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. 2015 Sep 30;5(4):2247–2337. doi: 10.3390/biom5042247

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© 2015 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

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Mass spectrum of dihydroxynonene urethane (HPLC plus MS) with fluorimetric detection. The preparation of the reduced HNE product 1,4-DHN was carried out in the following manner: reduction of HNE with sodium boronate; extraction with dichloromethane; drying by means of sodium sulphate; purification by TLC separation, detection with 10% phosphomolybdenic acid dissolved in ethanol, extraction from TLC plates with CH₂Cl₂. Identification of 1,4-DHN: Siliylation; GC (85 °C, with 10 °/min up to 150 °C, 50 m SE30 column—i.d. 0.32 mm, FID); formation of urethane derivative (1,4-dinaphthyl-2,3-nonenylurethane); (305, 350 nm). Preparation of radioactively labelled 1,4-DHN: Reduction of radioactively labelled HNE with sodium boronate; extraction with dichloromethane; purification by means of TLC and elution on Bond Elut C18 columns (35% methanol/water, v/v). Chemical ionization (CH₄). Peaks: m/e 144 Ar-NH₃⁺, m/e 187 Ar-NH-COOH, m/e 123 (M + 1)-(2 × 187); (M + 1) = 497 m/e [301].