Online preparation of complex biological samples prior To analysis by HPLC, LC/MS and/or protein sequencing
1995; Elsevier BV; Linguagem: Inglês
10.1016/s1080-8914(06)80035-9
ISSN1080-8914
Autores Tópico(s)Advanced Proteomics Techniques and Applications
ResumoComplex biological samples often require some degree of sample preparation prior to final analysis by HPLC, LC/MS, mass spectrometry, AAA, or protein sequencing due to sample matrix interferences. This chapter describes the online preparation of complex biological samples prior to analysis by HPLC, LC/MS and/or protein sequencing. On-line sample preparation techniques, such as the trap cartridge systems, offer distinct advantages of minimizing prep time and maximizing sample recovery over other conventional sample preparation protocols. Use of the RP salt removal/concentration trap cartridges provides a rapid means for volume reduction, salt elimination and/or buffer exchange, since the entire procedure takes place in less than 5 minutes. Removal of salts improves instrumental performance and data interpretation for mass spectrometry, AAA, and protein sequence analysis. Use of SDS removal trap cartridge eliminates interference with reversed-phase separations and avoids column fouling, which is commonly experienced when SDS is a part of the separation, while non-ionic detergent (NID) removal cartridges used in the multistep procedure offer efficient cleanup, in addition to concentration of samples. Future work is planned with these cartridges to see if they can be extended to other contaminants, such as PolyEthylene Glycol (PEG), and a trap cartridge for removal of non ionic detergents from peptide samples needs to be investigated.
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