Separation of amino acid enantiomers and chiral amines using precolumn derivatization with (+)-1-(9-fluorenyl)ethyl chloroformate and reversed-phase liquid chromatography

1987 ◽  
Vol 59 (8) ◽  
pp. 1191-1195 ◽  
Author(s):  
Stefan. Einarsson ◽  
Bjoern. Josefsson ◽  
Per. Moeller ◽  
Domingo. Sanchez
Keyword(s):  
Liquid Chromatography ◽  
Amino Acid ◽  
Reversed Phase ◽  
Ethyl Chloroformate ◽  
Chiral Amines ◽  
Precolumn Derivatization ◽  
Reversed Phase Liquid Chromatography ◽  
Amino Acid Enantiomers ◽  
Phase Liquid

scholarly journals Isolation and analysis of human parathyrin in parathyroid tissue and plasma Use of reversed-phase liquid chromatography

1981 ◽  
Vol 197 (2) ◽  
pp. 391-400 ◽  
Author(s):  
H P J Bennett ◽  
S Solomon ◽  
D Goltzman
Keyword(s):  
Liquid Chromatography ◽  
Amino Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Parathyroid Tissue ◽  
Reversed Phase ◽  
Resolving Power ◽  
Molecular Forms ◽  
Reversed Phase Liquid Chromatography ◽  
Phase Liquid

Reversed-phase liquid chromatography techniques have been used to extract and purify human parathyrin from parathyroid adenomas and to analyse the circulating forms of human parathyrin in plasma. Both the supernatant from tissue homogenates, and plasma were extracted with octadecylsilyl-silica (ODS-silica) in a batch procedure. Extracts were subjected to reversed-phase high-pressure liquid chromatography (h.p.l.c.) employing solvent systems composed of aqueous acetonitrile containing trifluoroacetic acid or heptafluorobutyric acid as hydrophobic ion-pairing reagents. The volatile solvents facilitated the radioimmunoassay, bioassay in vitro and amino acid analysis of column fractions and permitted monitoring for u.v. absorbance at 210nm. Isolated glandular parathyrin was found to be homogeneous by sodium dodecyl sulphate/urea/polyacrylamide-gel electrophoresis, to have an amino acid composition conforming to that of human parathyrin-(1-84)-tetraoctacontapeptide and to be bioactive in both renal adenylate cyclase and cytochemical bioassays. ODS-silica extraction permitted examination of large plasms samples by reversed-phase h.p.l.c., facilitating the resolution of the various circulating molecular forms of parathyrin according to their hydrophobic character. Because of its rapidity, excellent recovery and high resolving power, the methodology utilized is uniquely suited to the purification and analysis of parathyrin in tissues and body fluids.

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