initial purification step before HPLC analysis. Organic solvents, anion-exchange resins, and other solid phase extraction procedures have aU been used. For many systems, direct injection of urine onto the analytical column is a common practice,and samples are often merely diluted with a buffer to protect the HPLC system from contamination. Methods that analyze 5-HIAA without prior sample cleanup rely on the selectivity of the HPLC separation combined with fluorescence or electrochemical detection to provide the requisite specificity. 

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