Protein high pressure liquid chromatography

Biochemical analyses of 6-OHDA-injected animals revealed a 93 percent depletion of dopamine. The tissue was assayed using electrochemical detection following separation by high-pressure liquid chromatography (Felice et al. 1978). recorded as ng/mg protein in the nucleus accumbens and compared to control rats with sham lesions (sham=65.5 4.4, lesion=4.9 1.5 t(39)=23.4). A lesion was defined as complete if 75 percent or more of the dopamine was determined to be depleted from the nucleus accumbens compared to mean sham group values. 

The introduction and eventual commercialization of matrix-assisted laser desorption/ionization (MALDI) and electrospray (ESI) allowed biomarker status to be extended to proteins in 1996.15"17 With a few exceptions, ESI has been used in conjunction with extractions and high-pressure liquid chromatography (UPLC) interfaced with mass spectrometry. MALDI, on the other hand, has been widely adapted for rapid analysis of intact organisms, supported by bioinformatics.1819... 

Isolation of individual amino acids started about 1820 by 1904 all of the naturally occurring amino acids in proteins had been isolated except methionine (Mueller, 1922) and threonine (Rose, 1937). One of the earliest methods for the separation of amino acids was through the differential volatility of their methyl or ethyl esters (Emil Fischer, 1901). This approach led to the discovery of valine, proline, and hydroxyproline. [In the 1970s Fischer s method was modified for microanalysis of proteins, separating the amino acid esters by gas phase chromatography. Separation is now usually performed by hplc (high pressure liquid chromatography).]... 

G Seipke, H Mtillner, U Grau. High-pressure liquid chromatography (HPLC) of proteins. Angew Chem Int Ed Engl 25 535-552, 1986. 

AF Groen, R van der Vegt, MAJS van Boekel, OLAM de Rouw, H Vos. Case study on individual animal variation in milk protein composition as estimated by high-pressure liquid chromatography. Neth Milk Dairy J 48 201-212, 1994. 

Heme proteins 373 Heparin 803, 819, e9 binding site 805 Hepatitis C virus 381, 384, 391 Herbal extracts e277, e279 Heterogeneous catalysis 932 immunoassays 924 Hexadecanethiol 243 Hg 521, 523-524, 526 Hg-Au amalgam 236 High-pressure liquid chromatography (HPLC) 212... 

The determination (3) of physical and chemical properties such as solubility, stability, pKa, glassbinding and protein-binding of A -tetrahydrocannabinol 1 and correlated congeners were only possible after the development of new high pressure liquid chromatography (HPLC) techniques and the modification of gas liquid chromatography (GLC) technologies. 

The process of separation and quantitation of amino acids has been automated. In one automated method, a single cation exchange resin column separates all the amino acids in the protein hydrolysate. The analyzer is capable of detecting as little as 1-2 nmol of an amino acid and a complete analysis can be obtained in about 4 hours. In newer procedures, the complete analysis can be performed in about Ihour and permit detection of as little as 1-2 nmol of an amino acid. Picomole amounts of amino acids can be determined when the separated amino acids are coupled to fluorescent reagents such as o-phthalaldehyde. Amino acid separation and quantitation can also be accomplished by reverse-phase high-pressure liquid chromatography of amino acid derivatives—a rapid and sensitive procedure. 

Figure 3.6 High-pressure liquid chromatography (HPLC). Gel filtration by HPLC clearly defines the individual proteins because of its greater resolving power (1) thyroglobulin (669 kd), (2) catalase (232 kd), (3) bovine serum albumin (67 kd), (4) ovalbumin (43 kd), and (5) ribonuclease (13.4 kd). [After K. J. Wilson and T, D. Schlabach. In Current Protocols in Molecular Biology, vol. 2, suppl. 41, F. M. Ausubel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith, and K. Struhl. Eds. (Wiley, 1998), p, 10.14.1.]...

H2. Hancock, W. S., Bishop, C. A., Prestidge, R. L., Harding, D. R. K., and Hearn, M. T. W., Reversed-phase high-pressure liquid chromatography of peptides and proteins with ion-pairing reagents. Science 200, 1168-1170 (1978). 

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