title: Development of a Method for Measuring Phytanic Acid as a Lifestyle-related Disease Biomarker in Rat Serum Using Ultra-fast Liquid Chromatography-Ultraviolet Spectrophotometry Combined with a Modified 2-Nitrophenylhydrazine Derivatization Method.
abstract: A simple and rapid ultra-fast liquid chromatography-ultraviolet spectrophotometry (UFLC-UV) method combined with modified 2-nitrophenylhydrazine (2-NPH) derivatization was developed for determining phytanic acid (Phy) in rat serum. Serum Phy and heptadecanoic acid (the internal standard) were derivatized by 2-NPH at ambient temperature for 20 min and extracted in n-hexane. After extracting derivatized Phy (D-Phy) and derivatized IS from the reaction mixture, the extracts were separated with a YMC-Pack C8 column (150 × 3.0 mm i.d., S-3 μm) using an isocratic mobile phase comprised of acetonitrile:H2O (90:10; pH 4.4) at 0.5 mL/min. The detection wavelength was 228 nm. Linearity was observed over 1 - 20 μg/mL (r = 0.9997). The intra- and inter-day reproducibilities of D-Phy measurements were ≤13.0%. To our knowledge, this is the first report of the quantitative and qualitative measurement of serum Phy using 2-NPH derivatization and UFLC-UV. This method can be performed rapidly under mild conditions.
authors.affiliation: School of Pharmacy, Nihon University.
authors.forename: Aoi
authors.initials: A
authors.lastname: Miyamoto
authors.forename: Takahiko
authors.initials: T
authors.lastname: Aoyama
authors.forename: Masahiro
authors.initials: M
authors.lastname: Okamura
authors.forename: Noboru
authors.initials: N
authors.lastname: Fukuda
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ueno
authors.forename: Masanori
authors.initials: M
authors.lastname: Abe
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Matsumoto
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000097:blood / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004194:Disease; D008019:Life Style; D015394:Molecular Structure; D010659:Phenylhydrazines / Q000737:chemistry*; D010831:Phytanic Acid / Q000097:blood* / Q000737:chemistry; D051381:Rats; D013056:Spectrophotometry, Ultraviolet
chemical_list: D015415:Biomarkers; D010659:Phenylhydrazines; D010831:Phytanic Acid; C060574:2-nitrophenylhydrazine