食品科学 ›› 2019, Vol. 40 ›› Issue (22): 263-269.doi: 10.7506/spkx1002-6630-20181123-275

• 成分分析 • 上一篇    下一篇

超高效液相色谱-串联质谱法测定圆椒中甘油磷脂含量

何秀梅,藏志焕,陈晴,赵瑛博   

  1. (沈阳农业大学食品学院,辽宁 沈阳 110866)
  • 出版日期:2019-11-25 发布日期:2019-12-02
  • 基金资助:
    辽宁省大学生创新创业项目(201710157000262)

Determination of Phospholipids Contents in Bell Pepper by Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry

HE Xiumei, ZANG Zhihuan, CHEN Qing, ZHAO Yingbo   

  1. (College of Food Science, Shenyang Agricultural University, Shenyang 110866, China)
  • Online:2019-11-25 Published:2019-12-02

摘要: 建立圆椒中磷脂含量测定的超高效液相色谱-串联质谱方法。样品用异丙醇-氯仿-水体系作为提取溶剂,液液萃取法净化,采用HILIC亲水作用色谱柱(2.1 mm×100 mm,1.7 μm)体系分离磷脂,电喷雾电离-串联四极杆质谱测定。分别以磷脂酰胆碱(C28:0)、溶血磷脂乙醇胺(C14:0)、溶血磷脂胆碱(C14:0)、磷脂酰甘油(C28:0)、磷脂酰甘油酸(C28:0)、溶血磷脂丝氨酸(C16:0)为对照进行相对定量。实验对提取溶剂和净化方法进行选择,以抽提得到磷脂种类最多、含量最高为前处理方法,并对该方法进行适应性检验。结果表明,6 种磷脂在10~1 000 ng/mL之间线性良好,检出限为2~300 ng/mL,定量限为10~1 000 ng/mL。在125、250 ng/mL和375 ng/mL的添加水平上,回收率在68.06%~84.17%之间,相对标准偏差介于3.40%~9.60%之间。该方法为圆椒磷脂测定提供了可靠的平台,为基于膜脂代谢的圆椒冷害研究提供基础。

关键词: 圆椒, 磷脂, 超高效液相色谱-串联质谱

Abstract: An analytical method for the determination of phospholipids contents in bell pepper by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established. Samples were extracted with isopropanol-methanol-water and the extract was purified by liquid-liquid extraction, and then separated on a hydrophilic interaction liquid chromatography (HILIC) (2.1 mm × 100 mm, 1.7 μm) system before being analyzed by electrospray ionization tandem quadrupole mass spectrometry. Relative quantification was carried out with phosphatidylcholine (PC), lysophosphatidylamide ethanolamine (LPE), lysophosphatidylcholine (LPC), phosphatidylglycerol (PG), phosphatidyl amide glycerol (PA), and lysophosphatidyl amide serine (LPS) as reference materials, separately. The extraction solvent and the purification method were selected in the experiment. The pretreatment method that could extract the largest number and amount of phospholipids was used, and its applicability was tested. The results showed that good linearity was observed for the six phospholipids in the range of 10–1 000 ng/mL. The limits of detection (LODs) were 2–300 ng/mL, and the limits of quantitation (LOQs) were 10–1 000 ng/mL. At spiked levels of 125, 250, and 375 ng/mL, the recovery was 68.06%–84.17%, with relative standard deviations (RSDs) of 3.40%–9.60%. The method provides a reliable platform for the determination of phospholipids in bell pepper, and more broadly for research on cold injury based on membrane lipid metabolism.

Key words: bell pepper, phospholipid, ultra-high performance liquid chromatography-tandem mass spectrometry

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