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首页> 外文期刊>Journal of Oleo Science >Determination of sphingoid bases from hydrolyzed glucosylceramide in rice and wheat by online post-column high-performance liquid chromatography with O-phthalaldehyde derivatization
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Determination of sphingoid bases from hydrolyzed glucosylceramide in rice and wheat by online post-column high-performance liquid chromatography with O-phthalaldehyde derivatization

机译：在线柱后高效液相色谱-邻苯二甲醛衍生化法测定大米和小麦中水解的葡萄糖基神经酰胺中的类神经鞘碱基

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摘要

We developed a determination method for sphingoid bases using online post-column high-performance liquid chromatography (HPLC) with O-phthalaldehyde (OPA) derivatization. Good separation was achieved using a reversed-phase column and eluting with 50% acetonitrile containing formic acid and heptafluorobutyric acid. Using these conditions, an excellent linearity (R~2 > 0.999) was achieved using standard solutions of sphinganine (dl8:0), sphingosine (d18:1~(4t)), 4-hydroxy-sphinganine (t18:0), glucosylsphingosine (glc-d18:1~(4t)), and galactosylsphingosine (gal-d18:1~(4t)). Plant glucosylceramides were hydrolyzed with 1 M aqueous HC1 in methanol for 18 h at 90℃, followed by extraction of sphingoid bases with diethyl ether in preparation for analysis using the proposed HPLC conditions. The glc-d18:1~(4t) standard was also hydrolyzed and analyzed by HPLC using the same procedure, and the d18:1~(4t) peak obtained from the hydrolyzed glc-d18:1~(4t) standard was used as a reference for calculation. We also confirmed the applicability of this method to the analysis of sphingoid bases in rice and wheat, obtaining relative standard deviations of 8.0% for rice and 4.6% for wheat. The recoveries of spiked rice and wheat samples were 104% and 106%, respectively. Our proposed method enables the straightforward determination of sphingoid bases without expensive facilities, employing fluorescence detection of OPA derivatives.
机译：我们开发了一种使用在线柱后高效液相色谱（HPLC）和邻苯二甲醛（OPA）衍生化方法测定鞘氨醇碱的方法。使用反相柱并用含有甲酸和七氟丁酸的50％乙腈洗脱可实现良好分离。在这些条件下，使用鞘氨醇（dl8：0），鞘氨醇（d18：1〜（4t）），4-羟基-鞘氨醇（t18：0），葡糖基鞘氨醇的标准溶液可获得极好的线性（R〜2> 0.999） （glc-d18：1〜（4t））和半乳糖基鞘氨醇（gal-d18：1〜（4t））。在90℃下，将植物葡萄糖基神经酰胺在甲醇中用1 M HCl水溶液水解18 h，然后用乙醚萃取鞘氨醇碱，以准备使用建议的HPLC条件进行分析。还使用相同的方法对glc-d18：1〜（4t）标样进行水解和HPLC分析，并使用从水解的glc-d18：1〜（4t）标样获得的d18：1〜（4t）峰作为计算参考。我们还证实了该方法在分析水稻和小麦中鞘氨醇碱中的适用性，相对标准偏差分别为水稻8.0％和小麦4.6％。加标大米和小麦样品的回收率分别为104％和106％。我们提出的方法可通过荧光检测OPA衍生物，直接测定鞘氨醇碱基，而无需昂贵的设备。

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  • 来源
    《Journal of Oleo Science》 |2012年第12期|681-688|共8页
  • 作者

    Hirofumi Goto; Keiko Nishikawa; Noriko Shionoya; Makoto Taniguchi; Tomoji Igarashi;

  • 作者单位

    Japan Food Research Laboratories, Nagoya Branch, 4-5-13 Osu, Naka-ku, Nagoya, Aichi 460-0011, JAPAN,Mimasaka University 50 Kitazono-cho, Tsuyama, Okayama 708-8511, JAPAN;

    Japan Food Research Laboratories, Nagoya Branch, 4-5-13 Osu, Naka-ku, Nagoya, Aichi 460-0011, JAPAN;

    Japan Food Research Laboratories, Nagoya Branch, 4-5-13 Osu, Naka-ku, Nagoya, Aichi 460-0011, JAPAN;

    Mimasaka University 50 Kitazono-cho, Tsuyama, Okayama 708-8511, JAPAN;

    Japan Food Research Laboratories, Nagoya Branch, 4-5-13 Osu, Naka-ku, Nagoya, Aichi 460-0011, JAPAN;

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  • 正文语种 eng
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  • 关键词

    sphingoid base; glucosylceramide; post-column HPLC; O-phthalaldehyde derivatization;

    机译：鞘基;葡萄糖基神经酰胺柱后HPLC;邻苯二甲醛衍生化;

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