A new fluorometric method for the determination of pyridoxal 5′-phosphate

Satish K. Srivastava; Ernest Beutler

doi:10.1016/0304-4165(73)90223-7

A new fluorometric method for the determination of pyridoxal 5′-phosphate

Satish K. Srivastava, Ernest Beutler

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72 Scopus citations

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.

Original language	English (US)
Pages (from-to)	765-773
Number of pages	9
Journal	BBA - General Subjects
Volume	304
Issue number	3
DOIs	https://doi.org/10.1016/0304-4165(73)90223-7
State	Published - May 28 1973
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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title = "A new fluorometric method for the determination of pyridoxal 5′-phosphate",

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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T1 - A new fluorometric method for the determination of pyridoxal 5′-phosphate

AU - Srivastava, Satish K.

AU - Beutler, Ernest

PY - 1973/5/28

Y1 - 1973/5/28

N2 - 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.

AB - 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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A new fluorometric method for the determination of pyridoxal 5′-phosphate

Abstract

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