Extraction and high-performance liquid detection methods of beta-ecdysone in euphausia superba

An Antarctic krill and ecdysone technology, which is applied in the fields of food and chemical industry, can solve the problems of high price of enzyme-linked immunization, complicated extraction steps, low accuracy of results, etc., and achieves accurate and reliable peak area quantification, good detection effect, and detection limit. low effect

Inactive Publication Date: 2018-02-23
SHANDONG NORMAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] β-ecdysone was discovered for the first time in dew grass. Because of its rich content in plants, there are many researches on β-ecdysone in plants. For the extraction of β-ecdysone in plants, water boiling and ethanol extraction are mostly used. And through column chromatography or sherwood oil degreasing and purification, the extraction steps are complicated and time-consuming; and for the detection of β-ecdysone in plants, there are currently thin-layer scanning methods, radioimmunoassay methods, liquid chromatography, etc., as disclosed in CN103837615A A method for the simultaneous determination of plant ecdysterone substances by high performance liquid chromatography was developed. Xu Lirong disclosed the extraction, purification process and quality standard research of β-ecdysterone in Dewweed Grass. Wang Qiang et al. -The content determination of ecdysterone (thin-layer chromatography and high-performance li

Method used

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  • Extraction and high-performance liquid detection methods of beta-ecdysone in euphausia superba
  • Extraction and high-performance liquid detection methods of beta-ecdysone in euphausia superba

Examples

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Embodiment 1

[0037] Example 1: Extraction of Antarctic krill β-ecdysone and detection by high performance liquid chromatography

[0038] 1, the extraction of Antarctic krill β-molting hormone, the steps are as follows:

[0039] (1) Take the frozen Antarctic krill, thaw it in 5% normal saline for 3 hours, absorb the water from the thawed Antarctic krill, and cut it into pieces;

[0040] (2) Take shredded Antarctic krill (w) and methanol (v) with a ratio of 1:5-8, ultrasonically extract under the conditions of 35°C, 300w, and 40kHz, extract 3 times for 30min each time, and filter the extract , combining the 3 extracts;

[0041] (3) Place the above-mentioned filtrate in a rotary evaporator and evaporate to dryness, and the rotary evaporation temperature is 50° C. to obtain the crude extract of β-ecdysone;

[0042] (4) Completely dissolve the crude extract with 40ml of methanol, add an equal volume of n-hexane, ultrasonically mix, centrifuge at 3000rpm at 4°C for 30min, discard the supernata...

Embodiment 2

[0050] Example 2: Extraction and HPLC detection of Antarctic krill β-ecdysone

[0051] Frozen Antarctic krill, thawed in 5% normal saline, thawed for 3 hours, blotted dry, took 100g of thawed Antarctic krill, chopped the tissue, put it into a 1000ml beaker, added 500ml of methanol, and heated it at 25℃, 250w, 40kHz Ultrasonic heating and extraction in an ultrasonic cleaning machine, each extraction was 30min, repeated 3 times, the extracts were combined and filtered, and the filtrate was evaporated to dryness in a rotary evaporator at an evaporation temperature of 50°C to obtain a crude extract of β-ecdysone.

[0052] Completely dissolve the crude extract with 40ml of methanol, add an equal volume of n-hexane, mix it with ultrasound, centrifuge at 3000rpm at 4°C for 30min, discard the supernatant, repeat three times, put the lower layer solution in a -20°C refrigerator, take the supernatant, Impurities were removed, and the supernatant was evaporated to 3 ml in a rotary evapor...

Embodiment 3

[0055] Example 3: Extraction of Antarctic krill β-ecdysone and detection by high performance liquid chromatography

[0056] Frozen Antarctic krill, thawed in 5% normal saline, thawed for 3 hours, blotted dry, took 100g of thawed Antarctic krill tissue, cut it into pieces, put it into a 1000ml beaker, added 800ml of methanol, and heated it under 30℃, 300w, 40kHz ultrasonic waves Ultrasonic heating and extraction in a cleaning machine, each extraction for 30 minutes, repeated 3 times, combined extracts, and filtered, the filtrate was evaporated to dryness in a rotary evaporator at an evaporation temperature of 50°C to obtain a crude extract of β-ecdysone.

[0057] Completely dissolve the crude extract with 40ml of methanol, add an equal volume of n-hexane, mix it with ultrasound, centrifuge at 3000rpm at 4°C for 30min, discard the supernatant, repeat three times, put the lower layer solution in a -20°C refrigerator, take the supernatant, Impurities were removed, and the supernat...

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Abstract

The invention discloses extraction and high-performance liquid detection methods of beta-ecdysone in euphausia superba. Fresh frozen euphausia superba is taken as a material, subjected to unfreezing and tissue cutting treatment, ultrasonic extraction with methanol and degreasing with n-hexane and frozen at subzero 20 DEG C for removal of other impurities, and an extract containing beta-ecdysone isobtained; a high-performance liquid detection method is adopted for detecting the beta-ecdysone in the euphausia superba under the conditions of a UV/Visible detector, a YMC-PACK ODS-A250*4.6 mml.D.Schromatographic column, mobile phase methanol-water, isocratic elution and detection wavelength 248 nm, the peak area is tracked and integrated by Breeze software, and the content of the beta-ecdysone in the euphausia superba is obtained. The operation method is simple and quick and the result is accurate and reliable.

Description

technical field [0001] The invention relates to an extraction and high-efficiency liquid phase detection method of Antarctic krill β-molting hormone, belonging to the technical fields of food and chemical industry. Background technique [0002] β-ecdysterone, also known as β-ecdysterone, 20-hydroxyecdysterone; molecular formula is C 27 h 44 o 7 , with a molecular weight of 480.63 Daltons, is a light yellow crystalline powder. In animals and plants, β-ecdysone participates in the regulation of many aspects such as molting, reproduction, lifespan, diapause and immune response, and plays an important role in maintaining basic physiological functions. β-ecdysone acts on the human body to promote cell growth and division, accelerate collagen synthesis, stimulate dermal cell division, and promote skin regeneration. At the same time, a large number of animal experiments have proved that β-ecdysone can eliminate cholesterol in the body, promote protein, carbohydrate, and lipid m...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/14G01N30/88
CPCG01N30/06G01N30/14G01N30/88G01N2030/146G01N2030/8813
Inventor 韩香凝刘代成
Owner SHANDONG NORMAL UNIV
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