Sample preparation miniaturization

Specifically for triazines in water, multi-residue methods incorporating SPE and LC/MS/MS will soon be available that are capable of measuring numerous parent compounds and all their relevant degradates (including the hydroxytriazines) in one analysis. Continued increases in liquid chromatography/atmospheric pressure ionization tandem mass spectrometry (LC/API-MS/MS) sensitivity will lead to methods requiring no aqueous sample preparation at all, and portions of water samples will be injected directly into the LC column. The use of SPE and GC or LC coupled with MS and MS/MS systems will also be applied routinely to the analysis of more complex sample matrices such as soil and crop and animal tissues. However, the analyte(s) must first be removed from the sample matrix, and additional research is needed to develop more efficient extraction procedures. Increased selectivity during extraction also simplifies the sample purification requirements prior to injection. Certainly, miniaturization of all aspects of the analysis (sample extraction, purification, and instrumentation) will continue, and some of this may involve SEE, subcritical and microwave extraction, sonication, others or even combinations of these techniques for the initial isolation of the analyte(s) from the bulk of the sample matrix. 

During the last few years, miniaturization has become a dominant trend in the analysis of low-level contaminants in food and environmental samples. This has resulted in a significant reduction in the volume of hazardous and expensive solvents. Typical examples of miniaturization in sample preparation techniques are micro liquid/liquid extractions (in-vial) and solvent-free techniques such as solid-phase microextraction (SPME). Combined with state-of-the-art analytical instrumentation, this trend has resulted in faster analyses, higher sample throughputs and lower solvent consumption, whilst maintaining or even increasing assay sensitivity. 

HPLC is extremely useful in monitoring and optimizing industrial processes. Conventional process monitors measure only bulk properties, such as the temperature and pressure of a reactor, while HPLC permits continuous realtime monitoring of consumption of starting materials, product composition, and impurity profile. There are a number of new initiatives relevant to HPLC for process monitoring, including sample preparation, automation, miniaturization, and specialized detectors. 

Large volume injection Miniaturization of sample preparation... 

The use of robotics can be adopted also in sample preparation steps, in particular on-line SPE [7], This necessity is particular evident when small quantity of starting materials is available and the target molecules are present at low concentration levels. With the advent of miniaturization and automated procedures for samples handling, treatments and analysis, the lost of analytes due to a laboratory steps can be reduced. The reduction of analyte losses and the possibility to analyze even a total sample (no loss) leads to lower limits of detection (and consequently lower limits of quantification). Smaller volumes bring to obtain adequate sensitivity and selectivity for a large variety of compounds. In addition, on-line SPE requires low solvent consumption without the need to remove all residual water from cartridges, since elution solvents are compatible with the separation methods. 

Nanospray is a miniaturized version of electrospray. In the original setup of Wilm and Mann (8) it is utilized as an off-line technique using disposable, finely drawn (1 -gm tip), metallized glass capillaries to infuse samples at 10-30 nL/min flow rates. This allows more than 50 min analysis time with just a 1-pT sample. Due to the formation of much smaller droplets and the more effective ionization, there is often no need for LC separation, since the separation is accomplished in m/z or by MS/MS. However, limited reproducibility with respect to quantification and a more complex sample preparation can be seen as drawbacks. An on-line version for hyphenation with capillary and nano-LC as well as CE (slightly modified) is now commercially available. 

One drawback of capillary electrophoresis is the state of the capillary wall. Often, constituents of the buffer or analyte are absorbed on the sin-face, causing not only an irreproducible shift of EOF, but even the possibility of questionable binding isotherms. A lot of effort has gone into overcoming this problem. Capillaries with coated inner walls are now commercially available and capillary electrophoresis on chips of different materials is also under development now. Not only do these chips represent a miniaturized form of capillary electrophoresis, but this technique also enables the incorporation of such sample preparation steps as preconcentration and even PCR and immobilization of immunoreagents. It is not difficult to anticipate a very exciting development in this field, one with a high commercial impact. 

The use of small columns such as microbore liquid chromatographic columns, requiring smaller sample size, and computer-controlled solvent delivery and collection systems should lead to the development of fully integrated and automated cleanup systems. Small sample sizes facilitate miniaturization of sample preparation procedures, which in turn brings several benefits including reduced solvent and reagent consumption, reduced processing time, less demand for bench space, and ease of automation. 

Lab-on-a-chip is the miniaturization and integration of the complete set of devices used in separation science. It most important functions involve sample preparation, reactions, separations, and detection on a single chip. This arrangement was called p.-TAS by Manz in 1990 [1]. We call it nanoanalyses as the... 

Miniaturized LC/MS formats based on micromachined chip-based electrospray emitters and ionization sources on silicon (Schultz et al., 2000 Licklider et al., 2000 Ramsey and Ramsey 1997 Xue et al., 1997) and plastic (Vrouwe et al., 2000 Yuan and Shiea, 2001, Tang et al., 2001) microchips is a proactive approach for scale-down platforms. Various micromachining processes are used to fabricate these devices. These microanalytical technologies would create integrated sample preparation and LC/MS applications. The potential benefits of such a system include reduced consumption of sample/reagents, low cost, and disposability. 

As a corollary to this, more direct sample preparation procedures have been the pursuit of many scientists, who believe that miniaturization of analytical techniques can be a key solution to many of the unwanted drawbacks of LLE and SPE. Currently, several miniaturized extraction systems have been investigated, which are based primarily on utilizing downsized liquid, solid, or membrane extraction phases. 

Saito, Y., M. Nojiri, M. Imaizumi, Y. Nakao, Y. Morishima, H. Kanehara, H. Matsuura, K. Kotera, H. Wada, and K. Jinno. 2002. Polymer-coated synthetic fibers designed for miniaturized sample preparation process. J. Chromatogr. A 975 105-112. 

Saito, Y. and K. Jinno. 2003. Miniaturized sample preparation combined with liquid phase separations. J. Chromatogr. A 1000 53-67. 

Jinno, K., M. Ogawa, I. Ueta, and Y. Saito. 2007. Miniaturized sample preparation using a fiber-packed capillary as the medium. Trends Anal. Chem. 26 27-35. 

The microfluidic lab-on-a-chip has provided a platform to conduct chemical and biochemical analysis in a miniaturized format. Miniaturized analysis has various advantages such as fast analysis time, small reagent consumption, and less waste generation. Moreover, it has the capability of integration, coupling to sample preparation and further analysis. 

Other extensions that have been made include the construction of a biochip in which a reverse transcription PCRprocess can be carried out [384] and the integration of PCR with capillary electrophoresis [385], DNA microarray hybridization [386] and sample preparation [387,388]. The speed of analysis, the ease of integrating different functions and the relative low costs of micro-PCR production are important advantages of the miniaturized PCR technique that suggest that this bioorganic microreactor device will be thoroughly implemented in many analytical labs. 

Cell harvesters were developed to capture multiple samples of cells on membrane filters, wash away unincorporated isotopes, and prepare samples for liquid scintillation counting on special equipment developed to process and count multiple samples. Despite miniaturization and improvements in efficiency of this technique, the disadvantages of multiple liquid handling steps and increasing costs for disposal of radioactive waste materials severely limit its usefulness. Although specific applications require measuring DNA synthesis as a marker for cell proliferation, much better choices are available for detecting viable cell number for HTS. 

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