4种金橘黄酮含量测定及自由基清除活性研究

Study on the Content Determination and the Radical-Scavenging Activities of Four Flavonoids in Kumquat

建立4种金橘黄酮的一测多评含量测定方法(QAMS),并对其自由基清除活性进行评价。采用超高效液相色谱法(UPLC),通过QAMS测定4种金橘黄酮的含量;以1,1-二苯基-2-三硝基苯肼(DPPH)、2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸(ABTS)自由基清除法评价其自由基清除活性。结果显示:QAMS测得8批金橘样品中根皮素-3',5'-二-C-β-葡萄糖苷、4'-甲氧基-牡荆素-2''-O-α-L-鼠李糖苷、4'-甲氧基-异牡荆素-2''-O-α-L-鼠李糖苷、金橘苷含量范围分别为6.475~16.257,0.844~1.438,0.665~1.229和0.777~1.481 mg/g;其清除DPPH自由基的能力为4'-甲氧基-牡荆素-2''-O-α-L-鼠李糖苷>根皮素-3',5'-二-C-β-葡萄糖苷>4'-甲氧基-异牡荆素-2''-O-α-L-鼠李糖苷>金橘苷,清除ABTS自由基的能力为根皮素-3',5'-二-C-β-葡萄糖苷>4'-甲氧基-牡荆素-2''-O-α-L-鼠李糖苷>金橘苷>4'-甲氧基-异牡荆素-2''-O-α-L-鼠李糖苷。试验建立的一测多评法准确可靠,可用于金橘黄酮的质量评价,且4种金橘黄酮均具有一定的自由基清除活性,可为金橘的开发利用提供理论基础。

Establish a method of quantitative analysis of multi-components by single marker(QAMS)for simultaneous determination offour flavonoids in kumquat,and evaluate the radical-scavenging activities of these four flavonoids.Ultra high performance liquid chromatography(UPLC)was used to determine the contents of four flavonoids in kumquat by QAMS.The radical-scavenging activities were evaluated toward 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)radical.The results showed that the contents of phloretin-3’,5’-di-C-β-glucopyranoside,4’-methoxy-vitexin-2’’-O-α-L-rhamnopyranoside,4’-methoxy-isovitexin-2”-O-α-L-rhamnopyranoside and fortunellin in 8 batches of kumquats were determined by QAMS in the range of 6.475-16.257,0.844-1.438,0.665-1.229 and 0.777-1.481 mg/g,respectively.The DPPH radical-scavenging abilities were in order as follows:4’-methoxy-vitexin-2’’-O-α-L-rhamnopyranoside>phloretin-3’,5’-di-C-β-glucopyranoside>4’-methoxy-isovitexin-2’’-O-α-L-rhamnopyranoside>fortunellin,and the ABTS radical-scavenging abilities were in order as follows:phloretin-3’,5’-di-C-β-glucopyranoside>4’-methoxy-vitexin-2’’-O-α-L-rhamnopyranoside>fortunellin>4’-methoxy-isovitexin-2’’-O-α-L-rhamnopyranoside.The established method(QAMS)for simultaneous determination of four flavonoids in kumquat is accurate and reliable,and can be used for quality evaluation of flavonoids in kumquat.All of the four flavonoids have certain radical-scavenging activities,which can provide theoretical basis for the development and utilization of kumquat.