食品科学

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高效液相色谱-串联质谱法测定各生长期紫苏中酚类物质的含量

艾鑫卫,胡思平,龚姮姮,黄文邺,杨洁球,王 力,廖照江   

  1. 1.三峡大学生物与制药学院,湖北 宜昌 443000;2.宜昌人福口服固体制剂厂,湖北 宜昌 443000
  • 出版日期:2016-09-25 发布日期:2016-09-26
  • 通讯作者: 廖照江
  • 基金资助:

    三峡大学科学基金项目(KJ2013B070);宜昌市科技局自然基础科学研究与应用专项(A15-302-a05)

Quantitation of Phenolic Compounds in Perilla frutescens at Different Growing Stages Using HPLC-MS/MS

AI Xinwei, HU Siping, GONG Hengheng, HUANG Wenye, YANG Jieqiu, WANG Li, LIAO Zhaojiang   

  1. 1. College of Biological and Pharmaceutical, China Three Gorges University, Yichang 443000, China;
    2. Yichang Humanwell Oral Solid Dosage Plant, Yichang 443000, China
  • Online:2016-09-25 Published:2016-09-26
  • Contact: LIAO Zhaojiang

摘要:

目的:建立一种简单、快速、准确的高效液相色谱-串联质谱法,同时分离与检测紫苏叶、茎和根中酚类化合物含量,探讨紫苏各部位酚类物质的累积规律,为确定紫苏各部位最佳采收时期提供依据。方法:对紫苏进行人工栽培,样品分期采集,采集后的样品经乙醇提取,采用高效液相色谱-串联质谱法进行分析,对影响色谱、质谱的重要参数进行优化,通过对照标准物质的保留时间及质谱分析,确定相应的化合物;通过标准曲线法定量,以其中13 种酚类化合物为主要参考指标,观察它们在各部位的动态含量变化。结果:鉴定出化合物:咖啡酸、咖啡酸-3-O-葡萄糖苷、迷迭香酸、迷迭香酸-3-O-葡萄糖苷、木犀草素、木犀草素-7-O-葡萄糖苷、木犀草素-7-O-葡萄糖苷酸、木犀草素-7-O-二葡萄糖苷酸、芹菜素、芹菜素-7-咖啡酰基葡萄糖苷、芹菜素-7-O-二葡萄糖苷、野黄岑素-7-O-二葡萄糖苷、迷迭香酸甲酯,以上化合物中酚酸及其糖苷化合物含量呈先降低,至结果期急剧上升达到最大,黄酮及其糖苷化合物含量则递增至结果期达到最大。其中鉴定出迷迭香酸和木犀草素-7-O-葡萄糖苷酸分别在结果期达到最大,含量分别为21.41 mg/g和19.89 mg/g,结果期叶部位总酚类含量达到最大,含量为47.50 mg/g。结论:通过对紫苏各个部位不同采收时期黄酮、黄酮糖基化、酚酸的动态变化、总含量变化和部位成分分布的研究,对紫苏酚类资源的利用提供参考依据。

关键词: 紫苏, 酚类物质, 高效液相色谱-串联质谱法, 动态积累, 最佳采收时期

Abstract:

Purpose: To establish a simple, rapid, accurate high performance liquid chromatography-tandem mass
spectrometry (HPLC-MS/MS) for the simultaneous separation and determination of phenolic compounds in leaves, stems
and roots of Perilla frutescens and further to examine the accumulation patterns of phenolics in different plant parts during
the growing period for the purpose of determining the optimal harvest time for Perilla frutescens. Methods: Samples
of cultivated Perilla frutescens were collected at three different growing stages and extracted with ethanol for HPLCMS
analysis. The important chromatographic and mass spectrometric parameters were optimized. The identification of
phenolic compounds was done by retention time comparison with reference standards and quantitation was performed using
calibration curves. Moreover, changes in the contents of 13 major phenolic compounds in various parts of Perilla frutescens
at different growing stages were observed. Results: The following compounds were identified: caffeic acid, caffeic acid-3-
O-glucoside, rosmarinic-acid, rosmarinic-acid-3-O-glucoside, luteolin, luteloin-7-O-glucoside, luteloin-7-O-diglucuronide,
luteloin-7-O-glucuronide, apigenin, apigenin-7-O-caffeoyglucoside, apigenin-7-O-diglucuronide and scutellarein-7-Odiglucuronide.
Among these, the contents of rosmarinic acid and its glucoside derivatives decreased initially and then sharply
increased to the maximum at the fruiting stage, while the flavonoids and their glucoside derivatives gradually increased
until reaching the maximum at the fruiting stage. The contents of rosmarinic acid and luteloin-7-O-glucuronide in leaves at
the fruiting stage were the highest, reaching 21.41 mg/g and 19.89 mg/g, respectively. Furthermore, leaves at the fruiting
stage had the highest total phenol content of 47.50 mg/g among the various parts examined. Conclusions: The results of our
present work may provide a good reference for the utilization of Perilla frutescens as a phenolics resource.

Key words: Perilla frutescens, phenolics, high performance liquid chormatography-tandem mass spectrometry (HPLC-MS/MS), accumulation, optimal harvest time

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