Semi-micro HPLC of long chain fiatty acids with resonance raman detection.

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  • 共鳴Ｒａｍａｎ検出法による長鎖脂肪族カルボン酸のセミ・ミクロ高速液体クロマトグラフィー
  • キョウメイ ラマン ケンシュツホウ ニ ヨル チョウサ シボウゾク カルボンサ
  • 共鳴ラマン検出法による長鎖脂肪族カルボン酸のセミ・ミクロ高速液体クロマトグラフィー

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Abstract

A semi-micro HPLC of fatty acids coupled with a resonance Raman detection system is described. 4-Nitro-4'-N·methyl, N·β-hydroxy ethylaminoazobenzene (NHA) was synthesized as a labeling reagent for sensitive Raman detection. Fatty acids were activated with oxalyl chloride, followed by derivatizing with NHA reagent. Since these NHA esters in methanol exhibited their absorption maximum at 468 nm in visible region, resonance Raman effect could be obtained by 488.0 nm line of an Ar laser. HPLC separation of NHA esters of long chain fatty acids was accomplished on a μS-Finepak C18 (1.5φ×250 mm) and 95% acetonitrile in water as a mobile phase at a flow rate of 100 μl/min. Chromatograms were obtained by measuring the intensity of Raman scattering at 1343 cm-1 with 488.0 nm line of an Ar ion laser at 300 mW output. By this procedure, calibration curve (relative peak height for pentadecanoic acid as the internal standard) was linear in the concentration range of 5 ppm to 200 ppm for lauric acid. The relative standard deviation of relative peak height (n=5) was 4.4% at 20 ppm of lauric acid.

Journal

  • BUNSEKI KAGAKU

    BUNSEKI KAGAKU 37 (9), 456-459, 1988

    The Japan Society for Analytical Chemistry

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