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Method for extracting catechin small molecules in lotus root and application thereof

A technology of catechins and extraction methods, which is applied in the field of extraction of catechins small molecules, can solve the problems of organic solvent toxicity residues, difficulties in the development of medicines and health food, etc., achieve low cost, improve the immune level of the body, and use raw materials resource rich effect

Active Publication Date: 2010-09-08
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional organic solvent extraction method of total phenols in lotus root failed to clearly point out the structure of its antioxidant components, and due to the residual organic solvent toxicity, it brought difficulties to the development of drugs and health food
The process of extracting catechin small molecules from lotus root with water-ethanol system and the research on the biological activity of the extract have not been reported yet

Method used

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  • Method for extracting catechin small molecules in lotus root and application thereof
  • Method for extracting catechin small molecules in lotus root and application thereof
  • Method for extracting catechin small molecules in lotus root and application thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0029] 1. Extraction and purification of small molecule active substances from lotus root

[0030] (1) Take 10kg of fresh lotus root, pulverize, soak in 10L deionized water at 4°C overnight, and filter to remove the residue. Centrifuge at 5000rpm for 10min, take the supernatant and rotate to evaporate excess water, and freeze-dry at -45°C to obtain crude extract L;

[0031] (2) Weigh 1 g of crude extract L, dissolve it in deionized water, and load it on a 1.5 × 35 cm NKA column of weakly polar macroporous adsorption resin, purchased from Tianjin Nankai Hecheng Technology Co., Ltd., and the elution rate is 1 ml / min, first wash the impurity L1 with 200ml deionized water, then use 30% ethanol to obtain the sample L2, wash the column with 90% ethanol to remove the impurity L3, and then use deionized water to rinse and regenerate it can be used continuously;

[0032] (3) After the obtained sample L2 was freeze-dried, 100 mg was weighed and loaded onto a Sephadex column LH-20 of 1...

Embodiment 2

[0044] Antioxidant activity of small and medium catechins in lotus root

[0045] 1. Determination of antioxidant activity at the cellular level

[0046] Normal Kunming male mice were removed from their eyeballs and blood was collected in a centrifuge tube with 0.15M NaCl in an ice bath, and centrifuged at 2500 rpm for 10 min to separate red blood cells from plasma and broken cell membranes. And use 10 times the volume of pH7.4 PBS solution (10mM pH7.4Na 2 HPO 4 -NaH 2 PO 4 Buffer, 125mM NaCl) was washed twice, and finally complete red blood cells were obtained (Miki M, Tamai H, MinoM, et al. Free-radical chain oxidation of rat red blood cells by molecular oxygen and its inhibition by α-tocopherol. Arch Int Physiol Biochim Biophys 1987, 258: 373-380.). This analytical system uses peroxyl radicals to cause erythrocyte hemolysis (Sugiyama H, Fung K P. and Wu T W. Purpruogallinas an antioxidant protector of human erythrocytes against lysis by peroxyl radicals. J. Life Sci., 1...

Embodiment 3

[0065] Anti-HIV-1 Reverse Transcriptase (HIV-1RT) Activity of Small Molecular Catechins in Lotus Root

[0066] In a 1.5ml centrifuge tube, 20μl of samples with different concentrations, 20μl of RT (0.2ng / μl) and 20μl of template and nucleotide mixture were sequentially added to form a 60μl reaction system, which was reacted at 37°C for 1h (positive control: 20μl lysis buffer instead Sample; blank control: replace sample and RT with 40 μl lysis buffer). Transfer 60 μl of the reaction solution to a small well of the MTP plate, react at 37°C for 1 h, completely remove the residual liquid and wash with 250 μl of washing solution for 5 times, each wash for 30 s, discard the residual liquid, and add 200 μl of Anti-DIG-POD to each well. (200mU / ml), add membrane, and react at 37°C for 1h. After removing the residual liquid, wash 5 times with 250 μl of washing solution for 30 s each time, discard the residual liquid, add 200 μl of ABTS (about 1.3 mg / ml) to each well, and react at 15-2...

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Abstract

The invention provides a method for extracting catechin small molecules in lotus roots and an application thereof, wherein the method for extracting catechin small molecules in lotus roots obtains two purified products, gallocatechin grade and catechin through water extraction, macroporous resin adsorption and sephadex filtering. The gallocatechin grade and catechin extracted from lotus roots notonly have strong in vitro and intracellular antioxidant activity, but also have activity for efficiently removing active oxygen free radicals and HIV-resistant activity, which can improve the body immune level and prevent various diseases related to free radicals. The invention is characterized by antioxidation and HIV resistance; the raw materials are abundant and absolutely nontoxic with low cost and convenient extraction; and therefore, the invention is applicable to development of medicine or natural health care product.

Description

Technical field: [0001] The invention relates to an extraction method and application of catechin-like small molecules in lotus root. Background technique: [0002] Medical research has shown that reactive oxygen species are linked to many aging-related diseases, including tumors, diabetes, arteriosclerosis, cancer, asthma, myocardial ischemia-reperfusion, Alzheimer's disease, arthritis, cataracts, dermatitis, retinal damage, and liver damage At the same time, reactive oxygen species can also induce cell apoptosis. In recent years, studies have shown that oxidative damage is closely related to viral diseases such as AIDS. [0003] Vegetables that people eat every day contain many antioxidants. Such as: eggplant, cabbage, peppers, tomatoes, etc. have a high inhibition rate on DPPH free radicals; It is closely related to the total phenolic content. [0004] Lotus (Nulumbo nuciferra Gaertn), also known as Hibiscus, Lotus, Shuizhi, Yuhuan, June Chun, etc., is cultivated all ...

Claims

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Application Information

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IPC IPC(8): C07D311/62A61K31/353A61P39/06A61P31/12A23L1/30A23L33/105
Inventor 刘方江筠王常荣李宁陈芝惠张聃吕鹏云曲丽媛
Owner NANKAI UNIV
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