Optimization of the extraction technology of total coumarin from Viola yedoenis and its antioxidant and anti-tumor activities
- VernacularTitle:紫花地丁总香豆素提取工艺优化及其抗氧化和抗肿瘤活性考察
- Author:
Renqiang CHEN
1
,
2
;
Zhen LIU
1
;
Huiling HU
1
;
Linfeng LI
1
;
Chun ZHANG
1
Author Information
1. School of Pharmacy，Southwest Medical University，Sichuan Luzhou 646000，China
2. Dept. of Pharmacy，the Affiliated Hospital of Southwest Medical University，Sichuan Luzhou 646000，China
- Publication Type:Journal Article
- Keywords:
Viola yedoensis;
total coumarin;
extraction technology;
antioxidant effect;
anti-tumor effect
- From:
China Pharmacy
2022;33(20):2471-2476
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To optimize the extraction technology of total coumarin from Viola yedoensis and to evaluate its in vitro antioxidant and anti -tumor activities of its purified product . METHODS On the basis of single factor experiments ，the extraction technology of total coumarin from V. yedoensis was optimized by the response surface methodology ，using yield of total coumarin from V. yedoensis as response value ，solid-liquid ratio ，extraction time and ethanol volume fraction as variables . The verification tests were conducted . The antioxidant activity of purified product of total coumarin from V. yedoensis was evaluated by 1，1-diphenyl-2-picrylhydrazyl（DPPH）method，2，2′-azino-bis（3-ethylbenzothiazoline-6-sulfonic）acid ammonium salt （ABTS） method and 2-phenyl-4，4，5，5-tetramethylimidazoline-3-oxide-1-oxyl（PTIO）method. CCK-8 assay was used to determine the inhibitory effect of it on human cervical cancer cells HeLa ，human intrahepatic bile duct cancer cells HCCC -9810，human hepatocellular carcinoma cells HepG 2，human breast cancer cells MDA -MB-231 and human non -small cell lung cancer cells A 549. The apoptosis morphology of HCCC -9810 cells was detected by Hoechst 33258 staining，and lactate dehydrogenase （LDH）and superoxide dismutase （SOD）content were determined . RESULTS The optimal extraction technology included solid -liquid ratio was 25∶1（mL/g），extraction time was 1.9 h，ethanol volume fraction was 68%. Average yield of total coumarin from V. yedoensis was （21.10±0.17）mg/g，and RSD was 0.80%（n＝3），relative error of which with predicted value （20.79 mg/g）was 1.49%. The median inhibitory concentrations （IC50）of purified product of total coumarin from V. yedoensis to DPPH and ABTS free radicals were 21.54 and 17.11 μg/mL，which were slightly higher than that of ascorbic acid （15.21，12.45 μg/mL）. The IC 50 to PTIO free radicals was 1 422.01 μg/mL，which was far higher the that of ascorbic acid （37.20 μg/mL）. The IC 50 values of total coumarin from V. yedoensis to above 5 cells were （113.18±15.97），（51.17±1.72），（134.15±11.75），（171.83±15.27）and（93.38±4.65）μg/mL，respectively. Total coumarin from V. yedoensis induced the apoptosis of HCCC -9810 cells and increased the content of LDH ，as well as decreased the content of SOD . CONCLUSIONS The extraction technology of total coumarin from V. yedoensis is optimized successfully ；the purified product of obtained extract has good antioxidant and anti -tumor activities.
