Monolithic columns applications

Monolithic columns, formed from the co-polymerization of divinylbenzene and vinylbenzyl chloride or styrene, were observed to be resistant to bubble formation.11 Application of pressure in electrochromatography, discussed below, also reduces bubble formation. A massively parallel detector capable of scanning up to 1000 capillaries using planar confocal fluorescence has been used for DNA sequencing.1213 Recovery of fluorescence following pho-tobleaching has been used to measure DNA mobility in agarose gel.14... 

Rieux, L., Niederlander, H., Verpoorte, E., Bischoff, R. (2005). Silica monolithic columns Synthesis, characterisation and applications to the analysis of biological molecules. J. Sep. Sci. 28, 1628-1641. 

Obviously, the monolithic material may serve its purpose only if provided with a suitable surface chemistry, which depends on the desired application. For example, hydrophobic moieties are required for reversed phase chromatography, ionizable groups must be present for separation in the ion-exchange mode, and chiral functionalities are the prerequisite for enantioselective separations. Several methods can be used to prepare monolithic columns with a wide variety of surface chemistries. 

The newly developed monolithic-type column has also found application in the HPLC determination of wine phenolics. Red wine samples were filtered and injected into the column without any other pretreatment. Separations were performed in an ODS monolithic column (100 X 4.6mm i.d.) at 30 1°C. Solvent A was methanol-double-distilled water (2.5 97.5, v/v) at pH 3 with H3P04, and solvent B consisted of methanol-double-distilled water (50 50, v/v) at pH 3 with H3P04. Conditions of gradient elution were as follows 0-lQmin 100... 

Monolithic columns have been available for many years and continue to be a part of the high-throughput discussion. Chapter 4 provides a brief description of the structure of monolithic columns as well as a discussion of some of their advantages and disadvantages. In this section we will briefly discuss some of the applications developed as related to fast analyses and what chemists are hoping to achieve by using monoliths. 

CEC is a miniaturized separation technique that combines capabilities of both interactive chromatography and CE. In Chapter 17, the theory of CEC and the factors affecting separation, such as the stationary phase and mobile phase, are discussed. The chapter focuses on the preparation of various types of columns used in CEC and describes the progress made in the development of open-tubular, particle-packed, and monolithic columns. The detection techniques in CEC, such as traditional UV detection and improvements made by coupling with more sensitive detectors like mass spectrometry (MS), are also described. Furthermore, some of the applications of CEC in the analysis of pharmaceuticals and biotechnology products are provided. 

The emerging of CEC and the increased scientific work on the preparation of different phases, characterization, and applications of the CEC columns have given much credence to their future potentials in microseparations. The fabrication and availability of different phases for analysis with both particle-packed and monolithic columns give the technique a great future. This is because a variety of mechanisms can be exploited in the analysis and separation of compounds that could otherwise be difficult to analyze with HPLC or CE alone. The ease of coupling CEC to sensitive detectors such as mass spectrometers for enhanced sensitivity, structural elucidation, and characterization bestows the technique with great versatility. 

Even if MIP and BET are widely accepted regarding the characterization of HPLC stationary phases, they are only applicable to the samples in the dry state. In order to investigate the impact of polymerization time on the porous properties of wet monolithic columns, ISEC measurements of 200 jm I.D. poly(p-methylstyrene-co-l,2-bis(vinylphenyl)ethane) (MS/BVPE) capillary columns (prepared using a total polymerization time ranging from 45 min to 24 h) have been additionally evaluated (see Table 1.2 for a summary of determined e values). On a stepwise decrease in the time down to 45 min, the total porosity (St) is systematically increasing to about 30% in total (62.8% for 24 h and 97.2% for 45 min). This is caused by a simultaneous increase in the fraction of interparticulate porosity (e. ) as well as the fraction of pores (Cp). The ISEC measurements are in agreement with those of the MIP as well as BET analyses, as an increase in should be reflected in an increase in 8p and as the relative increase in the total porosity (caused by decreasing the polymerization time... 

Methacrylate monoliths have been fabricated by free radical polymerization of a number of different methacrylate monomers and cross-linkers [107,141-163], whose combination allowed the creation of monolithic columns with different chemical properties (RP [149-154], HIC [158], and HILIC [163]) and functionalities (lEX [141-153,161,162], IMAC [143], and bioreactors [159,160]). Unlike the fabrication of styrene monoliths, the copolymerization of methacrylate building blocks can be accomplished by thermal [141-148], photochemical [149-151,155,156], as well as chemical [154] initiation. In addition to HPLC, monolithic methacrylate supports have been subjected to numerous CEC applications [146-148,151]. Acrylate monoliths have been prepared by free radical polymerization of various acrylate monomers and cross-linkers [164-172]. Comparable to monolithic methacrylate supports, chemical [170], photochemical [164,169], as well as thermal [165-168,171,172] initiation techniques have been employed for fabrication. The application of acrylate polymer columns, however, is more focused on CEC than HPLC. 

---