Application of Second Order Derivative Spectroscopy in Determining Aromatic Amino Acid Content of Polypeptides in Size Exclusion HPLC with On-Line Photodiode Array Detection

1991 ◽  
Vol 29 (3) ◽  
pp. 91-97 ◽  
Author(s):  
D. E. H. Palladino ◽  
K. A. Cohen
Keyword(s):  
Amino Acid ◽  
Acid Content ◽  
Amino Acid Content ◽  
Size Exclusion ◽  
Photodiode Array Detection ◽  
Derivative Spectroscopy ◽  
On Line ◽  
Photodiode Array ◽  
Array Detection ◽  
Size Exclusion Hplc

scholarly journals Measurement of the acyl-CoA intermediates of β-oxidation by h.p.l.c. with on-line radiochemical and photodiode-array detection. Application to the study of [U-14C]hexadecanoate oxidation by intact rat liver mitochondria

1989 ◽  
Vol 262 (1) ◽  
pp. 261-269 ◽  
Author(s):  
N J Watmough ◽  
D M Turnbull ◽  
H S A Sherratt ◽  
K Bartlett
Keyword(s):  
Rat Liver ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Beta Oxidation ◽  
Reverse Phase ◽  
Photodiode Array Detection ◽  
Mitochondrial Fractions ◽  
On Line ◽  
Photodiode Array ◽  
Array Detection

The quantitative isolation of acyl-CoA esters of chain length C2-C17 from mitochondrial incubations and their analysis by reverse-phase radio-h.p.l.c. is described. Photodiode-array detection was used to characterize 2-enoyl-CoA esters. The chromatographic behaviour of all 27 intermediates of the beta-oxidation of hexadecanoyl-CoA is documented. Only C16, C14 and C12 intermediates were detected in uncoupled mitochondria oxidizing [U-14C]hexadecanoyl-CoA in the presence of fluorocitrate and carnitine, providing evidence for some organization of the enzymes of beta-oxidation [Garland, Shepherd & Yates (1965) Biochem. J. 97, 587-594; Sumegi & Srere (1984) J. Biol. Chem. 259, 8748-8752]. Rotenone increased concentrations of 3-hydroxyacyl-CoA and 2-enoyl-CoA esters and inhibited flux. These experiments provide the first direct unambiguous measurements of acyl-CoA esters in intact respiring rat liver mitochondrial fractions.

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