Abstract
Free radicals, including 1,1-diphenyl-2-picrylhydrazyl, mediate oxidative stress to cause many chronic diseases (including cardiovascular diseases, diabetes and cancer). The extract of traditional Tibetan medicine Lepidium latifolium L. (L. latifolium) was reported to have free radical inhibition ability. Therefore, a system method was established to separate the ethanol extract of L. latifolium to prepare two main antioxidant compounds. First of all, silica gel and a medium-pressure liquid chromatography tower were used for pre-treatment of the ethanol extract of L. latifolium to obtain the main antioxidant active component fraction 4 through online high-performance liquid chromatography-1,1-diphenyl-2-picrylhydrazyl (HPLC-DPPH) assay. Then, fraction 4-1 was obtained by one-dimensional preparation using Megres C18 chromatographic column, and two active compounds with IC50 values 59.9 and 71.3 μg/mL were obtained by two-dimensional preparation using Click XIon chromatographic column. Through the study of the chemical components and separation methods of L. latifolium, the combination of HPLC-DPPH assay and two-dimensional preparative liquid chromatography was realized, providing a reference for the separation of active compounds from L. latifolium.
Highlights
Oxidative stress is vital for organisms, in some given conditions, oxidative stress is inextricably linked to all kinds of diseases, such as Alzheimer, Parkinson, inflammation, aging, cancer, neuronal disorders, Diabetic Complications and cardiovascular diseases [1,2,3,4,5,6,7]
The experimental instrument of online HPLC-DPPH assay consists of two instruments, LC-16 and LC-10AD (Shimadzu Enterprise Management Co., Ltd., Kyoto, Japan), which were linked by a three-way links and a reaction coil
The two antioxidant flavonoids were not found by Yuan [10] who obtained nine antioxidant flavonoids from L. latifolium, so that the results of this study complemented the research on antioxidant compounds in L. latifolium
Summary
Oxidative stress is vital for organisms, in some given conditions, oxidative stress is inextricably linked to all kinds of diseases, such as Alzheimer, Parkinson, inflammation, aging, cancer, neuronal disorders, Diabetic Complications and cardiovascular diseases [1,2,3,4,5,6,7]. Antioxidants are divided into synthetic antioxidants and natural antioxidants. The most commonly used synthetic antioxidants are bhabutylated hydroxyanisole (BHA), butylated hydroxytolune (BHT), and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) etc [8]. They all have good antioxidant activity, but, due to environmental and economic concerns, research is more and more inclined towards natural antioxidants. Natural antioxidants include flavonoids [9]. (L. latifolium) was reported for its better antioxidant activity of ethyl acetate site [10] The plant Lepidium latifolium L. (L. latifolium) was reported for its better antioxidant activity of ethyl acetate site [10]
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