A kind of extraction and high performance liquid phase detection method of Antarctic krill β-ecdysterone
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
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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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