CAS Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
CAS Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, PR China.
Talanta. 2018 Apr 1;180:389-395. doi: 10.1016/j.talanta.2017.12.078. Epub 2017 Dec 25.
Lycii Fructus is widely cultivated in the Northwest China. It is well-known for its antiaging effect in traditional Chinese medicines (TCMs), but the effective components are not clear. In this work, the ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was used to study the antioxidant components of Lycii Fructus through analyzing the spectrum-effect relationship, and the positive correlation components with antioxidant activity were partially identified. The extractums of Lycii Fructus were adsorbed with macroporous resin, and then eluted with water and 30%, 60%, 90% ethanol in turn. The extract fraction eluted with 60% ethanol was determined as the best, and was taken for subsequent experiments. With the above separation method, UPLC fingerprints of thirty batches of Lycii Fructus (from different areas) were obtained, and thirty common peaks were selected through similarity analysis (SA). Combined with the data of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays, the spectrum-effect relationship was studied. The results showed that the main peaks with antioxidant activity were P14, P26, P8, and P21 for DPPH, and P26, P14, P21, and P19 for ABTS. Using the UPLC-MS/MS data, peaks P14, P19, P21, and P30 were respectively identified as chlorogenic acid, quercetin, kaempferol, and isorhamnetin, and then the results were confirmed through comparison with the standards and other references. Finally, their strong antioxidant activities were validated experimentally.
枸杞在中国西北地区广泛种植。在传统中药中，它以抗衰老作用而闻名，但有效成分尚不清楚。在这项工作中，通过分析谱效关系，采用超高效液相色谱串联质谱（UPLC-MS/MS）研究了枸杞的抗氧化成分，部分鉴定了与抗氧化活性呈正相关的成分。枸杞提取物用大孔树脂吸附，然后分别用水和 30%、60%、90%乙醇依次洗脱。用 60%乙醇洗脱的提取物部分被确定为最佳部分，并用于后续实验。采用上述分离方法，获得了 30 批（来自不同地区）枸杞的 UPLC 指纹图谱，并通过相似度分析（SA）选择了 30 个共有峰。结合 2,2-二苯基-1-苦基肼（DPPH）和 2,2'-联氮-双（3-乙基苯并噻唑啉-6-磺酸）二铵盐（ABTS）测定的数据，研究了谱效关系。结果表明，DPPH 法中具有抗氧化活性的主要峰为 P14、P26、P8 和 P21，ABTS 法中为 P26、P14、P21 和 P19。利用 UPLC-MS/MS 数据，分别鉴定峰 P14、P19、P21 和 P30 为绿原酸、槲皮素、山柰酚和异鼠李素，并通过与标准品和其他参考文献的比较进行了验证。最后，通过实验验证了它们的强抗氧化活性。
