Methods in Enzymology Vol.189 Retinoids, Part A: Molecular and Metabolic Aspects

This chapter describes the chemical synthesis of retinyl β-glucuronide (ROG) and retinoyl β-glucuronide (RAG). It discusses the extraction of ROG and RAG from human serum. The chapter discusses the analysis of ROG and Rag by high-performance liquid chromatography (HPLC) and characterization by mass spectrometry (MS). For the estimation of endogenous retinoid glucuronides and retinoic acid, but not retinol, in serum, an isocratic reversed-phase HPLC system is preferred. The isocratic method gives a stable baseline and facilitates accurate quantitation of these minor retinoids in serum. It is not uncommon that two compounds with different structures coelute during HPLC. Therefore, besides comparing the retention time of the compound under investigation with a standard compound, often it is necessary to confirm the identity of the compound by determining the mass spectrum. A satisfactory mass spectrum of methylated retinoic acid is obtained in either the electron-impact (EI) mode or chemical ionization (CI) mode using an appropriate reagent gas.

• Series: Methods in Enzymology (Book 189)
• Hardcover: 583 pages
• Publisher: Academic Press;
  
• Language: English
• ISBN-10: 0121820904
• ISBN-13: 978-0121820909
• Product Dimensions: 1.2 x 6.2 x 9 inches

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