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Development and application of gradi...

Link, Jason C.

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Development and application of gradient ultrahigh pressure liquid chromatography for separations of complex biological mixtures.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Development and application of gradient ultrahigh pressure liquid chromatography for separations of complex biological mixtures./
Author:	Link, Jason C.
Description:	187 p.
Notes:	Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6353.
Contained By:	Dissertation Abstracts International65-12B.
Subject:	Chemistry, Analytical. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3156171
ISBN:	0496169661

Development and application of gradient ultrahigh pressure liquid chromatography for separations of complex biological mixtures.
Link, Jason C.

Development and application of gradient ultrahigh pressure liquid chromatography for separations of complex biological mixtures. - 187 p.

Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6353.

Thesis (Ph.D.)--The University of North Carolina at Chapel Hill, 2004.

The combination of reversed phase microbore liquid chromatography (LC) and electrospray mass spectrometry (ESI-MS) has become a commonly used technique for analyzing biological samples. Traditional reversed phase separations utilizing 5 mum particles, however, typically do not provide high enough separation efficiencies to completely resolve complex biological mixtures, such as peptide digests. Reducing the particle size to 1.0 mum greatly increases the separation efficiency at the expense of a high pressure requirement. Instruments developed and evaluated by our lab allow for the use of such small particles at pressures up to 100,000 psi.

ISBN: 0496169661Subjects--Topical Terms:

586156
Chemistry, Analytical.

Development and application of gradient ultrahigh pressure liquid chromatography for separations of complex biological mixtures.
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245 1 0 $a Development and application of gradient ultrahigh pressure liquid chromatography for separations of complex biological mixtures.
300 $a 187 p.
500 $a Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6353.
500 $a Adviser: James W. Jorgenson.
502 $a Thesis (Ph.D.)--The University of North Carolina at Chapel Hill, 2004.
520 $a The combination of reversed phase microbore liquid chromatography (LC) and electrospray mass spectrometry (ESI-MS) has become a commonly used technique for analyzing biological samples. Traditional reversed phase separations utilizing 5 mum particles, however, typically do not provide high enough separation efficiencies to completely resolve complex biological mixtures, such as peptide digests. Reducing the particle size to 1.0 mum greatly increases the separation efficiency at the expense of a high pressure requirement. Instruments developed and evaluated by our lab allow for the use of such small particles at pressures up to 100,000 psi.
520 $a Two new gradient instruments were evaluated for their performance in gradient ultrahigh pressure liquid chromatography (UHPLC) separations. One of the systems tested utilized a dual syringe design for gradient generation. The other system required a separate LC pump for generation of a temporarily stored, preloaded gradient. This preloaded gradient was then pump at ultrahigh pressures into the system. Characteristics of both systems, as well as their advantages and disadvantages versus our laboratory built instrument, are presented.
520 $a In addition to new instruments in UHPLC, new porous, reversed phase particles were evaluated for peptide separations. Porous particles offer increased surface area when compared to traditional, nonporous particles used in UHPLC. Porous particles of approximately 1.5 mum in diameter were analyzed for their loading capacity when separating peptides under gradient UHPLC conditions. The results presented show that the loading capacity increase is not as great when compared to loading capacity investigations under isocratic conditions.
520 $a Finally, the application of UHPLC for the separation of intact proteins was studied. Reversed phase protein separations have typically been plagued with poor peak shape and increased sample carry-over. An E. coli lysate was prepared for analysis using gradient UHPLC. Separations using gradient reversed phase UHPLC showed increased peak capacities and reduced ghosting when analyzing intact proteins. Furthermore, investigations using reversed phase gradient UHPLC in a two-dimensional configuration has shown to further increase the resolving power when analyzing intact protein samples.
590 $a School code: 0153.
650 4 $a Chemistry, Analytical. $3 586156
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710 2 0 $a The University of North Carolina at Chapel Hill. $3 1017449
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790 1 0 $a Jorgenson, James W., $e advisor
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791 $a Ph.D.
792 $a 2004
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3156171