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用 HPLC-DAD-APCI-MS 法测定蒲公英中类胡萝卜素的含量及柱色谱法制备。

Determination of carotenoids in Taraxacum formosanum by HPLC-DAD-APCI-MS and preparation by column chromatography.

机构信息

Department of Food Science, Fu Jen University, Taipei 242, Taiwan.

出版信息

J Pharm Biomed Anal. 2012 Jul;66:144-53. doi: 10.1016/j.jpba.2012.03.035. Epub 2012 Mar 28.

Abstract

The objectives of this study were to determine the variety and content of carotenoids in Taraxacum formosanum, a traditional Chinese herb possessing vital biological activities, by developing an HPLC-DAD-APCI-MS method and a preparative column chromatographic method for carotenoid isolation. A total of 25 carotenoids were resolved within 66 min by employing a YMC C30 column and a gradient mobile phase of methanol-acetonitrile-water (79:14:7, v/v/v) and methylene chloride (100%) with flow rate at 1.0 mL/min and detection at 450 nm. All-trans-canthaxanthin was shown to be an appropriate internal standard for quantitation, with all-trans-β-carotene and its cis isomers present in largest amount (413.6 μg/g), followed by all-trans-violoxanthin and its cis isomers (209.5 μg/g), all-trans-lutein and its cis isomers (212.4 μg/g), all-trans-neoxanthin and its cis isomers (134.6 μg/g), antheraxanthin (16.5 μg/g), all-trans-β-cryptoxanthin and its cis isomers (5.8 μg/g), all-trans-zeaxanthin (3.6 μg/g) and neochrome (0.1 μg/g). For preparative chromatography, with a glass column containing 52 g of magnesium oxide-diatomaceous earth (1:3, w/w) as adsorbent, the carotenoid fraction was eluted with 300 mL of ethyl acetate with flow rate at 10 mL/min. Some more epoxides and cis isomers of carotenoids were generated during preparative column chromatography. Nevertheless, the carotenoids isolated from T. formosanum may be used as raw material for possible production of health food in the future.

摘要

本研究旨在通过建立高效液相色谱-二极管阵列检测-大气压化学电离-质谱联用(HPLC-DAD-APCI-MS)方法和制备柱色谱分离方法,对具有重要生物活性的传统中药蒲公英(Taraxacum formosanum)中的类胡萝卜素种类和含量进行研究。采用 YMC C30 柱和甲醇-乙腈-水(79:14:7,v/v/v)和二氯甲烷(100%)的梯度流动相,以 1.0 mL/min 的流速和 450nm 的检测波长,在 66min 内可分离出 25 种类胡萝卜素。全反式角黄素被证明是定量分析的合适内标,全反式-β-胡萝卜素及其顺式异构体的含量最高(413.6μg/g),其次是全反式 violaxanthin 及其顺式异构体(209.5μg/g),全反式叶黄素及其顺式异构体(212.4μg/g),全反式 neoxanthin 及其顺式异构体(134.6μg/g),花药黄质(16.5μg/g),全反式-β-隐黄质及其顺式异构体(5.8μg/g),全反式玉米黄质(3.6μg/g)和新黄质(0.1μg/g)。对于制备性色谱分离,采用含有 52g 氧化镁-硅藻土(1:3,w/w)的玻璃柱作为吸附剂,以 10mL/min 的流速用 300mL 乙酸乙酯洗脱类胡萝卜素馏分。在制备柱色谱分离过程中会产生一些环氧化物和类胡萝卜素的顺式异构体。然而,从蒲公英中分离出的类胡萝卜素可作为未来生产保健食品的原料。

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