古丽霞 傅咏梅 张蜀 林居逸 邓红 卢永美

中圖分类号 R284.2 文献标志码 A 文章编号 1001-0408（2021）18-2236-07

DOI 10.6039/j.issn.1001-0408.2021.18.10

摘 要 目的：建立猴耳环多酚的改良型双水相萃取体系。方法：以猴耳环多酚含量、提取效率和分配系数为指标，采用单因素实验筛选双水相萃取体系的料液比、乙醇质量分数和超声时间。制备猴耳环不同极性部位的双水相萃取液，以多酚含量和抗氧化活性[1，1-二苯基-2-三硝基苯肼（DPPH）、2，2′-联氨-双（3-乙基苯并噻唑-6-磺酸）二铵盐（ABTS）的半数抑制浓度（IC50）]为指标，结合多酚类成分与抗氧化活性的灰色关联度分析，筛选改良剂。考察含不同质量分数（0～90%）改良剂的改良型双水相萃取体系对猴耳环多酚提取的影响，并与传统提取工艺（超声提取和加热回流提取）进行比较。结果：最优双水相萃取体系为料液比1 ∶ 45，乙醇质量分数40%，硫酸铵质量分数11%，室温下超声提取20 min。3次验证实验中，猴耳环多酚含量、提取效率和分配系数分别为（28.35±1.01）%（RSD=3.56%，n=3）、（98.87±0.19）%（RSD=0.19%，n=3）和13.25±0.71（RSD=5.39%，n=3）。改良剂为乙酸乙酯。当乙酸乙酯-乙醇混合溶液中乙酸乙酯的质量分数为70%时，改良型双水相萃取体系提取所得萃取物中多酚含量和抗氧化活性与超声提取、加热回流提取比较差异均无统计学意义（P>0.05），但得率和转移率均显著高于超声提取、加热回流提取（P<0.05）。结论：成功建立提取得率和转移率优于传统提取方法的改良型双水相萃取体系，可一步提取猴耳环多酚。

关键词 猴耳环;多酚;改良型双水相萃取;抗氧化活性

Establishment of Modified Aqueous Two-phase Extraction System of Polyphenols from Archidendron clypearia

GU Lixia1，FU Yongmei2，ZHANG Shu1，3，LIN Juyi1，DENG Hong1，3，LU Yongmei1（1. Guangdong Provincial Key Laboratory of Advanced Drug Delivery Systems， Guangdong Pharmaceutical University， Guangzhou 510006， China; 2. Sinopharm Group Guangdong Medi-World Pharmaceutical Co.， Ltd.， Guangdong Foshan 528000， China; 3. Guangdong Provincial Engineering Center of Topical Precise Drug Delivery Systems， Guangdong Pharmaceutical University， Guangzhou 510006， China）

ABSTRACT   OBJECTIVE： To establish modified aqueous two-phase extraction （ATPE） system of polyphenols from Archidendron clypearia. METHODS： Taking the polyphenol content， extraction efficiency and partition coefficient of A. clypearia as indexes， the solid-liquid ratio， ethanol mass fraction and ultrasonic time of ATPE system were selected by single factor tests. The aqueous two-phase extracts from different polar parts of A. clypearia was prepared. The modifier was selected by taking polyphenol content and antioxidant activity [IC50 of 1，1-diphenyl-2-trinitrophenylhydrazine （DPPH）， 2，2′-hydrazine bis （3-ethylbenzothiazole-6-sulfonic acid） diammonium salt （ABTS）] as indicators， combined with the grey correlation analysis between polyphenols and antioxidant activity. The effects of modified ATPE system with different mass fraction （0-90%） of modifier on the extraction of polyphenols from A. clypearia were investigated and compared with traditional extraction technology （ultrasonic extraction and heating reflux extraction）. RESULTS： The optimal ATPE system included solid-liquid ratio of 1 ∶ 45， ethanol mass fraction of 40%，（NH4）2SO4 mass fraction of 11%，ultrasonic extraction time of 20 min， at room temperature. In 3 validation tests， polyphenol content， extraction efficiency and partition coefficient were （28.35±1.01）%（RSD=3.56%，n=3），（98.87±0.19）%（RSD=0.19%，n=3） and 13.25±0.71 （RSD=5.39%，n=3）， respectively. The modifier was ethyl acetate. When the mass fraction of ethyl acetate in the ethyl acetate-ethanol mixed solvent was 70%， there was no significant difference in the content and anti-oxidant activity of polyphenols from A. clypearia of modified ATPE （P>0.05）. The yield and transfer rate of it were significantly higher than those of ultrasonic extraction and heating reflux extraction （P<0.05）. CONCLUSIONS： The modified ATPE system with better extraction yield and transfer rate than the traditional extraction technology is successfully established， which can extract polyphenols from A. clypearia in one step.

KEYWORDS   Archidendron clypearia; Polyphenols; Modified aqueous two-phase extraction; Antioxidant activity

猴耳环Archidendron clypearia（Jack）Benth为含羞草科猴耳环属植物，主要分布在广东、海南、广西、湖南等地，是我国南方的特色中药材，具有清热解毒、凉血消肿、止泻、去湿敛疮的功效[1]。猴耳环的主要活性成分有黄酮类、黄烷类和有机酚酸类等多酚物质，具有抗炎、抗菌、抗病毒、降血糖等作用[2-6]。