A new fluorometric method for the determination of pyridoxal 5′-phosphate
A new fluorometric method using semicarbazide for the determination of pyridoxal and pyridoxal 5′-phosphate (PLP) in whole blood, red cells and plasma has been developed. Semicarbazide breaks the Schiff base of PLP and proteins by “trans-Schiffization” reaction and forms semicarbazone of PLP. The se...
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| Published in | Biochimica et biophysica acta Vol. 304; no. 3; pp. 765 - 773 |
|---|---|
| Main Authors | , |
| Format | Journal Article |
| Language | English |
| Published |
Netherlands
Elsevier B.V
28.05.1973
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0304-4165 0006-3002 1872-8006 |
| DOI | 10.1016/0304-4165(73)90223-7 |
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| Abstract | A new fluorometric method using semicarbazide for the determination of pyridoxal and pyridoxal 5′-phosphate (PLP) in whole blood, red cells and plasma has been developed. Semicarbazide breaks the Schiff base of PLP and proteins by “trans-Schiffization” reaction and forms semicarbazone of PLP. The semicarbazone of PLP emits strongly at 460 nm when excited at 380 nm. Several metabolic intermediates were tested for the possible interference. Only pyridoxal was found to interfere. The interference can be corrected since pyridoxal emits at 380 nm when excited at 320 nm. Using this method we found that rabbit red cells
in vivo are freely permeable to PLP. |
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| AbstractList | A new fluorometric method using semicarbazide for the determination of pyridoxal and pyridoxal 5′-phosphate (PLP) in whole blood, red cells and plasma has been developed. Semicarbazide breaks the Schiff base of PLP and proteins by “trans-Schiffization” reaction and forms semicarbazone of PLP. The semicarbazone of PLP emits strongly at 460 nm when excited at 380 nm. Several metabolic intermediates were tested for the possible interference. Only pyridoxal was found to interfere. The interference can be corrected since pyridoxal emits at 380 nm when excited at 320 nm. Using this method we found that rabbit red cells
in vivo are freely permeable to PLP. |
| Author | Beutler, Ernest Srivastava, Satish K. |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/4726857$$D View this record in MEDLINE/PubMed |
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| Cites_doi | 10.1016/S0021-9258(18)81375-8 10.1021/bi00911a007 10.1016/0009-8981(68)90012-0 10.5925/jnsv1954.3.183 10.1093/ajcp/46.2_ts.282 10.1016/0009-8981(62)90167-5 10.1016/S0021-9258(18)57129-5 10.1016/0003-9861(60)90252-6 10.1016/0003-2697(64)90139-3 10.1016/0003-2697(69)90248-6 10.1136/jcp.23.3.232 |
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| SubjectTerms | Animals Cell Membrane Permeability Chemical Phenomena Chemistry Erythrocytes - cytology Erythrocytes - metabolism Humans Kinetics Methods Pyridoxal Pyridoxal Phosphate - blood Pyridoxal Phosphate - isolation & purification Pyridoxal Phosphate - metabolism Rabbits Semicarbazides Semicarbazones - chemical synthesis Spectrometry, Fluorescence Trichloroacetic Acid |
| Title | A new fluorometric method for the determination of pyridoxal 5′-phosphate |
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