基于UPLC-Q-TOF-MS的黄精炮制前后低聚糖成分分析及抗氧化活性研究

UPLC-Q-TOF-MS Identifies Components of Oligosaccharides with Antioxidant Activity in Polygonati Rhizoma before and after Processing

目的:探究黄精酒制前后低聚糖的成分种类、变化情况,以及生、酒黄精低聚糖抗氧化活性。方法:采用超高效液相色谱串联四极杆飞行时间高分辨质谱对1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化后的低聚糖进行成分分析,采用苯酚-硫酸法测定低聚糖含量,采用1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS+)自由基清除法测定其体外抗氧化活性,利用秀丽隐杆线虫测定体内抗氧化活性。结果:在生、酒黄精低聚糖中同时检测到蔗糖、蔗果三糖和蔗果四糖3种低聚糖,蔗糖、蔗果三糖、蔗果四糖在炮制后减少,棉子糖、蜜二糖、蔗果五糖、蔗果六糖在炮制后未检测出,葡萄糖、果糖和半乳糖3种单糖含量增加;生、酒黄精低聚糖质量分数分别为78.17%、70.80%;生、酒黄精低聚糖对DPPH自由基的半数效应浓度(EC50)分别为2.12、0.41mg·mL^(-1)、ABTS+自由基的EC50分别为2.90、0.95 mg·mL^(-1);生、酒黄精低聚糖对秀丽隐杆线虫热应激和氧化应激损伤具有良好的缓解作用。结论:黄精经炮制后低聚糖成分含量发生变化;黄精低聚糖具有一定的体外和体内抗氧化作用,酒制后活性增强。以上结果可为黄精炮制前后作用改变的研究及对黄精的开发利用提供参考。

Objective:To explore the components,changes,and antioxidant activity of oligosaccharides in Polygonati Rhizoma before and after processing with wine.Methods:The composition of the oligosaccharides derivatized with 1-pheny-3-methyl-5-pyrazolone(PMP)was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight high-resolution mass spectrometry.The content of oligosaccharides was determined with the phenol-sulfuric acid method.The antioxidant activity of PMP-derivatized oligosaccharides in vitro was evaluated based on the 1,1-diphenyl-2-picrylhydrazine(DPPH)and diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate)(ABTS+)free radical scavenging activities,and that in vivo was determined in Caenorhabditis elegans.Results:Three oligosaccharides(sucrose,1-kestose and nystose)were detected in both raw and processed Polygonati Rhizoma.The content of sucrose,1-kestose,and nystose decreased after processing,and raffinose,melibiose,1F-fructofuranosylnystose,and kestohexaose were not detected after processing.The content of glucose,fructose,and galactose increased after processing.The content of oligosaccharides in raw and processed Polygonati Rhizoma was 78.17%and 70.80%,respectively.The median effective concentrations(EC50)of oligosaccharides in raw and processed Polygonati Rhizoma were 2.12 mg·mL^(-1) and 0.41 mg·mL^(-1) against DPPH free radicals and 2.90 mg·mL^(-1) and 0.95 mg·mL^(-1) against ABTS+free radicals,respectively.The oligosaccharides in both raw and processed Polygonati Rhizoma alleviated the heat stress and oxidative stress damage in C.elegans.Conclusion:The composition of oligosaccharides in Polygonati Rhizoma changed after processing.Polygonati Rhizoma oligosaccharides demonstrated antioxidant activity both in vitro and in vivo,and the activity was enhanced after Polygonati Rhizoma was processed with wine.The above results provide a basis for explaining the effect changes of Polygonati Rhizoma before and after processing and have reference significance for the development and utilization of Polygonati Rhizoma.