Methods in Enzymology Vol.280 Vitamins and Coenzymes, Part J

Discussion in 'Methods in Enzymology Book Series' started by lebinhanseo, Aug 1, 2016.

  1. lebinhanseo

    lebinhanseo New Member

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    This chapter determines sample preparation and analytical techniques appropriate for the determination of all forms of vitamin B6 in a biological sample. For the determination of all B6, compounds except pyridoxic acid 5'-phosphate (PIC-P), the fluorescence spectrophotometer is set at an emission wavelength of 390 nm with an excitation wavelength of 305 nm. For the determination of PIC-P, which is prepared from pyridoxal 5'-phosphate (PLP) by KCN treatment, the fluorescence measurement is carried out at an emission wavelength of 420 nm with an excitation wavelength of 320 nm. An isocratic elution system is adopted to quantitate B6 compounds fluorometrically, because the fluorescence quantum yield of a compound drastically changed, depending on the slight change in ionic strength and pH of the solvent. Two isocratic solvent systems are used for the determination of naturally occurring pyridoxal derivatives, in which most pyridoxal derivatives can be analyzed with nearly the same sensitivity.
    • Series: Methods in Enzymology (Book 280)
    • Hardcover: 496 pages
    • Publisher: Academic Press;
    • Language: English
    • ISBN-10: 0121821811
    • ISBN-13: 978-0121821814
    • Product Dimensions: 6 x 1.1 x 9 inches
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