A screening method and preparation method of Enteromorpha antioxidant

A technology for antioxidants and screening methods, applied in liquid solution solvent extraction, instruments, measuring devices, etc., can solve problems such as inaccurate results, changes in activity and quantity, and inability to accurately know antioxidant activity, achieving good reproducibility Effect

Active Publication Date: 2022-03-04
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when evaluating the antioxidant activity of natural product extracts with complex components, the biggest defect of this method is that it is impossible to accurately know which compound in the complex mixture has antioxidant activity, and it can only be known after a certain compound is separated and purified. Whether it has antioxidant activity, but in the process of separation and purification, some compounds will deteriorate due to decomposition, and their activity and amount will also change, so it is time-consuming and laborious, and the results are not accurate

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  • A screening method and preparation method of Enteromorpha antioxidant
  • A screening method and preparation method of Enteromorpha antioxidant
  • A screening method and preparation method of Enteromorpha antioxidant

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Sample extraction:

[0041]The dried strips of Enteromorpha enteromorpha (1 kg) were completely pulverized into powder, extracted with 3 L of 60% ethanol for 3 times, each time for 3 hours, and then the extract was filtered with a vacuum filter. All extracts were combined and concentrated to no alcohol smell, and then the concentrate was extracted with an equal amount of ethyl acetate. The ethyl acetate fraction was rotovapped to dryness at 55 °C to obtain 10 g of crude sample, which was stored in a refrigerator at 4 °C.

[0042] DPPH-HPLC detection:

[0043] The DPPH-HPLC method was used to quickly screen the free radical scavenging properties of natural products. After reacting with DPPH, the peaks on the HPLC spectrum decreased or disappeared, indicating that there might be antioxidant compounds, and the peaks that did not change were considered to have no antioxidant capacity. Enteromorpha enteromorpha sample (25 μL) was mixed with DPPH solution (40 μL, 0.4 mg / mL)...

Embodiment 2

[0062] The difference between this example and Example 1 is that the conditions for preparing the liquid phase are different, and the rest are the same. Use preparative liquid chromatography to analyze the isolate, liquid phase conditions: waters-RP C 18 Column (4.6×250mm), UV detection wavelength 280nm, column temperature: 25°C, flow rate: 3.0mL / min, injection volume: 200μL, mobile phase gradient elution with acetonitrile (A) and 0.2% formic acid aqueous solution (B), Gradient conditions were as follows: 0 min, 10% A; 0–5 min, 10–30% A; 5–15 min, 30–30% A; 15–40 min, 30–45% A; 40–45 min, 45–45% A; 45–46min, 45–10%A; 46–51min, 10–10%A.

[0063] The preparation liquid phase diagram of embodiment 2 is shown in Figure 4 .

Embodiment 3

[0065] The difference between this example and Example 1 is that the conditions for preparing the liquid phase are different, and the rest are the same. Use preparative liquid chromatography to analyze the isolate, liquid phase conditions: waters-RP C 18 Column (4.6×250mm), UV detection wavelength 280nm, column temperature: 25°C, flow rate: 3.0mL / min, injection volume: 200μL, mobile phase gradient elution with acetonitrile (A) and 0.2% formic acid aqueous solution (B), Gradient conditions were as follows: 0 min, 10% A; 0–5 min, 10–25% A; 5–45 min, 25–45% A; 45–46 min, 45–10% A; 46–51 min, 10–10% A. The preparation liquid phase diagram of embodiment 3 is shown in Figure 5 .

[0066] Examples 2 and 3 show that if the elution procedure of the preparative liquid chromatography affects the separation of the screened compound with antioxidant properties, only the elution procedure of Example 1 can be used to completely separate and obtain compound 1, Compound 4, Compound 5 and C...

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Abstract

The invention discloses a screening method and a preparation method of Enteromorpha antioxidants. The screening method is to obtain the Enteromorpha extract by performing ethyl acetate extraction on the alcohol extract of Enteromorpha, and use DPPH-HPLC to analyze the Enteromorpha extract. Antioxidant detection; gradient elution program in HPLC: 0min, 10%A; 0–10min, 10–40%A; 10–20min, 40‑55%A; mobile phase A is acetonitrile, mobile phase B The phase is 0.19-0.21% formic acid aqueous solution. The preparation method is to use liquid chromatography to separate and purify the screened compounds with antioxidant properties. The invention can screen and prepare polyphenol antioxidants from enteromorpha on-line in large batches, and the method is fast, intuitive, accurate, simple and easy, and has good reproducibility.

Description

technical field [0001] The invention relates to a screening method and preparation method of enteromorpha antioxidant. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] The oxidation reaction caused by free radicals is not only closely related to the occurrence of various diseases, but also causes food spoilage. Studies have found that most of the currently used antioxidants are synthetic antioxidants, which have many negative effects on human health. In order to find high-efficiency antioxidant substances, research on screening methods for antioxidant substances is very important. The development of natural plant antioxidants has broad prospects. my country's Enteromorph...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D11/04G01N30/02
CPCB01D11/04G01N30/02
Inventor 张进杰王宸杨文鸽徐大伦李超王岱杰
Owner NINGBO UNIV
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