基于UPLC-QE-Orbitrap-MS的米邦塔仙人掌茎果化学成分表征及其抗氧化活性评价

Research on Chemical Constituents and Antioxidant Capacity of Opuntia Milpa Alta and its Fruits Using UPLC-QE-Orbitrap-MS

关键词:  米邦塔仙人掌  超高效液相色谱-四级杆-静电轨道串联高分辨质谱法  化学成分  抗氧化

Key words:  Opuntia Milpa Alta  UPLC-QE-Orbitrap-MS  chemical constituents  antioxidant

摘要:

目的 建立超高效液相色谱串联四极杆静电场轨道阱质谱方法(UPLC-Q-Exactive Orbitrap MS),对米邦塔仙人掌的叶状茎及果实化学成分进行表征,并对其抗氧化活性进行考察,初步探讨成分与活性的关系。方法 选择UPLC HSS T3 C18 (2.1 mm×10 mm, 1.8 μm) 色谱柱,以乙腈-0.1% 甲酸水为流动相梯度洗脱,流速为 0.2 mL.min-1；采用加热电喷雾离子源(HESI),正、负离子模式分别进行一级、二级质谱数据采集；采用数据库和文献报道对米邦塔仙人掌叶状茎及果实的化学成分进行表征。采用DPPH、羟自由基(.OH)和超氧阴离子自由基(.O2-)清除能力测试方法,对米邦塔仙人掌叶状茎及果实的抗氧化活性进行考察。结果 从米邦塔仙人掌的叶状茎及果实中共表征出39种化合物,其中酚酸类22个,黄酮类13个,其他成分4个。米邦塔仙人掌叶状茎及果实均具有一定的抗氧化能力,米邦塔仙人掌果实对DPPH的清除能力强于叶状茎,而叶状茎对.OH清除能力稍强于果实,叶状茎皮和果皮清除能力相当,果汁对.O2-清除能力最强,米邦塔仙人掌的抗氧化活性与其富含的酚酸类成分密切相关。结论 本研究建立了一种快速、准确、可靠的米邦塔仙人掌茎果化学成分的鉴别方法,为米邦塔仙人掌资源的综合开发利用提供了科学依据。

Abstract:

ABSTRACT: OBJECTIVE To characterize the chemical constituents of the cladodes and fruits of Opuntia Milpa Alta by using ultra-high performance liquid chromatography-quadrupole electrostatic-Orbitrap mass spectrometry (UPLC-QE-Orbitrap MS), and investigate the antioxidant activity, and to explore the relationship between the constituents and biological activity. METHODS The UPLC HSS T3 C18 column (2.1 mm×10 mm, 1.8 μm) was used. Acetonitrile and water with 0.1% formic acid was used for gradient elution at a 0.2 mL·min-1 flow rateas. Heated electrospray ion source (HESI) was used for primary and secondary mass spectrometry data acquisition in positive and negative ion modes, respectively. Database and literature reports were used to characterize the chemical constituents of the cladodes and fruits of Opuntia Mipa Alta. DPPH radical, hydroxyl radical (·OH) and superoxide anion radical (·O2-) scavenging ability were used to investigate the antioxidant activities of the cladodes and fruits of Opuntia Milpa Alta. RESULTS A total of 39 compounds were characterized from the cladodes and fruits of Opuntia Milpa Alta, including 22 phenolic acids, 13 flavonoids and 4 other components. Both cladodes and fruits of Opuntia Milpa Alta have certain antioxidant capacity. The fruits of Opuntia Milpa Alta has better scavenging ability of DPPH than the cladodes, while the cladodes has a slightly stronger scavenging ability of ·OH than the fruits. The scavenging ability of ·O2- between cladode peels and fruit peels is not significantly different, and the juice has the strongest scavenging ability of ·O2- . The antioxidant activity of Opuntia Milpa Alta is closely related to its rich phenolic acids. CONCLUSION In this study, a rapid, accurate and reliable method was established to identify the chemical constituents of the cladodes and fruits of Opuntia Milpa Alta, which provided scientific basis for the comprehensive development and utilization of Opuntia Milpa Alta.